1. health and fitness
2. disease

The Graduate Student Association & The Graduate College present the 40th Graduate Research Education & Technology Symposium ABSTRACT BOOK KEEP CALM
&
RESEARCH ON
University of Oklahoma Health Sciences Center March 30, 2015 -­‐ April 2, 2015 1 2 FOREWORD The 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 students to showcase
their research and the scholarly experience of organization, interpretation, display, and
discussion of research results. Students may elect to present their work in either competitive or noncompetitive formats.
Travel awards are presented to winning competitive presenters for outstanding poster and oral
presentations in clinical sciences, human population, and bench research. A panel of judges
selects the winners according to clarity of the abstract, organization of the presentation, project
design, data analysis and quality of the presentation. Award winners receive travel grants from
$1,000 to $1,500 to attend a scientific meeting. This year the GREAT committee sponsored three workshops to help students prepare for
GREAT; the workshops assisted students with learning best practices for oral and poster
presentation techniques and abstract writing. Many thanks to Melissa Medina, EdD, Alix
Darden, PhD, and Sanjay Bidichandani, MBBS, PhD, who presented the Pre-GREAT
workshops. The GREAT Committee also wishes to thank our speakers; Dr. Chris Corbett, Dr. Elaine
Hamm, Dr. Kelley Dowd, and Manu Nair, Bioscience Round Table Speakers, Stephanie A.
Dunson, Career Development Speaker and Dr. Shai Silberberg, Keynote Speaker. The Graduate Student Association and the Graduate College wish to express their appreciation
to you for sharing this week with our student scholars as they pursue their professional goals.
We also express our thanks to the sponsors, judges, and companies that participated in the
Research Vendor Show. Special Acknowledgements The GREAT Committee wishes to express their appreciation to the University of Oklahoma
Health Sciences Center faculty, staff, and graduate students who have dedicated their time and
effort to make GREAT 2015 a success: Anne Pereira, PhD, Gillian Air, PhD, Katie Bryant,
PhD, the Graduate College staff, and all GSA members. GREAT Logo Design:
Maggie Mwoyosvi
3 Table of Contents
I.
Schedule of Events
II.
Alphabetized Listing of Presenters
III.
Graduate Student Oral Presentation Abstracts
IV.
Graduate Student Poster Presentation Abstracts
V.
Oklahoma School of Science and Mathematics (OSSM)
Poster Presentation Abstracts
VI.
Professional Student Poster Presentation Abstracts
VII.
Postdoctoral Fellow Oral Presentation Abstracts
4 5 GREAT Abstract Index
Student/Postdoc
Abstract #
Page #
Alam, Khondoker
54
Pharmaceutical Sciences
INDIRECT INHIBITION OF ORGANIC ANION TRANSPORTING POLYPEPTIDE (OATP)
1B1-MEDIATED TRANSPORT BY LYSOSOME INHIBITOR CHLOROQUINE
75
Ali, Quaisar
Department of Physiology
SECRETED KLOTHO AUGMENTS THE THERAPEUTIC POTENTIAL OF
MESENCHYMAL STEM CELLS FOR MONOCROTALINE INDUCED PULMONARY
ARTERIAL HYPERTENSION
124
100
Alomer, Reem
1
Cell Biology
DEFINING THE UNIQUE ROLES OF ESCO1 AND ESCO2 ACETYLTRANSFERASES
IN CHROMOSOME COHESION AND DNA REPAIR
21
Bastian, Anja
Physiology
NOVEL ANTICANCER COMPOUND AG311 TARGETS THE MITOCHONDRIAL
ELECTRONTRANSPORT CHAIN TO INDUCE BREAST CANCER CELL DEATH
2
22
Boominathan, Soorajnath
Oklahoma School of Science and Mathematics
A CELL SOMA ORGANELLE TRANSPORT FUNCTION FOR CAENORHABDITIS
ELEGANS CDK-5, SAD-1, AND SYD-2 IN MOTOR NEURON AXONS AND
DENDRITES AND ITS CORRELATION WITH LOCOMOTION PATTERNS
82
104
Boswell-Casteel, Rebba
Biochemistry & Molecular Biology
FUNCTIONAL CHARACTERIZATION OF A PURIFIED EQUILIBRATIVE
NUCLEOSIDE TRANSPORTER
3
23
Brame, Lacy
Biostatistics and Epidemiology
EXPOSURE TO ELECTRONIC-CIGARETTE AEROSOL EXTRACT INDUCES
SIGNIFICANT DNA DAMAGE
55
76
Brophy, Megan
Biochemistry & Molecular Biology
THE ROLE OF MACROPHAGE EPSINS IN THE REGULATION OF LRP-1
IN ATHEROSCLEROSIS
4
24
6 GREAT Abstract Index
Student/Postdoc
Bruxvoort, Christina
Biochemistry & Molecular Biology
HOW BCL-XL INTERACTS WITH TBID AT MITOCHONDRIA TO REGULATE
APOPTOSIS
Abstract #
56
Page #
77
Calhoun, Kaitlin
Pharmaceutical Sciences
IMMUNOLOGICAL DIFFERENCE BETWEEN TH1 AND TH2 DOMINANT
