University of Oklahoma Health Sciences Center Graduate College Graduate Research Education and Technology Symposium

Index

1. Foreword ...... 3

2. Schedule of Events ...... 4

3. Sponsors ...... 6

4. Alphabetical List of Presenters ...... 8

5. Oral Presentation Abstracts ...... 22 5.1. Graduate Students Oral Presentations ...... 23 5.2. Postdoctoral Fellows Oral Presentations ...... 55 5.3. Professional Students Oral Presentations ...... 70

6. Poster Abstracts ...... 74 6.1. Graduate Students - Competitive ...... 75 6.2. Graduate Students - Noncompetitive ...... 97 6.3. Postdoctoral Fellows - Noncompetitive ...... 106 6.4. Professional Students - Noncompetitive ...... 111 6.5. Oklahoma School of Science and Mathematics - Competitive ...... 123 6.6. OUHSC Core Facilities ...... 131

7. Flash Talk Presentations ...... 134 7.1. Graduate Students ...... 135 7.2. Postdoctoral Fellows ...... 138

The Graduate Research, Education and Technology (GREAT) Symposium originated in 1976 as a joint effort between the Graduate Student Association (GSA) and the Graduate College at the University of Oklahoma Health Sciences Center. The purpose of the symposium is to provide an opportunity for graduate students to showcase their research and to gain experience in the organization, interpretation, presentation, and discussion of research results. In recent years, sessions have been added to include postdoctoral fellows, professional students, and students from the Oklahoma School of Science and Mathematics (OSSM) who undertake research projects on the OUHSC campus. Thus, the GREAT symposium has become a robust representation of the diverse research performed on campus.

Participants may elect to present their work in either competitive or noncompetitive formats. For competitive presentations, a panel of judges select the most outstanding presentations in bench research, clinical sciences, or human population studies according to clarity of the abstract, organization of the presentation, project design, data analysis, and the overall quality of the presentation. Travel awards ranging from $1,000 to $1,500 are presented to winners of poster and oral sessions to attend a scientific meeting of their choosing. Flash Talk competitions were introduced this year with winners receiving cash prizes and certificates. Funding for the GREAT awards are provided by contributions from generous sponsors and from companies that participate in the Research Vendor Show.

This year the GREAT committee of the GSA organized three workshops to help students prepare for GREAT. These workshops helped students learn best practices for abstract writing and oral and poster presentation techniques. The GREAT committee thanks Dr. Melissa Medina, Dr. Alix Darden, and Dr. Deirdre Terrell, who presented the Pre-GREAT workshops.

The GREAT Committee also wishes to thank the GREAT Symposium speakers, including the Keynote Speaker, Dr. Steven Novella; the Roundtable Discussion Panelists: Dr. Thomas Kupiec, Dr. Lori Garman, Dr. Aaron Wendelboe, Dr. James Papin and Sherry Kurtz; the Cultural Awareness speaker, Carrie McClain; and the Career Development Speaker, Dr. Sanjay Bidichandani.

The GREAT Committee wishes to express their appreciation to the University of Oklahoma Health Sciences Center faculty, staff, mentors, postdocs and students who have dedicated their time and efforts to make GREAT a success. In particular, the committee thanks: Dr. Anne Pereira, Dr. Gillian Air, Dr. Michelle Staudt, Dr. Caleb Marlin, the Graduate College staff, GSA members and the Graduate Faculty Mentors and all of the participating students and postdocs. Special thanks also go to Dr. Caleb Marlin for the 2017 GREAT logo design.

The Graduate Student Association and the Graduate College wish to express their appreciation to you for sharing this week with our students and scholars as they pursue their professional goals. 3

University of Oklahoma Health Sciences Center Graduate College Graduate Research Education and Technology Symposium

College of Allied Health College of Dentistry College of Medicine College of Medicine Alumni College of Nursing College of Pharmacy College of Public Health Dean McGee Eye Institute Dunlap Codding Embassy Suites Office of the Provost Student Affairs Office of the Vice President for Research OKC Chamber OMRF OU Medical Center OU Physicians Phi Kappa Phi Stephenson Cancer Center Stillwater National Bank

Vendor Sponsors

University of Oklahoma Health Sciences Center Graduate College Graduate Research Education and Technology Symposium

Presenter ______Page #

Abraham, Rachel ...... 124 OSSM SYNTHESIS AND CHARACTERIZATION OF A NOVEL FLUORESCENT PROBE FOR TARGETING GASTRIN RELEASING PEPTIDE RECEPTOR EXPRESSING CANCER CELLS Abstract#: 93

Ahmed, Rebaz ...... 24 Pathology ROLE OF BRG1 (SMARCA4) IN THE RESISTANCE OF NON-SMALL CELL LUNG CANCER (NSCLC) TO TYROSINE KINASE INHIBITORS (TKIS) Abstract#: 1

Almatouq, Fatimah ...... 98 Allied Health Sciences QUANTITATIVE INVESTIGATION OF IMAGE ARTIFACT IN CT IMAGES OF A MULTIPLE DIODE ARRAY DETECTOR AND THEIR EFFECTS ON THE DOSE DISTRIBUTIONS CALCULATED WITH AAA AND ACUROS-XB ALGORITHMS Abstract#: 70

Amreddy, Narsireddy ...... 56 Pathology TUMOR-TARGETED DENDRIMER-BASED MULTIFUNCTIONAL NANOPARTICLE FOR BIO-CHEMODRUG DELIVERY AND IMAGING OF LUNG CANCER Abstract#: 32

Babu, Anish ...... 57 Pathology CHEMODRUG DELIVERY USING INTEGRIN-TARGETED CHITOSAN-PLGA HYBRID NANOPARTICLE FOR LUNG CANCER THERAPY Abstract#: 33

Bailey, Lisa ...... 99 Nursing DISRUPTIONS IN INFORMATION PROCESSING IN BREAST CANCER SURVIVORS Abstract#: 71

Baker, Quinn ...... 112 Neuroscience BLADDER HYPERPERMEABILITY INDUCES PERSISTENT VISCERAL PAIN: A NOVEL MECHANISM FOR VISCERAL ORGAN CROSSTALK Abstract#: 82

Baldwin, Jonathan ...... 25 Biostatistics and Epidemiology ENDOGENOUS TIME VARYING COVARIATES AND TIME VARYING CONFOUNDING ASSOCIATIONS: DEFINING AND ADJUSTING FOR THEM Abstract#: 2

Presenter ______Page #

Barrett, Zachary ...... 26 Communication Sciences and Disorders THE AMPI CLASSIFICATION OF THE DYSARTHRIAS Abstract#: 3

Bender, Dawn ...... 76 Cell Biology MECHANISMS OF ESCO-DEPENDENT COHESIN REGULATION Abstract#: 49

Branham, Kimberly ...... 77 Genetic Counseling POSSIBLE NOVEL EXTERIOR PHENOTYPE TO IDENTIFY INDIVIDUALS AT-RISK FOR LYNCH SYNDROME Abstract#: 50

Bressi, Rebekah ...... 78 Genetic Counseling ANALYSIS OF PARENTAL PERSPECTIVES REGARDING THE DIAGNOSTIC PROCESS FOR THEIR CHILD WITH AUTISM Abstract#: 51

Chandrasekar, Akansha ...... 125 OSSM REGULATION OF HIPPOCAMPAL GENE EXPRESSION ACROSS THE ESTRUS CYCLE Abstract#: 94

Colijn, Sarah ...... 27 Cell Biology EXCESSIVE PLASMIN ACTIVITY PROMOTES ENDOTHELIAL CELL NECROPTOSIS AND EMBRYONIC VASCULAR RUPTURE Abstract#: 4

Crowe, Alexandra ...... 79 Pharmaceutical Sciences ALTERED PHOSPHORYLATION OF OATP1B1 AND 1B3 IS ASSOCIATED WITH DECREASED TRANSPORT FUNCTION FOLLOWING CYCLOSPORINE A TREATMENT Abstract#: 52

Dang, Tuyen ...... 58 Neurosurgery DETERMINING THE ROLE OF XRN2 IN GLIOMA PROGRESSION Abstract#: 34

Presenter ______Page #

Dao, Hanh Dung ...... 80 Biostatistics and Epidemiology THE ASSOCIATION BETWEEN ENTERAL FEEDING AND NECROTIZING ENTEROCOLITIS BY ETIOLOGIC PATHWAY AMONG PREMATURE INFANTS IN OKLAHOMA 2001–2011 Abstract#: 53

Deen, Munim ...... 71 Biostatistics and Epidemiology ADVANCE DIRECTIVE USAGE BARRIERS IN OKLAHOMA AND POTENTIAL IMPROVEMENT Abstract#: 46

DeVette, Christa ...... 28 Microbiology and Immunology EVALUATION OF ANTI-TUMOR IMMUNITY IN THE MMTV-PYMT MOUSE Abstract#: 5

Dubon, Alejandra ...... 100 Nursing DEHYDRATION, DEMENTIA, AND DELIRIUM INTERACTIONS IN LONG-TERM INSTITUTIONAL SETTINGS Abstract#: 72

Edwards, Clara ...... 101 Nursing ANALYSIS OF BIOFEEDBACK IN TREATMENT OF PARKINSON’S DISEASE-ASSOCIATED DYSPHAGIA Abstract#: 73

Farasyn, Taleah ...... 81 Pharmaceutical Sciences DOWN-REGULATION OF OATP1B1 AND 1B3 TRANSPORT FUNCTION BY MTOR INHIBITORS EVEROLIMUS AND SIROLIMUS: POTENTIAL MECHANISM AND IMPLICATIONS IN OATP-MEDIATED DRUG-DRUG INTERACTIONS Abstract#: 54

Ford, Lance ...... 29 Biostatistics and Epidemiology A NON-LINEAR AND SEGMENTED REGRESSION APPROACH TO EXPLORING TRENDS OF COATED- PLATELET LEVELS IN SUBARACHNOID HEMORRHAGE PATIENTS Abstract#: 6

Frempah, Benjamin ...... 82 Pharmaceutical Sciences KERATINOCYTE-SPECIFIC DELETION OF THE IL-6RΑ EXACERBATES THE INFLAMMATORY RESPONSE DURING IRRITANT CONTACT DERMATITIS Abstract#: 55

Presenter ______Page #

Gaeta, Laura ...... 30 Communication Sciences and Disorders THE EFFECT OF SENSORINEURAL HEARING LOSS ON COGNITIVE ASSESSMENT SCORES Abstract#: 7

Gao, Siqi ...... 31 Cell Biology THE ROLE OF PLASMIN IN POSTNATAL HEPATIC VASCULAR INTEGRITY AFTER LIVER INJURY Abstract#: 8

Garcia, Victor ...... 32 Radiological Sciences A PROGRAM FOR FETAL DOSE ESTIMATION IN RADIOGRAPHIC EXAMINATIONS Abstract#: 9

Girton, Alanson ...... 33 Microbiology and Immunology IGG AND SAP SUPPORT INNATE IMMUNE RESPONSES TO BACILLUS ANTHRACIS PEPTIDOGLYCAN VIA DISTINCT AND REDUNDANT MECHANISMS. Abstract#: 10

Gopalakrishnan, Jaanam ...... 34 Pathology IDENTIFICATION OF SYSTEMIC LUPUS ERYTHEMATOSUS CAUSAL RISK VARIANT CANDIDATES SPANNING UBE2L3 HAPLOTYPE. Abstract#: 11

Gurley, Jami ...... 59 Ophthalmology/DMEI NEURORETINA-SPECIFIC CAVEOLIN-1 DEPLETION BLUNTS RETINAL INFLAMMATION: POTENTIAL ROLE OF ENHANCED TRAF3 PRODUCTION Abstract#: 35

Guthmiller, Jenna ...... 35 Microbiology and Immunology B CELL-INTRINSIC IFN-G SIGNALING DRIVES POLYCLONAL B CELL RESPONSES DURING EXPERIMENTAL PLASMODIUM INFECTION Abstract#: 12

Hadad, Niran ...... 36 Neuroscience CALORIC-RESTRICTION PREVENTS AGE-ASSOCIATED EPIGENETIC CHANGES IN THE BRAIN Abstract#: 13

Presenter ______Page #

Harpool, Kristyn ...... 37 Radiological Sciences A CLINICAL INVESTIGATION TO DETERMINE THE MOST BENEFICIAL RADIATION THERAPY TECHNIQUE FOR PANCREATIC CANCER Abstract#: 14

Hassink, Veronica ...... 102 Communication Sciences and Disorders AFRICAN AMERICAN ENGLISH IN EARLY CHILDHOOD: A COMPARISON OF DEVELOPMENTAL AND DIALECTICAL VARIATIONS IN GRAMMAR Abstract#: 74

Hatipoglu, Monolya Kukut ...... 107 Pharmaceutical Sciences INHALABLE MICROPARTICULATE SHETA2 NANOCRYSTALS FOR LUNG CANCER TREATMENT Abstract#: 78

Hilton, Jeremiah ...... 113 Biostatistics and Epidemiology ABNORMAL WEIGHTING OF MID AND HIGH-FREQUENCY BANDS IN SENSORINEURAL HEARING LOSS Abstract#: 83

Hoover, Christopher ...... 38 Biochemistry & Molecular Biology ROLE FOR NEURAL PODOPLANIN-ACTIVATED PLATELETS IN VASCULAR DEVELOPMENT IN THE BRAIN Abstract#: 15

Houson, Hailey ...... 39 Pharmaceutical Sciences IMAGING OF ISOPROTERENOL-INDUCED MYOCARDIAL INJURY WITH F-18-LABELED FLUOROGLUCARIC ACID IN A RAT MODEL Abstract#: 16

Hriscu, Josephine ...... 126 OSSM COMPARATIVE LINEAGE TRACING OF PDGFR ALPHA AND BETA IN LIMB BONE DEVELOPMENT Abstract#: 95

Jefferson, Keirah ...... 127 OSSM PHENOTYPE-BASED SCREENING OF FUNGAL METABOLITES AND EXTRACTS IN ZEBRAFISH IDENTIFIED MODIFIERS OF NOTOCHORD DEVELOPMENT Abstract#: 96

Presenter ______Page #

Johnson, Anthony ...... 60 Neuroscience SHINING THE LIGHT ON BRAIN CIRCUITS RESPONSIBLE FOR STRESS-INDUCED VISCERAL PAIN Abstract#: 36

Kadel, Sapana ...... 83 Microbiology and Immunology REGULATION OF LUNG RESIDENT TYPE II INNATE LYMPHOID CELLS (ILC2S) BY ERΑ Abstract#: 56

Kamal, Mohamed ...... 108 Stephenson Cancer Center PHENOTYPIC HETEROGENEITY OF CIRCULATING TUMOR CELLS IN BREAST CANCER Abstract#: 79

Kendall, Ethan ...... 40 Radiological Sciences GAMMA KNIFE PERFEXION TREATMENT PLANS FOR TRIGEMINAL NEURALGIA USING SECTOR- BLOCKED AND NON-SECTOR-BLOCKED SHOTS Abstract#: 17

Khan, Asher ...... 114 Pharmaceutical Sciences CHANGES IN HISTAMINE AND ITS HISTAMINE-3 RECEPTOR FOLLOWING A MILD TRAUMATIC BRAIN INJURY Abstract#: 84

Koldoff, Elizabeth ...... 84 Nursing WHY PARENTS PARTICIPATE IN EARLY INTERVENTIONS: PARENT PERSPECTIVE Abstract#: 57

Latorre, Rocco ...... 61 Neuroscience TARGETING THE RECEPTOR KINASE RET FOR THE TREATMENT OF INTESTINAL DISORDERS Abstract#: 37

Lee, Sang-Min ...... 62 Biochemistry & Molecular Biology A SINGLE N-ACETYLGLUCOSAMINE ON ASPARAGINE 130 IN THE CALCITONIN RECEPTOR EXTRACELLULAR DOMAIN SIGNIFICANTLY ENHANCES PEPTIDE HORMONE BINDING AFFINITY Abstract#: 38

Li, Yue ...... 41 Physiology DROSOPHILA TRANSMEMBRANE PROTEIN 214 REGULATES CARBOHYDRATE METABOLISM Abstract#: 18

Presenter ______Page #

Logan, Sreemathi ...... 63 Geriatric Medicine ROLE OF IGF-1 IN ASTROCYTE MITOCHONDRIAL METABOLISM AND BRAIN AGING Abstract#: 39

Long, Cassandra ...... 115 Biostatistics and Epidemiology MODELLING MUMPS TRANSMISSION USING OKLAHOMA OUTBREAK DATA: IS VACCINE EFFICACY LOWER THAN EXPECTED? Abstract#: 85

Long, Cassandra ...... 116 Biostatistics and Epidemiology SPORADIC SALMONELLOSIS CASES AND FOOD CONSUMPTION TRENDS IN OKLAHOMA Abstract#: 86

Longobardi, Sherri ...... 85 GPiBS SPECIFICITY OF SALIVARY GLAND-DERIVED MONOCLONAL ANTIBODIES FROM SJÖGREN’S SYNDROME PATIENTS USING HUMAN PROTEOME ARRAYS Abstract#: 58

Lutter, Mary ...... 103 Communication Sciences and Disorders AFRICAN AMERICAN ENGLISH DIALECT: EXAMINING THE DEVELOPMENT OF GRAMMATICAL FEAURES IN EARLY CHILDHOOD Abstract#: 75

Mahjabeen, Sanjida ...... 86 Pharmaceutical Sciences SHETA2 VAGINAL SUPPOSITORIES TO TREAT CERVICAL DYSPLASIA: INFLUENCE OF ESTROUS CYCLE ON DRUG ABSORPTION AND PHARMACODYNAMIC ENDPOINT Abstract#: 59

Malireddy, SaiSridhar ...... 72 Biostatistics and Epidemiology COST ANALYSIS OF ALLOWING ADDITIONAL TIME IN CLEANING CONTACT PRECAUTION ROOMS. Abstract#: 47

McGrew, Kaitlin ...... 87 Biostatistics and Epidemiology TRANSMISSION DYNAMICS OF COMMUNITY-ACQUIRED STAPHYLOCOCCUS AUREUS COLONIZATION AND INFECTION AMONG INJECTION DRUG USERS IN THE US Abstract#: 60

Presenter ______Page #

Morita, Yoshihiro ...... 109 Stephenson Cancer Center PEGYLATED THIOL-APTAMER FOR PREVENTION OF BREAST CANCER METASTASES Abstract#: 80

Motwani, Anita ...... 117 Biostatistics and Epidemiology AN INVESTIGATION INTO THE RELATIONSHIP BETWEEN INFLUENZA VACCINATION STATUS AND EDUCATION LEVEL FROM 2015 BEHAVIORAL RISK FACTOR SURVEILLANCE SYSTEM (BRFSS) Abstract#: 87

Motwani, Anita ...... 118 Biostatistics and Epidemiology INVESTIGATING THE HIGH SUCCESS OF TOBACCO ABSTINENCE IN WOMEN WITH CONTINGENCY MANAGEMENT INTERVENTION IN THE PREVAIL I STUDY Abstract#: 88

Odejimi, Ope ...... 128 OSSM WORK PERFORMANCE AND RESIDUAL LIMB VOLUME IN MEN WITH TRANSTIBIAL AMPUTATION OVER A 12-MONTH TIMEFRAME Abstract#: 97

Oh, Jackie ...... 129 OSSM IL-24 SUPPRESSES OSTEOPONTIN IN LUNG CANCER CELLS Abstract#: 98

Okong, Kuna Tiga ...... 119 Biostatistics and Epidemiology PREVALENCE OF NEUROCYSTICERCOSIS AMONG PATIENTS WITH SELECT NEUROLOGICAL DISORDERS IN SIXTY VILLAGES IN BURKINA FASO, 2011-2012 Abstract#: 89

Okong, Kuna Tiga ...... 120 Biostatistics and Epidemiology INFLUENZA MORBIDITY AND MORTALITY IN OKLAHOMA, 2013-2016 Abstract#: 90

Oluborode, Babawale ...... 73 Biostatistics and Epidemiology DOES ULTRASOUND APPEARANCE OF THE ENDOMETRIUM AFFECT PREGNANCY RATES IN ASSISTED REPRODUCTIVE TECHNOLOGY TREATMENT? Abstract#: 48

Presenter ______Page #

Onopiuk, Marta ...... 64 Cell Biology CONTROL OF PTH SECRETION BY THE TRPC1 ION CHANNEL Abstract#: 40

Panneerselvam, Janani ...... 65 Pathology IL-24 INHIBITS GLI1 AND INDUCES DNA DAMAGE IN LUNG CANCER CELLS Abstract#: 41

Park, Eunsun ...... 42 Communication Sciences and Disorders AN INVESTIGATION OF THE RELATIONSHIP BETWEEN EYE MOVEMENTS IN SILENT AND ORAL READING AND HYPOKINETIC DYSARTHRIA Abstract#: 19

Parks, Eileen ...... 43 Neuroscience THE IMPACT OF REDUCED NEUROSTEROIDS IN COGNITIVE AGING Abstract#: 20

Patel, Sneha ...... 130 OSSM DIFFERENTIATION OF RAT CARDIOMYOBLASTS H9C2 IN CULTURE Abstract#: 99

Pharaoh, Gavin ...... 44 Physiology MITOCHONDRIAL FUNCTION AND TRANSCRIPTIONAL REGULATION FOLLOWING DENERVATION- INDUCED SKELETAL MUSCLE ATROPHY Abstract#: 21

Porter, Tavanna ...... 88 Genetic Counseling FROM ADVERSITY TO ADVOCACY: A PARENTING CONTINUUM AFTER A CHILD’S DIAGNOSIS OF DOWN SYNDROME Abstract#: 61

Pruett, James ...... 89 Radiological Sciences MANUFACTURING COST EFFECTIVE HETEROGENEOUS PHANTOMS FOR USE IN RADIATION THERAPY Abstract#: 62

Presenter ______Page #

Quinn, James ...... 45 Microbiology and Immunology FOLLICULAR T HELPER CELLS AND T HELPER 17 CELLS COOPERATE IN PROMOTING DISEASE IN NEURO-INFLAMMATION Abstract#: 22

Rodden, Layne ...... 90 Neuroscience A CRISPR-CAS9 BASED APPROACH FOR TARGETED EPIGENETIC MODIFICATION IN FRIEDREICH ATAXIA Abstract#: 63

Royer, Derek ...... 66 Ophthalmology/DMEI ANTIBODY MEDIATES PROTECTION AGAINST VIRAL KERATITIS VIA COMPLEMENT FIXATION AND NEONATAL FC RECEPTOR TRAFFICKING. Abstract#: 42

Sapkota, Bishwa ...... 67 Pediatrics POST-GWAS FOLLOW-UP OF CANDIDATE GENES OF DIABETIC DYSLIPIDEMIA USING NGS AND FUNCTIONAL STUDIES IN ZEBRAFISH Abstract#: 43

Sapkota, Hem ...... 46 Cell Biology COHESION FATIGUE, A NOVEL SOURCE OF CHROMOSOMAL INSTABILITY Abstract#: 23

Sarwar, Zoona ...... 47 Biostatistics and Epidemiology ASSOCIATION OF DIFFERENT TOBACCO PRODUCTS WITH CARDIOVASCULAR DISEASE RISK PARAMETERS AND METABOLIC SYNDROME. Abstract#: 24

Satern, Marissa ...... 91 Genetic Counseling EXPERIENCES OF INDIVIDUALS ADJUSTING TO A RECENT DIAGNOSIS OF AUTOSOMAL DOMINANT RETINITIS PIGMENTOSA Abstract#: 64

Seshadri, Sudarshan ...... 68 Microbiology and Immunology ANTHRAX LETHAL TOXIN SUPPRESSES THE MAP KINASE PATHWAY TO DECREASE IL-22 PRODUCTION IN TYPE 3 INNATE LYMPHOID CELLS Abstract#: 44

Presenter ______Page #

Sharif, Rabab ...... 48 Cell Biology KERATOCONUS: DEVELOPMENT OF A NOVEL HUMAN CORNEAL COLLAGEN CROSS-LINKING 3-D IN VITRO MODEL Abstract#: 25

Shin, Young-Hwa ...... 49 Physiology A DOMINANT MUTATION IN RPE65, D477G, DELAYS DARK-ADAPTATION AND DISTURBS THE VISUAL CYCLE IN THE MUTANT KNOCK-IN MICE Abstract#: 26

Shrestha, Binu ...... 110 Microbiology and Immunology NKTFH CELLS STIMULATE IGG1 CLASS SWITCH AGAINST CARBOHYDRATE ANTIGEN Abstract#: 81

Siefert, Joseph ...... 50 Cell Biology DNA REPLICATION TIMING DURING DEVELOPMENT ANTICIPATES TRANSCRIPTIONAL PROGRAMS AND PARALLELS ENHANCER ACTIVATION Abstract#: 27

Sjoelund, Virginie ...... 132 Core Facilities OUHSC LABORATORY FOR MOLECULAR BIOLOGY AND CYTOMETRY RESEARCH Abstract#: 100

Smith, Chelsea L...... 51 Nutritional Sciences HEALTH CARE PROVIDER’S ROLE IN OBESITY PREVENTION AND HEALTHY DEVELOPMENT OF YOUNG AMERICAN INDIAN CHILDREN Abstract#: 28

Smith, Diane ...... 92 Nursing LIVED EXPERIENCES OF OKLAHOMA CAREGIVERS OF SPOUSES WITH FRONTOTEMPORAL DEMENTIA: A QUALITATIVE PILOT STUDY Abstract#: 65

Smith, Jacob ...... 104 Medical Imaging A DOSIMETRIC COMPARISON AND CLINICAL IMPLICATIONS OF PELVIC INSUFFICIENCY FRACTURES (PIF) IN POST-MENOPAUSAL WOMEN TREATED FOR UTERINE AND CERVICAL CANCERS WITH IMRT VS. 3DCRT Abstract#: 76

Presenter ______Page #

Srivastava, Akhil ...... 69 Pathology NANOSOME: A SMART DRUG CARRIER FOR TREATING LUNG CANCER Abstract#: 45

Starnes, Andrew ...... 121 Biostatistics and Epidemiology HOSPITAL ADMISSION STATUS AND FIRST-TIME DIAGNOSIS OF VENOUS THROMBOEMBOLISM IN A CONTEMPORARY POPULATION-BASED COHORT IN OKLAHOMA COUNTY Abstract#: 91

Terzyan, Simon ...... 133 Biochemistry & Molecular Biology THE LABORATORY OF BIOMOLECULAR STRUCTURE AND FUNCTION AT OUHSC Abstract#: 101

Tran, Hai ...... 105 Allied Health Sciences ASSESSING THE ROLE OF A CONTOURING ACCURACY PROGRAM IN STANDARDIZING MEDICAL DOSIMETRY STUDENTS' ANATOMICAL CONTOURING SKILLS. Abstract#: 77

Twum-Ampofo, Nana ...... 93 Allied Health Sciences CENTER OF PRESSURE AND MOVEMENT PROFICIENCY IN INFANTS WITH AND WITHOUT CP USING THE SELF INITIATED PRONE PROGRESSION CRAWLER-2 Abstract#: 66

Veirs, Kimberly ...... 94 Allied Health Sciences PREVALENCE OF DANCE-RELATED INJURY OF ADOLESCENT FEMALE BALLET DANCERS TRAINING EN POINTE: A SYSTEMATIC REVIEW Abstract#: 67

Wright, Anna ...... 95 Genetic Counseling ATTITUDES TOWARD CARRIER SCREENING IN FAMILIES AFFECTED BY MUCOPOLYSACCHARIDOSIS Abstract#: 68

Wright, Olivia ...... 122 Biostatistics and Epidemiology EXPLORING SELECTION BIAS DUE TO INADVERTENT USE OF A RISK FACTOR FORM IN A HEPATITIS C SCREENING STUDY. Abstract#: 92

Presenter ______Page #

Xu, Yifan ...... 52 Pathology MIR-1246 IS PRIMARILY DERIVED FROM U2 SMALL NUCLEAR RNA 1 (RNU2-1) AND HIGHLY ENRICHED IN PANCREATIC CANCER EXOSOMES Abstract#: 29

Yari, Hooman ...... 96 Pharmaceutical Sciences PROTEASOME ACTIVITY IS ELEVATED IN ILEAL TISSUE OF RATS WITH HEMORRHAGIC SHOCK Abstract#: 69

Zauner, Katherine ...... 53 Nutritional Sciences PRELIMINARY ANALYSIS OF THE EFFECT OF PREHABILITATION ON MUSCLE MASS, WEIGHT, AND DIETARY INTAKE STRATIFIED BY HAND GRIP STRENGTH IN PANCREATIC CANCER PATIENTS Abstract#: 30

Ziegler, Jadith ...... 54 Pathology XZ488 AS A NOVEL THERAPY AGAINST GLIOMAS Abstract#: 31

University of Oklahoma Health Sciences Center Graduate College Graduate Research Education and Technology Symposium

Abstract #1

ROLE OF BRG1 (SMARCA4) IN THE RESISTANCE OF NON-SMALL CELL LUNG CANCER (NSCLC) TO TYROSINE KINASE INHIBITORS (TKIS)

Rebaz Ahmad 1,4, Ranganayaki Muralidharan 1,3, Anupama Munshi 2,3, Rajagopal Ramesh 1,3,5. Departments of 1Pathology, 2Radiation Oncology, 3Stephenson Cancer Center, and 4Graduate Program in Biomedical Sciences, The University of Oklahoma Health Science Center, Oklahoma, 73104, USA.

Background: Epidermal growth factor receptor (EGFR) is overexpressed in several human cancers including lung cancer and has become an important molecular target for treatment. The occurrence of gain of function via activating mutations in the intracellular kinase domain (IKD) of EGFR provides signaling cues for cell proliferation, differentiation, and survival. Inhibition of EGFR using tyrosine kinase inhibitors (TKIs), (e.g. Gefitinib), is a well-known treatment for NSCLC. Unfortunately, a subset of patients acquire resistance to EGFR- TKI treatment over time warranting improved therapies. Recent studies indicate a role for BRG1 and other subunits of the large SWI/SNF complex in modulating the response to anticancer therapies including TKIs. However, little is known whether BRG1 status influences the response to EGFR-TKIs.

Methods: Cell sensitivity to Gefitinib (GF) was assessed in lung cancer cell lines with different BRG1 and EGFR status. Cells that were wild-type (wt) BRG1 and EGFR (H358); wt BRG1 and activated mutated EGFR (HCC827); mutant (mt) BRG1 and wt EGFR (A549) were used. The expression of BRG1, EGFR and pEGFR (Tyr1173) in the GF-treated cells were detected by western blotting. DMSO-treated cells were used as controls.

Results: Cell viability studies showed that the cell lines responded differently to GF. H358 was resistant, A549 demonstrated intermediate sensitivity and HCC827 was sensitive. Western bot results showed the upregulation of BRG1 while EGFR was downregulated in BRG1 wt/mt EGFR HCC827 cell lines treated with GF. In contrast, mt BRG1/wt EGFR cell line (A549) showed no change in EGFR expression levels when treated with GF.

Conclusion: The upregulation of BRG1 in GF-treated cell lines that are BRG1 wt suggests possible molecular interaction between EGFR and BRG1 and that BRG1 possibly influences resistance to EGFR-TKIs. Demonstrating a role for BRG1 in EGFR-TKI resistance will allow in new and improved combinatorial treatment strategies targeting both BRG1 and EGFR.

Funding: The study was supported in part by funds received from the Presbyterian Health Foundation Bridge Grant and the Experimental Therapeutics Seed Grant from Stephenson Cancer Center.