MOUSE STRAINS
IN A MODEL OF ICD
57
78
Chacko, Michael
Radiological Sciences
A DOSE CALCULATION ALGORITHM FOR DIAGNOSTIC IMAGING BEAMS BY
EMPIRICAL MODELING
58
79
Chen, Jiani
Genetic Counseling
MOLECULAR CYTOGENETIC CHARACTERIZATION OF SUBTELOMERIC
REARRANGEMENT
CASES BY PERFORMING CGH MICROARRAY
59
80
Chen, Qian
Physiology
DIFFERENTIAL ROLES OF VERY LOW-DENSITY LIPOPROTEIN RECEPTOR
SPLICE VARIANTS IN REGULATING CANONICAL WNT SIGNALING
5
25
Chin, Jane
Cell Biology
CGMP/PKG SIGNALING REGULATES ENDOPLASMIC RETICULUM
CALCIUM CHANNELS
IN CONE PHOTORECEPTORS
83
105
Clowdus, Emily
Cell Biology
THE ROLE OF THE DNA UNWINDING ELEMENT-BINDING PROTEIN (DUE-B)
IN VERTEBRATE DEVELOPMENT
60
81
Crowe, Alexandra
Pharmaceutical Sciences
ESTABLISHMENT OF AN IN VITRO CELL CULTURE MODEL FOR FUNCTIONAL
STUDY OF THE OATP1B1-V174A POLYMORPHISM
61
82
7 GREAT Abstract Index
Student/Postdoc
Cullin, Nyssa
Microbiology and Immunology
CHARACTERIZATION OF THE PUTATIVE MUREIN HYDROLASE LYTF IN
STREPTOCOCCUS SANGUINIS
Abstract #
6
Page #
26
Du, Mei
Physiology
TRANSGENIC MICE OVER-EXPRESSING SERUM RETINOL-BINDING PROTEIN
DEVELOP PROGRESSIVE RETINAL DEGENERATION THROUGH A
RETINOID-INDEPENDENT MECHANISM
7
27
Duong, Ngoc Quyen
Biostatistics and Epidemiology
ELECTRONIC HEALTH RECORD SYSTEM IMPLEMENTATION AND
MEANINGFUL USE CERTIFICATION AMONG OKLAHOMA HOSPITALS
62
83
Dvorak, Justin
Communication Sciences and Disorders
REFINING WORKING-MEMORY MODELS IN CONTINUOUS MAPPING VIA
RESPONSE-TIME ANALYSIS
8
28
Esteban Florez, Fernando
Department of Restorative Dentistry
SURFACE CHARACTERIZATION OF A NOVEL ANTIBACTERIAL DENTAL
ADHESIVE RESIN
101
125
Farasyn, Taleah
63
Pharmaceutical Sciences
RAPID DOWN-REGULATION OF ORGANIC ANION TRANSPORTING POLYPEPTIDE (OATP)
1B1 AND 1B3 TRANSPORT FUNCTION BY MAMMALIAN TARGET OF RAPAMYCIN (MTOR)
INHIBITOR SIROLIMUS
84
Ganapathy, Vengatesh
Department of Otorhinolaryngology
A UNIQUE DNA DAMAGE DETECTION ASSAY FOR EARLY DIAGNOSIS OF
OROPHARYNGEAL CANCER
102
126
Greuel, Regina
Health Promotion Sciences
PRIVATIZED HEALTHCARE POSSIBILITIES: A COMPARATIVE HEALTHCARE
PERSPECTIVE OF HEALTHCARE DELIVERY IN THE UNITED STATES UNDER
THE AFFORDABLE CARE ACT
64
85
8 GREAT Abstract Index
Student/Postdoc
Gurley, Jami
Biochemistry & Molecular Biology
ENDURANCE EXERCISE-DEPENDENT INCREASE IN SKELETAL MUSCLE
GLUT4EXPRESSION IN MICE OCCURS THROUGH A POST-TRANSCRIPTIONAL
MECHANISM
Abstract #
9
Page #
29
Gurung, Hem
Microbiology and Immunology
INFILTRATING IMMUNE CELLS DO NOT CONTRIBUTE TO HSV-1-INDUCED
CORNEAL NEOVASCULARIZATION
10
30
Guthmiller, Jenna
Microbiology and Immunology
IDENTIFICATION OF IMMUNO-REGULATORY CD138HI B CELLS DURING
MALARIA INFECTION
11
31
Hadad, Niran
Neuroscience
THE ROLE OF THE GLUCOCORTICOID RECEPTOR IN THE REGULATION OF
STRESS-INDUCED NOCICEPTION
65
86
Hannafon, Bethany
Department of Pathology
THE POTENTIAL OF EXOSOME MICRORNAS AS BIOMARKERS FOR BREAST
CANCER
103
127
He, Xuemin
Physiology
LOW-DENSITY LIPOPROTEIN RECEPTOR-RELATED PROTEIN 5 DRIVES RENAL
FIBROSIS VIA THE TGF-Β1 SIGNALING PATHWAY
12
32
Hebner, Allison
Genetic Counseling
THE UTILITY OF GENOMIC SEQUENCING IN ASSESSING GAMETE DONORS
USED IN DONOR ASSISTED CONCEPTIONS
13
33
Hermann, Kristina
14
Genetic Counseling
GENETIC COUNSELING GRADUATE PROGRAM WEBSITES AND THEIR INFLUENCE
ON PROSPECTIVE STUDENT APPLICATION DECISIONS
34
9 GREAT Abstract Index
Student/Postdoc
Hill, Claude
Biostatistics and Epidemiology
A COMPARISON BETWEEN BAYESIAN AND FREQUENTIST METHODS FOR
GEOSTATISTICAL ANALYSIS OF FINE PARTICULATE MATTER POLLUTION
Abstract #
15
Page #
35
Homco, Juell
Biostatistics and Epidemiology
USING BAYESIAN METHODS TO ESTIMATE THE PREVALENCE OF
INAPPROPRIATE EMERGENCY ROOM UTILIZATION IN THE SOONER
HAN MEDICAID POPULATION
16
36
Hopiavuori, Blake
Neuroscience
A NOVEL ROLE FOR VERY LONG CHAIN FATTY ACIDS IN BRAIN FUNCTION
17
37
Houson, Hailey
Pharmaceutical Sciences
COMPARING LIPOSOME-ENCAPSULATED OKN-007 TO NON-FORMULATED
OKN-007 IN TREATING GLIOMAS
66
87
Ibrahim, Mariam
Pharmaceutical Sciences
FORMULATION OF SHETA2 AS RESPIRABLE MICROPARTICLES FOR
TUBERCULOSIS TREATMENT
18
38
Iqbal, Henna
19
Microbiology and Immunology
CHARACTERIZATION OF A PUTATIVE TRANSLOCATION AND ASSEMBLY MODULE (TAM)
PROTEIN FROM BORRELIA BURGDORFERI.
39
Jackson, Rob
Biochemistry & Molecular Biology
REVERSAL OF DIET INDUCED-OBESITY BY ALTERING DIET COMPOSITION
RAPIDLY RESTORES INSULIN SENSITIVITY IN MICE
20
40
Janitz, Amanda
Biostatistics and Epidemiology
BENZENE AND CHILDHOOD ACUTE LEUKEMIA IN OKLAHOMA
21
41
Jilla, Anna
Communication Sciences and Disorders
YOGA AS A FORM OF VESTIBULAR REHABILITATION: A SYSTEMATIC
REVIEW WITH META ANALYSIS
67
88
10 GREAT Abstract Index
Student/Postdoc
Johnston, Sarah
Biostatistics and Epidemiology
DEMONSTRATION ON JOINPOINT: A STATISTICAL METHOD TO ANALYZE
TRENDS
Abstract #
68
Page #
89
Kaabinejadian, Saghar
Department of Microbiology and Immunology
A NOVEL MONOCLONAL ANTIBODY TO THE HLA OF CISPLATIN RESISTANT
OVARIAN CANCER CELLS
104
128
Kelley, Ryan
Cell Biology
RETBINDIN IS A NOVEL PHOTORECEPTOR-SPECIFIC PROTEIN AND A
MEMBER OF THE INTER-PHOTORECEPTOR MATRIX
22
42
Kevin, Weng
Oklahoma School of Science and Mathematics
TARGETED TREATMENT OF CANCER USING NEAR INFRA-RED LIGHT
84
106
Khan, Maaz
Cell Biology
EPIGENETIC CONTROL OF DNA REPLICATION THROUGH TICRR-BRD4
85
107
Kim, Seok ho
Department of Cell Biology
THE POLYCYSTIN COMPLEX MEDIATES WNT/CA2+ SIGNALING
105
129
Kroll, Chandra
Microbiology and Immunology
CONSEQUENCES OF HSV-1 INFECTION AND LATENCY VARY DEPENDING
ON THE TYPE OF TISSUE INFECTED WITHIN THE NERVOUS SYSTEM.
23
43
Kurdzo, Emily
Cell Biology
HOLDING HOMOLOGS TOGETHER: CENTROMERE-CENTROMERE
INTERACTIONS DURING MEIOTIC PROPHASE DEPEND ON THE
N- AND C-TERMINUS OF ZIP1
24
44
Lacroix, Jeffery
College of Medicine
CHARACTERIZING OCULAR SURFACE INFLAMMATION INDUCED BY
CERAMIDE SYNTHESIS INHIBITOR FTY720.
94
117
11 GREAT Abstract Index
Student/Postdoc
Lakhwani, Sudha
Biostatistics and Epidemiology
CANCER-TREATMENT ADHERENCE AND SENSITIVITY TO THE EMOTIONAL
TONE OF VERBAL COMMANDS
Abstract #
95
Page #
118
Lapolla, Suzanne
Biochemistry and Molecular Biology
MOLECULAR MECHANISM FOR TBID ACTIVATION OF BAX IN MEMBRANES
25
45
Larabee, Chelsea
26
Neuroscience
ABSENCE OF THE ANTIOXIDANT TRANSCRIPTION FACTOR NRF2
EXACERBATES OPTIC NEURITIS IN A MOUSE MODEL OF MULTIPLE SCLEROSIS
46
Lee, Sang-Min
Department of Biochemistry & Molecular Biology
CRITICAL INTERACTIONS OF PEPTIDE LIGANDS WITH THE EXTRACELLULAR
DOMAIN OF CALCITONIN AND AMYLIN RECEPTORS: IMPLICATIONS FOR
PEPTIDE RECOGNITION MECHANISMS OF CLASS B GPCRS
106
130
Leehan, Kerry
Pathology
SALIVARY GLAND FIBROSIS IS INCREASED IRRESPECTIVE OF AGE IN
PRIMARY SJÖGREN’S SYNDROME AND CORRELATES WITH CLINICAL
MEASURES OF DISEASE
27
47
Li, Hongliang
Department of Endocrinology
AT2R AUTOANTIBODIES BLOCK ANGIOTENSIN II AND
AT1R AUTOANTIBODY-INDUCED VASOCONSTRICTION
107
131
Li, Yue
Physiology
GENETIC SCREEN FOR GENES INVOLVED IN HIGH-SUGAR DIET-INDUCED
DIABETES MELLITUS
69
90
Lively, Kathryn
Nutritional Sciences
ASSOCIATIONS OF PARENTAL FEEDING PRACTICES, AND PARENT-CHILD
FOODPURCHASE INTERACTIONS IN GROCERY STORES: A PROPOSAL
70
91
12 GREAT Abstract Index
Student/Postdoc
Logan, Sreemathi
Department of Geriatrics
IGF-1 REGULATES LIPID BIOSYNTHETIC PATHWAYS IN HIPPOCAMPUS.