Abstract #2

ENDOGENOUS TIME VARYING COVARIATES AND TIME VARYING CONFOUNDING ASSOCIATIONS: DEFINING AND ADJUSTING FOR THEM

Jonathan Baldwin1, David Thompson1 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Science Center

Introduction: Longitudinal models containing time varying covariates (TVC) must be carefully constructed when endogenous associations may be present. An important example of an endogenous association is encountered when exposure variables are associated with previous outcomes, and are regarded as endogenous TVCs. Adjusting for ETVCs in longitudinal models can’t be accomplished by conventional means, but particular structural models that use inverse probability of treatment weighting (IPTW) are required. The goal of this study is to inform the audience about ETVCs and time varying confounding associations, and to describe the structure and performance of models that adjust for them.

Methods: A comprehensive literature review included searches for: endogenous covariates, time dependent confounding, inverse probability weighting, marginal structural models, and linear structural models. To examine statistical approaches for ETVC adjustment, we simulated a population of individuals among whom a covariate, moderate physical activity (MPA), has a known endogenous association with an outcome (BMI). 500 samples of 100 individuals were drawn from the simulated population, and linear structural models were constructed and compared to a naïve random intercept model which neglected endogenous associations.

Results: We compared models by examining mean estimates compared to known population value, standard deviation of the estimates, coverage of true values within 95% confidence intervals, and mean square error from the known population value. In addition, we compared models after removing confounding covariates to simulate situations encountered in observational studies where certain confounding variables are unmeasured.

Conclusion: Endogenous associations are often ignored in longitudinal analysis. This study describes construction of linear structural models with IVPW, and demonstrates the benefits and shortcomings of these models compared to common longitudinal methods.

Funding: None

Abstract #3

THE AMPI CLASSIFICATION OF THE DYSARTHRIAS

Zachary Barrett1, Frank R. Boutsen1, and Justin D. Dvorak1,2 1 Motor Speech and Prosody Research Laboratory, University of Oklahoma Health Sciences Center 2 Department of Biostatistics & Epidemiology, University of Oklahoma Health Sciences Center

Introduction: The dysarthrias have been classified using the auditory-perceptual method along the Mayo Classification system. Recently, investigations used prosodic matrices to see if they differentiate the dysarthrias. Liss et al. (2009) found that a combination of these matrices can distinguish the dysarthrias while Lowit (2014) reported that none of the matrices could do this. The present investigation evaluates the newly developed Acoustic Multidimensional Prosody Index (AMPI) when classifying the dysarthrias.

Methods: Audio recordings of the Grandfather Passage were taken from 57 participants and then digitized at 44,100 Hz with 16-bit resolution. Vowel nuclei were identified for a sample of the passage using SpeechMark, then vowel nuclei were confirmed and data was extracted in custom MATLAB® software. Measurements for each vowel included duration (ms), intensity (dB), and pitch (Hz). For each pair of groups, prosody indices were computed at the patient level following Dvorak, Boutsen, & Ding (submitted), with weight vectors optimized for AUC via grid search. Those weights which maximally separated each pair of groups were extracted, and the corresponding AUC was assessed for significance via permutation test with 1,000 iterations.

Results & Discussion: Overall, AUC of each binary classification was 80%, and ranged from 68% (flaccid vs. spastic; p=0.1820) to 93% (control vs. hypokinetic; p<0.0001). Of the 15 group comparisons, 10 were significant at α=0.05. Adjusting for multiple testing, the prosody index distinguished ataxic vs. hypokinetic (AUC=87%, p=0.0150), ataxic vs. spastic (AUC= 89%; p=0.0300), and control vs. hypokinetic (AUC=93%; p<0.0001). These results show that controls differ prosodically from dysarthria subtypes. Duration control is driving the differences in (C vs S) and pitch in (C vs F and C vs O) and intensity in (C vs R). Different acoustic dimensions differentiate different types from the controls. Subtypes also were shown to differ from one another (7/11 contrasts, or 64%)

Funding: None

Abstract #4

EXCESSIVE PLASMIN ACTIVITY PROMOTES ENDOTHELIAL CELL NECROPTOSIS AND EMBRYONIC VASCULAR RUPTURE

Sarah Colijn1,2, Kyle G. Ingram1,2, Matt Menendez2, Vijay Muthukumar2, Robert Silasi-Mansat2, Florea Lupu1,2, & Courtney T. Griffin1,2 1Department of Cell Biology, University of Oklahoma Health Sciences Center 2Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation

Necroptosis is a newly identified form of programmed cell death that mimics accidental necrosis in its morphology and in the release of pro-inflammatory molecules. Necroptosis is distinct from accidental necrosis because it requires a series of intracellular kinases that mediate plasma membrane destruction. Overexpression of one of these kinases, RIPK3, can cause spontaneous necroptosis in some cell types in vitro, however the mechanisms regulating RIPK3 expression in vivo are unknown. We found that mutating a chromatin-remodeling enzyme, CHD4, in the developing murine vasculature caused excessive plasmin activation, extracellular matrix (ECM) proteolysis, endothelial cell (EC) Ripk3 transcription, EC necroptosis, and vascular rupture. We are now exploiting our Chd4 mutant embryos and cell culture systems to study Ripk3 transcriptional regulation in ECs. We have shown that neither CHD4 itself nor matrix detachment directly regulates Ripk3 transcription in ECs. Our data show that plasmin, which is excessively activated in CHD4 mutant embryos, can induce Ripk3 transcription in vitro independently of CHD4 in a time-dependent manner. We have also established that this Ripk3 upregulation is a result of active transcription as opposed to improved transcript stability. Our current data suggest that plasmin directly activates Ripk3 transcription in ECs by cleaving PAR1/4, thus inducing PAR-related intracellular signaling. Our new knockdown data suggest that PAR1 may play an inhibitory role in Ripk3 transcription that is halted upon plasmin cleavage. Conversely, our knockdown data indicate that PAR4 activates Ripk3 transcription upon plasmin cleavage. This work has important implications for our mechanistic understanding of RIPK3-mediated EC necroptosis, which is detrimental to vascular integrity.

Funding: OMRF Barrett Scholarship, NHLBI, NIGMS, OCAST, and PHF Seed Grant

Abstract #5

EVALUATION OF ANTI-TUMOR IMMUNITY IN THE MMTV-PYMT MOUSE

Christa I. DeVette1, William Hildebrand1, Alana L. Welm2, 1University of Oklahoma Health Sciences Center, United States 2Huntsman Cancer Institute, United States

A number of new tools are available for characterizing T cell immunity to tumors. The FVB mouse represents one such emerging tool for cancer biologists – these mice exhibit clinically relevant breast cancer development and spontaneous metastases when transgenically expressing PyMT. Surprisingly, the class I MHC, which present tumor associated peptides for T cell recognition, remain uncharacterized in FVBs. To strengthen this mouse model as an immunologic tool, we combine a PyMT vaccine with immunoproteomics, creating a predictive algorithm for identification of peptide ligands presented by the FVB MHC class I molecules H2-Kq and -Dq. We then apply this algorithm to PyMT encoded peptide ligands that are presented to T cells by these MHC I molecules. A PyMT DNA vaccine leads to T cell mediated elimination of MMTV-PyMT tumors. To identify PyMT tumor peptides that are targeted by T cells, we generated an overlapping peptide library to test in vaccinated animals. Conformational epitopes were determined using soluble MHC. HeLa and 721.221 cells were transfected with PyMT and soluble H2-Kq/Dq. Cells secreting these MHC I were cultured in roller bottles, peptide-bearing MHC I were affinity purified, and presented peptides were analyzed by mass spectrometry. The peptide sequence data was used to develop a predictive algorithm to enable in silico candidate peptide searches. We are currently screening candidate PyMT peptides by γ-interferon ELISPOT using splenocytes from PyMT vaccinated mice. These ligands will be validated in MMTV-PyMT tumor-bearing mice treated with checkpoint inhibitors. Developing a PyMT tumor vaccine model in the FVB mouse will allow us to study poorly understood antigen specific responses in breast cancer.

Funding: NIAID/NIH Training Grant (T32AI007633) (CID) DOD Breast Cancer Research Program Innovator and Scholar Concept Award (AW)

Abstract #6

A NON-LINEAR AND SEGMENTED REGRESSION APPROACH TO EXPLORING TRENDS OF COATED- PLATELET LEVELS IN SUBARACHNOID HEMORRHAGE PATIENTS

Lance Ford1, Bappaditya Ray2, David Thompson1 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center 2Department of Neurology, University of Oklahoma Health Sciences Center

Introduction: Non-linear regression and segmented linear regression are statistical modeling techniques that can be used to make inferences about longitudinal data and detect evidence of change in linear trend. These modeling techniques are applied in the analysis of coated-platelet levels in patients who have experienced a subarachnoid hemorrhage. It is of interest to find a change of trend in coated-platelet levels and to compare groups by patient characteristics such as clinical outcomes and severity of presentation. This methodological approach can be generalized to other longitudinal studies.

Methods: Coated-platelet information was collected recurrently from 45 patients who experienced a subarachnoid hemorrhage. Non-linear regression was used to determine a change of trend in coated-platelet levels; as a result, an estimated day of transition is obtained. Segmented regression models were built to predict rates of change in coated-platelet levels over time and to test between-group differences in these rates before and after the day of transition. These groups were defined by outcomes such as delayed cerebral ischemia (DCI).

Results: Non-linear regression models detected a change in the trend of coated-platelet levels at day 3.75 (95% CI: 2.45, 5.05) after event. However, the non-linear models were unable to determine between-group differences. Because we specify, rather than estimate, the day of transition in the segmented model, it has more statistical power to estimate between-group differences in trend. For groups defined by DCI status, there was a significant difference in between-group rates of change before (p < 0.0001) and after (p < 0.0001) day of transition.

Conclusion: This study demonstrates, through analysis of coated-platelet levels in patients with subarachnoid hemorrhages, the limitations and strengths of using non-linear regression and segmented regression techniques when exploring changes of trend in longitudinal data.

Funding: OSCTR: U54GM104938

Abstract #7

THE EFFECT OF SENSORINEURAL HEARING LOSS ON COGNITIVE ASSESSMENT SCORES

Laura Gaeta1, Andrew John1, Carrie Ciro2, Carole Johnson1, Mary Hudson1, and Jo Azzarello3 1Department of Communication Sciences and Disorders, University of Oklahoma Health Sciences Center 2Department of Rehabilitation Sciences, University of Oklahoma Health Sciences Center 3College of Nursing, University of Oklahoma Health Sciences Center

Introduction: The interaction of audition and cognition has been of interest to researchers and clinicians, especially as the prevalence of hearing loss and cognitive decline increases with advancing age. A consequence of the interaction between hearing and cognitive change is erroneous interpretation of incorrect responses to unheard or misheard verbal questions. The purpose of this study was to examine the effects of aided and unaided sensorineural hearing loss (SNHL) on the Mini-Mental State Examination (MMSE), a common bedside cognitive screening instrument.

Methods: A 1:1 gender-matched case-control design was employed for this study. To date, 17 older adults (60 to 80 years old) with mild to moderately severe SNHL and hearing aids (cases) and nine young adults (18 to 35 years old) with normal hearing (controls) have completed all measures. Case participants were administered a recorded version of the MMSE in three counterbalanced conditions (without hearing aids, with hearing aids, and with a listening device) in background noise. Controls were administered a digitally altered version of the MMSE that had been filtered to reflect the hearing loss of the matched case. Performance on the MMSE was scored using standard criteria and response times to test items were measured.

Results: Data analysis is preliminary and will be complete by presentation. Initial findings show decreased audibility due to SNHL had a significant effect on MMSE scores. Additional analyses are in progress to determine whether these effects are equivalent for matched controls and whether there is an effect of hearing loss on response time for MMSE items.

Conclusion: Preliminary findings demonstrate that SNHL has a significant effect on MMSE score in otherwise cognitively intact older adults. Results support the use of amplification when administering cognitive and other assessments. These results may provide further evidence of the interaction between hearing loss, cognition, and aging.

Funding: None

Abstract #8

THE ROLE OF PLASMIN IN POSTNATAL HEPATIC VASCULAR INTEGRITY AFTER LIVER INJURY

Siqi Gao1, 2, Courtney Griffin1, 2 1Department of Cell Biology, University of Oklahoma Health Sciences Center 2Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation

The liver is a major internal organ with multiple biological functions. Our lab previously found that embryonic hepatic sinusoidal vessels are susceptible to rupture after midgestation if their surrounding extracellular matrix (ECM) is degraded by the protease plasmin. Our lab also has generated preliminary evidence that excessive plasmin activity facilitates programmed necrosis (necroptosis) in embryonic endothelial cells, which contributes to vascular rupture. These findings have led us to question the effect of excessive plasmin activity on postnatal hepatic vascular maintenance and integrity. We are using an acetaminophen (APAP) overdose mouse model to study the effects of plasmin on liver vasculature. APAP is one of the most widely used analgesics in the world and is the leading cause of acute liver failure when consumed at overdose levels. It has been reported that plasmin contributes to the liver damage induced by APAP overdose. However, the relationship between plasmin activity, APAP-induced vascular damage, and overall liver failure is understudied. The objective of this research is to identify mechanistic causes and effects of hepatic vascular damage following APAP overdose. We found 8- to 10-week-old male mice achieve significant liver toxicity and centrilobular hemorrhage within 6-24 hour after APAP overdose compared to saline-injected controls. We also found that plasmin activity is upregulated at 6 hr after APAP overdose. In our hands, use of the pharmacological plasmin inhibitor tranexamic acid or partial genetic reduction of plasminogen, the circulating zymogen of plasmin, ameliorates APAP-induced hepatic vascular injury. These studies are the first to demonstrate the time course of plasmin activation after APAP overdose and to suggest that partial reduction of plasmin activity stabilizes vascular integrity after APAP overdose.

Funding: None

Abstract #9

A PROGRAM FOR FETAL DOSE ESTIMATION IN RADIOGRAPHIC EXAMINATIONS

Victor L. Garcia, Weiyuan Wang

Purpose: Exposure to radiation can present potential risks and hazards to the fetal growth and development. Therefore, it is necessary to develop a program to estimate dose to the conceptus for diagnostic exams on various diagnostic radiographic systems throughout the OUHSC campus.

Methods: Calculations are done using the Percent Depth Dose (PDD) method. As per PDD, the quantities needed from the diagnostic exam are peak kilovoltage (kVp), milliampere-seconds (mAs), patient thickness, fetal depth, and X-ray field size at the image receptor, and the half value layer (HVL) and exposure per milliampere- seconds (mR/mAs), which are found in annual reports for individual systems. Field size and exposure are calculated at the location where the x-ray field enters the patient. Using the previous values, the percent of dose at fetal depth can be determined to calculate dose.

Results: The program calculates the fetal dose as functions of kVp, mAs, patient thickness, fetal depth, and field size. Calculations in the program account for anterior-posterior and lateral exposures to the patient.

Conclusion and Future Work: Radiation risk is considered negligible under 50 mGy and significantly increased above 150 mGy. Using the calculations, risk from fetal exposure to radiation can accurately be assessed for patients. Further research will continue to develop the program to estimate fetal dose for other imaging modalities, such as computed tomography (CT), fluoroscopy, mammography, etc., as well as extending the program to dose estimations for pediatric and adult patients.

Funding: Radiological Sciences Department

Abstract #10

IGG AND SAP SUPPORT INNATE IMMUNE RESPONSES TO BACILLUS ANTHRACIS PEPTIDOGLYCAN VIA DISTINCT AND REDUNDANT MECHANISMS.

Alanson Girtona,b, Narcis Popescub, Ravi Kesharic, and K. Mark Coggeshallb a, Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Blvd., BMSB 1053, Oklahoma City, OK, 73104, United States. b, Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, 825 NE 13th St, Oklahoma City, OK, 73104, United States. c, Cardiovascular Biology, Oklahoma Medical Research Foundation, 825 NE 13th St, Oklahoma City, OK, 73104, United States.

Pulmonary infection by Bacillus anthracis leads to a rapid onset of systemic disease that progresses to overwhelming bacteremia and severe septicemia in both animal models and human patients. Sepsis pathophysiology is driven by interactions between the innate immune system and pathogen-associated molecular patterns (PAMPs). One such PAMP is peptidoglycan (PGN). Recently, we have sought to understand the mechanisms that govern immune cell responses to PGN. We showed that responses by human monocytes, neutrophils, platelets and the complement system to PGN require human serum and that human IgG supports these responses. These data are consistent with a model in which naturally occurring anti-PGN IgG opsonizes PGN for recognition and response by human immune cells and the classical complement pathway. However, we found no correlation between anti-PGN IgG titer and monocyte inflammatory responses to PGN. The lack of correlation suggests that either other opsonins are present in human serum or that PGN is recognized in other ways. We sought to determine if TLR2, a member of the toll-like receptor family and proposed PGN receptor, could support monocyte responses to B. anthracis PGN. We found murine bone marrow-derived macrophages failed to respond to B. anthracis PGN while human monocytes responded well in the presence of human serum constituents, leaving the possibility that other opsonins are present in human serum. We have found that the novel peptidoglycan recognition protein serum amyloid P (SAP) can bind PGN and support monocyte TNFα responses to PGN. These responses to PGN relied on phagocytosis and Src family kinase signaling, consistent with Fcgamma receptor-mediated phagocytosis. However, unlike IgG, SAP was unable to induce complement activation in the presence of PGN. These data suggests that IgG and SAP support innate immune responses through both distinct and redundant mechanisms. Therapies targeting these interactions may prove beneficial for sepsis.

Funding: 5U19AI062629-14

Abstract #11

IDENTIFICATION OF SYSTEMIC LUPUS ERYTHEMATOSUS CAUSAL RISK VARIANT CANDIDATES SPANNING UBE2L3 HAPLOTYPE.

Jaanam Gopalakrishnan1,3, Shaofeng Wang2, Satish Pasula3, Ajay Nair3, Mandy Wiley3 and Patrick M Gaffney1,3. 1Department of Pathology, University of Oklahoma Health Science Center, Oklahoma City, OK, USA. 2Bethune Institute of Epigenetic Medicine, The First Hospital of Jilin University, Changchun, China. 3Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA.

Systemic Lupus Erythematosus (SLE) is a prototype systemic autoimmune disease. Genome-wide association studies (GWAS) have fine mapped a 67kb risk haplotype that spans the UBE2L3 gene to confer disease risk in SLE. UBE2L3 encodes an E2 ubiquitin-conjugating enzyme (UbcH7) involved in the ubiquitin proteasome pathway. The UBE2L3 risk haplotype is found to significantly increase UBE2L3 expression. Despite the power of GWAS in identifying disease susceptibility loci, major challenges lie ahead in identifying the precise causal variants responsible for these statistically significant associations. In this study, we aim to identify the potential SLE causal variant candidates among the single nucleotide polymorphisms(SNPs) spanning UBE2L3 risk haplotype. Methods: Five variants within the UBE2L3 risk haplotype that overlap the chromatin accessible regions (ATAC peaks) in T or B lymphocytes or monocytes from SLE patients were selected for screening causal variant candidates. Electrophoretic Mobility Shift Assays (EMSA) were performed to evaluate the differential binding ability of non-risk and risk alleles of individual variants to nuclear proteins. Nuclear proteins were extracted from T lymphocytes (Jurkat) with or without PMA/Ionomycin stimulation for 2hrs. Results: Variants rs40490, rs12484550, rs3747093 showed increased binding of nuclear protein complexes to the risk alleles compared to non-risk when incubated with Jurkat cell nuclear extracts with or without stimulation, whereas variant rs9621715 demonstrated increased binding to the risk allele only in the un-stimulated nuclear extract. Variant rs5998599 showed decreased binding to the risk allele compared to the non-risk in nuclear extracts with or without stimulation. Our results indicate that 4 out of the 5 variants tested have increased binding of nuclear proteins to the risk allele compared to non-risk either with or without stimulation. Conclusion: Increased binding observed in four variants suggests enhanced binding of transcription factors to the risk allele and might account for increased UBE2L3 expression contributing to SLE risk.

Funding: NIH RO1 - ARO63124, P30 - GM110766

Abstract #12

B CELL-INTRINSIC IFN-G SIGNALING DRIVES POLYCLONAL B CELL RESPONSES DURING EXPERIMENTAL PLASMODIUM INFECTION

Jenna J. Guthmiller1, Rosemary L. Pope1, Michelle L. Ratliff2, Alisha Chitrakar1, Carol F. Webb1,2, Demin Wang3, Noah S. Butler1,4 1Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 2 Department of Medicine, Division of Rheumatology, Immunology, and Allergy, University of Oklahoma Health Sciences Center, Oklahoma City, OK 3Blood Research Institute, BloodCenter of Wisconsin, Milwaukee, WI 4Department of Microbiology, University of Iowa, Iowa City, IA

Plasmodium infections and malarial disease remain global health crises. Antibodies are essential for limiting disease severity and for promoting parasite control/clearance. However, antibody responses are short-lived and often non-neutralizing, leaving individuals susceptible to reinfections and persistent parasitemia. Human and murine models show that Plasmodium infections drive polyclonal B cell activation, non-specific antibody responses, and hyperglobulinemia. However, the B cell subsets producing non-specific antibodies, the longevity of these responses, and the factors that drive polyclonal B cell responses are not known. Here we report that experimental Plasmodium yoelii infection drives the expansion of short-lived plasmablasts that are largely directed against irrelevant, non-Plasmodium-specific, antigens. Plasmablasts are polyclonally activated and secrete short-lived antibodies against non-parasite derived foreign- and self-antigens. Strikingly, polyclonal B cell activation and non-specific antibody responses were CD4 T cell-dependent and magnified by B cell intrinsic- IFN-g signaling. Collectively, our data identify that Plasmodium infection and IFN-g-mediated inflammation drive the expansion of short-lived, non-specific plasmablasts. Our data corroborate accumulating data supporting that the induction of plasmablast responses may be a mechanism of Plasmodium immune evasion and a poor prognostic of protective immunity against malaria.

Funding: This work was supported by grants from the American Heart Association (16PRE27660002 to J.J.G.), the National Institutes of Health/National Institute of Allergy and Infectious Diseases (R01AI125446 and R01AI127481 to N.S.B.), and the National Institutes of Health/National Institute of General Medical Sciences under Grant 8P20GM103447.

Abstract #13

CALORIC-RESTRICTION PREVENTS AGE-ASSOCIATED EPIGENETIC CHANGES IN THE BRAIN

Niran Hadad1,2, Dustin R. Masser2,3, David R. Stanford2,3, Archana Unnikrishnan2,4, Arlan Richardson2,4, Willard M. Freeman1,2,3,4 1Oklahoma Center for Neuroscience, 2Reynolds Oklahoma Center on Aging, 3Department of Physiology, 4Department of Geriatric Medicine

Brain aging is characterized by cognitive decline and increased risk to neurodegenerative disease development. In the CNS, epigenetic mechanisms are vital to proper cellular function and memory formation. Aberrant epigenomic control, specifically in the methylome, is evident with aging and age-related disease. Anti-aging treatments, such as caloric restriction (CR), increase neurogenesis and induce expression of neuroprotective genes in the aged brain. However, the mechanisms underlying these changes remain unknown. In this study, we investigated whether CR, the most proven anti-aging treatment, prevents age-related changes in DNA methylation. To determine the effect of CR on age-related differential methylation in the brain, hippocampal tissue was collected from young (3M) and old (24M) mice fed ad lib diet and 24M old mice calorie restricted from 3M to 24M of age. Hippocampal DNA was extracted and used for bisulfite oligonucleotide capture sequencing to determine DNA methylation levels, genome-wide, at a base-specific resolution. A large number of differentially methylated CpGs (dmCGs) were evident with aging, of which 34% were prevented by CR. CR specific dmCGs were also evident. dmCGs with both age and CR were enriched in CpG island-shelves and gene bodies. Age- related dmCGs unaffected by diet were enriched in H3K4me1 while CR prevented age dmCGs were enriched in H3K27me3. Age-dmCGs affected genes were over-represented in in inflammatory pathways but this was not observed in age-matched CR animals. Genes affected by CR-dmCGs and age-dmCGs, although being different sets of genes, were often co-enriched in similar pathways. Our findings demonstrate for the first time that caloric restriction prevents age-induced changes in DNA methylation in the brain. DNA methylation changes induced by CR were also independent of age, suggesting CR in part counter-balance the aging process. The prevention of age-dmCGs by CR highlights the prominent role of DNA methylation as a regulator of the aging process.

Funding: Reynolds Oklahoma Center on Aging, Oklahoma Nathan Shock Center of Excellence in the Biology of Aging (P30AG050911) and OCNS translational seed grant

Abstract #14

A CLINICAL INVESTIGATION TO DETERMINE THE MOST BENEFICIAL RADIATION THERAPY TECHNIQUE FOR PANCREATIC CANCER

Kristyn Harpool, Erich Schnell, Salahuddin Ahmad, Terence Herman, Tania De La Fuente. The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma. U.S.A.

Introduction: The Department of Radiation Oncology strives to obtain advancements in technology for improving the treatment of cancer with radiation therapy. Previously, Intensity Modulated Radiation Therapy (IMRT) was considered the most advanced treatment technique available but Volumetric Modulated Arc Therapy (VMAT) has now emerged. Assuming it is a better form of treatment, there is an increasing demand for the VMAT technique to treat patients; therefore, it was of interest to study potential advantages of VMAT compared to IMRT, and specifically, when creating a treatment plan for borderline resectable pancreatic cancer lesions. This investigation was based on dosimetry and radiobiology models.

Methods: Twenty pancreatic cancer treatment plans were analyzed. Original plans were re-planned and calculated with Varian Eclipse treatment planning system using VMAT or IMRT to create paired data sets. IMRT plans utilized 6-9 fields and VMAT plans used two arc fields. Both techniques used 6 MV beams and prescription dose of 4950 cGy in 18 fractions. Plan evaluations used Conformity Index (CI) and normal tissue (kidneys, liver, spinal cord, and bowel) doses analyzed with QUANTEC guidelines; and radiobiology analysis using the Uncomplicated Tumor Control Probability (UTCP). All comparisons had two-tail’s student t-test (P< 0.05 considered statistically significant).

Results: The VMAT resulted in 6.5%, 9.5%, and 18.5% higher mean doses to the right and left kidneys and liver, and 7.5% and 0.4% lower doses to the bowel and spinal cord, respectively. Though CI was comparable with 1.21 (IMRT), 1.18 (VMAT), their differences were significant (p=0.03). UTCPs were comparable with 0.92 (IMRT) and 0.91 (VMAT) but not significant (p = 0.17).

Conclusions: The VMAT technique did not result in a significantly superior form of treatment at providing target coverage and sparing normal structures. For good clinical practice, determination of the treatment technique should be based on individual cases.

Funding: None

Abstract #15

ROLE FOR NEURAL PODOPLANIN-ACTIVATED PLATELETS IN VASCULAR DEVELOPMENT IN THE BRAIN

Christopher Hoover, Lijun Xia Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center and the Oklahoma Medical Research Foundation

Vascular development within the central nervous system (CNS) progresses through extremely active sprouting angiogenesis. Nascent vascular sprouts at the angiogenic front are sensitive to bleeding, but the mechanisms that maintain their integrity are not fully understood. The O-glycoprotein podoplanin (PDPN) and its platelet receptor C-type lectin-like receptor 2 (CLEC-2) are required for vascular integrity in the developing CNS, but the defects resulting in hemorrhages in Pdpn-/- and Clec-2-/- embryos and the mechanism of PDPN-CLEC-2- mediated platelet signaling are unknown. We hypothesize that this interaction is crucial for supporting structural stability of the developing nascent vasculature in the CNS. To test this, we analyzed vascular development in the CNS and found that KOs had significant decreases in sprout ingression prior to bleeding. Two-photon microscopy revealed that the lower diencephalon had abnormal vascular development with large dilated sprouts. Quantification of the diameter of nascent primary sprouts showed significantly more were dilated in KO embryos. These sprouts were packed with red blood cells and a source of hemorrhaging. Semi-thin section imaging found an irregular extracellular matrix (ECM) and transmission electron microscopy revealed unstable endothelial cells of nascent sprouts in Pdpn-/- CNS. CLEC-2-activated platelets secrete the molecule sphingosine-1-phosphate (S1P), which is known to counteract VEGF induced permeability and maintain vascular integrity. We treated pregnant dams with a potent S1PR1 agonist and found that it significantly decreased bleeding severity in Pdpn- /- embryos, though large aneurisms were observed on histology. RNAseq was performed on E10.5 WT and Pdpn- /- CNS to evaluate gene expression. Differential expression analysis revealed increased immune response, ECM component expression, and VEGF signaling in KO. Staining for collagen IV showed incomplete coverage on sprouts and PDPN was highly expressed in areas without it. This work reveals a possible role for neural PDPN as an ECM component to maintain sprout integrity in the CNS.

Funding: OMRF Pre-Doctoral Scholarship

Abstract #16

IMAGING OF ISOPROTERENOL-INDUCED MYOCARDIAL INJURY WITH F-18-LABELED FLUOROGLUCARIC ACID IN A RAT MODEL

Hailey A. Houson BS, Gregory N. Nkepang PhD, Andria F. Hedrick MS, and Vibhudutta Awasthi PhD Department of Pharmaceutical Sciences, University of Oklahoma Health Science Center, 1110 N. Stonewall Avenue, Oklahoma City, OK 73117 (USA)

Positron emission tomography (PET) of myocardial infarction (MI) by infarct avid imaging has the potential to reduce the time to diagnosis and improve diagnostic accuracy. The objective of this work was to synthesize 18F- labeled glucaric acid (FGA) for PET imaging of isoproterenol (ISO)-induced cardiomyopathy in a rat model. Methods: We synthesized 18F-FGA by controlled oxidation of 18F-fluorodeoxy glucose (FDG), mediated by 4- acetamido-2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO) in presence of NaBr and NaOCl in highly-buffered reaction conditions. Uptake of 18F-FGA was ascertained in normal and necrotic H9c2 myoblasts, where necrosis was induced with H2O2-treatment. Biodistribution and circulation kinetics of 18F-FGA were assessed in mice. The potential of 18F-FGA to image myocardial damage was investigated in a rat model of ISO-induced cardiomyopathy. ISO-induced myocardial injury was verified at necropsy by tissue staining and plasma cardiac troponin levels. Results: Synthesis of radiochemically pure 18F-FGA was accomplished by a 5 min, one step oxidation of 18F-FDG. Compared to normal H9c2 cells, the uptake of 18F-FGA in H2O2-treated cells was greater than 4 fold higher (p < 0.05). Biodistribution studies showed rapid elimination from the body (ke = 0.83 h-1); the major organ of 18F-FGA accumulation was kidney. In the rat model, ISO-treatment resulted in significant increase in cardiac troponin. PET images showed that the hearts of ISO-treated rats accumulated significant amounts of 18F-FGA, whereas healthy hearts showed negligible uptake of 18F-FGA. Target-to-nontarget contrast for 18F-FGA accumulation became significantly more pronounced in 4 h images as compared to images acquired 1 h post- injection. Conclusion: 18F-FGA can be easily and quantitatively synthesized from ubiquitously available 18F-FDG as a precursor. Resultant 18F-FGA avidly accumulates in ISO-induced myocardial injury and enables high resolution imaging. Direct imaging of myocardial infarct has a potential to complement current diagnostic techniques employed for evaluation of acute MI which afflicts over 700,000 Americans yearly.

Funding: Sandra K and David L Gilliland Chair in Nuclear Pharmacy

Abstract #17

GAMMA KNIFE PERFEXION TREATMENT PLANS FOR TRIGEMINAL NEURALGIA USING SECTOR- BLOCKED AND NON-SECTOR-BLOCKED SHOTS

Ethan Kendall, Ozer Algan, Salahuddin Ahmad

Purpose: To compare the quality of treatment plans for patients with trigeminal neuralgia (TN) using sector- blocked and non-sector-blocked Gamma Knife (GK) shots.