Abstract #
108
Page #
132
Ma, Hongwei
Department of Cell Biology
INHIBITION OF THYROID HORMONE RECEPTOR PROTECTS CONE
PHOTORECEPTORS IN RETINAL DEGENERATION
109
133
Mahjabeen, Sanjida
Pharmaceutical Sciences
FORMULATION OF A SUBLINGUAL TABLET CONTAINING THE
IPAD-IPAB-FUSION PROTEIN, AS A NOVEL VACCINE CANDIDATE
AGAINST SHIGELLOSIS
71
92
Marlin, Matthew
Biochemistry & Molecular Biology
DISTINCT BIOCHEMICAL CHARACTERISTICS BETWEEN TWO HOMOLOGOUS
ENDOSOMAL RABS
28
48
Martinez, Sydney
Biostatistics and Epidemiology
QUITLINE UTILIZATION AND OUTCOMES AMONG TOBACCO USERS
WITH A GENERAL EDUCATIONAL DEVELOPMENT (GED) DIPLOMA
29
49
Maskey, Dipak
Department of Cell Biology
CELL CYCLE-DEPENDENT UBIQUITYLATION AND DESTRUCTION OF NDE1 BY
CDK5-FBW7 REGULATES CILIARY LENGTH
110
134
Masser, Dustin
Physiology
AGE-RELATED COMMONALITIES AND SEXUAL DIMORPHISMS OF DNA
METHYLATION IN THE MOUSE HIPPOCAMPUS
30
50
Mata, Storm
Biochemistry and Molecular Biology
DEVELOPMENT OF A STANDARDIZED HLA-E ASSAY PROCEDURE
86
108
Matthews, Kathryn
Genetic Counseling
VISUALIZATION OF RETINOIC ACID SIGNALING IN THE DEVELOPING
CHICK EYE
31
51
13 GREAT Abstract Index
Student/Postdoc
McCullor, Kimberly
Graduate Program in Biomedical Sciences
STREPTOCOCCUS PYOGENES CHROMOSOMAL ISLAND SPYCIM1 AND
PROPHAGE SF370.1 BURST SIZE
Abstract #
72
Page #
93
McDonald, Katherine
Oklahoma School of Science and Mathematics
THE ROLES OF ESCO1 AND ESCO2 IN MITOSIS
87
109
McGrew, Kaitlin
Biostatistics and Epidemiology
RACIAL/ETHNIC DISPARITIES IN COLORECTAL CANCER DIAGNOSIS AND
SURVIVAL: AN ANALYSIS OF OKLAHOMA CENTRAL CANCER REGISTRY DATA
73
94
Merritt, Breanca
32
Health Promotion Sciences
ADDRESSING ADULT PHYSICAL ACTIVITY THROUGH PUBLIC SAFETY POLICIES:
A REGIONAL AND RACIAL/ETHNIC ANALYSIS FROM 2006-2013
52
Mowls, Dana
Biostatistics and Epidemiology
UNHEALTHY BEHAVIORS AMONG CANCER SURVIVORS
33
53
Mowls, Dana
Biostatistics and Epidemiology
HEPATITIS C VIRUS TRANSMISSION AMONG INJECTION DRUG USERS
74
95
Mwoyosvi, Maggie
Cell Biology
VISION AND HEARING LOSS ASSOCIATED WITH USHER SYNDROME TYPE 2A
34
54
Myers, Jamie
Communication Sciences and Disorders
PREVALENCE OF TINNITUS AND NOISE INDUCED HEARING LOSS IN
OKLAHOMA DENTISTS
96
119
Nguyen, Tanya
Clinical and Administrative Sciences
ACCEPTABILITY AND FEASIBILITY OF AN AFRICAN-AMERICAN HEALTH &
HERITAGE NUTRITION INTERVENTION AMONG AFRICAN-AMERICANS
WITH TYPE 2 DIABETES
97
120
Oh, Sangphil
Department of Cell Biology
A NOVEL ONCOGENIC ENZYME, RCL, PROMOTES BREAST TUMORIGENESIS
111
135
14 GREAT Abstract Index
Student/Postdoc
Okere, Samantha
Physiology
COMPOUND H INCREASES ANTI-AGING PROTEIN KLOTHO EXPRESSION AND
ATTENUATES ARTERIAL STIFFENING AND HYPERTENSION
Abstract #
88
Page #
110
O'Mealey, Gary
Cell Biology
UNCONSTRAINED KEAP1 RESTRICTS STRESS-INDUCED MITOCHONDRIAL
CLUSTERING
35
55
Orock, Albert
Neuroscience
ROLE OF REDUCED SYNAPTOBREVIN 2 LEVELS IN AGE-RELATED
COGNITIVE DECLINE
36
56
Pahwa, Sonia
Pharmaceutical Sciences
PRE-INCUBATION POTENTIATES THE INHIBITION POTENCY OF
DASATINIB AND VEMURAFENIB TOWARD OATP1B1-MEDIATED TRANSPORT
112
136
Panneerselvam, Janani
Department of Pathology
IL-24 REGULATES AKT BY INHIBITING THE HIGH MOBILITY GROUP (HMG)
A1/MIR222 SIGNALING NODE IN LUNG CANCER CELLS
113
137
Park, Eunsun
Communication Sciences and Disorders
THE EFFECT OF “SPEAK-OUT!®” VOICE THERAPY ON PROSODY IN
PERSONS WITH PARKINSON’S DISEASE
37
57
Parker, Kimberly
Rehabilitation Sciences
IMPROVING PERCEPTION OF MEANINGFUL ACTIVITY IN A GROUP OF
LOW-INCOME, OLDER ADULTS: A PILOT STUDY
98
121
Patel, Maulin
Cell Biology
NOVEL PROTEIN ADTRP NEGATIVELY REGULATES WNT SIGNALING
38
58
Patterson, Andrea
Microbiology and Immunology
TARGETING TUMORS LIKE A T CELL: EVALUATION AND TARGETING OF
HLA-PRESENTED MIF IN OVARIAN CANCER
39
59
15 GREAT Abstract Index
Student/Postdoc
Abstract #
Pham, Timothy
40
Pharmaceutical Sciences
THE ABCS OF CARDIOVASCULAR DISEASE AND STROKE PREVENTION:
PHARMACIST AND PAYER PERSPECTIVES ON SERVICE PROVISION AND PAYMENT
Page #
60
Prusator, Dawn
Neuroscience
AMYGDALA-MEDIATED MECHANISMS OF VISCERAL PAIN IN ADULTHOOD
FOLLOWING EARLY LIFE STRESS
41
61
Rahman, Kazi
Microbiology and Immunology
MAMMALIAN GLYCOGENIN-1 RELATED GLYCOSYLTRANSFERASE IS
IMPORTANT FOR TOXOPLASMA PROLIFERATION
42
62
Ramani, Vijay
Pharmaceutical Sciences
TLR4-INTERACTING SPA4 PEPTIDE SUPPRESSES NLRP3-INFLAMMASOME
AGAINST LPS AND ATP STIMULI.