Methods: In this retrospective study we evaluated thirteen cases of TN. Treatments were planned with GammaPlan v10. All MRI contour sets and prescription doses (range 75-80 Gy) used for sector-blocked (mixing blocked collimators with 4 mm collimator diameters) treatment plans were reused to generate non-sector-blocked (all sectors 4mm) treatment plans. A paired t-test was used to compare treatment plans. A single radiation oncologist evaluated all treatment plans

Results: The average volumes of normal brain tissue receiving 8, 5, 2, and 1 Gy for sector-blocked vs. non- sector-blocked treatment plans were respectively (1.36±0.32 vs. 1.28±0.40 cc, p=0.02), (2.76±0.67 vs. 2.49±0.79 cc, p<0.01), (12.05±2.89 vs. 10.16±3.15 cc, p<0.01), and (43.58±13.07 vs. 32.57±12.74 cc, p<0.01). Sector- blocked plans had a slightly higher average maximum dose to the normal brain tissue than non-sector-blocked treatment plans (47.2±16.0 vs. 46.5±16.9 Gy, p=0.65). When compared to the corresponding sector-blocked treatment plan, the average volumes of brainstem receiving 8, 5, 2, and 1 Gy were 17%, 17%, 25%, and 52% higher in non-sector-blocked treatment plans, respectively. The average maximum doses to the brainstem for sector-blocked vs. non-sector-blocked treatment plans were (37.0±22.8 vs. 41.0±23.3, p<0.01). When compared to non-sector-blocked treatment plans, the average volumes of ipsilateral temporal lobe receiving 8, 5, 2, and 1 Gy were respectively 4%, 14%, 33%, and 38% higher in sector-blocked treatment plans. The mean beam-on times for sector-blocked vs. non-sector-blocked treatment plans were (57.8±12.8 vs. 37.4±6.5 minutes, p<0.01).

Conclusions: Treatment plans utilizing sector-blocking were capable of achieving greater brainstem sparing than those using non-sector-blocked shots. This came at the cost of higher doses to the cerebellum and temporal lobes, and also a significant increase in treatment time.

Funding: None

Abstract #18

DROSOPHILA TRANSMEMBRANE PROTEIN 214 REGULATES CARBOHYDRATE METABOLISM

Yue Li1, Hong Bao1, Weidong Wang2,Hui-Ying Lim1 1Department of Physiology, University of Oklahoma Health Sciences Center 2Department of Medicine, Section of Endocrinology, University of Oklahoma Health Sciences Center

Drosophila is a well-established and genetically tractable model that shares many of the same metabolic and energy-sensing pathways with vertebrates. In a systemic screen for genes that could alter the high sugar diet (HSD)-induced increase in systemic glucose levels, we identified Drosophila transmembrane protein 214 (dTMEM214) which when haploinsufficient, could protect against the increase in systemic glucose levels under HSD condition. dTMEM214 is predicted to be a multipass-membrane protein that so far is not known to be involved in carbohydrate metabolism. We found that flies with RNAi-mediated knockdown (KD) of dTMEM214 throughout the whole body were more resistant to HSD-induced gain in systemic glucose levels compared to the controls, further supporting a role of dTMEM214 in carbohydrate metabolism. We also observed that one of the tissues dTMEM214 is highly expressed in is the intestine, leading to the hypothesis that dTMEM214 acts in the intestine to regulate carbohydrate metabolism. To test this, we performed dTMEM214 KD specifically in the intestine and detected significantly reduced systemic and circulating sugar levels as well as sugar storage in these flies compared to the control flies. We further postulated that dTMEM214 could be involved in dietary glucose uptake in the enterocytes. To test this hypothesis, we examined whether targeted KD of dTMEM214 in the intestine, as well as deficiency in Drosophila glucose transporter 1 (dglut1), a functional equivalent of human glut4, could affect glucose uptake in the enterocytes. We found that both the intestine- specific dTMEM214 KD flies, as well as the dglut1 heterozygous flies, exhibited an attenuated ability in enterocyte glucose uptake, suggesting that like dGLUT1, dTMEM214 could serve a role in glucose uptake into the enterocytes. r. Future direction is to study how dTMEM214 could regulate dGLUT1-mediated in glucose uptake into the enterocyte. Our work could provide novel insights into the regulation of carbohydrate metabolism and diabetes pathogenesis.

Funding: 5R01HL128455-02

Abstract #19

AN INVESTIGATION OF THE RELATIONSHIP BETWEEN EYE MOVEMENTS IN SILENT AND ORAL READING AND HYPOKINETIC DYSARTHRIA

Eunsun Park, MA1, Frank R. Boutsen, PhD1 & Justin D. Dvorak, PhD1,2 1Motor Speech and Prosody Research Laboratory, Dept. of Communication Sciences and Disorders 2Dept. of Biostatistics and Epidemiology University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

Introduction: A majority of individuals with Parkinson’s disease (PD) manifest hypokinetic dysarthria, which is characterized by dysprosody. Some also experience reading difficulties and impaired eye movements. An objective approach to evaluate the association between acoustic correlates of prosody and eye-movement behavior during silent and oral reading in individuals with PD is lacking. Purpose: The goal of this study is to examine eye movements and prosody planning during silent and oral reading in individuals with mild to moderate PD, as compared to age-matched healthy individuals.

Methods: 13 participants with mild to moderate PD and 17 healthy participants read 36 sentences under silent and oral conditions. Each reading sentence consisted of short (10) or long (16) syllables and contained one of two possible target words with one or two stressed syllables. Independent variables of interest included task condition; number of lexical stress; sentence lengths; “Word Reading” subtest scores; and background information such as gender; age; PD duration; and dysarthria severity. Eye-movement variables and speech variables were measured as dependent variables. All statistical tests were conducted in SAS 9.4.

Results: During both oral and silent reading conditions, participants with PD demonstrated significantly longer first-fixation duration and average fixation durations. Participants in both groups descriptively demonstrated longer dwell duration on the target word in the oral reading condition as compared to the silent reading condition. Longer EVS was associated with higher word frequency (p=0.01) but not group (p=0.26) or number of stressed syllables (p=0.08). Participants with PD had significantly slower speech rate than controls (p=0.039).

Conclusion: Participants with PD require additional cognitive processing time to encode and decode the target word during silent and oral reading. Objective evaluation of prosody and behavioral aspects of eye movement characteristics during reading may provide valuable evidence to predict presence and early-stage PD.

Funding: Parkinson Foundation of Oklahoma

Abstract #20

THE IMPACT OF REDUCED NEUROSTEROIDS IN COGNITIVE AGING

Eileen Parks1,2, Dr. William E. Sonntag1,2,3. 1Oklahoma Center for Neuroscience, 2Reynolds Oklahoma Center on Aging, 3Department of Geriatric Medicine

A large percentage of the population experiences cognitive decline with age, which includes impairments in memory, processing speed, and executive function. Allopregnanolone (ALLO) is a neurosteroid that decreases with age and in various models of neurodegenerative diseases where deficits in cognitive function are evident. However, mechanisms by which reduced ALLO contributes to age-related cognitive decline has not been investigated. My hypothesis is that the decline in ALLO with age contributes to reduced neurogenesis and impairments in learning and memory in aged mice. Cortical brain samples from young (3M), middle (12M) and old (24M) male C57BL/6 mice were analyzed by LC/MS for neurosteroids. The effects of ALLO treatment on neurogenesis were assessed in old male mice subcutaneously injected with 10 mg/kg ALLO or vehicle, followed by 100 mg/kg EdU to label proliferating neural stem cells. Hippocampal-dependent learning and memory was tested in the Radial Arm Water Maze (RAWM). Newborn neurons labelled with EdU were visualized by IHC using frozen hippocampal sections. ALLO levels were significantly reduced at both middle and old age compared to young animals. Performance in the RAWM (Pathlength to target, number of entries (errors) and duration in incorrect arms) decreased with age across all 3 measures and significantly improved in ALLO-treated aged mice. When normalized to young, aged mice had approximately 20% surviving newborn neurons. ALLO-treatment tripled the number of surviving newborn neurons (60%), partially rescuing the age-related deficit in neurogenesis. The age-related decline in ALLO is likely a contributing factor in learning and memory impairments with age. Importantly, a single injection of ALLO was sufficient to partially restore neurogenesis and improve learning and memory in older animals. Future experiments will elucidate the cellular mechanisms responsible for the decline in ALLO, which is dependent on a network of enzymes that synthesize both ALLO and other steroid metabolites.

Funding: Reynolds Oklahoma Center on Aging, ROCA pre-doctoral fellowship

Abstract #21

MITOCHONDRIAL FUNCTION AND TRANSCRIPTIONAL REGULATION FOLLOWING DENERVATION- INDUCED SKELETAL MUSCLE ATROPHY

Gavin Pharaoh1,2, Kavithalakshmi Sataranatarajan1,2, Constantin Georgescu3, Niran Hadad4,5, Bumsoo Ahn2, Rojina Ranjit2, Kaitlyn Riddle2, Bill Freeman1,4,5, Holly Van Remmen1,2,5,6 1 Physiology Department, University of Oklahoma Health Sciences Center, 2Aging & Metabolism Research Program, Oklahoma Medical Research Foundation, 3Arthritis & Clinical Immunology Research Program, Oklahoma Medical Research Foundation, 4Reynolds Oklahoma Center on Aging, University of Oklahoma Health Sciences Center, 5Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, 6Oklahoma City VA Medical Center.

Age-related declines in muscle mass and function reduce quality of life. The mechanisms underlying these changes during aging are still not defined, but loss of innervation and increased mitochondrial reactive oxygen species (ROS) generation are proposed to play a key role. The goal of this study was to measure the changes in gene expression that occur following loss of innervation. Mice were exposed to sciatic nerve transection on one limb, sham surgery on the other, and then sacrificed at 0.5, 1, 2, 4, 7, and 14 days post-transection. The gastrocnemius muscle, innervated by the sciatic nerve, was collected for RNA isolation and measurement of ROS generation in isolated mitochondria (fluorometer) and permeabilized muscle fibers (Oroboros O2K). Gastrocnemius mass was significantly reduced at 4 days post-transection and was reduced 42% by 14 days. ROS production was significantly increased at 4 days post-transection, and the elevated ROS production was correlated with the decrease in muscle mass (r2=0.3246, p=0.0008). RNAseq analysis showed that approximately 450 transcripts were differentially regulated post-transection with most changes occurring at 7 and 14 days. The biological processes of the differentially regulated transcripts at 12-24 hours and 2-4 days primarily involve stress responses, while those occurring at 7 and 14 days are characterized by a disruption in homeostasis of metabolic, cellular, and contractile functions. Four transcripts (Arpp21, Gadd45a, Gdf5, and Myog) were upregulated at all time points and are involved in calcium signaling, growth arrest, and stimulation of muscle reinnervation. The motifs for the transcription factors Mef2a-d were significantly enriched in the promoters of differentially regulated transcripts beginning at 4-7 days suggesting a fiber type shift to slow-twitch. Loss of innervation causes muscle atrophy, increased ROS production, an acute stress response, a chronic dysregulation of metabolic, cellular, and contractile homeostasis, and transcriptional induction of a fiber type shift.

Funding: 1R01AG050676-01A1 Defining the Relative Roles of Pre- and Post-Synaptic Events in the Initiation and Progression of Sarcopenia. John and Mildred Carson Oklahoma Medical Research Foundation Pre-doctoral Scholarship.

Abstract #22

FOLLICULAR T HELPER CELLS AND T HELPER 17 CELLS COOPERATE IN PROMOTING DISEASE IN NEURO-INFLAMMATION

James L. Quinn1,2, Agnieshka Agasing1,2, Rose Ko2, Robert C. Axtell2 1Department of Microbiology and Immunology, University of Oklahoma HSC, Oklahoma City, OK 73104, 2Oklahoma Medical Research Foundation, Oklahoma City, OK 73104

Both T-cells and B-cells are implicated in the pathology of multiple sclerosis, but how these cells cooperate to drive disease activity remains unclear. Recent studies using experimental autoimmune encephalomyelitis (EAE) demonstrated that the T helper 17 (TH17) pathway is correlated with increased numbers of ectopic B-cell follicles in the CNS. As follicular T helper (TFH) cells are mediators of B-cell response, our hypothesis was that TH17 cells and TFH cells cooperate to drive pathogenic B-cell responses in the CNS and contribute to the severity of EAE. Therefore, the main objective of this project was to examine the role of TFH cells in EAE induced by the transfer of myelin-specific TH17 cells (TH17 EAE). At the peak of TH17 EAE, we saw approximately 20% of CD4+ cells in the CNS were CXCR5+PD1+ TFH cells. Furthermore, the number of TFH cells in the CNS was positively correlated with the number of infiltrating B cells and disease severity. We next transferred myelin- specific TH17 cells from congenic CD45.1+ mice into CD45.2+ recipient mice to determine whether the TFH cells were donor or recipient-derived. We observed that the TFH cells were CD45.2+ recipient-derived whereas IL-17 producing cells were CD45.1+ donor-derived. We also found that these TFH cells increased within the CNS tissue over the course of disease development. We next used CD4+ Cre mice crossed with BCL6flox/flox mice to generate mice deficient for a TFH population. These mice were used as recipient mice in TH17 EAE and we found that these TFH-deficient mice had significantly reduced EAE compared to wild type recipient mice. These results show that TH17 cells migrate to the CNS to initiate neuro-inflammation in EAE and mediate of a second wave of CNS-infiltrating TFHs that facilitate a B-cell response in the CNS and contributes to disease severity.

Funding: T32 Immunology Training Grant, NIH-NINDS, Merck-KgAA

Abstract #23

COHESION FATIGUE, A NOVEL SOURCE OF CHROMOSOMAL INSTABILITY

Hem Sapkota1, 2, Emilia Wasiak 1, John Daum 1 and Gary Gorbsky 1, 2 1 Cell Cycle and Cancer Biology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 2 Dept. of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK

Errors during segregation of the replicated genome process lead to genome instability, a cause of developmental defects and cancer. In cells delayed at metaphase, chromatids of individual chromosomes can undergo an unscheduled and asynchronous separation termed cohesion fatigue. All cell types examined undergo cohesion fatigue though the rates of chromatid separation can be strikingly varied. However, molecular mechanism and consequences of cohesion fatigue are unknown. Therefore, this study aims to identify the pathways involved in cohesion fatigue as well as its role in generating chromosomal abnormalities. Using low concentrations of Nocodazole and Taxol, strength of mitotic spindle pulling force was altered. In Hela cells partial destabilization of mitotic microtubules (MTs) delayed cohesion fatigue. In contrast, stabilizing MTs accelerated the process. By temporarily extending early mitosis facilitated by the disruption of MTs, cells were able to continue the prophase pathway. Cells with longer pre-arrest had significantly higher proportion of fatigued cells when subjected to metaphase arrest. In addition, deletion of Stromal Antigen 2 (SA2), a component of the core cohesin complex and a recruiter of positive regulators of cohesin in HCT116 cells resulted in accelerated cohesion fatigue. Similarly, depletion of cohesion establishment enzyme ESCO2 in HeLa cells led to faster cohesion fatigue. Finally, LLCPK and HeLa cells were transiently arrested in metaphase by MG132 then released to proceed into anaphase. Fixed cell analysis and live cell imaging of transiently metaphase-arrested cells showed time dependent increase in segregation defects. Our studies indicate that cohesion fatigue is a multifactorial, progressive event, in which microtubule pulling forces, integrity of the cohesin complex, and activity of the prophase pathway all influence the rate at which unscheduled chromatid separation occurs in cells delayed at metaphase. Cohesion fatigue, even at initial stages, leads to chromosome segregation defects.

Funding: R01GM111731 to Gary J Gorbsky and OMRF Predoctoral scholarship to Hem Sapkota

Abstract #24

ASSOCIATION OF DIFFERENT TOBACCO PRODUCTS WITH CARDIOVASCULAR DISEASE RISK PARAMETERS AND METABOLIC SYNDROME.

Zoona Sarwar, Nasir Mushtaq Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center

Introduction: Despite a substantial decrease in cigarette smoking prevalence, use of other tobacco products including smokeless tobacco (ST), cigar, and use of more than one tobacco product – polytobacco (PT) has increased in the recent years. Association of cigarette smoking with cardiovascular disease (CVD) is well documented, however, there are inconsistent findings related to CVD and use of other tobacco products. Aim: Purpose of this study is to examine the association between other tobacco products use and CVD risk parameters

Methods: Data from 28,240 participants of National Health and Nutrition Examination Survey (NHANES) 2003- 2014 were used. Exclusive ST users, exclusive cigar smokers, and PT users were compared with exclusive cigarette smokers and nontobacco users. Association of tobacco use categories with CVD risk parameters and metabolic syndrome (MS) were examined by univariate and multiple logistic regression analysis. Weighted analyses were performed and a level of 0.05 was used for statistical significance.

Results: 20.1% of the participants were exclusive cigarettes smokers, whereas 2.1% were exclusive ST users and 1.5% were exclusive cigar smokers. PT use was reported by 1.1%. 32.0% of the participants had hypertension, 29.4% had high cholesterol, 68.8% were overweight or obese, and19.5% of the participants had MS. Male exclusive ST users were 1.67 (95%CI:1.25, 2.25) times more likely to have hypertension compared to exclusive cigarette smokers when adjusted for other variables. Similarly, these odds were significantly higher when ST users were compared to nontobacco users (OR 1.30, 95%CI:1.39, 3.76). Odds of CVD risk parameters and MS among PT users were not significantly different from those of cigarette smokers.

Conclusion: The study results indicate positive association of ST use and hypertension. Findings suggest that other tobacco products are not safer substitute for cigarettes. Future research is warranted to further examine the effect of other tobacco products on CVD.

Funding: None

Abstract #25

KERATOCONUS: DEVELOPMENT OF A NOVEL HUMAN CORNEAL COLLAGEN CROSS-LINKING 3-D IN VITRO MODEL

Rabab Sharif1, Jesper Hjortdal2, Henrik Sejersen2, Dimitrios Karamichos1,3 1Department of Cell Biology , University of Oklahoma Health science Center, Oklahoma City, Oklahoma, USA 2Department of Ophthalmology, Aarhus University Hospital, Aarhus C, Denmark 3Department of Ophthalmology, Dean McGee Eye Institute, University of Oklahoma Health Science Center, Oklahoma City, Oklahoma, USA

Purpose: Keratoconus (KC) is a progressive corneal dystrophy, represented by biomechanical corneal instability, and severe vision impairment. Despite the large number of studies, but no existing animal model, the exact underlying KC pathobiology remains unclear. We utilized our established 3-D in vitro KC model to study cellular and molecular mechanisms of the disease following radiation induced corneal collagen crosslinking (CXL). This 2016 FDA approved application is the only current aetiopathogenic mechanism that promises to halt KC progression, for certain KC-affected groups. In this study we take the lead in determining the cellular and molecular effects of pre/post CXL, which surprisingly, have not been previously investigated.

Methods: Primary Human corneal fibroblasts (HCF) and keratoconus (HKC) cells were seeded on polycarbonate membranes and cultured in a 10% FBS EMEM medium stimulated with 0.5 mM of Vitamin C. At the end of week 4 both assembled 3-D constructs were exposed to UVA light (wavelength 365 nm) at a 3 cm distance for 3 minutes with an irradiance of 3mW/cm2. Cell viability was determined using live/dead cell staining technique. All samples were analyzed using western blotting (WB) to quantify changes in the expressions of key proteins LOX, COL-I,-III,-V, and the transcriptional SMAD proteins, between HCFs and HKCs.

Results: Our data indicates small but significant percentage of apoptotic KC cells following CXL. WB analysis revealed a reduced expression of the regulatory SMAD 6 (p<0.0001) in untreated HKCs, contrary to SMAD 3 that showed to be downregulated in treated HKCs (p=0.0226), suggesting a dysregulation in the SMAD downstream signaling pathway following CXL. In addition, a significant increase in LOX expression (p< 0.0001) was observed in HKCs post-CXL compared to controls.

Conclusion: Our 3-D model provided excellent grounds to study CXL effects on the corneal stromal microenvironment. In addition, this in vitro CXL system is promising to establish optimized treatment modalities in patients suffering from KC.

Funding: EY023568 (NIH/NEI)

Abstract #26

A DOMINANT MUTATION IN RPE65, D477G, DELAYS DARK-ADAPTATION AND DISTURBS THE VISUAL CYCLE IN THE MUTANT KNOCK-IN MICE

Younghwa Shin1, Gennadiy Moiseyev1, Dibyendu Charkraborty1, Jian-xing Ma1 1Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, U.S.A.

Purpose: RPE65 is an indispensable component of the retinoid visual cycle in vertebrates, through which the visual chromophore 11-cis-retinal is generated to support normal vision. Various inherited blinding conditions in humans such as Leber’s congenital amaurosis and retinitis pigmentosa are attributed to either homozygous or compound heterozygous mutations in RPE65. Here we investigated the D477G missense mutation, an unprecedented dominant-acting mutation of RPE65 identified in patients with autosomal dominant retinitis pigmentosa.

Methods: We generated a D477G knock-in mouse and characterized its phenotypes. Expression of RPE65 was examined at both the transcriptional and translational levels by real-time PCR and western blot analysis, respectively. Visual function was accessed by electroretinography, and retinoid profile was analyzed by HPLC. Retinal morphology was assessed by immunohistochemistry, light microscopy and transmission electron microscopy.

Results: RPE65 protein levels were decreased in the heterozygous knock-in mice, however, visual function assessed by electroretinography was normal. Fully dark-adapted retinoid profile was comparable between wild- type and heterozygous knock-in mice as well. However, kinetics of 11-cis-retinal regeneration following light exposure was significantly slower in the heterozygous knock-in mice compared to wild-type and RPE65 heterozygous knock-out mice. Furthermore, the heterozygous knock-in mice exhibited lower A-wave recovery compared to wild-type mice following photo-bleaching, suggesting a delayed dark-adaptation.

Conclusion: Taken together, these observations suggest that D477G acts as a dominant-negative mutant of RPE65 and delays the regeneration of the chromophore. The knock-in mice provide a useful model for further understanding of the pathogenesis of this mutant of RPE65 and for the development of therapeutic strategies.

Funding: NIH grants EY012231, EY018659, EY019309, EY016507, and GM104934; and Oklahoma Center for the Advancement of Science and Technology grant HR-13-076.

Abstract #27

DNA REPLICATION TIMING DURING DEVELOPMENT ANTICIPATES TRANSCRIPTIONAL PROGRAMS AND PARALLELS ENHANCER ACTIVATION

Joseph C. Siefert1,2, Constantin Georgescu4, Jonathan D. Wren4,5, Amnon Koren3, Christopher L. Sansam1,2 1Cell Cycle and Cancer Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA 2Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA 3Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA 4Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK 73104, USA 5Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA

In dividing cells, DNA replication occurs in a precise order, but many questions remain regarding the mechanisms of replication timing establishment and regulation. We now have generated genome-wide, high-resolution replication timing maps throughout zebrafish development. Unexpectedly, in the rapid cell cycles preceding the midblastula transition, a defined timing program was present that predicted the initial wave of zygotic transcription. Replication timing was thereafter progressively and continuously remodeled across the majority of the genome, and epigenetic changes involved in enhancer activation frequently paralleled developmental changes in replication timing. Strikingly, the long arm of chromosome 4 underwent a dramatic developmentally regulated switch to late replication during gastrulation, reminiscent of mammalian X chromosome inactivation. This study reveals that replication timing is dynamic and tightly linked to epigenetic and transcriptional changes throughout early zebrafish development. These data provide insight into the regulation and functions of replication timing and will enable further mechanistic studies.

Funding: OCASCR, NIH

Abstract #28

HEALTH CARE PROVIDER’S ROLE IN OBESITY PREVENTION AND HEALTHY DEVELOPMENT OF YOUNG AMERICAN INDIAN CHILDREN

Chelsea L. Smith1, Devon Walker1, Kelly Kerr1, Lancer Stephens2,9, Amber Anderson3, Julie Seward4,9, Ashley Weedn5, Marshall Cheney6, Audrey Jacob7, Michelle Key8 and Susan B. Sisson1 1Department of Nutritional Sciences, University of Oklahoma Health Sciences Center 2Department of Health Promotion, University of Oklahoma Health Sciences Center 3Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center 4Southern Plains Tribal Health Board 5 Department of Pediatrics, University of Oklahoma Health Sciences Center 6Department of Health and Exercise Science, University of Oklahoma 7Choctaw Nation of Oklahoma 8Chickasaw Nation Department of Health 9Oklahoma Shared Clinical and Translational Resources

Introduction: Early childhood obesity is a public health concern with its implications for obesity later in life and many chronic diseases. American Indian (AI) children have higher rates of childhood obesity than the national average. Health care providers (HCP)s are an essential component of working with children and families in preventing childhood obesity. The outlooks and perceived roles of HCPs related to obesity prevention in AI children are not fully understood. The purpose of this study is advance understanding of how HCP perceive their role in the healthy development of young American Indian children.

Methods: This qualitative study involved 20 HCPs who served AI children between the ages of 3-5 years old. Structured individual interviews were conducted with the HCPs involving their role in obesity prevention and healthy development of the children they serve. Five researchers developed a codebook, coded transcripts and developed themes from the interviews. Of the 20 HCPs, 50% worked in outpatient setting and 40% were a part of a tribally affiliated practice.

Results: There were four main themes that arose from the interviews. 1. HCPs noted that family health was just as important as the child’s health in obesity prevention. 2. HCPs recognized the importance of public policy and had policy level concerns regarding nutrition. 3. HCPs felt they view health habits of young children differently than parents. HCPs felt that 3-5 years old is an important time for change though some parents felt children could grow out of poor health habits early on in life. 4. HCPs felt a holistic approach was necessary since obesity influences one’s social and emotional health.

Conclusion: HCPs felt working with families towards the total health of the child may help adopt healthy habits early on and contribute to the healthy development of young American Indian children.

Funding: Presbyterian Health Foundation; Harold Hamm Diabetes Center

Abstract #29

MIR-1246 IS PRIMARILY DERIVED FROM U2 SMALL NUCLEAR RNA 1 (RNU2-1) AND HIGHLY ENRICHED IN PANCREATIC CANCER EXOSOMES

1Yi-Fan Xu, 1Bethany N. Hannafon, 2Russell G. Postier, 1Wei-Qun Ding 1Department of Pathology, 2Department of Surgery, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, OK 73104, USA

Background: microRNAs (miRNAs) are short non-coding RNAs that bind to mRNAs and negatively regulate gene expression. miR-1246 was identified in 2008 and its precursor was mapped to human chromosome 2 (2q31.1). However, the mature miR-1246 sequence is identical to a fragment of RNU2-1. High levels of the RNU2-1 fragment in serum and miR-1246 in plasma exosomes have been reported in pancreatic cancer patients, yet the origin of the detected small RNA remains controversial.

Methods: An immortalized normal human pancreatic ductal cell line (hTERT-HPNE) and a human pancreatic cancer cell line (PANC-1) were used as in vitro models. Plasma were collected from patients with localized pancreatic cancer and matched healthy subjects. Exosomes were isolated from the cell culture medium and plasma by ultracentrifugation. Next Generation small RNA sequencing was used to profile the miRNA contents of hTERT-HPNE and PANC-1 cells and exosomes. qRT-PCR was performed to confirm RNU2-1 and miR-1246 expression. CRISPR-Cas9 knockout of miR-1246 precursor was performed in PANC-1 cells. Over-expression of miR-1246 and RNU2-1 in PANC-1 cells was achieved by transfection of previously established plasmid vectors.

Results: Bioinformatics analysis revealed that the mature miR-1246 sequence is identical to a fragment of RNU2- 1. miR-1246 was the most enriched miRNA in PANC-1 exosomes, but barely detectable in PANC-1 or hTERT- HPNE cells. The pre-miR-1246 was undetectable, yet RNU2-1 expression was evident, both in PANC-1 cells and PANC-1 exosomes. Knockout or overexpression of miR-1246 in PANC-1 cells did not alter exosome miR- 1246 levels. However, over-expression of RNU2-1 increased exosome miR-1246 expression. Elevated miR- 1246 was confirmed in patient plasma exosomes as compared to control plasma exosomes.

Conclusion: Our data indicate that the mature miR-1246 is primarily derived from RNU2-1 and highly enriched in pancreatic cancer exosomes. The cellular mechanisms and biological consequences of the miR-1246 enrichment in pancreatic cancer exosomes merit further investigation.

Funding: Presbyterian Health Foundation and Oklahoma Shared Clinical and Translational Resources

Abstract #30

PRELIMINARY ANALYSIS OF THE EFFECT OF PREHABILITATION ON MUSCLE MASS, WEIGHT, AND DIETARY INTAKE STRATIFIED BY HAND GRIP STRENGTH IN PANCREATIC CANCER PATIENTS

Katherine Zauner RD/LD1; Leah Hoffman PhD, RD/LD, CNSC2; Elizabeth Hile PhD, PT, NCS, CLT3 1Department of Nutritional Sciences student, University of Oklahoma Health Sciences Center, RD/LD; 2 Department of Nutritional Sciences, University of Oklahoma Health Sciences Center; 3 Cancer Rehabilitation Science Program, OUHSC Stephenson Cancer Center

The purpose of this study is to describe the impact of nutrition education during prehabilitation on dietary intake, weight, and percent fat-free mass for patients undergoing pancreatic surgery and determine if these outcomes change based on a participant’s hand grip strength. Methods: A total of 20 participants will be enrolled for this study at the Stephenson Cancer Center in Oklahoma City Oklahoma. Participants will be aged >30, either newly diagnosed or following neoadjuvant therapy, and deemed eligible for a PD. For this study, participants will be assessed both at baseline and at approximately one month post-surgery. Hand grip strength will be measured at baseline using a JAMAR Hydraulic hand dynamometer and participants will be classified as either at or above average hand grip strength or below average hand grip strength. At both time points the participants will be measured by BIA analysis using a Tanita TBF-310GS Total Body Composition Analyzer to measure weight, fat mass, and fat free mass. Participants will also be asked to complete a 24-hour food recall. All participants will receive an exercise intervention and nutrition education that was created specifically for this study. Change in muscle mass and weight will be assessed using repeated measures ANOVA. 24-hour recalls will be assessed by FoodWorks version 11 software and described as descriptive statistics only. Results: This study is looking at preliminary results for a larger study being completed at the Stephenson Cancer Center. It is hypothesized that there will be a difference between patients who have above average hand grip strength at baseline and those who are below average hand grip strength at baseline when comparing muscle mass and weight along with being able to describe dietary intake. This study is currently in place; data collection should be completed in March.

Funding: None

Abstract #31

XZ488 AS A NOVEL THERAPY AGAINST GLIOMAS

Jadith Ziegler1,4, Debra Saunders4, Nataliya Smith4, Megan Lerner3, Anja Bastain2, Lora Bailey-Downs2, Michael Ihnat2, Aleem Gangjee5, and Rheal A.Towner1,2,4 1Department of Pathology, 2Department of Pharmaceutical Sciences, College of Pharmacy, 3Department of Surgery Research Laboratory, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA, 4Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA, and 5Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, USA

High-grade gliomas such as glioblastomas (GBM) present a deadly prognosis following diagnosis and very few effective treatment options. Here, we investigate if the small molecule XZ488 can be an effective therapy against GBM, with both anti-angiogenic, as well as tubule inhibitory properties, using a human G55 glioma xenograft model in nude mice. From in vitro studies, we report that XZ488 incubation reduced cell viability in G55 and HMEC-1 cells more so than TMZ treatment. In vivo investigations indicated that XZ488 therapy helped reduce tumor volumes (p<0.0001), prolong life (p<0.01), increase tumor perfusion (p<0.01), as well as decrease microvessel density (MVD) (p<0.05), compared to untreated mice or mice treated with non-specific IgG, in the G55 xenograft model. AG488 therapy was compared to TMZ and anti-VEGF (bevacizumab) therapies. Animal survival and tumor volume changes for XZ488 were comparable to TMZ or anti-VEGF therapies, however XZ488 was found to be more effective in decreasing tumor-related vascularity (perfusion and MVD). XZ488 is a potential novel therapy against high-grade gliomas, which could either be used alone, or in conjunction with other anticancer drugs.