43
63
Rampuria, Pragya
Microbiology and Immunology
NKT CELLS MEDIATED ENHANCEMENT OF HUMORAL IMMUNITY AGAINST
CLOSTRIDIUM DIFFICILE TOXIN B
44
64
Reagan, Alaina
Neuroscience
CAVEOLIN-1 PROMOTES RETINAL DAMAGE RESPONSE FOLLOWING ACUTE
ISCHEMIA REPERFUSION
45
65
Richardson, Whitney
Biostatistics and Epidemiology
A SYSTEMATIC REVIEW OF RISK FACTOR DIFFERENCES BETWEEN
ETIOLOGICAL PATHWAYS IN NECROTIZING ENTEROCOLITIS IN NEONATES
75
96
Sapkota, Bishwa
Department of Pediatrics
GENOME-WIDE ASSOCIATION STUDY OF CIRCULATING 25(OH) VITAMIN D
LEVELS IDENTIFIES A NOVEL LOCUS ASSOCIATED WITH VITAMIN D
DEFICIENCY
99
122
16 GREAT Abstract Index
Student/Postdoc
Schalo, Ian
Pharmaceutical Sciences
THALIDOMIDE BLOCKS SYMPTOMS OF PTSD AND CO-MORBID PAIN IN AN
ANIMAL MODEL OF PTSD
Abstract #
76
Page #
97
Seshadri, Sudarshan
Department of Microbiology and Immunology
ANTHRAX LETHAL TOXIN SUPPRESSES IL-22 PRODUCTION IN THE TYPE 3
INNATE LYMPHOID CELLS
114
138
Shrestha, Binu
Microbiology and Immunology
IDENTIFICATION OF BORRELIA BURGDORFERI NOVEL INTEGRAL OUTER
MEMBRANE PROTEINS.
77
98
Siefert, Joseph
Cell Biology
THE ROLE OF RIF1 AND THE REPLICATION TIMING PROGRAM IN
VERTEBRATE DEVELOPMENT
46
66
Sneed, Kayle
Communication Sciences and Disorders
PROCESSING OF FACIAL EMOTIONS IN HEALTHY ADULTS
47
67
Song, Caron
Oklahoma School of Science and Mathematics
NEURONAL AND MICROGLIAL CELLS DIFFER IN THEIR SENSITIVITY TO
GLUTAMATE TOXICITY
89
111
Stuck, Michael
Cell Biology
RDS GLYCOSYLATION; A WINDOW INTO THE DIFFERENTIAL ROLE OF RDS IN
RODS AND CONES.
48
68
Templeton, Amanda
Pathology
A NOVEL MISSENSE MUTATION IN THE EXTRACELLULAR DOMAIN OF
THE PDGFRA GENE INDUCES FUNCTIONAL CONSEQUENCES IN VIVO
78
99
17 GREAT Abstract Index
Student/Postdoc
Truong, Dat
Oklahoma School of Science and Mathematics
LA POSITIVE, RO60 NEGATIVE SUBSET OF PRIMARY SJÖGREN’S SYNDROME
IS A REALITY
Abstract #
90
Page #
112
Ward, Julie
Microbiology and Immunology
ARID3A-MEDIATED IFNA EXPRESSION IN SYSTEMIC LUPUS
ERYTHEMATOSUS B CELLS
49
69
Wei, Tao
Health Promotion Sciences
IS MARRIAGE A BIG HEADACHE --BEING LESBIAN IN CHINA
79
100
Wiafe, Kobby
Oklahoma School of Science and Mathematics
STAPHYLOCOCCUS AUREUS CONTRIBUTES TO THE PATHOGENESIS OF
ENDOGENOUS BACTERIAL ENDOPHTHALMITIS BY DISRUPTION OF THE
TIGHT JUNCTIONS BETWEEN RETINAL PIGMENT EPITHELIAL CELLS OF
THE BLOOD RETINAL BARRIER
91
113
Wilkerson, Joseph
Cell Biology
SPHINGOLIPID SIGNALING IN CORNEAL NEOVASCULARIZATION.
50
70
Yari, Hooman
Pharmaceutical Sciences
IN VITRO COMPARISON OF TWO NASAL DELIVERY DEVICES TO ADMINISTER
DRY POWDERS
80
101
Zander, Ryan
Microbiology and Immunology
CO-INHIBITORY AND CO-STIMULATORY CROSSTALK REGULATES
HELPER T CELL DIFFERENTIATION AND HUMORAL IMMUNITY DURING
PLASMODIUM INFECTION
51
71
Zhang, Binyi
Oklahoma School of Science and Mathematics
EPSIN IS REQUIRED FOR DISHEVELLED STABILITY AND WNT SIGNALING
ACTIVATION IN COLON CANCER DEVELOPMENT
92
114
18 GREAT Abstract Index
Student/Postdoc
Zhang, Nan
Biochemistry & Molecular Biology
REPLACEMENT OF 1.4-KB PROMOTER OF MURINE SELP GENE WITH ITS
HUMAN COUNTERPART PARTIALLY RECAPITULATES REGULATION OF
HUMAN SELP GENE IN VIVO
Abstract #
52
Page #
72
Zhao, Benjamin
Biochemistry & Molecular Biology
MOLECULAR CLONING AND CHARACTERIZATION OF A TYROSINE
PHOSPHATASE FROM MONOSIGA BREVICOLLIS
81
102
Zhu, Julie
93
Oklahoma School of Science and Mathematics
IMPROVING TIME-EFFICIENCY FOR EVALUATING THE PITUITARY GLAND UNDER
SEMI-AUTOMATED METHOD
115
Ziegler, Jadith
Pathology
ELTD1 AND SLIT3 AS NOVEL ANTIBODY THERAPIES AGAINST GLIOMA
BIOMARKERS
53
73
Zou, Jun
Department of Pharmaceutical Sciences
INTRACELLULAR SURVIVAL OF E. FAECALIS IN MACROPHAGES
115
139
Zulliger, Rahel
Department of Cell Biology
SNARES INTERACT WITH RDS/ROM-1 DURING CONVENTIONAL AND
UNCONVENTIONAL OUTER SEGMENT TARGETING
116
141
19 Section III. Graduate Student Oral Presentation Abstracts #1 -­‐ #53 20 Abstract #1
DEFINING THE UNIQUE ROLES OF ESCO1 AND ESCO2 ACETYLTRANSFERASES IN CHROMOSOME
COHESION AND DNA REPAIR
Reem Alomer1,2, Katherine McDonald3, Susannah Rankin2,1 1University of Oklahoma Health Sciences Center, 2Oklahoma
Medical Research Foundation, 3Oklahoma School of Science and Mathematics
Sister chromatid cohesion is a central process in maintaining genomic integrity through its roles in ensuring equal
chromosome segregation and DNA repair. Cohesion is mediated by a protein complex called cohesin. Cohesin plays
critical roles in DNA repair, chromatin structure organization, and gene expression regulation. Cohesin is thought to be
differentially modified during these different functions. One modification is mediated by the Eco family of
acetyltransferases. In yeast, Establishment of Cohesion (Eco1) acetylates subunits of cohesin during DNA replication or in
G2 in response to DNA damage. This acetylation is indispensable for proper cohesin function. Interestingly, vertebrates
express two homologs of Eco1, called Esco1 and Esco2. Both enzymes are required for proper mitotic cohesion and some
studies suggest they are both important for DNA repair. Their specific contributions to cohesin function during the DNA
damage response are not known. Our goal is to delineate the mechanisms that specify Esco1 and Esco2 unique functions,
and to characterize the crosstalk between cohesin regulation and the DNA damage response. In this study, we
characterized the phenotypes conferred by depletion of Esco1 and Esco2, both separately and together. We assessed both
mitotic cohesion and kinetics of DSB repair. In complementary experiments, we characterized the phenotypes of patientderived cells that are genetically deficient in Esco2 function. Collectively, our data suggest that Esco1 and Esco2 make
unique contributions to replication-dependent and damage-induced cohesion. Because DNA repair pathways are critical to
both the development and drug sensitivity of tumors, the Esco enzymes provide uniquely attractive chemotherapeutic
targets. Funding: R01GM101250
Funding: R01GM101250
21 Abstract #2
NOVEL ANTICANCER COMPOUND AG311 TARGETS THE MITOCHONDRIAL ELECTRON TRANSPORT
CHAIN TO INDUCE BREAST CANCER CELL DEATH
Anja Bastian1, Lora C. Bailey-Downs3, Jessica E. Thorpe2, Aleem Gangjee4, Kenneth Humphries5, Michael A. Ihnat2,3
1
Department of Physiology, University of Oklahoma Health Sciences Center, 2Department of Pharmaceutical Sciences,
University of Oklahoma College of Pharmacy, 3DormaTarg, Inc, Oklahoma City, OK 73104, 4Division of Medicinal
Chemistry, Graduate School of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA, 5Oklahoma Medical
Research Foundation
Chemotherapeutic agents that target mitochondria are emerging as an appealing strategy for the development of new
anticancer therapies. The inhibition of the mitochondrial electron transport chain (ETC) provides an alternate and
selective mechanism for killing cancer cells, bypassing upstream apoptosis-inducing pathways. The mechanism is based
on the fact that cancer cells have higher mitochondrial membrane potential rendering them inherently susceptible to
increased superoxide generation through the respiratory chain. In addition pharmacological inhibition of complex I and III
at the ubiquinone-binding site could further increase superoxide production. Collectively, this mitochondrial sensitivity
represents a promising approach to selectively kill cancer cells. The purpose of this study was to investigate the
mitochondria-associated cell death of AG311, which is a novel anticancer compound designed by our group. AG311 has
been shown to induce rapid mitochondrial depolarization resulting in necrotic cell death in a cancer cell-selective manner.
In two mouse orthotopic breast cancer models (MDA-MB-435 and 4T1), AG311 significantly reduced tumor volume by
85% and 81%, respectively (n=4-6), with no apparent systemic toxicity. First, upregulation of the mitochondrial ETC by
culturing MDA-MB-435 cells in galactose media sensitized cells to AG311-induced cell death. Mitochondrial oxygen
consumption (XFe96 analyzer) drastically decreased by 62.9% (±12.9, n=8) in response to AG311 (7.5 µM). The effect of
AG311 on mitochondrial ETC complexes was determined by measuring NADH oxidation (complex I), ubiquinol
oxidation (complex III-IV) or cytochrome c oxidation (complex IV). We found that AG311 inhibited complex I and III
activity, but not complex IV. Further kinetic assays suggested that AG311 competitively inhibited ubiquinone binding.
Finally, treatment with AG311 generated superoxide production (MitoSOX), while treatment with an antioxidant (lipoic
acid) partially prevented AG311-induced cell death. In summary, the present results indicate ubiquinone as a likely target
for AG311-induced selective cancer cell death through generation of mitochondrial superoxide.
Funding: CLNOPCOP20100311PMAI1
Funding: Research reported in this study was supported an Institutional Development Award (IDeA) from the National
Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103639, Oklahoma
Center for the Advancement of Science grant HR11-046 (to F.A.H.), OUHSC College of Medicine Alumni Association
seed grant (to F.A.H.), and American Heart Association predoctoral fellowship 13PRE17040024 (to R.C.C-B.).
22 Abstract #3
FUNCTIONAL CHARACTERIZATION OF A PURIFIED EQUILIBRATIVE NUCLEOSIDE TRANSPORTER
Rebba C. Boswell-Casteel, Jennifer M. Johnson, and Franklin A. Hays Department of Biochemistry and Molecular
Biology, University of Oklahoma Health Sciences Center and Stephenson Oklahoma Cancer Center, Oklahoma City, OK
Equilibrative nucleoside transporters (ENTs) are major pharmaceutical targets responsible for modulating the efficacy of
more than 30 FDA/EMA approved drugs that treat an expansive range of disease states (e.g., pancreatic cancer, acute
myeloid leukemia, non-Hodgkin lymphoma). In fact, expression levels of human ENT1 have been linked to prolonged
survival for pancreatic cancer patients receiving gemcitabine treatment – a nucleoside analog transported by ENTs.