Funding: Funding was provided by the Oklahoma Medical Research Foundation (RAT), College of Pharmacy startup funds (MAI), NIH, NCI, R01 CA136944 (AG).

Abstract #32

TUMOR-TARGETED DENDRIMER-BASED MULTIFUNCTIONAL NANOPARTICLE FOR BIO- CHEMODRUG DELIVERY AND IMAGING OF LUNG CANCER

Narsireddy Amreddy1,3, Anish Babu1,3, Ranganayaki Muralidharan1,3, Akhil Srivastava1,3, Janani Panneerselvam1,3, Rheal Towner4, Anupama Munshi2,3, Rajagopal Ramesh1,3,5 Departments of 1Pathology, 2Radiation Oncology, 3Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma 73104, USA; 5Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.

Introduction: The effectiveness of chemotherapy is often limited by its poor distribution to tumor cells. Further, the response of tumor cells to chemotherapy is not known unless subjected to imaging. In the clinic, monitoring of response to therapy is delayed and offers limited options to physicians in making treatment decisions in switching therapy. Therefore, the availability of technologies that can efficiently deliver drugs to tumors and simultaneously monitor response to therapy will provide opportunities in providing the best treatment decisions. In the present study we developed and tested tumor-targeted multifunctional dendrimer-based nanoparticles (tt- MDNP) that can co-deliver biologic/chemotherapeutic in combination with a magnetic resonance imaging (MRI) contrast agent using lung cancer as a model.

Methods: PEI functionalized PAMAM dendrimer was used to encapsulate cisplatin (chemodrug), and a siRNA targeted against the RNA binding protein, HuR (biologic). Subsequently, the MRI contrast agent Gadolinium (Gd) was covalently conjugated to functionalized dendrimer nanoparticles and finally decorated with folic acid for tumor-targeted drug delivery to cancer cells overexpressing the folate receptor (FR). The tt-MDNP was tested against lung cancer (H1299 and A549) and normal (MRC-9) cells in vitro. Further, the imaging capabilities of tt- MDNP was determined in H1299 cells by MRI.

Results: Cell uptake studies showed FR expressing cancer cells had high uptake of tt-MDNP compared to normal cells and efficient silencing of HuR. Further, tt-MDNP-mediated co-delivery of siHuR and CDDP produced greater therapeutic efficacy in terms of cell growth inhibition and HuR gene expression in lung cancer compared to individual agent treatments. Finally, excellent contrast imaging was observed by MRI in tt-MDNP-treated H1299 cells compared to MDNP-treated cells.

Conclusions: Our tt-MDNP study results demonstrate co-delivery of therapeutics and imaging agents is feasible and can be used for combinatorial imaging and therapy. In vivo evaluation of tt-MDNP is warranted prior to advancing to clinical testing.

Funding: The study was supported in part by grants received from the National Institutes of Health R01 CA167516 (RR), an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences (P20 GM103639) of the National Institutes of Health, and by funds received from the Presbyterian Health Foundation Seed Grant, Presbyterian Health Foundation Bridge Grant, Stephenson Cancer Center Seed Grant and Jim and Christy Everest Endowed Chair in Cancer Developmental Therapeutics, OUHSC

Abstract #33

CHEMODRUG DELIVERY USING INTEGRIN-TARGETED CHITOSAN-PLGA HYBRID NANOPARTICLE FOR LUNG CANCER THERAPY

Anish Babu1,4, Narsireddy Amreddy, 1,4 Ranganayaki Muralidharan,1,4 Allshine Chen, 2,4 Yan D. Zhao, 2,4 Anupama Munshi, 3,4 Rajagopal Ramesh1,4,5* Department of 1Pathology, 2Biostatistics and Epidemiology, 3Radiation Oncology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; 4Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; 5Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.

Introduction: The non-specific, primarily dose-dependent toxicity of chemotherapeutics toward normal cells is a continuing problem in cancer therapy. However targeted nanoparticle-based drug delivery shows promise in overcoming this challenge. This study aimed to develop a targeted nanoparticle based on PLGA and chitosan by exploring RGD-integrin αvβ3 receptor interaction for the delivery of paclitaxel (PTX) towards lung cancer cells and evaluate its efficiency. Aiming for primary therapeutic outcome, studies often explore integrin (αvβ3) receptor targeted chemotherapeutic delivery in tumor neovasculature endothelial cells. However, their tumor cell targeting efficiency and their impact in normal cells with integrin (αvβ3) receptor expression have seldom been reported. Therefore we also sought to understand the effect of RGD-based targeted nanoparticle drug delivery in normal lung fibroblasts expressing integrin αvβ3 and in broncho-epithelial cells with negligible integrin αvβ3 expression.

Methods: RGD-modified chitosan was synthesized and coated onto PTX-PLGA nanoparticles prepared using a double emulsion method. We carried out comparative cell uptake, cell viability, cell cycle arrest, apoptosis, and western analysis for apoptotic proteins to evaluate our targeted nanoparticle system in integrin (αvβ3) receptor expressing lung cancer and normal cells.

Results: The PTX-PLGA-CSNP-RGD system showed increased cell uptake, triggered enhanced apoptosis, and induced G2/M cell cycle arrest and more overall cytotoxicity than its non-targeted counterpart, but had therapeutic efficiency comparable to free PTX. The cell uptake and therapeutic efficiency of PTX-PLGA-CSNP- RGD depended on integrin αvβ3 expression levels in NSCLC cells. However, the increased cell uptake of PTX- PLGA-CSNP-RGD observed in integrin-expressing fibroblasts cells did not induce significant toxicity. Broncho- epithelial cells showed negligible sensitivity to PTX-PLGA-CSNP-RGD cytotoxicity, at drug concentrations equivalent to those used in cancer cells. Cisplatin (CDDP) was used to confirm the findings with PTX.

Conclusion: Our results demonstrate that PLGA-CSNP-RGD is a promising nanoplatform for targeted chemotherapeutic delivery to lung cancer, sparing normal cells from drug toxicity.

Funding: National Institutes of Health R01 CA167516 (RR),Institutional Development Award (IDeA) from the National Institute of General Medical Sciences (P20 GM103639) of the National Institutes of Health, Presbyterian Health Foundation Seed Grant, Presbyterian Health Foundation Bridge Grant, Stephenson Cancer Center Seed Grant, and Jim and Christy Everest Endowed Chair in Cancer Developmental Therapeutics, The University of Oklahoma Health Sciences Center.

Abstract #34

DETERMINING THE ROLE OF XRN2 IN GLIOMA PROGRESSION

Tuyen Dang1, Julio Morales1,2 1Department of Neurosurgery, University of Oklahoma Health Science Center 2 Stephenson Cancer Center, University of Oklahoma Health Science Center

Introduction- Glioblastoma is a highly aggressive brain cancer with a 4 to 7% five-year survival rate. A possible cause for the poor survival rate is the presence of radiation/chemotherapy resistant invasive neoplastic cells. We find that expression of XRN2, a 5’ to 3’ exonuclease, is elevated in glioblastoma and may contribute to therapy resistance and cancer cell migration.

Methods- For both cell survival and migration assays, glioblastoma cells were labeled with H2B-GFP. LN229, U251, and U87 glioblastoma cell lines were used for these studies. Cell survival were determined by GFP signal during treatment with Cisplatin or Etoposide. Signals were recorded at 0 hour, 24 hour, 48 hour, and 72 hour post-treatment. Cell motility was determined by live cell imaging on a Zeiss LSM 710 confocal (OMRF). Images were taken at 30 minute intervals for at least 6 hours. Before imaging, cells were transfected with 50 nM of siRNA for 72 hours and fed with growth media supplemented with 100 ng/ml EGF. Tracking was done based on the H2B-GFP signal using Trackmate free software (FIJI).

Results- XRN2 expression correlated with cell survival upon treatment with Cisplatin and Etoposide. LN229, which has a higher expression than U251, were more resistant to Cisplatin and Etoposide as compared to U251. Resistance can be seen through dose response and time course. XRN2 was also required for cell migration in 2D. In both U87 and U251, knockdown of XRN2 through siRNAs, resulted in reduction of cell migration and speed as compared to control.

Conclusions- XRN2 expression is correlative to cell survival of glioblastoma cells upon radiation and chemotherapy exposures. XRN2 is required for neoplastic cell motility. Radiation and chemotherapy resistances and cell migration are the first steps of cancer metastasis. XRN2 shows promise to be a lynchpin to resistance and invasion progression in glioma disease.

Funding: None

Abstract #35

NEURORETINA-SPECIFIC CAVEOLIN-1 DEPLETION BLUNTS RETINAL INFLAMMATION: POTENTIAL ROLE OF ENHANCED TRAF3 PRODUCTION

Jami Gurley1, Daniel J. Carr1,2 Stefanie M. Hauck3, and Michael H. Elliott1 1Department of Ophthalmology/Dean McGee Eye Institute 2Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center 3Research Unit Protein Science, Helmholtz Center Munich, Germany

Introduction: Uncontrolled immune responses in the eye lead to ocular tissue dysfunction and vision loss. Previously, we showed that global Caveolin-1 (Cav1) depletion in mice modulates retinal inflammation and immune cell recruitment in response to ocular lipopolysaccharide (LPS). This study evaluated effects of local, neuroretinal Cav1 ablation on LPS-induced retinal inflammation and also investigated potential mediators of inflammatory protection in neuroretinal Cav1-depleted animals.

Methods: Neuroretina-specific Cav1 knockout (Chx10-KO) mice were generated by Chx10-driven Cre-mediated recombination. Cav1 deletion efficiency/specificity and TNF receptor-associated factor 3 (TRAF3) levels were assessed by immunohistochemistry, western blotting, and mass spectrometry. Association of TRAF3 with detergent-resistant “raft” fractions from Chx10-KO and control retinas was evaluated by western blotting. Chx10- KO and littermate controls were challenged with intravitreal LPS (0.5 µg/eye) or vehicle. Twenty-four hours post- challenge, retinal cytokine/chemokine (Cxcl1/KC, Ccl2/MCP-1, IL-6) production and immune cell infiltration were measured by multiplex protein suspension array (n=4) and flow cytometry (n=7-9), respectively. Data were represented as mean ± SEM and analyzed by Student’s t-test or 2-way ANOVA.

Results: Chx-10 mediated recombination in retinal Müller glia and neuronal cells (but not in retinal pigmented epithelium or vasculature) depleted neuroretinal Cav1 >70%. Compared to controls, Chx10-KO animals had significantly decreased retinal inflammatory cytokines/chemokines (~30-80%; p<0.05) and infiltrating immune cells (~75-80%; p<0.01) following LPS challenge. TRAF3 protein levels were increased 2.7-fold in Ch10-KO retinas compared to controls (p < 0.001). TRAF3 co-fractionated with Cav1 in detergent-resistant membrane fractions from control retinas but was predominantly detergent-soluble in Chx10-KOs indicating that TRAF3’s association with “rafts” depends on Cav1.

Conclusions: Our results suggest that neuroretinal Cav1 plays a key role in modulating local innate inflammatory responses in the retina and support a role for Cav1 in modulating TRAF3 protein levels. This is consistent with the novel hypothesis that Cav1 inflammatory modulation is mediated through control of TRAF3 levels.

Funding: NIH R01EY019494, NIH core grant EY021725, and an unrestricted grant from Research to Prevent Blindness, Inc.

Abstract #36

SHINING THE LIGHT ON BRAIN CIRCUITS RESPONSIBLE FOR STRESS-INDUCED VISCERAL PAIN

Anthony C. Johnson1, Casey O. Ligon1, Rocco Latorre1, Beverley Greenwood-Van Meerveld1,2,3 1Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center 2Department of Physiology, University of Oklahoma Health Sciences Center 3Oklahoma City Department of Veterans Affairs Medical Center

Background: Optogenetics identifies brain circuits controlling behaviors in conscious animals by using light to control neuronal function, offering a novel tool to study brain-gut axis regulation of visceral sensitivity. Evidence suggests that a connection between the central nucleus of the amygdala (CeA) and the bed nucleus of the stria terminalis (BNST) modulates colonic sensitivity. The goal of this study was to test the hypothesis that directly activating the CeA-BNST pathway, via optogenetic stimulation, will increase colonic sensitivity.

Methods: In male rats, CeA neurons were bilaterally infected with viral vectors expressing fluorescent proteins (control), halorhodopsin (inhibitory), or channelrhodopsin (excitatory). Bilateral fiber optic cannulas were implanted on the BNST. After 8-10 weeks, the response to graded (20-60 mmHg), isobaric colorectal distension (CRD) was measured as the number of abdominal contractions to CRD in freely moving rats, with and without laser stimulation of CeA fibers at the BNST. Immunohistochemistry and histology were used to evaluate vector expression and neuronal integrity. A repeated measure two-way ANOVA with Tukey’s post-hoc analysis was used to compare the change in abdominal contractions (delta) induced by laser illumination (mean±standard error).

Results: We found that infected neurons were healthy and the vector was correctly expressed. Neither laser illumination alone nor stimulation of halorhodopsin at the BNST changed colonic sensitivity (delta: 20 mmHg: 1.0±0.3/2.0±0.6; 40 mmHg: 1.7±0.3/5.3±1.4; 60 mmHg: 1.5±0.4/2.7±1.6, n=6/group). In contrast, laser stimulation of channelrhodopsin significantly increased colonic sensitivity (delta: 20 mmHg: 11.0±2.0, P<0.01; 40 mmHg: 11.8±2.6, P<0.05; 60 mmHg: 13.2±3.7, P<0.001, n=6), indicating that activation of CeA terminals at the BNST promotes colonic hypersensitivity.

Conclusions: In a freely moving rat, optogenetic activation of the CeA-BNST pathway induced colonic hypersensitivity. This is the first study demonstrating that optogenetics can be used to investigate the brain-gut axis, providing a new tool to investigate circuitry that modulates visceral pain.

Funding: VA Merit Grant (BX002188), Veterans Research and Education Foundation Seed Grant, OCNS Translational Neuroscience Research Initiative Seed Grant

Abstract #37

TARGETING THE RECEPTOR KINASE RET FOR THE TREATMENT OF INTESTINAL DISORDERS

Rocco Latorre2, Ehsan Mohammadi2, John Russell4, Hilary Eidam4, Sanjay Kumar4 and Beverley Greenwood- Van Meerveld123 1 VA Medical Center, Oklahoma City, OK 73104 2 Oklahoma Center for Neuroscience, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104 3 Department of Physiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104 4 Virtual Proof of Concept, Discovery Performance Unit (VPoC DPU), GlaxoSmithKline, King of Prussia, PA 19406

Background: The presence of the transmembrane receptor kinase RET in the enteric nervous system (ENS) suggests that RET may contribute to intestinal function modulation. Cholinergic nerves in ENS, through the release of acetylcholine (ACh), play a critical role in the control of colonic function. We hypothesized that a RET kinase-mediated mechanism may regulate colonic ion transport (CIT) and smooth muscle contractility (SMC) through modulation of cholinergic nerves. We employed in vitro and in vivo approaches to investigate whether CIT and SMC evoked by ACh-mediated mechanisms were attenuated by a novel RET kinase inhibitor (GSK106).

Methods: In isolated rat colonic tissue mounted in Ussing chambers, increases in CIT were assessed electrophysiologically via measurements of short circuit current (Isc) evoked by nerve stimulation (EFS at 64 Hz), or pharmacologically with carbachol (EC50 3.8 µM) and bethanechol (EC50 42.1 µM). In freely moving rats, we assessed the effect of a RET kinase inhibition on propulsive motility via quantification of fecal pellet output (FPO) induced by the acetylcholinesterase inhibitor neostigmine (0.1 mg/kg i.p.).

Results: In vitro GSK106 (100 nM) attenuated the mean increase in Isc induced by EFS (79±5.1 to 65.4±6.1 μA/cm2, P<0.001) or carbachol (109.5±4.7 to 68.8±3.3 μA/cm2, P<0.0001) but not bethanechol. In vivo acute administration of neostigmine produced a profound prokinetic effect compared to vehicle controls as measured by increased FPO (11±0.1 vs. 4.5±0.2 pellets, P<0.0001). GSK106 at 10 mg/kg given orally for 3.5 days BID significantly reduced the neostigmine prokinetic effect (11±0.1 to 8.2±0.2 pellets, P<0.01).

Conclusion: RET kinase-mediated mechanisms regulate colonic function by maintaining the neuronal cholinergic phenotype and enabling ACh-evoked CIT and SMC. We suggest that modulating the cholinergic control of the colon via a RET kinase inhibitor may represent a novel target for the treatment of intestinal disorders associated with increased secretion and accelerated GI transit.

Funding: GlaxoSmithKline Grant

Abstract #38

A SINGLE N-ACETYLGLUCOSAMINE ON ASPARAGINE 130 IN THE CALCITONIN RECEPTOR EXTRACELLULAR DOMAIN SIGNIFICANTLY ENHANCES PEPTIDE HORMONE BINDING AFFINITY

Sang-Min Lee1, Jason M. Booe1, Virginie Sjoelund2, and Augen A. Pioszak1 1Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center 2Proteomics division of the Laboratory for Molecular Biology and Cytometry Research, University of Oklahoma Health Sciences Center

Introduction: The peptide hormone calcitonin (CT) regulates calcium homeostasis and bone remodeling through the calcitonin receptor (CTR) and a peptide drug targeting CTR is a therapeutic for bone disorders. Although several reports examined the calcitonin interaction mechanisms for CTR, the effects of receptor N-glycosylation on peptide hormone binding are controversial and remain as an unresolved issue. Here, we define the role of CTR N-glycosylation in peptide binding using purified human CTR extracellular domain (ECD) and deglycosylation enzymes.

Methods: HEK293T cells, HEK293S GnTI- cells, and E.coli were used to produce fully glycosylated, core- glycosylated, and N-glycan-free CTR ECD, respectively. Peptide binding was measured with a fluorescence polarization/anisotropy assay. Two deglycosylation enzymes were used: PNGase F for complete removal of N- glycans and Endo H for core glycan trimming to one N-acetylglucosamine (GlcNAc) residue. CTR ECD N- glycosylation site mutants were produced to examine critical sites for peptide binding.

Results: Consistent with the peptide affinity decrease shown with N-glycan-free CTR ECD produced from E.coli, PNGase F-catalyzed removal of CTR ECD N-glycans decreased peptide affinity ~ 10-fold. However, Endo H- trimming of the CTR ECD N-glycans to single GlcNAc residues had no effect. CTR ECD N-glycosylation site N73Q and N125Q mutants had little effect, whereas an N130Q mutant dramatically reduced peptide affinity. CTR ECD N73Q/N125Q double mutant bearing core N-glycans only at N130 retained wild-type peptide affinity that was not altered by Endo H-trimming to a single GlcNAc, but a complete removal of N-glycans by PNGase F diminished peptide affinity ~ 10-fold. Mass spectrometry confirmed a single GlcNAc at N130 of the Endo H- treated CTR ECD.

Conclusion: Our results indicate that a single GlcNAc residue at N130 of CTR ECD is sufficient and responsible for the significant enhancement of peptide hormone affinity, thus providing clear evidence for the role of N- glycans in CTR function.

Funding: NIH grant R01GM104251 and a training grant from Harold Hamm Diabetes Center.

Abstract #39

ROLE OF IGF-1 IN ASTROCYTE MITOCHONDRIAL METABOLISM AND BRAIN AGING

Sreemathi Logan1,2, Alexander Yeganeh2, Julie Farley1, Daniel Owens1, William E. Sonntag1,2 1Dept. of Geriatric Medicine, 2Oklahoma Center for Neurosciences University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA

A decline in mitochondrial function and biogenesis and an increase in reactive oxygen species (ROS) have been reported as an important determinant of aging. With advancing age there is a concomitant reduction in circulating levels of insulin-like growth factor-1 (IGF-1) that is closely associated with brain aging and neurodegeneration. IGF-1 has been reported to protect many tissues and cell-types against oxidative stress, including neurons and astrocytes. In this study, we hypothesized that reduction of IGF-1 signaling negatively impacts astrocyte function, via impairment in energy and redox homeostasis that ultimately contributes to oxidative stress and cognitive decline. We used both igfrf/f and igf-1f/f mice and AAV-mediated CRE expression driven by tissue-specific promoters to reduce circulating IGF-1 or IGFR expression in astrocytes. Learning and memory was assessed in the radial arm water maze (RAWM) and gene expression determined via qRT-PCR. Mice with circulating IGF-1 deficiency showed significant impairments in learning compared to controls in the radial arm water maze for spatial memory task. Analysis of the hippocampus of circulating IGF-1 deficient mice showed increases in antioxidant (peroxiredoxin, Aldh2) proteins by mass spectrometry suggesting underlying oxidative stress. Knock- down of IGF-1R in cultured primary astrocytes (~30% reduction) significantly reduced the energy charge (ATP/AMP ratio) compared to controls, while the total mitochondrial copy number was unaffected. NADH oxidase (complex 1 subunit) activity was increased in astrocytes with IGF-1 signaling deficiency. Mitochondrial sirtuin activity and levels of sirtuin substrates, SOD1 and SOD2, were increased in astrocytes with IGFR knockdown suggesting compensatory mechanisms of oxidative stress. These results suggest that IGF-1 positively regulates astrocyte mitochondrial metabolism and age-related decline in IGF-1 levels alters mitochondrial redox status thereby increasing oxidative stress. Thus, targeting mitochondrial metabolism in astrocytes may be a potential avenue for therapy to avert or delay cognitive deficits in advanced aging.

Funding: AG37847 Reynolds postdoctoral fellowship Oklahoma Nathan Shock Center Career Development Award

Abstract #40

CONTROL OF PTH SECRETION BY THE TRPC1 ION CHANNEL

Marta Onopiuk1, Bonnie Eby2, Vasyl Nesin1, Peter Ngo1, Maria Luisa Brandi3, Wenhan Chang4, Mary Beth Humphrey1,5, Kai Lau2, Leonidas Tsiokas1. 1Department of Cell Biology, University of Oklahoma Health Sciences Center, Okalhoma City, OK73104, USA 2Department of Medicine, Division of Nephrology, University of Oklahoma Health Sciences Center, Okalhoma City, OK73104, USA 3Universita degli Studi di Firenze, CTO-SOD Malattie del Metabolismo Minerale e Osseo, Florence, Italy 4Department of Medicine, UCSF Endocrinology and Metabolism, UCSF, San Francisco, CA, USA 5Department of Medicine, Division of Rheumatology, Immunology, and Allergy, University of Oklahoma Health Sciences Center, Okalhoma City, OK73104, USA *equal contribution

Introduction: Hyperparathyroidism, one of the most common endocrine disorders, can develop from parathyroid adenomas, hyperplasia, secondary to severe renal disease, or due to genetic conditions such as in Familial Hypocalciuric Hypercalcemia (FHH). A cardinal feature of all these conditions is abnormally high serum levels of parathyroid hormone (PTH) and calcium. PTH secreted from parathyroid gland (PTG) is responsible for increasing the level of calcium in the serum. Increased level of extracellular calcium activates the calcium sensing receptor (CaSR) which leads to inhibition of PTH production and secretion. However, the mechanism by which activation of CaSR leads to the suppression of PTH secretion in the PTG is not completely understood. In this study, we focus on the role of TRPC1 in mediating the CaSR-induced suppression of PTH.

Methods: Our studies contain in vivo experiments, where parameters of serum and urine were compared between Trpc1-/- and Trpc1+/+ mice. Ex vivo PTH secretion was measured on freshly isolated parathyroid glands. In vitro experiments, PTH secretion and single cell Ca2+ imaging were conducted on the rat stable PTG cell line (PTH-C1).

Results: Trpc1-/- mice have elevated levels of serum calcium and PTH and decreased urinary calcium excretion. PTG from Trpc1-/- mice show increased overall PTH secretion and calcium set point. Overexpression of TRPC1 in the stable PTH-C1 cell line suppresses PTH secretion, while downregulation of TRPC1 leads to inability of these cells to suppress PTH secretion in high extracellular concentration. PTH-C1 cells depleted of endogenous TRPC1 show reduced Ca2+ influx in response to activation of CaSR.

Conclusion: These data suggest that TRPC1 functions downstream of CaSR in the regulation of Ca2+ influx, which in turn can control the secretion of PTH from the PTG and further, they identify Trpc1-null mice as a new mouse model for human FHH accompanied with elevated PTH level.

Funding: Support was provided by NIAMS and PHF Bridge funding

Abstract #41

IL-24 INHIBITS GLI1 AND INDUCES DNA DAMAGE IN LUNG CANCER CELLS

Janani Panneerselvam1,4, Meghna Mehta2,4, Alshine Chen3,4, Yan D. Zhao3,4, Anupama Munshi2,4, and Rajagopal Ramesh1,4,5 Departments of 1Pathology, 2Radiation Oncology, 3Biostatistics and Epidemiology, 4Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; 5Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.

Introduction: The major cause of lung cancer-related deaths is drug resistance, and metastasis. Advances made in treatment strategies have not improved the overall five-year survival of patients diagnosed with lung cancer. Therefore, new and more effective treatments are warranted. GLI1 is a transcriptional factor and downstream target of the Sonic Hedgehog signaling (sHH) pathway. GLI1 is overexpressed in lung cancer and is associated with tumor progression. Further, GLI1 plays a role in drug resistance by modulating the DNA damage response (DDR) pathways making it an attractive molecular target for lung cancer treatment. Here we investigated whether interleukin (IL)-24 can inhibit GLI1 and its associated DDR pathway in human lung cancer cells.

Methods: Human H1299 lung tumor cell line, stably transfected with a tetracycline-inducible plasmid vector carrying the IL-24 gene was used. Cells were induced to express IL-24 protein by addition of doxycycline (Dox; 1µg/ml). The expression levels of GLI1 and its downstream molecular targets as a consequence of IL-24 expression were analyzed in H1299 cells. The inhibitory effect of IL-24 on sHH signaling was determined by RT- qPCR, western blot, luciferase reporter assay, immunofluorescence assay and Comet assay.

Results: IL-24 treatment reduced the mRNA and protein expression of GLI1 and its downstream targets (Zeb1 and Zeb2). GLI1 reporter assay demonstrated IL-24 regulated GLI1 at the transcriptional level. Further, GLI1 inhibition significantly inhibited the ATM-mediated DDR pathway resulting in increased DNA damage as evidenced by γ-H2AX foci and Comet assay. Finally, attenuation of GLI1 expression by IL-24 increased caspase- 3 and PARP activity resulting in cancer cell apoptosis.

Conclusions: Our study provides (i) evidence that IL-24 treatment induces DNA damage by regulating GLI1 expression and (ii) offers an opportunity to test combinatorial therapies of IL-24 with GANT61, a GLI1 inhibitor for lung cancer.

Funding: Funding provided by Presbyterian Health Foundation Seed Grant, Presbyterian Health Foundation Bridge Grant, Jim and Christy Everest Endowed Fund in Cancer Developmental Therapeutics and from the National Institutes of Health (NIH) –R01 CA102716

Abstract #42

ANTIBODY MEDIATES PROTECTION AGAINST VIRAL KERATITIS VIA COMPLEMENT FIXATION AND NEONATAL FC RECEPTOR TRAFFICKING.

Derek J. Royer,1 Daniel J.J. Carr,1,2 1Department of Ophthalmology, University of Oklahoma Health Sciences Center 2Department of Microbiology & Immunology, University of Oklahoma Health Sciences Center

Introduction: Antibody effector function is a major determinant in the clinical outcomes of monoclonal immunotherapies and most licensed vaccines. However, antibody biodistribution is a critical barrier to the success of such clinical interventions in the eye. Here, we explore the mechanisms regulating antibody diffusion and effector function in the cornea following infection with herpes simplex virus type 1 (HSV-1).

Methods: Antibody concentrations in healthy, mock-infected, and HSV-1 infected murine cornea lysates were evaluated by suspension array on at 24-48 hours post infection (PI). Mice were immunized with an experimental HSV-1 vaccine termed “0ΔNLS.” HSV-1 infected corneas from naive and immunized mice were evaluated at 48 hours PI by confocal microscopy to evaluate tissue localization of the neonatal Fc receptor (FcRn) expression, complement component 3 (C3) deposition, and viral antigen. Flow cytometry was used to profile specific cell populations for FcRn expression. Mice deficient in C3 were utilized to confirm the impact of complement on antibody-mediated viral clearance.

Results: Antibody concentrations sharply increased in the corneas of mock-infected and HSV-1-infected mice (p < 0.05) concomitant with increased expression of FcRn in the corneal stroma and epithelium. Intracellular deposition of processed C3 colocalized with HSV-1 antigen and was observed exclusively in the epithelium of vaccinated animals. Moreover, viral shedding from the corneas of C3-/- mice was prolonged compared to wildtype.

Conclusion: The classical complement pathway and neonatal Fc receptor (FcRn) were identified as important factors in vaccine-mediated protection against herpes simplex virus type 1 (HSV-1) keratitis.

Funding: This work was supported by National Institutes of Health (NIH) Grants R01 AI053108, P30 EY021725, and T32 EY023202. Additional support was provided by an unrestricted grant from Research to Prevent Blindness.

Abstract #43

POST-GWAS FOLLOW-UP OF CANDIDATE GENES OF DIABETIC DYSLIPIDEMIA USING NGS AND FUNCTIONAL STUDIES IN ZEBRAFISH

Bishwa Sapkota1, Megan Malone-Perez2, Anil Singh3, Huda Mussa4, Duane Goins5, Chiara Borga2, Paul Whitby4, Christopher Sansam5, J Kimble Frazer2, Chinthalapally Rao3, Dharambir Sanghera1, Department of Pediatrics, 1Section of Genetics, 2Section of Pediatric Hematology-Oncology, 4Section of Infectious Diseases, University of Oklahoma Health Sciences Center, 3Center for Cancer Prevention and Drug Development, Hem-Onc Section, PC Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma; 5Cell Cycle & Cancer Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA.

Dyslipidemia is a well-known risk factor for cardiovascular disease and a principal cause of mortality in individuals with type 2 diabetes (T2D). Despite the high heritability (50-80%) of lipid traits, previous genome-wide association studies (GWAS) have only been able to account for a fraction of this heritability (<10%) in genes involved in lipid metabolism. In this study, we aim to identify the rare functional variants in known candidate genes for diabetic dyslipidemia. We performed targeted sequencing of 14 known candidate genes (~215 kb) for 940 diabetic dyslipidemia individuals [572 cases with high serum triglycerides (TG) (>150 mg/dl) and 368 controls with low TG (<100 mg/dl)] from the Asian Indians Diabetic Heart Study. Of the 2361 high-quality variants analyzed, 953 variants (40%) were unique to high TG cases and 321 variants (13.6%) were unique to controls. Further analysis of variants within the GCKR gene using the Combined Multivariate and Collapsing methods revealed clustering of 13 functionally damaging and deleterious rare mutations near Fructose Binding site and Glucokinase (GCK) Binding sites at the sugar isomerase domains. The GCKR encodes glucokinase regulatory protein that regulates GCK (a known T2D gene) by forming a complex, which plays a role in the control of blood glucose homeostasis. The lead variant with a missense mutation of Serine/Asparagine was restricted to individuals with high TG. More than 60% of the carriers were diabetic and 90% of carriers had high TG (ranging from 182 mg/dl to 560 mg/dl). However, this variant was absent in a large (n=48,689) multiethnic Exome Aggregation Consortium (ExAC) dataset. We have designed the transgenic zebrafish (Danio rerio) with human GCKR to test the phenotypic effects of functionally disruptive variants and evaluate their metabolic consequences in vivo. Taken together, our initial results suggest the potential for detecting novel pathways in diabetes linked with hypertriglyceridemia.