However, the molecular mechanism and chemical determinants of ENT-mediated substrate transport, and the atomic
resolution topology of ENTs, remains a mystery. The current studies are focused on defining the molecular basis for how
therapeutics interact with this class of integral membrane proteins. Function Unknown Now 26 (FUN26) is a yeast
ortholog of the human equilibrative nucleoside transporter (ENT) family. FUN26 was expressed and purified to
homogeneity and incorporated into proteoliposomes for functional analysis. A fundamental element of defining
mechanism is the identification of mutations that significantly alter protein function. Gain-of-function mutations have
been identified that significantly alter ENT substrate specificity by allowing the transport of nucleotides. An ab initio
structural model was generated and suggests the mutations play a role in substrate binding and conformational
switching/gating of the protein. Additionally, FUN26 has positional sensitivities for nucleoside substrates, which appear
to alter substrate selectivity. Defining the molecular transport mechanism and the chemical properties that govern
substrate transport will facilitate the development and tuning of novel therapeutics that selectively utilize isoform-specific
transporter functions of ENTs to gain access to intracellular targets. Therefore, targeting the transport mechanism of ENTs
through rational drug design will benefit multiple disease states requiring pharmaceutical intervention.
Funding: This work is supported by the NIH COBRE award (P20GM103639), OCAST (HR11-046), and AHA
predoctoral fellowship (13PRE17040024).
23 Abstract #4
THE ROLE OF MACROPHAGE EPSINS IN THE REGULATION OF LRP-1 IN ATHEROSCLEROSIS
Megan L. Brophy1,2, Yunzhou Dong1, Kandice L. Tessneer1, Satish Pasula1, Hoogeun Song1, Xiaofeng Cai1, Xiaolei Liu1,2,
Baojun Chang1, Hao Wu1, Klaus Ley3, Hong Chen1,2 1Cardiovascular Biology Research Program, OMRF, Oklahoma
City, Oklahoma 2Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center,
Oklahoma City, OK
Background Epsins are a family of ubiquitin-binding endocytic clathrin adaptors. We recently published that endothelial
epsins function as critical regulators of tumor angiogenesis by controlling VEGF signaling (JCI, 2012; ATVB, 2013). Our
goal is to define the novel role of epsins in macrophages in regulating atherogenesis. Methods and Results We engineered
mice with specific deletion of epsins in myeloid cells (MF-DKO). Strikingly, MF-DKO mice on ApoE-/- background fed
western diet significantly reduced atherosclerotic lesion formation and foam cell accumulation. In macrophages, epsin
deficiency did not alter LDL scavenger receptors, CD36, Lox1 or SRB1, or reverse cholesterol transport proteins, ABCA1
or ABCG1, but did significantly reduce Lucifer Yellow pinocytosis, indicating a major defect in lipid uptake. Epsin
deficiency did decrease total and surface protein levels of LRP-1, a protein with anti-inflammatory and antiatherosclerotic properties. Oil Red O staining of isolated ApoE-/-/M¿-DKO macrophages showed little lipid
accumulation, suggesting a mechanism in which epsin deficiency impairs foam cell formation. In addition, epsin 1 and
LRP-1 interact in macrophages. Furthermore, this interaction is abolished in the absence of epsin’s UIM domain and LPS
treatment increases LRP-1 ubiquitination, suggesting that epsin promotes the ubiquitin-dependent internalization of LRP1. Epsin deficiency also significantly suppressed the pro-inflammatory M1 macrophage phenotype found in plaques and
increased the anti-inflammatory macrophage phenotype, thus suggesting an important pro-inflammatory role for epsins in
macrophages. Our finding implicates epsin as a potential therapeutic target for atherosclerosis treatment. Conclusions We
demonstrate epsins promote atherogenesis by potentiating foam cell formation and maintaining pro-inflammatory
macrophages within the atherosclerotic plaque, thus suggesting epsins as a novel therapeutic target to combat
atherogenesis.
Funding: AHA Predoctoral Fellowship #15PRE21400010 (M.B.), R01 HL118676 (H.C.), AHA 0835544N (H.C.), AHA
12SDG8760002 (Y. D.)
24 Abstract #5
DIFFERENTIAL ROLES OF VERY LOW-DENSITY LIPOPROTEIN RECEPTOR SPLICE VARIANTS IN
REGULATING CANONICAL WNT SIGNALING
Qian Chen1, 3, Yusuke Takahashi2, 3, Kyungwon Lee3, Kazuhiro Oka4 and Jian-xing Ma1,3 1Department of Physiology,
2
Department of Medicine, 3Harold Hamm Diabetes Center, The University of Oklahoma Health Sciences Center,
Oklahoma City, OK; 4Department of Medicine, Baylor College of Medicine, Houston, TX.
Purpose: Very low-density lipoprotein receptor (VLDLR) is a multi-ligand receptor and has diverse roles beyond
lipoprotein metabolism. Our previous studies have shown that VLDLR negatively regulates Wnt signaling in the retina.
VLDLR has two major splice variants, variant I (VLDLRI) and variant II (VLDLRII). The present study was undertaken
to investigate the potential differential roles of VLDLR splice variants in the regulation of Wnt signaling. Methods: The
expression of VLDLRI and VLDLRII was analyzed by RT-PCR and Western blot analysis. The Wnt signaling activities
were evaluated by luciferase assay or Western blot analysis. Levels of the shed VLDLR extracellular domain (sVLDLRN) and full-length VLDLR in cell culture model were measured by Western blot analysis, and the ratio of sVLDLR-N to
full-length VLDLR was determined by densitometry. The levels of sVLDLR-N in the serum were measured by
ELISA. Results: Most of the examined tissues expressed both VLDLRI and VLDLRII, while the retina expressed only
VLDLRII. VLDLRII showed a stronger inhibitory effect on Wnt signaling in the retina of VLDLR-/- mice compared with
VLDLRI. sVLDLR-N was detected in the interphotoreceptor matrix of bovine eyes, the serum from human and mouse,
and conditioned medium (CM) from cells expressing VLDLR. The ratio of shed/full-length VLDLR was dramatically
higher in cells expressing VLDLRII than that of cells expressing VLDLRI. Moreover, sVLDLR-N was capable of
inhibiting Wnt signaling. CM from cells expressing VLDLRII exhibited a more potent Wnt inhibitory effect than those
from cells expressing VLDLRI. The Levels of sVLDLR-N in the serum of diabetic mice were significantly decreased
compared to non-diabetic control mice. Conclusion: VLDLR splice variants have differential regulatory effects on Wnt
signaling due to the differential extracellular domain shedding, and the shed VLDLR extracellular domain may represent a
novel mechanism for Wnt signaling regulation in the retina.