Funding: This study was supported by NIH grants-R01DK082766 (NIDDK) and NOT-HG-11-009 (NHGRI), and NHLBI’s R218 (U.S. Federal Government contract # HHSN268201100037C).

Abstract #44

ANTHRAX LETHAL TOXIN SUPPRESSES THE MAP KINASE PATHWAY TO DECREASE IL-22 PRODUCTION IN TYPE 3 INNATE LYMPHOID CELLS

Sudarshan Seshadri1, David Allan2, James Carlyle2, and Lauren A. Zenewicz1 1Department of Microbiology and Immunology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK. 2 Department of Immunology, University of Toronto, Toronto, Ontario, Canada.

Anthrax, the deadly disease caused by Bacillus anthracis, is a concern for humans, as it can be used as a bioweapon. B. anthracis requires a secreted virulence factor called lethal toxin to gain entry into the host by disrupting the barrier at mucosal surfaces such as the skin, gut and lungs. Type-3 innate lymphoid cells (ILC3) are recently described innate lymphocytes which lack specific antigen receptors and are involved in barrier maintenance and host defense by secreting the cytokine IL-22. IL-22 induces proliferation and production of antimicrobial proteins from epithelial cells and thus aids in host defense, barrier maintenance and tissue repair. As disruption of barrier is required for B. anthracis dissemination in the host, we hypothesized that the pathogen modulates the function of ILC3s, particularly IL-22 secretion. In this study we show that lethal toxin decreased IL-23 stimulated IL-22 production in murine and human innate lymphocytes without affecting cell death, suggesting conserved actions in mice and humans. Biochemical studies demonstrated that lethal toxin decreased the level and activation of MAP kinases, implicating a role of these pathways in IL-23 induced IL-22 production in ILC3s. Pharmacological inhibition or small interference RNA mediated suppression of p38 MAPK decreased IL-22 production in ILC3s. Our study elucidates a novel role of p38 MAPK in IL-23 induced IL-22 production in ILC3s. Our data suggest that B. anthracis can modulate the function of ILC3s by suppressing the MAPK pathway to favor its pathogenesis.

Funding: Pilot project funding, Subaward No. A195F under U19 AI062629-10 to K. Mark Coggeshall, Ph.D. (OMRF), National Institutes of Health

Abstract #45

NANOSOME: A SMART DRUG CARRIER FOR TREATING LUNG CANCER

Akhil Srivastava1,4, Narsireddy Amreddy1,4, Anish Babu1,4, Janani Panneerselvam1,4 Meghna Mehta3,4, Ranganayaki Muralidharan1,4 Alshine Chen2,4, Yan D. Zhao2,4, Natascha Riedinger5, Shaorong Liu6,4, Si Wu6,4, Anupama Munshi3,4, and Rajagopal Ramesh 1,4,7 Departments of 1Pathology, 2Biostatistics and Epidemiology, 3Radiation Oncology, 4Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma , USA; 5Boone Pickens School of Geology, Oklahoma State University, Stillwater, OK, USA, 6Department of Chemistry and Biochemistry, University of Oklahoma, Norman, OK, USA. 7Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma USA.

Introduction: Effective management of lung cancer by chemotherapy requires efficient delivery of anticancer therapeutics to tumor site with minimal damage to normal cells and tissues. Gold nanoparticles (GNPs) can incorporate various therapeutics however, application of GNP- based therapy is limited. Nano-sized cellular vesicles like exosomes (Exo) can ferry GNP-therapeutic complexes without causing any particle aggregation or immune responses. In the present study, we describe the development and testing of a novel smart Exo-GNP- based therapeutic delivery system -‘nanosomes’- for lung cancer therapy. The nanosomes consists of GNPs conjugated to Doxorubicin (Dox) or Cisplatin (CDDP) by a pH-cleavable bond that is physically loaded onto the exosomes (Exo-GNP-Rx). The nanosomes were tested against human lung cancer cells in vitro.

Methods: Purified exosomes were loaded with drug-conjugated GNPs to create “nanosomes”. The therapeutic efficacy of nanosomes containing Dox or CDDP was individually assessed in H1299 and A549 non-small cell lung cancer cells, and normal MRC9 lung fibroblasts. Efficacy was assessed by cell viability, western blotting, fluorescence microscopy, and COMET assays. Additionally, the benefit of nanosomes over conventional Dox on cardiotoxicity was assessed using human coronary artery smooth muscle (HCASM) cells.

Results: Nanosomes exhibited enhanced rate of drug release under acidic conditions. Further, cell uptake studies showed successful uptake of the nanosomes by the recipient cells and the cell viability assays demonstrated that nanosomes exhibit preferential cytotoxicity towards cancer cells and have minimal activity on non-cancerous cells. Molecular studies showed nanosome-mediated cytotoxicity involved ROS-mediated DNA damage. Finally, free Dox but not nanosomes exhibited cardiotoxicity on HCASM cells.

Conclusions: Our study results mark the establishment of an amenable drug delivery vehicle and highlight the advantages of a natural drug carrier that demonstrates reduced cellular toxicity and efficient delivery of therapeutics to cancer cells.

Funding: The study was supported in part by a grant received from the National Institutes of Health (NIH) R01 CA167516, an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences (P20GM103639) of the NIH, and by funds received from the Stephenson Cancer Center Seed Grant, Presbyterian Health Foundation Seed Grant and Chapman Foundation.

Abstract #46

ADVANCE DIRECTIVE USAGE BARRIERS IN OKLAHOMA AND POTENTIAL IMPROVEMENT

Munim H. Deen MPH CPH (Dept. of Biostatistics and Epidemiology), Elizabeth Wickersham MD (Department of Family and Preventive Medicine)

Advance Directive (AD) forms detail a patient’s end-of-life healthcare wishes; to be acted upon when or if the patient is unable to communicate and/or is medically unable to make his or her own decisions. Completed, signed, and witnessed AD forms are binding legal documents whose proper use reduces provider ambiguity, family anguish, and unnecessary medical care. Each state has its own AD form, generally written from a legal perspective. The current AD form in Oklahoma is written at a Flesch-Kincaid reading level of 15.5, corresponding to ’15.5th grade’. Twenty percent of Oklahoma adults read at or below a fifth-grade level. Also, the current Oklahoma AD form is not logically organized. We believe these are major contributory factors to very low levels of AD completion in Oklahoma – which are as low as 5%. We believe that a simpler, more readable, and more logical AD form would lead to higher rates of completion in Oklahoma. As the initial part of a pilot study comparing AD completion rates between the current Oklahoma AD form and a simpler alternative (the Five Wishes document), primary-care practices statewide were surveyed about interest in improving AD implementation, frequency of AD-related discussions, perceived barriers to increased AD-related discussions, willingness to participate in the pilot study, and reasons for not participating in the pilot study. Demographic and geographic information was also collected. All respondents were primary-care physicians. Results showed that 88% of respondents were interested in improving AD implementation. Time constraints were the main barrier to increased AD-related discussions (73%); other reasons included ‘not sure where to start’ and ‘lack of knowledge’. Also, potential time constraints, administrative hurdles, and potential cost were all reported barriers to participation in the pilot study. Nevertheless, several practices were willing to participate in the subsequent pilot study.

Funding: Oklahoma Shared Clinical and Translational Resources

Abstract #47

COST ANALYSIS OF ALLOWING ADDITIONAL TIME IN CLEANING CONTACT PRECAUTION ROOMS.

SaiSridhar Malireddy1, Aaron M. Wendelboe1*, Shari Kinney2, Ann Chou3, Daniel Pryor2, Linda Salinas4, Sue Kim5 1. Department of Biostatistics and Epidemiology, College of Public Health, University of Oklahoma Health Sciences Center 2. Department of Health Administration and Policy, College of Public Health, University of Oklahoma Health Sciences Center 3. Department of Family and Preventive Medicine, College of Medicine, University of Oklahoma Health Sciences Center 4. Department of Internal Medicine, College of Medicine, University of Oklahoma Health Sciences Center 5. University of Southern California

Introduction: We aimed to evaluate the benefits of allowing extra time for hospital housekeepers to terminal clean contact isolation rooms. Although the Association for Healthcare Environment, recommends 40-45 minutes to terminally clean these rooms effectively, it is typical for housekeepers to be expected to conduct the cleaning in ≤30 minutes to reduce waiting time among patients ready for admission and maximize hospital revenue.

Methods: We developed a cost model in MS Excel to examine the potential cost savings per room if 15 minutes were added to terminal cleaning time for contact isolation rooms. We included fixed parameters like the cost of the emergency department (ED) going on diversion and the cost of room cleaning and variable parameters such as the percent change in risk reduction, the cost of infection, and the incremental probability of the ED going on diversion. Our basic model estimated the costs associated with preventing Clostridium difficile, and our sensitivity analyses included the additional benefit of reducing the risk of transmission of vancomycin-resistant enterococcus (VRE) and methicillin-resistant Staphylococcus aureus (MRSA).

Results: Using the midpoint values, cost savings from preventing transmission of C. difficile by allowing an extra 15 minutes of terminal cleaning time is $513 per room. That translates into an annual savings of $232.4 million based on current incidence rates of C. difficile transmission in hospital facilities. These savings increase up to $884 per room when we include savings from preventing transmission of MRSA and VRE. The probability of the ED going on divert was an important parameter in the model, but EDs would have to go on divert 47% more than modeled to no longer be cost effective.

Conclusion: The extra 15 minutes of cleaning time both decreases the burden of hospital acquired infections and also increases the cost savings for the hospital.

Funding: None

Abstract #48

DOES ULTRASOUND APPEARANCE OF THE ENDOMETRIUM AFFECT PREGNANCY RATES IN ASSISTED REPRODUCTIVE TECHNOLOGY TREATMENT?

Babawale Oluborode1, Jennifer Peck 1, Heather Burks2, LaTasha Craig2 1Department of Biostatistics and Epidemiology, College of Public Health, University of Oklahoma Health Sciences Center 2 Section of Reproductive Endocrinology & Infertility, Department of Obstetrics & Gynecology, College of Medicine, University of Oklahoma Health Sciences Center

Introduction: Implantation of an embryo within the endometrial cavity depends on receptivity of the endometrium. Without direct measures of receptivity, endometrial thickness on ultrasound (US) is a commonly used measure. Endometrial appearance (trilinear, semi-trilinear, unilinear) is noted on US, but it is unclear if this is associated with outcomes of assisted reproductive technology (ART) treatment. Therefore, this study aimed to assess whether endometrial pattern, independent of endometrial thickness, is associated with pregnancy in ART cycles.

Materials and Methods: We conducted a retrospective cohort study of all ART cycles from 2008 to 2014 at OU Physicians Reproductive Medicine. Endometrial appearance and thickness was assessed by US before beginning intramuscular progesterone. Our primary outcome was clinical intrauterine pregnancy. Secondary outcome was live birth. Risk ratios (RR) and 95% confidence intervals (CI) were calculated using a cluster- weighted generalized estimating equation method to estimate Poisson regression models with robust standard errors to account for within-couple correlation among repeated cycles.

Results: There were 1034 ART cycles with embryo transfers (778 fresh, 256 frozen) among 695 women. Trilinear patterns were most common (80.0% of cycles) followed by semi-trilinear (16.2%) and unilinear (3.8%) patterns. The pregnancy rate per embryo transfer was 56.0% for fresh and 54.3% for frozen cycles. Semi-trilinear (RR:0.89 95%CI:0.74,1.06) and unilinear (RR:1.15 95%CI:0.94,1.42) endometrial patterns were not significantly associated with pregnancy rates in all ART cycle when compared to trilinear appearance, controlling for age, endometrial thickness, serum estradiol, number of embryos transferred, stimulation protocol, cycle type, ovarian reserve, smoking, day of transfer, body mass index, and assisted hatching. Results were similar when analyzed separately for fresh and frozen cycles and when evaluating associations with livebirth.

Conclusion:Endometrial appearance does not significantly affect treatment outcome in ART cycles. Hence, it may not be cost-effective to cancel ART cycles solely because of the ultrasound endometrial pattern.

Funding: This project is supported by the Oklahoma Shared Clinical and Translational Resource Institute NIGMS U54 GM104938.

University of Oklahoma Health Sciences Center Graduate College Graduate Research Education and Technology Symposium

Abstract #49

MECHANISMS OF ESCO-DEPENDENT COHESIN REGULATION

Dawn Bender1,2, Eulalia Maria Lima Da Silva1, Jingrong Chen1, Susannah Rankin1,2 Program in Cell Cycle and Cancer Biology, Oklahoma Medical Research Foundation1, Department of Cell Biology, Oklahoma University Health Science Center2, Oklahoma City, OK 73104

Cohesin is a heterotetrameric complex that tethers sister chromatids together, promotes normal chromosome organization, and mediates DNA repair. The association of the cohesin complex with chromatin is tightly regulated. Aberrant cohesin regulation leads to developmental disorders called cohesinopathies and also contributes to malignancies in somatic cells. The mechanism by which cohesin-regulated chromosome organization and gene expression occur is not well understood. The Eco family of cohesin acetyltransferase enzymes modify the Smc3 subunit of cohesin, and are essential for the establishment of cohesion between sister chromatids by stabilizing the interaction of cohesin with chromatin. Vertebrates express two distinctly different Eco enzymes, called Esco1 and Esco2. Recently, we have shown that Esco1 and Esco2 have distinct roles in cohesin regulation: Esco2 promotes establishment of sister chromatid cohesion during DNA replication, while Esco1 plays a critical role in regulating chromosome structure throughout interphase including G1 when Esco2 is not expressed. If Esco1 is important for chromosome structure, how is its activity controlled? Esco1 is the target of DNA damage signaling, suggesting that damage-dependent modifications alter Esco1’s influence on chromosome organization via the cohesin complex. We are utilizing live cell microscopy to investigate the signaling events that control interaction of Esco1 with chromatin, with attention to those relevant to DNA repair and cell cycle progression. Phosphomutations in this region alter chromatin-binding dynamics, consistent with changes in Esco1 binding during the DNA damage response. We have also shown that the extreme N-terminus of Esco1 promotes association of the protein with chromatin throughout the cell cycle, suggesting that Esco1 activity may contribute to epigenetic memory during cell division. In current experiments, we are investigating the role Esco1 on cohesin stability in interphase cells. With these experiments, we will gain a fuller understanding of how cohesin is controlled to ensure proper chromosome dynamics.

Funding: This work was supported by NIH R01GM101250 to SR and the Oklahoma Center for Adult Stem Cell Research

Abstract #50

POSSIBLE NOVEL EXTERIOR PHENOTYPE TO IDENTIFY INDIVIDUALS AT-RISK FOR LYNCH SYNDROME

Kimberly A. Branham1, Susan Hassed2, Christopher Aston2, Erin Youngs2 1Graduate College, University of Oklahoma Health Sciences Center 2Department of Pediatrics, University of Oklahoma Health Sciences Center

Fordyce granules (FG) are ectopic sebaceous glands that appear as whitish spots on areas of the mouth. Previous studies have shown a high prevalence of FG (87% to 100%) in Lynch syndrome (LS) gene mutation carriers. This project surveyed 377 individuals diagnosed with LS and 226 without the diagnosis (the control group) to evaluate the association of FG with LS and if there is a link with specific LS genes. All participants completed a survey asking about presence of FG; the LS group also completed questions about their LS diagnosis. Statistical analysis consisted of frequency comparisons using Chi-square statistics. The overall prevalence of FG in individuals with LS was 42% compared 20% in the control group. However, the difference was primarily in women (44% in LS vs. 18% in controls, p < 0.001) whereas in men there was no difference (30% in LS vs. 29% in controls). There was no significant difference in FG prevalence based on which LS gene mutation was present. This study supports the hypothesis that presence of FG is associated with LS, although the observed prevalence rates were much lower than previous literature estimates, which used smaller research groups (4-15 participants). This is the first study to suggest that the association only holds true in women, suggesting that FG can serve as a novel exterior phenotype to identify women with a possible LS mutation. The high sensitivity seen in previous studies (100%) is not seen in this study (44% in women), but the specificity is high (82%). Thus, this study supports the previous recommendation that FG, easily seen with the naked eye, can be used as a population-wide screening tool. Individuals identified with a LS gene mutation could then be placed on a cancer surveillance plan to decrease morbidity and mortality.

Funding: None

Abstract #51

ANALYSIS OF PARENTAL PERSPECTIVES REGARDING THE DIAGNOSTIC PROCESS FOR THEIR CHILD WITH AUTISM

Rebekah Bressi1, Erin Youngs1, Michael Anderson2, Ami Bax3, Susan Hassed1 1 Genetics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 2 Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 3 Developmental and Behavioral Pediatrics, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma

Introduction: Autism Spectrum Disorder (ASD) can be a difficult diagnosis to receive and support can help parents and families come to terms with the diagnosis (Hennel et al., 2016). Parents and families experience emotional ups and downs when their child is diagnosed with autism and there is little research available on their personal experiences with the diagnostic journey (Hennel et al, 2016). Genetic counseling is vastly underutilized by those who have Autism Spectrum Disorder (ASD) and can lead to deficits in care for patients their families (Vande Wydeven et al. 2012). This study examined parent and family dynamics following the diagnosis of Autism Spectrum Disorder through predominantly qualitative online surveys.

Methods: Guardians and adult family members of people with ASD were asked about their family member (with the diagnosis of ASD) to gain insight into perceptions of the diagnostic journey. Responses were analyzed using grounded theory and coded.

Results: Three primary themes were identified that spanned multiple survey questions and participant responses: (1) most parents noted a lack of adequate resources and support at the time of diagnosis, (2) there is a vast need for transitional and adult services for individuals with autism, and (3) genetic counseling and genetic testing could be helpful to families for their child with autism. These themes stress the importance of supporting families at the time of diagnosis and throughout life.

Conclusion: Overall, parents were dissatisfied with their experiences surrounding their child's diagnosis of autism. Areas where a genetic counselor can function to potentially increase the overall satisfaction were identified.

Funding: None

Abstract #52

ALTERED PHOSPHORYLATION OF OATP1B1 AND 1B3 IS ASSOCIATED WITH DECREASED TRANSPORT FUNCTION FOLLOWING CYCLOSPORINE A TREATMENT

Alexandra Crowe1, Khondoker Alam1, Sonia Pahwa1, and Wei Yue1 1Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center

Inhibition of hepatic transporters OATP1B1 and 1B3 is an important determinant of statin-induced myopathy. Cyclosporine A (CsA) causes known drug-drug interactions with OATP substrate statins; however, mechanism(s) underlying such interactions remain unknown. We previously reported that OATP1B3 is a phosphorylated protein and OATP1B1 is putatively phosphorylated. CsA has been reported to alter the phosphorylation and function of proteins. The current study aimed to determine the effects of CsA on phosphorylation status and plasma membrane localization of OATP1B1 and 1B3. CsA pretreatment effects on OATP1B1 and 1B3 expression and transport function were determined in HEK293-FLAG-tagged and -GFP- tagged stable cell lines expressing OATP1B1 or OATP1B3. CsA effects on the phosphorylation status of FLAG- OATP1B1 and -1B3 were determined using P32-orthophosphate labeling. After labeling, cells were treated with CsA or control and subsequently immunoprecipitated with FLAG and used for autoradiography and immunoblot. Effects of CsA on plasma membrane localization of OATP1B1 and 1B3 were determined in GFP-tagged cells via live-cell imaging. CsA pretreatment (1µM, 1h) significantly decreased OATP1B1- and 1B3-mediated [3H]E217βG accumulation to 0.05 ± 0.09 and 0.32 ± 0.07 fold of control, respectively. Compared to co-incubation alone, including a 1h pre-incubation with CsA decreased the IC50 values against OATP1B1 and 1B3. CsA treatment increased phosphorylation levels of OATP1B1 and 1B3 by 1.5 ± 0.03 and 1.5 ± 0.07 fold, respectively, while leaving total protein levels unchanged. Confocal live-cell imaging demonstrated that GFP-OATP1B1 and –1B3 are localized on the plasma membrane, independent of CsA pretreatment. CsA pretreatment significantly downregulated the transport function and increased phosphorylation of OATP1B1 and OATP1B3, without affecting plasma membrane levels. This is the first study reporting that OATP1B1 is phosphorylated and that a therapeutic drug modulates the phosphorylation status of OATP1B1 and 1B3. This provides novel insight into underlying mechanism(s) behind CsA-mediated downregulation of OATP1B1 and 1B3 transport function.

Funding: Supported by NIH R01 GM094268 and GM094268-06S1 (WY)

Abstract #53

THE ASSOCIATION BETWEEN ENTERAL FEEDING AND NECROTIZING ENTEROCOLITIS BY ETIOLOGIC PATHWAY AMONG PREMATURE INFANTS IN OKLAHOMA 2001–2011

Hanh Dung Dao1, M.S., Aaron Wendelboe1, Ph.D. 1University of Oklahoma Health Sciences Center, College of Public Health, Department of Biostatistics and Epidemiology

In the US, necrotizing enterocolitis (NEC) has an overall hospitalization rate of 110/100,000 livebirths. It is a multifactorial disease, with the strongest risk factors being prematurity and low birth weight. Although infectious and ischemic etiologic pathways have been well described, the relationships of potential risk factors have not been differentiated by etiologic pathway. We aimed to determine if the association of enteral feeding as a risk factor for NEC differed by the infectious or ischemic pathways. This was a case-control study among premature infants (≤ 36 gestational week-old) in the neonatal intensive care unit at the Children’ Hospital at OU Medical Center during 2001–2011. We used the modified Bell’s criteria to define case status. Stage I (suspected) NEC cases were excluded. Cases were assumed ischemic unless there was evidence of infection (primarily using positive blood and stool cultures). Exposure measurement was the number of days on enteral feeding. Controls were premature infants without a diagnosis of NEC and matched by gestational age group and quarter of birth. We used conditional logistic regression to estimate odds ratios (OR) and 95% confidence intervals (CI) and adjust for gestational age, birth weight group, maternal drug use, and premature rupture of membranes. There were 86 cases and 242 controls. Among the cases, there were 39 infectious and 47 ischemic cases. The adjusted OR was 1.42 (95% CI: 1.05–1.93, p=0.025). The OR for infectious cases was 1.45 (95% CI: 0.96–2.18, p=0.079) and the OR for ischemic cases was 1.36 (95% CI: 0.86–1.93, p=0.185). Enteral feeding was significantly associated with case status, but when stratified by etiologic pathway, there was insufficient statistical power to determine if the differences in point estimates was real or caused by random variation. Additional analyses to further investigate confounding and effect measure modification are ongoing.

Funding: None

Abstract #54

DOWN-REGULATION OF OATP1B1 AND 1B3 TRANSPORT FUNCTION BY MTOR INHIBITORS EVEROLIMUS AND SIROLIMUS: POTENTIAL MECHANISM AND IMPLICATIONS IN OATP-MEDIATED DRUG-DRUG INTERACTIONS

Taleah Farasyn1, Sonia Pahwa1, Xueying Wang3, Pengyue Zhang3 , Khondoker Alam1, Kai Ding2, Lang Li3, and Wei Yue1 1Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73117 2Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73117 3Center for Computational Biology and Bioinformatics, Indiana University School of Medicine, Indianapolis, IN 46202 Can you confirm the name of co-author in your lab?

Introduction: Hepatic organic anion transporting polypeptides (OATP) 1B1 and 1B3 transport many clinically important drugs and are important determinants of drug-drug interactions (DDIs). Unexpected DDIs have been reported stating that patients developed severe rhabdomyolysis following concurrent administration of statins and mTOR inhibitors. The mechanism of such DDIs is unknown. The current study was designed to determine effects of mTOR inhibitors on OATP transport function and evaluate the potential of these drugs to cause OATP- mediated DDIs.

Methods: Substrate accumulation and IC50 values of mTOR inhibitors toward the inhibition of OATP1B1 and - 1B3 were determined with or without pre-incubation in HEK293-OATP1B1 and -1B3 cells. The Cmax/IC50 and R-Values were calculated based on the FDA draft guidance for OATP-mediated DDI studies. Pharmacoepidemiologic studies were conducted using patient data from the FDA Adverse vents Reporting System to determine whether concurrent usage of SIR or EV with metabolically stable statins leads to higher myopathy risk. Confocal imaging was conducted in HEK293-GFP-OATP1B1 or -1B3 cells to determine the effects of EV and SIR on OATP membrane localization.

Results: OATP-mediated substrate transport decreased following pretreatment with EV and SIR. The pretreatment condition also decreased IC50 values toward OATP1B1 and -1B3. Only pre-treatment with EV results in R-values greater than 1.25 for both OATP1B1 and 1B3 with all substrates tested. Pharmacoepidemiological studies indicated that concurrent usage of EV, but not SIR, with statins leads to increased myopathy risk. Confocal imaging indicates neither mTOR inhibitors affected OATP membrane localization. Inhibition of both mTOR complexes using a dual inhibitor does not affect transport function, suggesting that mTORC2 signaling may be involved.

Conclusions: Following pre-treatment, mTOR inhibitors rapidly down-regulate OATP transport function. Both In vitro and pharmacoepidemiological studies support that EV has the potential to cause OATP-mediated DDIs with statins. The current study provides novel insights into predicting OATP-mediated DDIs.

Funding: Supported by NIH R01 GM094268

Abstract #55

KERATINOCYTE-SPECIFIC DELETION OF THE IL-6RΑ EXACERBATES THE INFLAMMATORY RESPONSE DURING IRRITANT CONTACT DERMATITIS

Benjamin Frempah1, JoLerin Chastain1, Kaithlin Calhoun1,Tayler Schartz1, and Randle Gallucci1 1Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center.

Introduction: Irritant Contact Dermatitis (ICD) is an inflammatory response of the skin to harmful stimuli. ICD is the most common occupational skin disorder with significant impact on the quality of life of affected individuals. Following exposure to irritants, keratinocytes, the predominant cells in the epidermis of the skin, initiate the inflammatory cascade by producing a myriad of cytokines that includes IL-6. Our laboratory has recently reported that IL-6 deficiency actually exacerbates skin inflammation in a murine model of ICD. The goal of the present study is to examine the function of the IL-6 receptor alpha in the pathogenesis of ICD.

Methods: We utilized a keratinocyte-specific knock-out mouse model of the IL-6 receptor alpha (Il6rαf/f; K14-Cre), to investigate the role of IL-6Rα in keratinocytes during ICD by using a chemical model of ICD. Mice were exposed to two well-known occupational irritants JP-8 jet fuel, Benzalkonium chloride, or acetone (control) for a period of seven days. Dermatitis lesions were collected from each mouse genotype and inflammatory cytokine protein expression profile was examined by multiplex assays (Luminex). In addition, immune cell influx was characterized by immunohistochemistry (IHC).

Results: Lesional skin from Il6rαf/f; K14-Cre mice had significantly increased expression of inflammatory cytokines such as IL-1α, IL-1b, TNF-α, and IFN-Â¿ as compared to control littermates. Furthermore, IHC data shows that immune cell influx into lesional skin was higher in Il6rαf/f; K14-Cre mice relative to litter mate control.

Conclusion: Our results indicate that IL-6Rα function in keratinocytes confers a protective effect during ICD. These results suggests that the previously reported protective effect of IL-6 during ICD might be mediated by the IL-6 receptor alpha. Overall, the IL-6/IL-6R axis confers a protective effect during ICD and could be explored in the therapy of ICD.

Funding: NIH-NIOSH/CDC, R01 (09/01/2013-08/31/2017) "The role of IL-6 receptor in irritant dermatitis

Abstract #56

REGULATION OF LUNG RESIDENT TYPE II INNATE LYMPHOID CELLS (ILC2S) BY ERΑ

S. Kadel1,2, E. Ainsua-Enrich1, I. Hatipoglu1, S. Turner1, S. Singh1, S. Kovats1. Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation1, University of Oklahoma Health Sciences Center2, Oklahoma City, OK

Introduction: Asthma prevalence and severity is greater in females after puberty, suggesting the role of sex hormones and their signaling. Estrogen Receptor alpha (ERα) is expressed by immune cells and plays an important role in their development and function. Immune cells important for the pathogenesis of asthma include the lung resident type II innate lymphoid cells (ILC2s), which produce type II cytokines such as IL-5. IL-5 production by lung ILC2s is crucial for eosinophil recruitment to lungs. The main objective of this study was to determine if estrogen and ERα signaling regulate lung ILC2 numbers and function.

Methods: We used two approaches to test the role of estrogen and ERα signaling in lung ILC2s. First, we analyzed ILC2s in global ERα-/- mice. Second, we studied wild-type (WT) males and females to identify sex- differences in lung ILC2s.

Results: ERα-/- mice harbored significantly lower numbers of lung ILC2s compared to WT. Lung ILC2s in ERα- /- mice were less capable of producing IL-5, resulting in lesser recruitment of eosinophils into lungs. Notably, females had a greater frequency of lung ILC2s capable of producing IL-5. Furthermore, lung ILC2s could be subdivided into two populations based on the marker KLRG1, a receptor for E-cadherin. The KLRG1– population of lung ILC2s was significantly higher in WT females compared to WT males and ERα-/- mice. The KLRG1– ILC2s were capable of producing IL-5 and arose mainly after reproductive age in WT females, suggesting that adult estrogen levels promote development of this functional ILC2 subset.

Conclusions: Here we demonstrate for the first time that estrogen signaling via ERα regulates the number and function of lung ILC2s. We also show a sex difference in male and female lung ILC2s. Taken together, these data may help to explain the female predisposition for asthma after puberty.

Funding: OMRF Don Capra Predoctoral Fellowship, NIH NHLBI, PHF seed funding

Abstract #57

WHY PARENTS PARTICIPATE IN EARLY INTERVENTIONS: PARENT PERSPECTIVE

Elizabeth Koldoff1, Barbara Holtzclaw1, Thubi Kolobe2 1Fran and Earl Ziegler College of Nursing, University of Oklahoma Health Sciences Center 2Department of Rehabilitation Sciences, University of Oklahoma Health Sciences Center

Introduction: Premature infants born with very low birth weight (VLBW) are at risk for developmental disabilities and studies show that interventions that are initiated early (early interventions) may lead to better health and higher functional status. Despite availability, only half of parents with VLBW infants participate in these early interventions. The purpose of this study was to explore the factors that influence parents of VLBW infants to be involved in early interventions.

Methods: We used a grounded theory design. Data collection was done through semi-structured interviews with participants (n=12) from a larger interventional study. Inclusion criteria was 1) parent or caregiver of a child who is age 1 to 3 years of age, and 2) child is eligible for/involved in an early intervention programs. Parent experiences sought to answer the question, “Why do parents participate in early interventions?” Analysis was performed using NVivo, version 11. Concepts and major themes were compared and contrasted.

Results: Preliminary analysis revealed several emerging themes. Facilitators of participation include information about the intervention, trust in the health care provider, and parent mentoring. Support from experts in childhood disability is highly valued from parents with infants born with VLBW. Parents report that engagement in early interventions can be an emotional process ranging from hope to skepticism to distress. Parents desire that their child will have optimal health and function, yet they struggle with implementing therapies that impede on childhood quality of life.

Conclusions: The results suggest that a key factor to improve participation in early interventions may be a dual approach to identification and treatment. Parents are the experts of their own child yet they acknowledge the need for experts of childhood disability. The best outcomes for infants with VLBW may occur when practitioners and parents are closely aligned as partners in care.

Funding: 2016-2018 Jonas Nurse Leaders Scholar, 2014-2016 Women and Child Health Initiative Scholar

Abstract #58

SPECIFICITY OF SALIVARY GLAND-DERIVED MONOCLONAL ANTIBODIES FROM SJÖGREN’S SYNDROME PATIENTS USING HUMAN PROTEOME ARRAYS

Sherri Longobardi1,2, Christina Lawrence2, Kristi A. Koelsch2,3, Ken Smith2, Jonathan Wren2, Michelle L. Joachims2, Linda F. Thompson2, Lida Radfar4, Astrid Rasmussen2, Kathy L. Sivils2, R. Hal Scofield2,3 and A. Darise Farris2 1Graduate Program in Biological Sciences, University of Oklahoma Health Sciences Center (OUHSC); 2Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation; 3Section of Endocrinology and Diabetes, OUHSC; 4Department of Oral Diagnosis and Radiology, OUHSC; Oklahoma City, Oklahoma 73104.

INTRODUCTION: Sjögren’s syndrome (SS) is a rheumatic autoimmune disease characterized by lymphocytic infiltrations in the lacrimal and salivary glands and the presence of anti-nuclear autoantibodies to the ubiquitous Ro and La nuclear antigens, leading to severe dry mouth and eyes, dental caries and oral infections, pain, and debilitation. However, autoimmunity to these ubiquitous antigens fails to explain selective involvement of exocrine glands. We hypothesize that unknown salivary gland antigens are targets of the autoimmune response in SS.

METHODS: Recombinant human monoclonal antibodies (mAb) derived from the salivary gland plasmablasts of five individuals meeting the 2002 American European consensus criteria for primary SS were tested for binding to human proteome microarrays containing >15,500 GST- and 6xhis-tagged recombinant human proteins representing >75% of the human proteome spotted in duplicate (HuProt 3.0 arrays, CDI). Each array was probed with a mAb pool (5 - 10 mAbs/pool) derived from one or two subjects. As controls, separate arrays were probed with secondary antibody alone and pools of mAbs specific for rabies, anthrax, and S. pneumoniae vaccines. The mean and standard deviation (SD) of the foreground to background ratio of each spot was calculated. Binding ≥ 6 SD above the mean of all non-control proteins on the array was considered positive.

RESULTS: A total of 49 proteins including Ro and La antigens were bound by patient mAbs but not by irrelevant mAbs or secondary antibody alone. Furthermore, of 46 novel non-Ro/non-La antigens identified, 3 antigens were shared on at least 2 arrays.

CONCLUSION: Three novel, non-Ro/La antigens shared between two or more patients were identified. Further studies will assess expression of these proteins in salivary gland tissue, determine whether these antibodies cross-react with or arise independently of the Ro/La specificities, and determine whether these specificities occur in serum, associating with disease activity or diagnosis.

Funding: National Institutes of Health (1P50 AR060804) and Oklahoma Center for the Advancement of Science and Technology (HR 16-055-01)

Abstract #59

SHETA2 VAGINAL SUPPOSITORIES TO TREAT CERVICAL DYSPLASIA: INFLUENCE OF ESTROUS CYCLE ON DRUG ABSORPTION AND PHARMACODYNAMIC ENDPOINT

Sanjida Mahjabeen1, Manolya Kukut Hatipoglu1, Stanley D. Kosanke2, Doris M. Benbrook3, Lucila Garcia- Contreras1 1College of Pharmacy, 2Department of Pathology, 3Department of Obstetrics and Gynecology The University of Oklahoma Health Sciences Center, Oklahoma City, OK

Purpose: We demonstrated that vaginal administration of SHetA2 as a suppository achieved cervix concentrations that were >100-fold the therapeutic concentration. However, large variability was observed among mice receiving the same dose, at the same time point. In this study, we evaluated the influence of the stage of estrus cycle on the extent of drug absorption and reduction in cyclin D1.

Method: FVB female mice received SHetA2 (15mg/kg) in a suppository by the vaginal route. Untreated mice and those receiving suppository without drug (placebo) were used as controls. Mice were euthanized at 0.5, 1, 4, 8h after administration and gynecological tissues collected. A piece of the uterus was used to prepare hematoxylin- eosin slides to determine the stage of estrus cycle in each animal. Drug content in tissues was determined by HPLC. Cyclin D1 expression was determined by double-antibody sandwich ELISA.

Results: Overall, drug concentrations in all gynecological tissues were above the therapeutic concentration at all time points, with the highest concentrations observed in the cervix at 0.5h (Cmax-cervix= 54.84±17.64 µg/g). As in our previous study, significantly large variability (standard-deviation, SD) was observed among mice from same time point, with the smallest SD=30% at 0.5h and the largest SD=100% at 1 and 4 h. Histological analysis revealed that the highest absorption was in the diestrus stage and the lowest absorption at the estrus stage. Both, SHetA2 and placebo suppository induced approximately 70% reduction in cyclin D1 in cervix compared to untreated controls.

Conclusion: The extent of drug absorption in gynecological tissues after suppository administration appears to be significantly influenced by the stage of the estrus cycle. However, regardless of the stage, drug concentrations at the cervix were above therapeutic level (≥4µM) at all times. Cyclin D1 expression was not affected by these changes.

Funding: Stephenson Cancer Center Grant in Women's Cancer Research, NIH Cervical Cancer Research Grant R01 CA200126

Abstract #60

TRANSMISSION DYNAMICS OF COMMUNITY-ACQUIRED STAPHYLOCOCCUS AUREUS COLONIZATION AND INFECTION AMONG INJECTION DRUG USERS IN THE US

Kaitlin McGrew, RN, MS, Hélène Carabin, DVM, MSc, PhD, Department of Biostatistics & Epidemiology, The University of Oklahoma Health Sciences Center, Oklahoma City, OK

Introduction: Staphylococcus aureus, a gram-positive bacterium, acts as both commensal flora and a pathogen. While injection drug users (IDUs) have increased risks of S. aureus colonization and infection, no transmission dynamics models of community-acquired (CA) S. aureus in this population have not been developed. This project aims at developing a transmission dynamics model of S. aureus colonization and skin and soft tissue infections (SSTIs) among IDUs in the US.

Methods: The model was structured as a Susceptible-Colonized-Infected-Susceptible (SCIS) frequency- dependent deterministic compartmental model including both Methicillin-Resistant S. aureus (MRSA) and Methicillin-Susceptible S. aureus (MSSA) infection and colonization compartments. Co-infection and co- colonization were not modelled. 1.5% and 28.5% of new IDUs entered the model colonized with MRSA or MSSA. Heterogeneity in injection behaviors was assessed in sensitivity analyses. The impact of a syringe services program (SSP) with 100% access was evaluated.

Results: The overall prevalences of colonization and infection were estimated at 34.3% and 20.5%. Prevalences of colonization and infection were higher for MSSA (30.3% and 18.4%) than for MRSA (3.9% and 2.1%), which agrees with the distribution of MSSA and MRSA estimated from observational studies. SSPs reduced the rate of needle sharing events from 16 to 6.4 per month and reduced the overall prevalence of colonization to 43% and of infection to 7%.

Conclusion: Assuming the true prevalence of SSTIs among IDUs is 30-40% and a majority of SSTIs among IDUs are caused by S. aureus, the output provides reasonable estimates. The estimated effect of SSPs reduced the prevalence of disease similar to modeling studies of other pathogens among IDUs. Future models should include heterogeneity in colonization characteristics and injection behaviors as well as additional data for calibration to better assess the impact of alternative control strategies.

Funding: None

Abstract #61

FROM ADVERSITY TO ADVOCACY: A PARENTING CONTINUUM AFTER A CHILD’S DIAGNOSIS OF DOWN SYNDROME

Tavanna R. Porter1, Noel Jacobs2, Michael Anderson3, Susan Hassed2 1Graduate College, University of Oklahoma Health Sciences Center 2Department of Pediatrics, University of Oklahoma Health Sciences Center 3Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center

Introduction: On the whole, parents rise to meet the needs of their children but when a child has special needs there are additional challenges to overcome. Down syndrome affects an estimated 3,000 to 5,000 newborns each year and 250,000 families in the United States. Yet little is known about the transition from learning of a child’s diagnosis of Down syndrome to becoming an advocate for their care. The specific aim of this study was to survey parents of a child who has Down Syndrome and identify commonalities among families during this transition.

Methodology: This study surveyed parents of children with Down syndrome to assess the experience of receiving their child’s diagnosis, explore the challenges they face, and survey the resources they utilize. Responses were analyzed for thematic content and coded by theme. The Family Empowerment Scale (FES) was administered and comparisons were made between high and low total score respondents.

Results: The results show low total FES respondents tend to have younger children whereas high FES respondents have older children. High FES respondents report a more negative diagnosis experience whereas low FES respondents report a more neutral or factual diagnostic experience. Both groups report difficulties with the school system as one challenge they face. More low FES respondents report challenges with medical and therapy services.

Conclusions: Findings can assist genetic counselors in facilitating the transition from adversity to advocacy for families of a child with special needs.

Funding: None

Abstract #62

MANUFACTURING COST EFFECTIVE HETEROGENEOUS PHANTOMS FOR USE IN RADIATION THERAPY

James Pruett B.S., Daniel Johnson Ph.D

Introduction: Current anthropomorphic phantoms utilized in radiation oncology clinics do not represent the highly variable anatomy of a typical treatment population and are quite expensive. 3D printers in combination with carefully selected materials may be able to account for any patient anatomy while also reducing the cost associated with existing manufactured phantoms. The goal of this study is to investigate if 3D printing a heterogeneous phantom is a viable means of reproducing patient anatomy by attempting to “copy” an existing phantom from a computed tomography (CT) simulation with 3D printing technology.

Method: Specific materials were selected to represent various tissues within the human body by considering their respective Hounsfield units (HU) which were acquired via CT simulation. A shell of an existing phantom was scanned using CT and replicated using a 3D printer. The shell was then filled with selected materials representing bone, lung, and soft tissues. Various radiotherapy treatments were applied to both the 3D printed and the original phantoms using film dosimetry. Dose distributions captured by the film were compared using gamma analysis tests for corresponding treatment plans in the treatment planning system as well as the distributions captured from the control phantom.

Results: Our study showed that the manufacturing of a 3D printed anthropomorphic phantom is likely cost effective but not necessarily time effective. The results achieved from gamma analysis varied according to the quality of the radiation used in our treatment plans, yet there is some potential for 3D printing to be used for patient specific quality assurance.

Conclusion: This study demonstrates that 3D printing anthropomorphic phantoms from CT simulation is a possibility and that it can be relatively cost effective when compared to existing phantoms.

Funding: None

Abstract #63

A CRISPR-CAS9 BASED APPROACH FOR TARGETED EPIGENETIC MODIFICATION IN FRIEDREICH ATAXIA

Layne N. Rodden1, Yogesh K. Chutake1, 2, Sanjay I. Bidichandani1, 2, 3 1Oklahoma Center for Neuroscience, 2Department of Pediatrics, 3Department of Biochemistry and Molecular Biology University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104

Friedreich ataxia (FRDA) is a genetic disease caused by an expanded GAA repeat mutation in intron 1 of the FXN gene. The GAA expansion causes epigenetic silencing of the FXN promoter. Using next-generation sequencing, we have identified a region of DNA hypermethylation within intron 1 of the FXN gene in FRDA. While the promoter is unmethylated, the hypermethylation lies just outside of the CpG island within a region known as the CpG island shore (CGI shore). DNA methylation in CGI shores has been shown to negatively regulate gene expression. We hypothesize that DNA hypermethylation in the FXN CGI shore contributes to epigenetic silencing of the FXN gene in FRDA. To test this hypothesis, we are using a CRISPR-based system consisting of deactivated Cas9 tethered to either DNA methyltransferase 3A (DNMT3A) or ten eleven translocation 1 (TET1) to modify the levels of DNA methylation specifically at the FXN locus. These fusion proteins are targeted specifically to the CGI shore of the FXN gene via nine different guide RNAs (gRNAs). HEK293T cells were transfected with dCas9-DNMT3a and various gRNAs targeting the FXN CGI shore as well as the promoter region of BACH2, as a positive control. Methylation-sensitive PCR, high-resolution melting assays, and bisulfite sequencing showed increased DNA methylation at the BACH2 promoter region, as well as in the FXN CGI shore. Ongoing experiments with patient-derived cell lines and dCas9-TET1 constructs will further explore if DNA hypermethylation seen in FRDA is a critical “off switch” for the FXN gene.

Funding: This research was funded by grants from the National Institutes of Health and the Muscular Dystrophy Association.

Abstract #64

EXPERIENCES OF INDIVIDUALS ADJUSTING TO A RECENT DIAGNOSIS OF AUTOSOMAL DOMINANT RETINITIS PIGMENTOSA

Marissa Satern, Susan Hassed1, Ryan Blucker1, R. Michael Siatkowski2, Tammy Yanovitch2, Christopher Aston1 1Department of Pediatrics, University of Oklahoma Health Sciences Center 2Dean McGee Eye Institute

Background: Retinitis pigmentosa (RP) describes a group of ocular disorders involving retinal pigment deposits and degeneration that lead to progressive, irreversible vision loss. The autosomal dominant (AD) type tends to have variable penetrance and a mid-life onset. There is no known cure for RP. Numerous studies have found that individuals with vision loss experience a variety of negative psychological outcomes and major life changes. These changes can be trying at mid-life, when work and social life can be paramount for individuals. Additionally, the progressive nature creates ongoing stress. Consequently, individuals with ADRP have unique psychosocial needs. The majority of research in this area has focused on the psychological experiences of individuals with age-related vision loss. Fewer studies have examined the experiences of individuals with inherited vision loss, and none have focused on individuals with ADRP. These individuals often attend genetic counseling shortly after diagnosis.

Objective: The present study sought to explore the experiences of adults recently diagnosed with ADRP and determine factors that might affect their ability to adjust to living with the condition. Methods: Participants took an online, multi-part survey to examine genetics knowledge, coping, and quality of life, and explore adjustment experiences through open-ended questions.

Results: Qualitative analysis revealed that individuals experience many emotions in response to their diagnosis of ADRP, and cope with the effects of progressive vision loss in a variety of ways. Comparisons will be made between subgroups of individuals and their coping styles.

Conclusions: These results reinforce the importance of exploring the emotions and experiences of individuals with ADRP during the genetic counseling session, recognizing that every individual has a unique experience. Identifying factors that affect adjustment will allow for more informed counseling of this population that will aid in meeting the needs of each individual, ultimately improving quality of life over time.

Funding: None

Abstract #65

LIVED EXPERIENCES OF OKLAHOMA CAREGIVERS OF SPOUSES WITH FRONTOTEMPORAL DEMENTIA: A QUALITATIVE PILOT STUDY

Diane Smith, MSN, Donald W. Reynolds Predoctoral Scholar 2015-2017 Fran and Earl Ziegler College of Nursing, Janet Wilson, Ph.D., RN, FAAN, Primary Investigator, Kathleen Buckwalter, Ph.D., RN, FAAN, Mentor, Donald W. Reynolds Center for Geriatric Nursing Excellence, University of Oklahoma Health Sciences Center

Purpose: Explore lived experiences of Oklahoma caregivers of spouses with frontotemporal dementia (FTD). Background/Significance: FTD onset usually occurs before age 65, lasting an average of 7-13 years. Although caregiver literature is abundant, little is known about specific needs of FTD caregivers. Few strategies are available to enhance quality of life for this vulnerable group. Theoretical framework/Rationale: Because of the exploratory nature of this pilot study no theoretical framework was utilized. Concepts were defined to underpin the study. Caregiver is a spouse providing care to a person with FTD. Stressors are relationship changes and care recipient problematic behaviors. Mediators of stressors are coping and social support. Intrapsychic strains are interpersonal gains and losses.

Methods: A qualitative exploratory descriptive approach was employed. Participants were interviewed individually. Conversations were engaged through the researcher-designed interview guide. Four participants met inclusion criteria: spousal caregiver of a person with FTD and age 45-85. Interviews were transcribed from audiotapes and checked for accuracy. Content analysis was confirmed through triangulation with the sub- investigator, primary investigator, and participants.

Results: Findings were limited because small sample size. Two themes emerged that aligned with concept of stressors: FTD diagnostic and disease process. Ambiguous loss aligned with concept of intrapsychic strains. However, mediators available to these caregivers were insufficient in that effective coping skills were unknown and social support was minimal. The significant theme of caregiver abuse perpetrated by the care recipient did not conform to the conceptual definitions.

Conclusions: Findings from this pilot study, although premature, illuminate next steps for future research. Traditional conceptual models may not be fully applicable to FTD caregivers. Research is needed to understand intimate partner violence against FTD spousal caregivers. Support services, effective coping, healthy grieving, and time of symptom onset to time of accurate diagnosis of FTD should be investigated in this population.

Funding: Donald W. Reynolds Center for Geriatric Nursing Excellence, Fran and Earl Ziegler College of Nursing, University of Oklahoma Health Sciences Center

Abstract #66

CENTER OF PRESSURE AND MOVEMENT PROFICIENCY IN INFANTS WITH AND WITHOUT CP USING THE SELF INITIATED PRONE PROGRESSION CRAWLER-2

Nana Twum-Ampofo,1 , Amanda Porter1, Laura Rauh1 , Andrew Fagg2 , Thubi Kolobe1 University of Oklahoma Health Sciences Center, Dept. of Rehab Science, Oklahoma City, OK1. University of Oklahoma, Schools of Biomedical Engineering and Computer Science, Norman, OK2

Introduction: Children with or at risk for cerebral palsy (CP) are often unable to achieve prone locomotion partly due to difficulty in shifting their center of pressure (COP) which therefore inhibits movement and advancement of arms and legs during propulsion. The purpose of this study was to examine the trend of COP displacements and their relationship to prone locomotion proficiency. The Self-Initiated Prone Progression Crawler (SIPPC-2), offered the opportunity to investigate the distribution of COP in real time.

Methods: Thirty-three, 4 month old infants were assigned to 2 groups; high risk (HR); n=15 and low risk for CP (LR); n=18 and trained twice a week, on the SIPPC, over twelve weeks. Weight bearing information on forward, backward, right and left side pressures was measured in real time from load cells near the edges of the SIPPC platform. We coded and analyzed frequencies of COP distributions, for all participants, the HR and LR groups, at the beginning of training (T1), 6 weeks (T2), and 12 weeks (T3) later.

Results: All the participants indicated consistently higher forward (p<0.002) and right side (p<0.001) COP distributions compared to the backward and left side pressures, across time. Between T2 and T3, there was a decrease in forward displacement (p=0.002) and an increase in backward displacement (p= 0.874). Similar results were noted for the HR group. The differences in frequencies of COP distributions between the HR and LR groups were statistically significant, at the left (p=0.001) and right (0.028) sides at T2.

Conclusions: The results indicate that infants placed more weight on their chest initially, compared to the other parts of the body. A propensity for the infants to lean towards the right side more than the left may be related to location of toys during training but merit further investigation using video data.

Funding: Funding for this study was partially supported by the National Institute of Health grant # HD061678,

Abstract #67

PREVALENCE OF DANCE-RELATED INJURY OF ADOLESCENT FEMALE BALLET DANCERS TRAINING EN POINTE: A SYSTEMATIC REVIEW

Kimberly P. Veirs1, Carol P. Dionne1 Department of Rehabilitation Sciences, College of Allied Health, University of Oklahoma Health Sciences

Introduction: In sports medicine research, robust models, such as the baseball pitch count model, have been established for training young athletes to remain safe whilst active in sport in order to minimize injury. Different from the sports medicine scope of investigation, dance medicine research predominantly focuses on epidemiology of injury. The purpose of this systematic review was to inform dance medicine researchers and clinicians on the 1) prevalence of dance-related injury of the adolescent female ballet dancer and 2) methods to assist with injury prevention.

Methods: The extent to which the prevalence of injury has changed since initial investigation in 1969, potential factors related to the unchanging injury rate of adolescent female ballet dancers, gaps in the literature, and areas in need of future study were documented.

Results: Sixteen peer-reviewed studies were included in this systematic review. Fifty-five to 95 percent of dancers surveyed reported at least one injury per year during their careers. The rate of injury among pre- professional dancers per 1,000 dance exposures (DE=one training or performance session) was calculated in five studies. The number of injuries ranged between 1.09 and 3.21 per 1,000 DEs. Injury rates varied according to many factors: level of training, demographics (e.g. age and gender), muscle strength, flexibility, and joint range of motion (ROM). A majority of injuries involving the adolescent ballet dancer are caused by overuse of the foot and ankle and develop about the time when the average dancer commences pointe work at 11-years of age.

Conclusion: Comparisons of the reported injury rates reveal that the rate of injury of the female ballet dancer has not changed since the late 1960’s when the prevalence of dance injuries was first studied. A positive association was made across several studies between number of DE hours and number of injuries.

Funding: None

Abstract #68

ATTITUDES TOWARD CARRIER SCREENING IN FAMILIES AFFECTED BY MUCOPOLYSACCHARIDOSIS

Anna Wright1, Susan Hassed1, Erin Youngs1, Michael Anderson2 1Department of Pediatrics, Section of Genetics 1Department of Biostatistics & Epidemiology

Mucopolysaccharidosis (MPS) is a group of disorders that are considered to be rare and are characterized by severe involvement of multiple organ systems often along with reduced life expectancy. MPS is predominantly an autosomal recessive disorder- with the exception of MPS type II which segregates in an X-linked fashion. Although carrier testing is widely believed to enhance reproductive autonomy of prospective parents, the practice also raises important ethical questions. This study evaluates the attitude of parents and families affected by MPS toward carrier testing, as well their perception of reproductive options. A questionnaire was distributed to parents and family members affected by MPS to determine the attitudes toward carrier testing, explore future reproductive intentions, and their perception of reproductive options. Surveys were administered via the National MPS Society. The study population was recruited from the National MPS Society and comprised 113 participants (78 of which were parents). The results from this study demonstrate unique attitudes to carrier testing in this population. We found that 78% of participants believed that preconception carrier testing for the MPS group of disorders should be offered to all individuals. In addition, participants did not feel that carrier testing would devalue the lives of people with MPS or lead to an increased number of pregnancy terminations. Motivations for carrier testing include informing at-risk relatives and for providing recurrence information. When identifying barriers to carrier testing, participants primarily quoted access and cost of testing as preventing them from pursuing testing. Findings from our study suggest that participants are in favor of carrier testing being widely available; however, participants identify important barriers and emotional impacts of carrier testing.

Funding: None

Abstract #69

PROTEASOME ACTIVITY IS ELEVATED IN ILEAL TISSUE OF RATS WITH HEMORRHAGIC SHOCK

Hooman Yari, Vibhudutta Awasthi Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center

Introduction: Gut injury characterized by loss of tight junctions (TJ) is the most important determinant of the poor clinical outcome in hemorrhagic shock (HS). HS has been attributed to cause more than 150,000 death/year in the US. To explain the rapid loss of TJs post-HS, we hypothesized that HS perturbs proteasome function in the intestine. The objectives of this work were to determine 1) the effect of HS on proteasome activity and 2) if EF24, a diphenyldihaloketone, modulates proteasome in ileum of HS rats. We have previously shown that EF24 is highly effective in restoring TJ barrier in HS (Yadav, et al 2014 JPET 351: 413-422).

Methods: A rat model of constant volume HS was utilized in an experimental design involving control, HS, and HS+EF24 groups. HS was induced by withdrawing 45% blood via an indwelling femoral artery catheter. EF24 treatment (0.4 mg/kg) was given at 15 min post-HS and rats were euthanized 6 h post-HS to collect ileum. Trypsin-, chymotrypsin- and caspase-like proteasome activities were measured in ileal tissue homogenates by a fluorogenic peptide-based assay. We also determined IC50 of EF24 by incubating purified 26S proteasome with various concentrations of EF24.

Results: Trypsin-, chymotrypsin-, and caspase-like activities were increased significantly in the ileal tissue of HS rats (p<0.001). EF24 reduced the HS-induced proteasome activity to basal levels. IC50 values for proteasome inhibition by EF24 were found to be 466 µM and 152 µM for trypsin-like and chymotrypsin-like, respectively.

Conclusion: Increased proteasome activity plays a role in the HS-induced gut injury, and the perturbed proteasome function is normalized by EF24 treatment. The high IC50 value of EF24 suggests that its effect is most likely dependent on an allosteric mechanism rather than competitive inhibition of catalytic sites present in 20S core of proteasome.

Funding: National Institute of Health RO1 HL104286

Abstract #70

QUANTITATIVE INVESTIGATION OF IMAGE ARTIFACT IN CT IMAGES OF A MULTIPLE DIODE ARRAY DETECTOR AND THEIR EFFECTS ON THE DOSE DISTRIBUTIONS CALCULATED WITH AAA AND ACUROS-XB ALGORITHMS

F Almatouq, D DiGiacinto , V Grantham, J Baldwin , S Anderson, Department of Allied Health Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK; Department of Medical Imaging and Radiation Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK; Graduate College, University of Oklahoma Health Sciences Center, Oklahoma City, OK.

Introduction: Image artifacts change the values of Computed Tomography (CT) numbers and thus affect the accuracy of dose calculation with heterogeneity corrections on CT images. Two-dimensional multiple-diode- array detectors provide robust dosimetric tools for dose verification by measurement of dose calculated by different dose calculation algorithms used in the treatment planning of the radiation therapy. These devices are imaged first with the CT-imager that is used in patient CT simulation. The CT images of the diodes have strong image artifacts that invalidate the CT-number values. The goal of this project is to investigate quantitatively the variations in CT numbers due to image artifacts in CT images of a multiple-array-diode detector (Mapcheck2) and their effects on dose distributions calculated by the anisotropic-analytical-algorithm (AAA) and Acuros-XB dose calculations algorithms.

Materials and Methods:The dose distributions were calculated using two dose calculation algorithms: AAA and Acuros-XB with the heterogeneity correction on/off using the CT-numbers from the helical CT images of MapCheck2. The dose distributions for the different algorithms were calculated using the same optimized-fluence maps and compared with the dose distributions measured with MapCheck2.

Results: The mean value of the CT-numbers of the MapCheck2 varied from 63 Hounsfield units (HU) in the solid- water-phantom to 1600 HU near the layer that included the diodes because of image artifacts. The CT-numbers varied over a wide range of -970 HU in air to 3000 HU in metal. The dose distributions calculated with AAA with heterogeneity correction agreed well with the measured dose distributions by MapCheck2 and passed the gamma test (95%,3%,3mm). The dose distribution calculated with Acuros-XB considering heterogeneity correction of the CT numbers of MapCheck2 were on average 7%higher than the corresponding dose distribution calculated with AAA. The dose deviations between Acuros-XB and AAA ranged from 3% to 13% due mainly to image artifacts.

Funding: None

Abstract #71

DISRUPTIONS IN INFORMATION PROCESSING IN BREAST CANCER SURVIVORS

Lisa Bailey1, Melissa Craft1, Barbara Carlson1 1College of Nursing, University of Oklahoma Health Sciences Center

Disruptions in information processing [DIPs] are characterized by lapses in memory, processing speed, and attention. Commonly reported in women who have undergone treatment for breast cancer, DIPs are often unpredictable, and have a negative impact on family, work, and community activities. In this study, we used journaling to undercover the impact of DIPs on family, work, and community activities in a convenience sample of 15 older breast cancer survivors (age 64.3 ± 9.3). Women at least 24 months post-chemotherapy were recruited from oncology clinics and support groups. All completed a brief cognitive screen (Montreal Cognitive Assessment – MOCA) and entered their thoughts into a journal for 4 days, 20 minutes/day about their experiences of cognitive problems since completing active treatment. Sixty seven percent (n=10) had MOCA scores suggestive of mild cognitive impairment (21-25 points). Content analyses of the journals describe DIPs as both pervasive and unpredictable. Most subjects reported disruptions in memory (n=12, 80%), but disruptions in processing speed (n=10, 67%) and attention (n=7, 47%) were also mentioned. Disruptions in memory and attention negatively impacted their ability to make decisions, plan, and organize. Associated with lower MOCA scores (r=-.58), DIPs led to feelings of embarrassment and insecurity in performing their role in the family/home, work, and community. Future studies will use mixed methods to further explore the terms older breast cancer survivors use to characterize DIPs, the impact of DIPs on role maintenance, and their perceptions of factors that contribute to incidence and severity of DIPs, including strategies they find effective to support information processing.

Funding: Donald W. Reynolds Center of Geriatric Nursing Excellence

Abstract #72

DEHYDRATION, DEMENTIA, AND DELIRIUM INTERACTIONS IN LONG-TERM INSTITUTIONAL SETTINGS

Alejandra Dubon1 College of Nursing, University of Oklahoma Health Sciences Center

Older adults with impaired cognition are at increased risk for morbidity and mortality. Adequate fluid intake can become compromised when signaling mechanisms become suppressed. Purpose: The purpose of this evidence-based literature review was to explore dehydration indicators, preventative strategies, and treatment options to reduce delirium. Search Strategy: Databases included in the search: CINAHL, Health Source: Nursing/Academic Edition, Library, Information Science & Technology Abstracts, MEDLINE, and article bibliographies. Initial MeSH terms were “dehydration” and “dementia” published in English, resulting in 359 articles. Search terms such as: “long term care”, “institutionalized”, “cognitively impaired”, “delirium”, “hydration”, and “fluid intake” were paired with original MeSh terms, yielding 228 articles. Results of Literature Search: Excluding studies of elder neglect and abuse, acutely hospitalized, feeding tubes, dysphagia, end of life, and duplicate titles produced a final sample of 27 articles. Levels of evidence were based on the American Association of Critical Care Nurses (AACN) evidence-level system. Synthesis of Evidence: The potential for dehydration among older adults is high due to physiological changes, underlying comorbidities, and secondary effects of medications. The problem may go undetected and with adverse outcomes. With impaired cognition, the risk of mortality is doubled for older adults experiencing delirium over those without cognitive deficits. Although assessment strategies for recognizing delirium among persons with dementia can be effective, differentiating delirium from other conditions with similar symptoms remains problematic in long-term institutional settings. Managing delirium among this cohort is challenging in terms of implementing successful universal practices and outcomes. Implications for Practice: Strategies to prevent dehydration in cognitively impaired older adults in long-term settings could be a leading factor in reducing delirium. Knowledge gaps reveal research is needed in three key domains: prevention, management, and treatment. Effective practices and outcomes for persons experiencing dehydration, dementia, and delirium requires addressing these three domains.

Funding: D.W. Reynolds Predoctoral Scholar from the Donald W. Reynolds Foundation

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Abstract #73

ANALYSIS OF BIOFEEDBACK IN TREATMENT OF PARKINSON’S DISEASE-ASSOCIATED DYSPHAGIA

Clara Edwards RN MSN, Karen Rose PhD RN FGSA FAAN

Parkinson’s Disease is the second most common neurodegenerative disorder in the United States. Most individuals with Parkinson’s Disease will be diagnosed with Parkinson’s Disease-Associated Dysphagia (PD-D), increasing the risk with poor health outcomes. There are a variety of treatment options, including deep brain stimulation, medications, muscle training, altered diets, external stimulators, and biofeedback. Biofeedback mechanisms have shown to have therapeutic value in the individual with Parkinson’s Disease, yet this intervention is not used frequently. The purpose of this review was to analyze the literature regarding the efficacy and future research implications of biofeedback’s use with the PD-D individual. PUBMED, MEDLINE, and CINHAL were searched through 2016. MeSh terms "Parkinson(s)," "dysphagia," "swallowing," "deglutition," "elderly" "aged" "geriatric.” Articles regarding medication, intubation, oncology, radiation, or surgically-induced dysphagia were excluded. All primary articles written in English using biofeedback as a treatment method were included. A total of four articles were included. Biofeedback methods using sEMG (two), verbal ques (one), and video-assisted technologies (one) were found to improve swallow efficiency in PD-D. When used along with other treatments of PD-D, biofeedback is found to further increase swallow efficiency. Although the limited literature supports biofeedback as beneficial in PD-D, further analysis is warranted. Biofeedback methods range in cost, availability, and ease of use. The next step in this line of study is in further validating biofeedback’s application in PD-D in delaying onset of PD-D and establishing a relationship between skill retention of biofeedback and other treatments. To reinforce the use of this promising strategy, innovation is required to decrease cost, improve the ability to be used in the home and public environments, and simplify user requirements.

Funding: Reynolds Scholar 2016-2018

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Abstract #74

AFRICAN AMERICAN ENGLISH IN EARLY CHILDHOOD: A COMPARISON OF DEVELOPMENTAL AND DIALECTICAL VARIATIONS IN GRAMMAR

Veronica Hassink, OUHSC Denise Finneran, OUHSC Maura Moyle, Marquette University

African American English (AAE) is comprised of its own rules based on variations from Standard American English (SAE). Some of the features seen in AAE children’s speech are also characteristic of early developmental errors. The AAE features that overlap with common developmental errors (e.g., plural –S omission) will be referred to as AAE-DEV. The features not typical of developmental errors will be referred to as AAE- ONLY. Numerous studies have shown that as children who speak African American English progress through school, specific features are more commonly used even though overall dialect use drops. In this study we examine AAE in young children for dialect feature use with age. We do this by looking at feature characteristics for differential patterns of development. The purpose is to examine how AAE features are used across preschool age groups by comparing features that overlap with developmental errors and features that do not. Language samples were taken from 113 African American children ranging in age from 32 to 76 months. At the time of data collection, most children were living in low-income communities where AAE was prevalent. The language samples were transcribed and coded for AAE features, and a dialect density measure was calculated for each. We will calculate DDM in words for the AAE-DEV and AAE-ONLY features for each child. A correlational analysis will examine age, AAE-DEV, and AAE-ONLY. We propose that the overlap of AAE-DEV features and developmental errors will result in a decrease in AAE-DEV features across ages. It is not expected that the AAE- ONLY features will differ significantly across ages given that code switching is most notable starting in the early elementary years. Results will help us better understand the relationship between developmental errors and AAE use in early childhood.

Funding: U. S. Department of Education, Office of Elementary and Secondary Education, Early Reading First Program Grant S359B08008, NIH, NIGMS (Grant 1 U54GM104938), an IDeA-CTR to OUHSC, and the Gullatt endowment at OUHSC

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Abstract #75

AFRICAN AMERICAN ENGLISH DIALECT: EXAMINING THE DEVELOPMENT OF GRAMMATICAL FEAURES IN EARLY CHILDHOOD

Mary Lutter, OUHSC Denise Finneran, OUHSC Maura Moyle, Marquette University

African-American English (AAE) is a rule-governed dialect of the English language with grammatical, phonological and semantic differences from Standard American English (SAE). These grammatical differences can be further subdivided into syntactic, which represent grammar at the whole-word level and morphological, grammar at the part-word level. Numerous studies have shown that as children who speak AAE progress through school, their use of dialect features decreases. However, past research has combined the morphological and syntactic features when examining these grammatical differences across age. The purpose is to further examine how AAE features develop when comparing those that are syntactic (e.g., I BEEN going; referred to as SYN) versus morphological (e.g., omitted plural –S; referred to here as MORPH). Subjects were 113 African-American children ranging in age from 32 to 76 months. The narrative language samples of these children were analyzed and their dialect density measure (DDM) was calculated. First, the current research involved a two-tailed Spearman’s Rho correlational analysis examining DDM and the age of the child. Additionally, we will calculate DDM in words for the SYN and MORPH features for each child. We will run a correlational analysis with age, SYN, and MORPH values. The initial analysis demonstrated a significant negative correlation in age and total DDM (r= -.273, p=.002). In our follow-up analysis, it is hypothesized that the MORPH features will show a significant decrease while the SYN features will not, because more MORPH features appear to have greater overlap with developmental errors than SYN features. Also, children typically focus on whole-word learning before bound morphemes, which might result in greater use of certain SYN features with age. The current research replicated the negative correlation between age and total DDM. The current research also hopes to extend the findings to understand the individual components of syntactic and morphological features.

Funding: Funding provided by the U. S. Department of Education, Office of Elementary and Secondary Education, Early Reading First Program Grant S359B08008, NIH, NIGMS (Grant 1 U54GM104938), an IDeA- CTR to OUHSC, and the Gullatt endowment at OUHSC.

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Abstract #76

A DOSIMETRIC COMPARISON AND CLINICAL IMPLICATIONS OF PELVIC INSUFFICIENCY FRACTURES (PIF) IN POST-MENOPAUSAL WOMEN TREATED FOR UTERINE AND CERVICAL CANCERS WITH IMRT VS. 3DCRT

Jacob Smith, Stacy Anderson, Jonathan Baldwin Department of Medical Imaging and Radiation Sciences, University of Oklahoma Health Sciences Center

Purpose/hypothesis: To quantify volumetric dose differences in pelvic bones when comparing intensity- modulated radiation therapy (IMRT) to 3-D conformal radiation therapy (3DCRT) when treating uterine and cervical cancers in post-menopausal women. Pelvic insufficiency fractures (PIF) are often overlooked, not found in CT images, or misdiagnosed as metastasis. It is a seemingly rare condition, though found in individuals whose bone density is already low. Menopause, along with radiotherapy, are known to cause decreased bone density which results in a high incidence of PIF in women with uterine and cervical cancers.

Methods: Twenty computed tomography (CT) data-sets from uterine and cervical cancer patients were used to generate an IMRT and a 3DCRT plan for each set. Volumetric dose calculation was completed by analyzing the contoured sacrum, sacroiliac (SI) joint, pubic symphysis (PS), and pubic rami on the dose-volume histogram (DVH). Dose mean, median, standard deviation, minimum, and maximum descriptive statistics were computed and reported for each pelvic structure among each type of treatment plan and dose volume. An alpha of 0.05 was used for significance.

Results: There is evidence that sacral, SI joint, PS, and pubic rami mean dose (p<0.0001), 10% dose volume (p=0.0319 - p<0.0001), 40% dose volume (p<0.0001), and 70% dose volume (p<0.0001) is lower with IMRT plans compared to conformal plans.

Discussions/Conclusions: All evidence supported a higher significant radiation dose difference in 3DCRT than IMRT in the 10% volume, 40%, volume, 70% volume, and mean dose for the sacrum, SI joint, PS, and pubic rami. An increase in radiation dose to a structure correlates to an increased risk of radiation-induced injuries, such as PIFs.

Funding: None

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Abstract #77

ASSESSING THE ROLE OF A CONTOURING ACCURACY PROGRAM IN STANDARDIZING MEDICAL DOSIMETRY STUDENTS' ANATOMICAL CONTOURING SKILLS.

Hai Tran1,2,3, Stacy Anderson1,2, Kari Boyce1,2, Jonathan Baldwin1,2 1Department of Allied Health Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK; 2Department of Medical Imaging and Radiation Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, OK; 3Graduate College, University of Oklahoma Health Sciences Center, Oklahoma City, OK.

Introduction: Treatment planning in radiation therapy begins with contour multiple organs which are adjacent to tumor cells and later used to quantify the amount of radiation dose delivered. It is critical to accurately contour organs at risk to limit the dose to normal tissues. Recently, a cloud-based contour accuracy program (CAP) is introduced in effort of improving contour accuracy in treatment planning. This study assesses the role of the new program in medical dosimetry education curriculum.

Methods: Subjects were students (n=26) enrolled in the medical dosimetry and radiation therapy programs at the College of Allied Health. During the first session, participants identified and outlined the requested structures using their own sectional anatomy knowledge. Students watched pre-recorded video demonstrations by experts in radiation oncology explaining how they would identify and delineate the same structures. In the second session, students re-contoured the same structures. At the end of each session (pre- and post-education module), contour accuracy was measured using StructSure and Dice Coefficient scores. Using SAS 9.3, pre- and post-education scores were analyzed using paired t-tests or Wilcoxon sign rank tests of sample medians, as appropriate.

Results: The study found significant differences between pre- and post-education scores. Median StructSure scores for the glottis and brainstem contoured structures respectively were 85.2 points (95%CI: 80.1, 86.9, p<0.0001) and 72.5 points (95%CI: 55.5, 74.7, p<0.0001) higher after education compared to pre- education. Median Dice Coefficient scores for the glottis and brainstem were 0.73 points (95%CI: 0.62, 0.80. p<0.0001) and 0.63 points (95%CI: 0.52, 0.73, p<0.0001) higher post-education than pre-education.

Conclusion: Students’ performance was significantly improved using the CAP program. This study demonstrated a positive role for the new contouring software program in medical dosimetry education.

Funding: None

105

106

Abstract #78

INHALABLE MICROPARTICULATE SHETA2 NANOCRYSTALS FOR LUNG CANCER TREATMENT

Manolya Kukut Hatipoglu1, Sanjida Mahjabeen1, Doris M. Benbrook1,2 and Lucila Garcia-Contreras1 1Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center, Oklahoma city, OK, USA 2Department of Obstetrics and Gynecology, University of Oklahoma Health Sciences Center, Oklahoma city, OK, USA

Introduction: Treatment of lung cancer by chemotherapy is associated with severe side effects. Thus, alternative approaches to improve the treatment of lung cancer are needed. SHetA2 is capable to kill cancer cells without harming normal cells. We developed a micropaticulate SHetA2 nanocrystal (NCs) formulation for pulmonary administration to maximize concentrations at the site of drug action and enhance the therapeutic efficacy of SHetA2.

Methods: SHetA2 NCs were prepared by the bottom-up approach. The process was optimized to yield NCs with the smallest possible geometric diameter (dg) using Design of Experiments (DoE) software. The parameters optimized by the DoE were drug concentration in the organic phase, sonication power, sonication time and ultrasonication time. Morphology of the NCs was evaluated by Scanning Electron Microscopy (SEM). The batch of NCs with the smallest dg was used to manufacture NCs into inhalable microparticles by spray drying. The parameters optimized by the DoE were feed concentration, temperature and mannitol ratio. Three different formulations were obtained: SHetA2 NCs alone, SHetA2-NCs-mannitol 50% and SHetA2-NCs-mannitol 10%. The morphology of the microparticles was evaluated by SEM.

Results: Optimized SHetA2 NCs were rectangular prism shaped crystals with dg between 157nm and 710nm and size distribution (GSD) between 1.54 and 2.00. SHetA2 NCs with dg= 157 and GSD= 2.0 were produced with 60 min of sonication time, SHetA2 concentration of 75mg/mL, 20 min of ultrasonication time and 10W sonication power. This SHetA2 NC formulation was selected to be spray dried to produce microparticles. The best microparticle formulation for pulmonary delivery was SHetA2-NCs-mannitol 50%, having an average volume diameter, dv=2.53µm±0.08, 95% respirable fraction and manufacturing yield of 35%.

Conclusion: The resulting inhalable SHetA2 microparticulate NCs are suitable for pulmonary delivery to mice to evaluate pharmacokinetics and preliminary efficacy in a lung cancer mouse model.

Funding: Supported by a Seed grant from the Presbyterian Health Foundation.

107

Abstract #79

PHENOTYPIC HETEROGENEITY OF CIRCULATING TUMOR CELLS IN BREAST CANCER

Mohamed Kamal1, Smita Adhikari2, Macall Leslie1, Roy Zhang3, Crista Horton4, Beverly Talbert5, Wajeeha Razaq6, Takemi Tanaka1, 3

Introduction: Circulating Tumor Cells (CTCs) have attracted significant attention as a new class of "liquid biopsy". Since circulation is an interface between the primary tumor and metastatic organs, CTCs are recognized as a critical metastasis driver. Currently, the number of CTCs present in blood (> 5 CTCs in 7.5 mL) is prognostic of metastatic disease in breast cancer. However, the distinct CTC phenotypes critical for organ metastasis remain to be elucidated. Transcriptomic analysis of single CTCs revealed the heterogeneity of CTC populations and classified them into 3 different subgroups with epithelial-, mesenchymal-, and stem-like signatures. These studies concluded that the prevalence of CTCs with stem-like or mesenchymal properties is associated with shorter survival. We investigated the phenotypic heterogeneity of breast CTCs in the context of stem-like and EMT characteristics at a single cell level using a marker-independent, unbiased size-based enrichment method.

Methods: CTCs were fixed and isolated from 17 invasive breast cancer patients using the Rarecell® system. Fixed CTCs were used for multiplexed immunofluorescence to investigate the expression of epithelial (CK and EPCAM), mesenchymal (Vimentin and a-smooth muscle actin (αSMA)), and stem-like markers (CD44 and ALDH1A1).

Results: CTCs were identified based on their morphological and phenotypic characteristics. The CTCs were further characterized based on their EMT and stem-like signatures to understand the degree of phenotypic heterogeneity.

Conclusion: Multiplex immunofluorescence staining of CTCs collected using a size-based enrichment approach enabled us to Characterize the phenotypic heterogeneity of CTCs, providing critical information on a subset of CTCs integral to metastasis and possibly associated with prognosis in breast cancer patients.

Funding: PHF Grant

108

Abstract #80

PEGYLATED THIOL-APTAMER FOR PREVENTION OF BREAST CANCER METASTASES

Yoshihiro Morita1, Mohamed Kamal1, Shin-Ae Kang1, Roy Zhang2, Ganesh L. R. Lokesh3, Thivi Varatharasa3, Sukyung Woo4, Daniel Zhao5, Macall Leslie1, Wajeeha Razaq1, David G. Gorenstein3,6, David E. Volk3, Takemi Tanaka1,2 University of Oklahoma Health Sciences Center, Stephenson Cancer Center1, University of Oklahoma Health Sciences Center, School of Medicine2, University of Texas Health Science Center at Houston3, University of Oklahoma Health Sciences Center, School of Pharmacy4, University of Oklahoma Health Sciences Center, School of Public Health5, AM Biotechnologies6

Hematogenous metastasis is mediated by E-selectin expressed on pre-metastatic inflamed vascular endothelium cells. Previously, we developed an antagonistic 73 mer thiolated aptamer against E-selectin (ESTA), which contains 3 independent stem-loops (loop-1, -2, and -3). Our data demonstrated that ESTA effectively prevents lung metastases of CD44high metastatic breast cancer cells with no overt toxicity. However, the short circulation half-life of ESTA remained a significant challenge for intravenous administration. Therefore, we aimed to develop the second generation of ESTA with improved pharmaceutical properties and ease of bulk production. A serial deletion of one or two stem-loops revealed the essential role of loop-1 and the additional supportive role of loop-2 for E-selectin binding and functional blockade of shear resistant adhesion. Subsequently, 55 mer ESTA-7 containing loop-1 and -2 was identified as the minimal structure for the blockade of E-selectin-mediated shear resistant adhesion. For the improvement of pharmaceutical parameters, different sizes of methoxy-polyethylene glycol (5-20 kDa) were chemically conjugated to 5’-end of ESTA7. The conjugation of PEG size up to 10kDa did not hamper the adhesion blockade of ESTA 7 in vitro; however 20 kDa PEG ablated its activity. Additionally, the elimination half-life of ESTA7-conjugated with 10 kDa PEG (ESTA7- p10) was significantly improved compared to parental ESTA. Lastly, in vivo study validated the function of PEGylated ESTA7. A single intravenous injection of ESTA7-p10 prevented lung, brain, and bone metastases of MDA-MB-231 cells without activation of the complement system. In conclusion, the second-generation E-selectin thio-aptamer at a size of 55 mer with 10kDa PEG conjugation, ESTA7-p10, effectively inhibited the shear- resistant adhesion and development of hematogenous metastases of CD44high breast cancer cells, highlighting the potential clinical application of ESTA7-p10 for metastasis prevention.

Funding: This work was supported by the Department of Defense (W81XWH-11-1-0238 to T.T), the National Institutes of Health (1R01CA160271-01A1 to T.T. and NCI U54CA151668 to D.G.G), the Welch Foundation (AU- 1296 to D.G.G.), the American Cancer Society (IRG-08-060-04), and the Pennsylvania Breast Cancer Coalition (T.T).

109

Abstract #81

NKTFH CELLS STIMULATE IGG1 CLASS SWITCH AGAINST CARBOHYDRATE ANTIGEN

Binu Shrestha, Gillian A. Lang, and Mark L. Lang Department of Microbiology and Immunology University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104

Natural Killer T follicular helper (NKTfh) cells are a functional subset of CD1d-restricted NKT cells that expand in response to the CD1d-binding glycolipid ligand α-galactosylceramide (α-GC). NKTfh cells are characteristically CD4+/CD44hi/PD-1+/CXCR5+/Bcl6+ and IL-21-secreting, and they are thought to uniquely interact with B cells responding to both T-dependent protein and -independent carbohydrate antigens. Interaction between NKTfh cells and B cells may promote enhanced antibody secretion, memory B cell generation, and longevity of plasma cells, and therefore be exploited to generate better vaccines. In this study, we tested the hypothesis that NKTfh cells contribute to the B cell response to the model carbohydrate antigen NP-conjugated Ficoll and the Clostridium difficile polysaccharide II (PSII) antigen. Immunization of B6 mice with co-administered NP-Ficoll and α-GC resulted in selective Ig class-switch, such that NP-specific IgG1 was produced. This was in contrast to the IgM and IgG3 responses stimulated by NP-Ficoll alone. NKTfh cells were then ablated by crossing Cd4Cre mice to Bcl6fl/fl mice in which Bcl6 exons are flanked by Cre-sensitive LoxP regions. The lack of NKTfh cells in Cd4Cre/Bcl6fl/fl mice resulted in the specific loss of the IgG1 response to NP-Ficoll plus α-GC, with intact IgM and IgG3 responses. This directly demonstrated that NKTfh cells provided help to carbohydrate-specific B cells. Further experiments are currently underway to isolate the C. difficile surface polysaccharide (PSII) and determine whether NKTfh cells are able to stimulate PSII-specific B cells to class switch to IgG1. If this is the case, then NKTfh cells may be harnessed for the development of vaccines based on immunogenic bacterial surface polysaccharides that stimulate the production of several Ig classes and subclasses and a wider range of protective effector functions in vivo.

Funding: R21 AI125708

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111

Abstract #82

BLADDER HYPERPERMEABILITY INDUCES PERSISTENT VISCERAL PAIN: A NOVEL MECHANISM FOR VISCERAL ORGAN CROSSTALK

Quinn Baker, Ehsan Mohammadi, Casey Ligon, Beverley Greenwood-Van Meerveld

Introduction: The comorbidity between interstitial cystitis (IC) and irritable bowel syndrome (IBS) involves visceral organ cross sensitization. Previously we showed that infusion of protamine sulfate (PS) into the bladder increased bladder and colonic permeability and heightened colonic sensitivity within 24 hrs. Here we investigate the hypothesis that PS into the bladder causes persistent visceral organ hypersensitivity via activation of specific neuronal populations in the spinal cord.

Methods: Bladder hyperpermeability was induced in female Sprague Dawley rats through the transurethral infusion of PS; control rats received a sham infusion. Bladder and colonic permeability were assessed in vitro 1, 3, or 5 days post PS infusion via transepithelial electrical resistance (TEER). Bladder and colonic sensitivity were assessed in vivo 1, 3, or 5 days post PS infusion. Referred bladder hyperalgesia was measured using von Frey filaments (0.16-15g) applied to the suprapubic region and quantified via the frequency of withdrawal responses. Colonic sensitivity was assessed as the visceromotor behavioral response (VMR) to graded pressures (0- 60mmHg) of isobaric colorectal distension (CRD) and quantified as the number of abdominal contractions. Neuronal activity in the spinal cord was assessed via immunofluorescence of phosphorylated extracellular signal- regulated kinase (pERK).

Results: PS significantly decreased bladder TEER (P<0.001) and colonic TEER (P<0.001) as compared to controls. Five days post bladder PS, rats exhibited bladder hyperalgesia (P<0.001) and colonic hypersensitivity (P<0.0001). PS treatment enhanced pERK expression within the dorsal horn of the spinal cord for up to 5 days (P<0.0001).

Conclusions: In response to a single infusion of PS into the bladder, our data highlights a persistent increase in i) permeability and pain sensitivity of the bladder and colon and ii) neuronal activity in the spinal cord. These findings advance our understanding of the mechanisms of visceral organ crosstalk and highlight the comorbidity between IC and IBS.

Funding: None

112

Abstract #83

ABNORMAL WEIGHTING OF MID AND HIGH-FREQUENCY BANDS IN SENSORINEURAL HEARING LOSS

Jeremiah Hilton, Blas Espinoza-Varas

Owing to reduced afferent input from high characteristic-frequency hair cells, steep-slope high-frequency sensorineural hearing loss (“Steep HF loss”) reorganizes the tonotopic maps in the central auditory nervous system: map expansion is observed in the mid frequencies (MF) and map contraction in the high frequencies (HF). We studied how this reorganization affects the relative weighting of MF and HF information. Listening to complex tones consisting of a barely-audible MF and a clearly-audible HF sinusoid, participants were asked discern simultaneous increments in the frequency (DF) of MF and in the duration (DT) of HF; the sensation level was 1.5-8.0 dB for the MF sinusoid and 25-39 dB for the HF sinusoid. We hypothesized that, in steep-HF-loss participants, the perceptual weight would be greater for the MF sinusoid impinging on the cortical map-expansion region than for the HF sinusoid impinging on the map-contraction region. In contrast, in normal-hearing or flat- hearing loss participants, the perceptual weighting would be greater for the clearly-audible HF than for the barely- audible MF sinusoid. In the discrimination task, the 1st, 2nd, and 3rd observation interval presented a standard complex tone followed by comparisons 1 and 2. Either the 2nd or 3rd interval presented DF in the MF sinusoid, DT in the HF sinusoid or both DF and DT, and participants to reported which comparison differed from the standard. The psychometric functions relating correct probability, P(C), to DF with DT as parameter, and vice versa, upheld the hypothesis: P(C) increased sharply with DF in the steep-HF-loss group but remained virtually at chance in the other two groups. P(C) increased with DT in all groups, but the parametric effect of DF was seen only in the steep-HF-loss group. Compared to one with flat hearing loss or with normal hearing, the steep- HF-loss participant processes more efficiently the mid- than the high-frequency information.

Funding: Presbyterian Health Foundation

113

Abstract #84

CHANGES IN HISTAMINE AND ITS HISTAMINE-3 RECEPTOR FOLLOWING A MILD TRAUMATIC BRAIN INJURY

Asher Khan1, Matt Baier3, Amarachi V. Oleru1, Alexander C. Edwards1, Megan R. Lerner2, Hibah O. Awwad1,4 1Department of Pharmaceutical Sciences, 2Department of Surgery, University of Oklahoma Health Sciences Center, 3Oklahoma School of Science and Mathematics, 4Oklahoma Center for Neuroscience

An estimated 1.7 million people sustain a traumatic brain injury (TBI) annually in the United States, of which approximately 52,000 are fatal cases (Centers for Disease Control and Prevention). Since there are currently no FDA-approved pharmacological agents for TBI treatment, identifying the neurochemical pathways in the brain that follow TBI is critical in the emergence of novel treatments. Histamine is one such endogenous neurotransmitter that modulates memory, pain, and sleep. Studies have shown mixed results linking histamine with protective responses to ischemic brain injury, prompting further question as to the specific role of histamine in TBI pathology. As a neurotransmitter, histamine acts through four receptor subtypes, including the Histamine- 3 receptor (H3R). H3R is a presynaptic autoreceptor localized in the central nervous system (CNS) that has been identified as a candidate therapeutic target for various neuronal diseases. The goal of this study was to quantify changes in histamine and its H3R receptor following mild TBI. Stereotaxic surgery (craniotomy and impact to left cortex) was performed to induce mTBI and sham rats received a craniotomy only. Rat brains were extracted 24 hr post-TBI, left and right sensory cortex (SC) and motor cortex (MC) were dissected and homogenized separately. Levels of H3R and injury proteins were quantified using immunoblotting analysis. Higher H3R levels were detected on the ipsilateral MC and SC compared to the contralateral side of mTBI rats. H3R and the astrocytic marker, glial fibrillary acidic protein (GFAP) increased in a similar pattern in injured rats on day 1 post-injury compared to naïve and sham rats. Histamine levels are currently being measured using fluormetric high performance liquid chromatography (HPLC). Mild TBI induced an increase in H3R expression levels in the MC and SC and that correlates with TBI-induced cortical astrogliosis detected in these brain regions.

Funding: This work was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number 8P20GM103447 and an OU College of Pharmacy Seed grant to HOA.

114

Abstract #85

MODELLING MUMPS TRANSMISSION USING OKLAHOMA OUTBREAK DATA: IS VACCINE EFFICACY LOWER THAN EXPECTED?

Cassandra Long1, Jolianne Stone2, Lacey Kovar2, Helene Carabin1, Laura A. Beebe1, Laurence Burnsed2 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Science Center 2Acute Disease Service, Oklahoma State Department of Health

Introduction: Beginning September 2016, Oklahoma has been experiencing an ongoing mumps outbreak. We utilized a transmission dynamics model to compare vaccination coverage among Oklahoma school-aged children and published vaccine efficacy (VE) of the mumps component of the measles mumps rubella (MMR) vaccine to predict duration of the outbreak.

Methods: A mathematical transmission dynamics model was constructed using Berkeley Madonna to predict mumps transmission based on Oklahoma vaccination rates, vaccine efficacy and basic reproductive rate parameters. The model is a modified SEIR (Susceptible – Exposed – Infectious – Recovered) model using two separate reservoirs for the outbreak-affected population and the general population. Outbreak data was obtained from the Oklahoma State Department of Health and compared to expected cases based on the model at five week intervals using chi-square analysis for goodness of fit.

Results: MMR vaccination coverage of school-aged children in Oklahoma is approximately 92.6%. Under the assumption that 2-dose mumps vaccine efficacy is 88%, and using a basic reproductive number of 5.4, the current level of vaccination should provide herd immunity and limit transmission of the disease. However, MMR vaccination levels in the outbreak-affected population and VE had to be modified in order to fit the model to the observed epidemic curve. The model fit through week 20 suggested a 58% VE with the primary affected population having only 65% vaccination coverage.

Conclusion: Oklahoma is experiencing higher than expected transmission of mumps virus. Based on mathematical modelling, we expect continued spread and a secondary peak of cases later this year. Further evaluation is needed to understand whether this outbreak is due to a lower than expected VE, or other factors affecting VE in the current outbreak-affected population.

Funding: Epidemiology and Laboratory Capacity for Infectious Diseases Cooperative Grant

115

Abstract #86

SPORADIC SALMONELLOSIS CASES AND FOOD CONSUMPTION TRENDS IN OKLAHOMA

Cassandra Long1, Lacey Kovar2, Amanda Shoemate2, Laura A. Beebe1, Laurence Burnsed2 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Science Center 2Acute Disease Service, Oklahoma State Department of Health

Introduction: Salmonella is a common source of foodborne illness in Oklahoma. Food consumption surveys are utilized to generate hypotheses during cluster investigations; however, the most recent national food consumption survey, FoodNet, was conducted during 2006-2007. Because food consumption trends change over time, several public health agencies including Oklahoma have recently begun food history interviews of sporadic Salmonella cases for case-case comparisons during cluster investigations. We compared interview responses from 2016 case interviews to FoodNet data to determine which food consumption trends have changed in the past decade.

Methods: We attempted to complete food consumption history interviews with each eligible sporadic Salmonella case reported in Oklahoma during 2016 (N=785). Eligible cases had GI symptoms and were not associated with an outbreak. We obtained information regarding food consumption 7 days prior to diarrhea onset. Proportion of food exposures were compared to FoodNet responses using a two proportion z-test. Food category proportions were tested using a chi-square analysis.

Results: Two hundred ninety-five Salmonellosis cases were interviewed in 2016. Foods commonly reported by respondents included poultry (64.7%), beef (61.9%), milk (54.2%), eggs (56.9%), chips (46.7%) and bananas (44.7%). Uncommon food histories included seafood (18.4%), fresh herbs (14.8%), and seeds (13.7%). One in four Oklahomans consumed no fruit or vegetables (24.1%). Foods with the most similar exposure patterns to FoodNet included raw milk (z=.96), ground beef (z=1.09), and lettuce (z=1.66). Some foods such as sushi, which are uncommonly consumed in Oklahoma, differed greatly from the FoodNet survey (z=-91.73).

Conclusion: The most common types of foods consumed by sporadic Salmonella cases in Oklahoma include meats and dairy products; whereas low levels of fruits and vegetables, seafood, seeds, and fresh herbs were reported. Results from this analysis reveal differences in food consumption trends between sporadic Oklahoma Salmonella cases and the last national food consumption population survey.

Funding: Epidemiology and Laboratory Capacity for Infectious Diseases Cooperative Grant

116

Abstract #87

AN INVESTIGATION INTO THE RELATIONSHIP BETWEEN INFLUENZA VACCINATION STATUS AND EDUCATION LEVEL FROM 2015 BEHAVIORAL RISK FACTOR SURVEILLANCE SYSTEM (BRFSS)

Anita Motwani1, Sara Vesely1 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center

Introduction: Vaccination was one of the ten great public health achievements of the 20th century. With general public perception regarding vaccination becoming tumultuous in recent years, the influenza vaccine in particular provides an additional barrier in compliance when compared to other vaccinations due to the necessity for individuals to receive it annually. This study investigated the association between influenza vaccination status and education level among respondents of the 2015 Behavioral Risk Factor Surveillance System (BRFSS) survey.

Methods: Data from the 2015 BRFSS survey was utilized to investigate the association between influenza vaccination status and education level. Data was collected for BRFSS through a nationwide telephone survey that collected state information regarding respondent’s demographics, risk behaviors, chronic health conditions, and use of preventative services. Data was analyzed with Statistical Analysis Software (SAS).

Results: Respondents who were vaccinated were more likely to be female (p<0.0001), white (p<0.0001), and have a higher income (p<0.0001) than respondents who were not vaccinated. Nationwide, the lowest prevalence of influenza vaccination was among respondents with some high school education at 41.98% (p<0.0001). The highest prevalence of influenza vaccination was among respondents who were college graduates at 53.75% (p<0.0001). When stratified by age group, there was a positive association between influenza vaccination status and education level (p<0.0001).

Conclusions: Among 2015 BRFSS survey respondents, there was an positive association between influenza vaccination status and education level. When stratified by age group, influenza vaccinaiton prevalence increased by education level within each age category. There was an overall increase in vaccination prevalence by age group. Investigation of the factors that affect influenza vaccination prevalence can aid public health initiatives to increase vaccination compliance.

Funding: None

117

Abstract #88

INVESTIGATING THE HIGH SUCCESS OF TOBACCO ABSTINENCE IN WOMEN WITH CONTINGENCY MANAGEMENT INTERVENTION IN THE PREVAIL I STUDY

Anita Motwani1, Darla Kendzor2 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center 2Department of Family and Preventative Medicine, University of Oklahoma Health Sciences Center

Introduction: In tobacco cessation studies, women often have poorer cessation outcomes compared to men. In the PREVAIL I study, women randomized to tobacco cessation intervention with contingency management (CM) had the highest rates of cessation abstinence success when compared to men in CM, men in usual care (UC), and women in UC. Conversely, women in UC had the lowest rates of cessation abstinence success. This study aims to investigate the factors that contribute to the large difference in tobacco abstinence success between women in the intervention and control groups.

Methods: Data from the PREVAIL I study was utilized to investigate the association between gender and success in contingency management intervention for tobacco abstinence in economically disadvantaged smokers. Data was collected at the Tobacco Cessation Clinic at Dallas County, Texas, safety net hospital between August 2011 and April 2013. Association between measures was evaluated with chi-square test. Statistical analyses were performed with SAS statistical software (University Edition, SAS Institute, Cary, NC, USA).

Results: Women in CM were more likely to remain abstinent to tobacco past the quit date week 1 (p=0.0337), week 2 (p=0.0095), week 3 (p=0.0204), and week 4 (p=0.0246). Women who were tobacco abstinent at week 1 past quit date were more likely to have a GED or high school diploma (p=0.2972) and be African American (0.1748).

Conclusions: The identification of factors associated with cessation success in women in CM will aid in the development of effective tobacco cessation for a group that has been shown to have poor cessation outcomes. The cost of CM intervention in the PREVAIL I study was $63.40 per participant. When compared to the large cost of treating a chronic smoker, the low cost of this intervention coupled with its success is encouragement to investigate this success further.

Funding: None

118

Abstract #89

PREVALENCE OF NEUROCYSTICERCOSIS AMONG PATIENTS WITH SELECT NEUROLOGICAL DISORDERS IN SIXTY VILLAGES IN BURKINA FASO, 2011-2012

Kuna Okong1, Athanase Millogo2, Rabou Cisse3, Ida Sahlu4, Veronique Dermauw5, Pierre-Marie Preux6, Marie- Paule Boncoeur-Martel6, Pierre Dorny7, Andrea Winkler8, Rasmané Ganaba9, Vivien Richter10, and Hélène Carabin1 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center 2Department of Internal Medicine, Sourou Sanou University Hospital Center, Bobo-Dioulasso, Burkina Faso 3Department of Radiodiagnosis and Medical Imagery, Yalgado Ouedraogo University Hospital Center, Ouagadougou, Burkina Faso 4 Department of Epidemiology, Brown University School of Public Health, 121 South Main Street, Providence, Rhode Island, USA, 02903 5 Institute of Tropical Medicine Antwerp, Nationalestraat 155, 2000 Antwerp, Belgium 6 INSERM, Univ. Limoges, CHU Limoges, UMR 1094, Tropical Neuroepidemiology, Institute of Neuroepidemiology and Tropical Neurology, GEIST, F-87000 Limoges, France 7 Institute of Tropical Medicine Antwerp, Nationalestraat 155, 2000 Antwerp, Belgium 8 Institute of Health and Society, Faculty of Medicine, University of Oslo, Norway, Department of Neurology, Technical University of Munich, Germany 9 Agence de Formation de Recherche et d'Expertise en Santé pour l’Afrique (AFRICSanté), 773, rue Guillaume Ouedraogo, Bobo Dioulasso, Burkina Faso 10 Department of Diagnostic and Interventional Radiology, University Hospital of Tübingen, Tübingen, Germany

Introduction: Neurocysticercosis (NCC) is considered the most frequent preventable cause of epilepsy in low income countries. The proportion of NCC among people with epilepsy has been estimated in clinical settings or small-scale community settings in Sub-Saharan Africa but no study has estimated this proportion among people with severe chronic headaches (SCH). The aim of this study is to estimate the prevalence of NCC among residents of 60 villages in Burkina Faso suffering from epilepsy and/or severe chronic headaches (SCH).

Methods: Baseline cross-sectional data collected between February 2011 and January 2012 from a large randomized community-based controlled trial were included. A screening questionnaire for epilepsy and SCH was used among 4,970 participants in 60 villages. Among 681 participants who screened positive, 623 were seen by a physician to confirm their diagnoses. An additional 257 participants who were asymptomatic were examined by the physician identify cases missed by screening. Of all 327 villagers with physician-confirmed symptoms, CT-scans were done on 260, of which 253 were available. NCC was defined as the presence of at least one definitive or highly suggestive lesion of NCC on the CT-scan. Prevalence Ratios (PR) and their 95% Confidence Intervals (95% CI) were calculated using SAS version 9.3.

Results: Lesions of NCC were present in 22.6% (95%CI: 15.6%-30.9%) of the 124 participants with epilepsy, of whom 21 also had SCH, and 12.1% (95%CI: 6.8-19.4%) of the 116 participants with only SCH. The prevalence of NCC was higher among those with epilepsy than those with SCH only (PR=1.78; 95%CI: 1.04-3.37).

Conclusion: This is the first large-scale study community-based study to not only estimate the prevalence of NCC among patients with epilepsy and SCH in Burkina Faso but also to report the prevalence of NCC among individuals with SCH in Sub-Saharan Africa.

Funding: This work was supported by the National Institute of Neurological Disorders and Stroke and the Fogarty International Center at the US National Institutes of Health under the Brain in the Developing World: Research across the life span program (R01NS064901)

119

Abstract #90

INFLUENZA MORBIDITY AND MORTALITY IN OKLAHOMA, 2013-2016

Kuna Okong1, Kendra Dougherty2, Laurence Burnsed2, and Helene Carabin1 1 Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center 2 Acute Disease Service, Oklahoma State Department of Health

Introduction: Multiple sources of independent data are used by the Oklahoma State Department of Health (OSDH) to describe influenza activity and severity including medically attended outpatient visits with influenza- like illness (ILI), laboratory confirmed influenza-positivity from sentinel healthcare partners as well as case-based reports of influenza-associated hospitalizations and deaths. We evaluated data from 2013 through 2016 to describe the last three influenza seasons in Oklahoma, as well as identify trends among the multiple data sources.

Methods: Influenza-associated hospitalizations and deaths were reported from Oklahoma hospitals and the state medical examiner. Sentinel providers’ reports were used to calculate the weekly percentage of all outpatient visits due to ILI. The percent influenza positivity was estimated as the proportion of ILI patients’ samples positive for the influenza virus submitted to laboratories performing influenza testing. Positive samples were then sent to the Oklahoma Public Health Laboratory (HL) for strain characterization. The mean percent ILI was estimated as the average proportion of ILI outpatient visits to sentinel providers during weeks when the percent influenza positivity was above 10%.

Results: During the 2013-14, 2014-15 and 2015-16 influenza seasons, there were 1385, 2326, and 565 hospitalizations, and 73, 115 and 16 deaths reported, respectively. The predominant circulating influenza strains identified by the Public Health Laboratory during these three seasons were A 2009 H1N1, H3N2, and 2009 H1N1. The average weekly proportions of patients seen for ILI were 3.9%, 5.1%, and 2.8%, the percent influenza positivity was 5.4%, 7.1% and 4.1% and the mean percent ILI was 6.7%, 8.2% and 3.0% for the 2013-14, 2014- 15 and 2015-16 seasons, respectively.

Conclusions: All sources of data agreed that the 2014-15 season during which the Influenza A (H3N2) predominated was more severe. Different sources ranked the three seasons identically for frequency and severity, although the measure of their relative importance varied.

Funding: None

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Abstract #91

HOSPITAL ADMISSION STATUS AND FIRST-TIME DIAGNOSIS OF VENOUS THROMBOEMBOLISM IN A CONTEMPORARY POPULATION-BASED COHORT IN OKLAHOMA COUNTY

Andrew Starnes1, Justin Dvorak1, Aaron Wendelboe1, Micah McCumber1, Janis Campbell1, Nimia Reyes2, Michele Beckman2, Gary E. Raskob1 1Department of Biostatistics and Epidemiology, University of Oklahoma College of Public Health 2National Center on Birth Defects and Developmental Disabilities, Centers for Disease Control and Prevention (CDC)

Background: More than 500,000 hospitalizations each year in the United States have an associated discharge diagnosis of venous thromboembolism (VTE) and morbidity and mortality are known to be significant. However, the proportion of inpatient versus outpatient diagnoses are ensuing management are uncertain. We aimed to measure incident VTE diagnoses and whether hospital admission followed or preceded diagnosis.

Methods: In collaboration with the Centers for Disease Control and Prevention, a population-based surveillance system was established in Oklahoma County, OK from April 1, 2012–March 31, 2014. Active surveillance involved reviewing all related imaging studies (e.g., chest computed tomography and compression ultrasound) from each inpatient and outpatient facility in the county. Diagnosis, treatment, and risk factor data were collected on all case-patients. Patients with a history of previous VTE or cancer were excluded from this analysis.

Results: Initial analysis identified 1,341 patients with imaging-confirmed VTE. Among patients with deep vein thrombosis (DVT) only (n=873), 42% were never admitted, 24% were diagnosed on or before admission, and 34% were diagnosed after admission. Among patients with pulmonary embolism (PE) only (n=256), 6% were never admitted, 67% were diagnosed on or before admission, and 27% were diagnosed after admission. Among patients with both PE and DVT (n=212), 5% were never admitted, 73% were diagnosed on or before admission, and 22% were diagnosed after admission. Of those diagnosed after admission (n=415), only 73% had received prophylaxis.

Conclusion: The majority of incident VTE cases (n=926; 69%) were diagnosed in outpatient settings. There is a marked difference in proportion of DVT vs. PE cases managed without admission. Continued research is indicated to determine if more patients could be managed without hospital admission and why 27% diagnosed after admission did not receive prophylaxis.

Funding: Centers for Disease Control and Prevention (CDC)

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Abstract #92

EXPLORING SELECTION BIAS DUE TO INADVERTENT USE OF A RISK FACTOR FORM IN A HEPATITIS C SCREENING STUDY.

Olivia Wright1, Hélène Carabin1, Mary Williams1, David Gahn2, Jorge Mera3, Whitney Essex3, and Doug Drevets4 1Department of Biostatistics and Epidemiology, University of Oklahoma Health Sciences Center 2Cherokee Nation Public Health 3Infectious Disease Division, Cherokee Nation Health Service 4Infectious Disease, College of Medicine, University of Oklahoma Health Sciences Center

Introduction: The Cherokee Nation Health Service (CNHS) is currently implementing a hepatitis C virus (HCV) elimination program that aims to screen all users of CNHS between 20 and 69. A risk factor form (RFF) was implemented at the time of screening, as this information was not available in the electronic medical records. Providers may have used the RFFs as a tool to decide whom to screen, possibly leading to selection bias. The goal of this study was to determine if CNHS patients aged 20 to 69 with known HCV risk factors were screened at a higher proportion compared to those without risk factors.

Methods: Screening status was examined by HCV risk factors as reported on the RFFs that included ever had tattoos or body art, ever use of intravenous drugs (IVDU), ever been in prison, blood transfusion before 1992, and ever lived with a person with HCV. A chi-square of independence test was used to determine the statistical significance of the presence of risk factors and being screened. The 95% confidence interval for the proportions and the cumulative incidence ratios was determined.

Results: Most risk factors were associated with an increased likelihood of getting screened with the cumulative incidence ratios and 95% confidence intervals as ever IVDU 1.13 (1.10, 1.17), ever been in prison 1.05 (1.04,1.07), blood transfusion before 1992 1.23 (1.20, 1.27), and ever living with a person with HCV 1.13 (1.10, 1.16); however, ever had a tattoo or body art was significantly associated with not being screened 0.91 (0.90, 0.93).

Conclusion: Presence of most risk factors were associated with being screened with the exception of history of tattooing or body art in this population. It is conceivable the REF was incorrectly being used as a screening tool.

Funding: Gilead Foundation

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Abstract #93

SYNTHESIS AND CHARACTERIZATION OF A NOVEL FLUORESCENT PROBE FOR TARGETING GASTRIN RELEASING PEPTIDE RECEPTOR EXPRESSING CANCER CELLS

Rachel Abraham1, Gopal Pathuri2, Hariprasad Gali2 1Oklahoma School of Science and Mathematics, 2Department of Pharmaceutical Sciences, College of Pharmacy, The University of Oklahoma Health Sciences Center, Oklahoma City, OK 73117, USA

Introduction: Gastrin Releasing Peptide (GRP) receptors are expressed with high densities on the surface of several types of cancer cells including prostate, breast, small cell lung, pancreatic, cervical, and colorectal cancers. Bombesin (BBN) is a 14 amino acid peptide that is an amphibian analog of human GRP that binds with high affinity and specificity to the GRP receptors. BBN(7-14) sequence is known to be essential for highly specific GRP receptor binding. The objective of this research was to synthesize a novel BBN(7-14) conjugate containing fluorescein isothiocyanate (FITC) for use in the quantification of GRP receptor expression on cancer cells by flow cytometry and fluorescence microscopy.

Methods: A BBN(7-14) analog containing a polyethylene glycol linker (PEG), PEG2-BBN(7-14), was synthesized by an in-house developed automated peptide synthesizer using traditional fmoc-based solid-phase peptide synthesis method. Following the synthesis, the peptide was purified using reverse-phased high performance liquid chromatography (HPLC) and characterized by electrospray mass spectrometry (ES-MS). Purified PEG2- BBN(7-14) was then reacted with FITC in the presence of diisopropylethylamine in dimethylformamide at room temperature overnight to obtain FITC-PEG2-BBN(7-14) conjugate. The conjugate was purified by HPLC and characterized by ES-MS and fluorescence spectroscopy.

Results: Purified PEG2-BBN(7-14) and FITC-PEG2-BBN(7-14) conjugates were obtained with a yield of 40.5% and 77.2% respectively. Mass spectral analyses were consistent with the molecular weights calculated for each conjugate. Fluorescence emission spectrum of FITC-PEG2-BBN(7-14) conjugate displayed an emission maximum at 518 nm confirming the presence of FITC.

Conclusion: A novel FITC-PEG2-BBN(7-14) conjugate was successfully synthesized and characterized.

Funding: The University of Oklahoma College of Pharmacy

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Abstract #94

REGULATION OF HIPPOCAMPAL GENE EXPRESSION ACROSS THE ESTRUS CYCLE

Akansha Chandrasekar1, Laura Otalora2, Benjamin Wronowski2, Willard Freeman2 1 Oklahoma School of Science and Mathematics 2 Department of Physiology, University of Oklahoma Health Sciences Center

Introduction: Brain aging and age-related changes across molecular and cellular processes likely contribute to the many neurological disorders and functional deficits evident with aging. Previously, we have identified sex divergences (i.e., sex differences that only become apparent with aging) in hippocampal neuroinflammation. A likely cause for age-related sex divergences are the hormonal changes that occur with reproductive senescence. However, the basic understanding of sex hormone regulation of brain gene expression is very limited. Therefore, by examining hippocampal gene expression across the estrus, diestrus, and proestrus phases of the estrus cycle, we sought to define the regulatory effects of sex hormones on gene expression and thereby determine if sex hormones are responsible for sex divergences with aging.

Methods: Female C57BL6 mice (3 months old) were monitored for estrus cycle phase for 3 weeks by vaginal lavage and cytology. Hippocampi were collected from mice at estrus, diestrus, and proestrus (n = 9/group). RNA was isolated from hippocampal samples (n = 4/group) and subjected to strand specific RNA sequencing (RNA- Seq). The resulting data was trimmed, aligned, and normalized followed by differential expression analysis (ANOVA, SNK) between the 3 groups. Differentially expressed genes were analyzed for cellular origin, pathways and processes, and compared to previously identified age-related sexually divergent genes.

Results: As expected, hippocampal gene expression varied across the estrus cycle. Differentially expressed genes from a variety of cellular processes were evident, including neuroinflammation. Estrus cycle responsive genes were enriched for microglial specific transcripts and demonstrated some commonalities with age-related sexually divergent genes.

Conclusion: These data support our hypothesis as exemplified by opposite regulation of mRNAs during proestrus, when estrogen levels are highest, as compared to age-related changes, when estrogen levels decline. These findings also provide novel insights into the previously underappreciated dynamism of brain gene expression across the estrus cycle.

Funding: This work was supported by the Donald W. Reynolds Foundation, the National Institute on Aging (P30AG050911), National Eye Institute (R01EY021716, R21EY024520) and Oklahoma Center for Advancement of Science and Technology (HR14-174).

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Abstract #95

COMPARATIVE LINEAGE TRACING OF PDGFR ALPHA AND BETA IN LIMB BONE DEVELOPMENT

Josephine Hriscu, Oklahoma School of Science and Mathematics

Platelet-Derived Growth Factor (PDGF) is an important mitogen for mesenchymal cells in the developing embryo. PDGF receptor alpha (PDGFRa) is required for skeleton development, but a role for PDGFRb is unknown. To understand the differential contribution of PDGFRa+ vs. PDGFRb+ mesenchymal cells to limb bone development, we performed lineage tracing with knockin mice expressing Flp under control of the endogenous PDGFR genes. The limbs from newborn mice were analyzed for the expression of a Flp/Frt-dependent Tomato reporter gene. PDGFRa lineage tracing broadly labeled cells of the newborn skeleton, including both bone and cartilage. In contrast, PDGFRb lineage tracing labeled a more restricted population of cartilage and bone cells, suggesting that PDGFR may have a more specialized role in skeleton development.

Funding: None

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Abstract #96

PHENOTYPE-BASED SCREENING OF FUNGAL METABOLITES AND EXTRACTS IN ZEBRAFISH IDENTIFIED MODIFIERS OF NOTOCHORD DEVELOPMENT

Keirah M. Jefferson1, Imelda T. Sandoval2, Richard Glenn C. Delacruz2, Kevin Boyd2, Ana Gabriel2, David A. Jones2 1Oklahoma School of Science and Mathematics, Oklahoma City, OK, 73104 2Department of Functional and Chemical Genomics, Oklahoma Medical Research Foundation, Oklahoma City, OK, 73104

Fungi are known to contain biochemically active compounds that have yet to be studied. Thus, fungal extracts are highly likely to contain compounds that could be used for novel drug development. One highly effective method of accomplishing this is phenotype-based screenings on zebrafish because the zebrafish physiology is highly similar to human physiology. In fact, 71% of human proteins are orthologous to zebrafish proteins (MacRae and Peterson, 2015). Further, zebrafish embryos are small and develop rapidly outside the body; these make zebrafish more convenient and efficient for studying large populations, such as in a drug screening. We screened fungal extracts and metabolites and on 7-hour post-fertilization (hpf) embryos. We observed the embryos for 72 hours and found several phenotypes, including undulated notochord. Of the 384 crude extracts and 253 pure compounds we screened, 1 pure compound (Leptosin D) and 3 semi pure extracts had the undulated notochord phenotype. We focused on the undulated notochord phenotype because it has been previously associated with copper deficiency (Mendelsohn et al., 2006; Sandoval et al., 2013). We successfully rescued the wavy notochord phenotype of one of the semi pure fungal extracts with exogenous copper, which suggests that the extract can diminish copper availability. To further confirm the copper deficient phenotype, we did whole-mount in situ hybridization and RT-PCR on several genes associated with copper. Phenotype-based screening on zebrafish revealed bioactive compounds that cause copper deficiency. The discovery of novel copper chelators could be utilized to treat diseases related to copper dysregulation, particularly Wilson’s disease.

Funding: Oklahoma Medical Research Foundation

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Abstract #97

WORK PERFORMANCE AND RESIDUAL LIMB VOLUME IN MEN WITH TRANSTIBIAL AMPUTATION OVER A 12-MONTH TIMEFRAME

Carol P. Dionne, PT, DPT, PhD, OCS, Cert MDT1,3; William J.J. Ertl, MD2,3; and Jonathan D. Day, MA, CPO2,3

Introduction: A major concern in rehabilitation is that otherwise healthy men with transtibial amputations (TTA) are nonetheless prone to experiencing injury due to work-related activity (WRA). This study’s investigators were trying to understand WRAs, perceived exertion, and anthropometric changes over time and how this knowledge can help working men with TTA. The purpose of this study is to examine and compare gait performance, other WRAs, perceived exertion, and residuum measures in men with unilateral TTA.

Methods: Each subject was studied twice, twelve months apart. Investigators hypothesized that: 1) subjects would demonstrate a comparative asymmetric gait over the 1-year time period, and that 2) workers with TTA would present less capacity in lift and carry testing, report a higher exertion, and display a decrease decline in residuum measures over the 1-year period. The primary site of the study was in the College of Allied Health at the University of Oklahoma Health Sciences Center. Self-paced walk, brisk walk, floor -to-knuckle lift, and 25- foot carry were the independent variables analyzed. The protocol was to examine and compare WRA performance, gait asymmetry, perceived exertion, and residuum anthropometrics. Subjects were chosen based on factors such as physical dimensions, emotional readiness, and prosthetic features.

Results: The results show small difference in distances walked and residuum anthropometrics. Yet, greater exertion was reported at visit two than in visit one, participants were able to lift more weight at visit two, and displayed an increase in asymmetry in self-paced walk between visits one and two. Despite improved lift and anthropometric stability, participants demonstrated worsened cadence asymmetry in the self-paced walk and increased exertion in the brisk-paced walk.

Conclusion: The investigators concluded that working men with TTA may benefit from continual intervention to reduce cadence asymmetry and perceived exertion in the hopes of potentially minimizing residuum injury risk.

Funding: Oklahoma Center for the Advancement of Science and Technology Health Grant HR15-17

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Abstract #98

IL-24 SUPPRESSES OSTEOPONTIN IN LUNG CANCER CELLS

Jackie Oh1, Janani Panneerselvam2,4, Anupama Munshi3,4, and Rajagopal Ramesh2,4,5 1Oklahoma School of Science and Mathematics; Departments of 2Pathology, and 3Radiation Oncology, 4Stephenson Cancer Center, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; 5Graduate Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.

Introduction: Osteopontin (OPN) is a phosphoglycoprotein expressed in numerous tissues in the body, and most abundant in the bones. OPN is overexpressed in various cancers, including lung cancer, and has been shown to promote tumor cell proliferation, survival, drug resistance, and metastasis. Furthermore, OPN plays a major role in bone lysis and resorption in cancer. Previous studies have shown that IL-24 functions as an anticancer drug and has been clinically tested for treatment of human cancers. IL-24 suppresses multiple signaling pathways of cancer cells, causing tumor cell killing and inhibiting tumor angiogenesis and metastasis. In the present study we investigated whether IL-24 can modulate OPN expression and its signaling in lung cancer thus reducing metastasis.

Methods: Expression of OPN and its correlation with overall survival (OS) in lung adenocarcinoma (LUAD) was determined using the Cancer Genome Atlas (TCGA) database. For in vitro studies, a human H1299 lung tumor cell line stably transfected with tetracycline-inducible plasmid vector that expressed exogenous human IL-24 on addition of doxycycline (Dox; 1µg/ml) was used. Additionally, transient transfection studies were conducted using H1299 and A549 cell lines. The inhibitory activity of IL-24 on OPN expression and its downstream targets was determined by ELISA and western blotting.

Results: TCGA dataset analysis of LUAD demonstrated high OPN expression in primary lung tumor samples than normal tissues (p<0.0001). Further, Kaplan-Meier (KM) survival plot showed lung cancer patients with high OPN gene expression had low OS rate (P =5.8e-06). In vitro studies showed expression of IL-24 in H1299 cells markedly inhibited OPN expression that was associated with reduction in OPN-regulated proteins such as pAKT, ROCK1 and ROCK2.

Conclusion: Our study demonstrates IL-24 inhibited OPN expression in lung cancer cells, which is critical for cancer progression and metastasis.

Funding: Funding provided by Presbyterian Health Foundation Seed Grant, Presbyterian Health Foundation Bridge Grant, and Jim and Christy Everest Endowed Fund in Cancer Developmental Therapeutics and from the National Institutes of Health (NIH) –R01 CA102716

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Abstract #99

DIFFERENTIATION OF RAT CARDIOMYOBLASTS H9C2 IN CULTURE

Sneha Patel (1), Hailey A. Houson (2), Vibhudutta Awasthi (2) 1: Oklahoma School of Science and Mathematics, 1141 N Lincoln Blvd, Oklahoma City, OK 73104 2: Department of Pharmaceutical Sciences, University of Oklahoma Health Science Center, 1110 N. Stonewall Avenue, Oklahoma City, OK 73117

H9c2 cells are cardiomyoblasts derived from the aorta of an embryonic rat. The mature myocytic form of H9c2 cells is widely used as representation of heart muscle cells in drug development studies. H9c2 myoblasts can be differentiated into mature cardiomyocytes by altered culture conditions. The objective of my work was to validate the differentiation protocol of using culture medium containing low fetal bovine serum (FBS) as reported by Pagano et al (J Cell Physiol 2004, 198: 408). Evidently, maturation of cardiomyoblasts into cardiomyocytes is characterized by slower proliferation rate and multi-nucleation. Methods We investigated H9c2 maturation by determining the cell proliferation rate (bromodeoxyuridine or BrdU uptake assay) and by light microscopic examination of 4',6-diamidino-2-phenylindole (DAPI)-stained cells . H9c2 cells were grown in three conditions in a 96 well plate. The first group was serum starved in 1% FBS media, the second group was grown in 10% FBS media for six days, and the third group was plated 1 day before the initiation of the assay. BrdU was added to the medium and allowed to incorporate for 18 h before detection by BrdU enzyme-linked immunoassay. For light microscopy, the cells were stained with DAPI to visualize the nuclei. Results The analysis of the BrdU uptake revealed a decrease of BrdU incorporation in the cells grown under low FBS conditions. H9c2 cells grown for the same time in 10% FBS medium showed no change in BrdU uptake. Microscopy further validated the BrdU observations, as the cells grown in 1% FBS formed multinucleated elongated structures, characteristic of mature cardiomyocytes. Conclusions The results showed that H9c2 cardiomyobalsts can be easily transformed into their mature myocytic phenotype by lowering the FBS content in culture medium. We plan to further use these cells in research on in vitro models of myocardial infarction or oxidative stress.

Funding: Sandra K and David L Gilliland Chair in Nuclear Pharmacy

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Abstract #100

OUHSC LABORATORY FOR MOLECULAR BIOLOGY AND CYTOMETRY RESEARCH

Virginie Sjoelund, Ph.D.1, J. Acquaviva, III, M.S.1, Jenny Gipson1, Jim Henthorn1, Huaiwen Wang1, Allison F Gillaspy, Ph.D.1,2 1OUHSC College of Medicine, Core Lab, OKC, OK 73104, 2OUHSC Department of Microbiology and Immunology, OKC, OK 73104

The goal of the Laboratory for Molecular Biology and Cytometry Research (LNBCR) is to facilitate high-quality, state-of-the-art research by providing expertise and access to technology for a variety of specialized biomedical research applications. It is a full-service, research-oriented, institutionally-managed core dedicated to enhancing research and providing education and training to all users. The LMBCR assists investigators in all aspects of research, including initial project consultation, assistance with experimental design, training in protocols and instrumentation usage, sample preparation, and assistance with analysis and interpretation of results. The core also assists manuscript preparation and grant submission for applications that require heavier usage of specialized technology or techniques

Funding: None

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Abstract #101

THE LABORATORY OF BIOMOLECULAR STRUCTURE AND FUNCTION AT OUHSC

Simon Terzyan and Blaine Mooers

We would like you to know about the services in protein purification and structural studies available to OUHSC in the Protein Expression Production (PEP) Facility and the Laboratory of Biomolecular Structure and Function (LBSF). Dr Simon Terzyan (BRC 406) manages both facilities. He has 25 years of experience in protein science and crystallography (co-author on 40+ papers, 40+ structures in the Protein Data Bank). The PEP has shakers for bacterial growth and chromatography equipment for non-structural biology labs to make pure protein on the milligram scale. The pure protein can be used for biological, biophysical, or crystallographic studies. Training in the use of this equipment is available from Dr. Terzyan, or he can do this work on a fee-for-service basis. Dr. Terzyan can do standard molecular modeling tasks (e.g, design mutants, compare structures). He can make images of protein structures for your grant applications and manuscripts. For advanced molecular modeling projects (e.g., homology modeling, structure-based drug design), the LBSF has a Molecular Modeling Unit run by Dr. Timothy Mather. This unit has a workstation with four GPUs optimized to run four independent molecular dynamics simulations. Dr. Terzyan can also provide training in dynamic light scattering, protein crystallization, X-ray data collection (the LBSF has a new X-ray generator with two detectors), and structure determination, refinement, and analysis. He can also provide these services on a fee-for-service basis. Dr. Terzyan serves as an interface with the OU Crystallization facility in Norman and the Physical Biochemistry Laboratory in the Department of Biochemistry and Molecular Biology (BMB). The latter facility has CD spectroscopy, microcalorimetry, and microscale thermophoresis instruments. The LBSF is a Vice President of Research Core Facility and a core facility of the Oklahoma COBRE in Structural Biology. The PEP is supported by the BMB.

Funding: Vice President of Research Program Oklahoma COBRE in Structural Biology

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University of Oklahoma Health Sciences Center Graduate College Graduate Research Education and Technology Symposium

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DeVette, Christa Microbiology and Immunology TUMOR IMMUNOLOGY: NOVEL DISCOVERIES USING MOUSE MODELS OF BREAST CANCER FT Title#: 1

Gaeta, Laura Communication Sciences and Disorders IN ONE EAR AND OUT THE OTHER: HOW HEARING LOSS IMPACTS RESPONSES TO COGNITIVE SCREENING QUESTIONS FT Title#: 2

Garcia, Victor Radiological Sciences PATIENT RADIATION DOSE ESTIMATION PROGRAM FT Title#: 3

Guthmiller, Jenna Microbiology and Immunology IMMUNOREGULATION OF B CELL RESPONSES DURING EXPERIMENTAL PLASMODIUM INFECTION FT Title#: 4

Harpool, Kristyn Radiological Sciences A CLINICAL INVESTIGATION TO DETERMINE THE MOST BENEFICIAL RADIATION THERAPY TECHNIQUE FOR PANCREATIC CANCER FT Title#: 5

Kadel, Sapana Microbiology and Immunology IT'S ALL ABOUT SEX: ESTROGEN SIGNALING IN LUNG ILC2S FT Title#: 6

Kendall, Ethan Radiological Sciences GAMMA KNIFE PERFEXION TREATMENT PLANS FOR TRIGEMINAL NEURALGIA USING SECTOR- BLOCKED AND NON-SECTOR-BLOCKED SHOTS FT Title#: 7

Muhammad, Fauziyya GPiBS “FROM MICE TO MEN - ' EXPLOITATION OF A NOVEL PATHWAY IN SUPPRESSION OF AUTOIMMUNE UVEITIS TO PREVENT BLINDNESS' FT Title#: 8

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Shin, Young-Hwa Physiology A BALANCING ACT OF THE RETINOID VISUAL CYCLE: SHEDDING LIGHT ON HUMAN BLINDNESS FT Title#: 9

Wickersham, Elizabeth Clinical and Translational Science ADVANCE DIRECTIVES: HOW TO HAVE THE LAST WORD FT Title#: 10

Ziegler, Jadith Pathology XZ488 AS A NOVEL THERAPY AGAINST GLIOMAS FT Title#: 11

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Johnson, Anthony Neuroscience THE BLACK BOX OF CHRONIC PAIN: SHINING LIGHT ON NOVEL MECHANISMS FT Title#: 12 Latorre, Rocco Neuroscience KEEP CALM... REDUCE STRESS AND NEUROINFLAMMATION FT Title#: 13 Panneerselvam, Janani Pathology IL-24 INHIBITS GLI1 AND INDUCES DNA DAMAGE IN LUNG CANCER CELLS FT Title#: 14 Seshadri, Sudarshan Microbiology and Immunology ANTHRAX TOXIN DECREASES THE FUNCTION OF AN IMMUNE CELL: BREACHING BARRIERS AND CRIPPLING HOST DEFENSE FT Title#: 15

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