Funding: NIH grants (EY018659, EY012231, EY019309, GM104934), a JDRF grant (2-SRA-2014-147-Q-R), OCAST
grants (HR12-103 and HR13-076), a AHA grant (14PRE20460229).
25 Abstract #6
CHARACTERIZATION OF THE PUTATIVE MUREIN HYDROLASE LYTF IN STREPTOCOCCUS SANGUINIS
Nyssa Cullin and Jens Kreth Department of Microbiology and Immunology, University of Oklahoma Health Sciences
Center, Oklahoma City, Oklahoma 73104
Introduction: Streptococcus sanguinis, a commensal organism in the oral cavity, is an early oral biofilm colonizer known
to produce hydrogen peroxide (H2O2) in concentrations inhibitory to other members of the oral biofilm. Furthermore, the
production of H2O2 itself plays a major role in the development of biofilms due to the induction of extracellular DNA
(eDNA) release, although the mechanism is not known. It is hypothesized that the putative competence-associated murein
hydrolase LytF in S. sanguinis is involved in the release of eDNA; thus, the objective of this study was to investigate the
regulation and activity of LytF in S. sanguinis. Methods: Expression of lytF was examined using qRT-PCR after induction
of competence with 0.5µg/ml species-specific competence stimulating peptide (CSP) at mid-log phase. Next, lytF mutants
were created using overlap-extension PCR to investigate the role of LytF in competence and eDNA release. Competencerelated genes comCDE and comX were examined using qRT-PCR and overall competence assayed in transformation
studies. eDNA release from growing cultures was measured using qRT-PCR and compared to a standard curve.
Results: lytF expression was induced over 100-fold with addition of CSP. The expression of genes comCDE was slightly
decreased in the lytF mutant, which corresponded to a decrease in transformation efficiency as compared to wildtype.
Expression of comX was not changed. eDNA release did not appear to differ between wildtype and lytF mutant.
Conclusion: lytF was confirmed to be competence-associated. eDNA release did not change as initially expected in the
lytF mutant; thus, LytF does not seem to be involved in eDNA release. The change in transformation efficiency indicates
LytF may play a role in DNA uptake. This warrants further investigation into the localization of LytF, its effect on DNA
uptake machinery, and biofilm formation.
Funding: NIH NIDCR: R01 DE021726
26 Abstract #7
TRANSGENIC MICE OVER-EXPRESSING SERUM RETINOL-BINDING PROTEIN DEVELOP PROGRESSIVE
RETINAL DEGENERATION THROUGH A RETINOID-INDEPENDENT MECHANISM
Mei Du, Krysten Farjo Department of Physiology, University of Oklahoma Health Sciences Center, OKC, OK
Introduction: Serum retinol-binding protein (RBP4) is the sole transport protein for retinol in the blood, and was recently
recognized as an adipokine that contributes to insulin resistance and type 2 diabetes. We previously demonstrated that
RBP4 induces inflammation in human retinal capillary endothelial cells through a retinol-independent mechanism,
indicating increased RBP4 may contribute to pathogenesis of diabetic retinopathy. Yet the effect of increased levels of
RBP4 on the retina has not been studied. In the current study, we use transgenic mice over-expressing RBP4 (RBP4-Tg)
to evaluate the physiologic effects of serum RBP4 elevation on the retina. Methods: RBP4-Tg mice with 10-fold increase
in serum RBP4 levels and wild-type controls from 1 to 9 months of age were used in this study. Electroretinography
(ERG) and histological analyses were performed to assess retinal function and structure, respectively. Retinal retinoid
levels were quantified by high performance liquid chromatography (HPLC). Proinflammatory cytokine expression was
measured by quantitative RT-PCR and western blotting. Results: RBP4-Tg mice maintain normal body mass, blood
glucose, triglyceride, and insulin levels. RBP4-Tg mice showed progressive retinal dysfunction and degeneration,
characterized by a predominant loss of inner retinal neuron photoresponse and cell number. RBP4-Tg mice have
photoreceptor ribbon synapse deficiency detectable by 1-month of age, and subsequent rod and cone bipolar cell loss
detectable by 6-month of age. HPLC analyses revealed normal ocular retinoid and bis-retinoid levels in RBP4-Tg mice,
suggesting that retinal degeneration occurs through a retinoid-independent mechanism. RBP4-Tg mice have early-onset
retinal microglia activation and increased expression of interleukin-18 mRNA and protein in retina, which indicates
neuroinflammation is an underlying mechanism of retinal degeneration. Conclusion: These studies reveal that RBP4-Tg
mice develop retinal neuroinflammation and neurodegeneration in the absence of retinal vascular pathology, obesity,
dyslipidemia, and hyperglycemia.
Funding: Funding: American Heart Association #13BGIA16920097 and Institutional Development Award from National
Institute of General Medical Sciences #P20GM104934.
27 Abstract #8
REFINING WORKING-MEMORY MODELS IN CONTINUOUS MAPPING VIA RESPONSE-TIME ANALYSIS
Justin D. Dvorak1, Derick D. Deweber1,2, Frank R. Boutsen1 1 Motor Speech and Prosody Research Laboratory, College
of Allied Health, OUHSC 2 Full Function Rehabilitation, Oklahoma City, OK
Introduction: Response time (RT) has been shown to be non-normally distributed and represents a sum of cognitive
processes and executions; however, it is often measured and analyzed as a monolithic entity. Assessment of working
memory models in lexical mapping experiments, such as Baddeley’s (2000) phonological loop, often relies on RT and
accuracy analyses to support claims of subsystem influence; however, recent modeling techniques suggest that RT in twoalternative forced-choice tasks can be separated into multiple cognitively meaningful sub-components. Critically,
different response strategies may emerge when presentation modalities (audio vs. visual) are changed or combined.
Methods: 15 Native Spanish speakers (8 female, 7 male) completed a auditory/visual continuous-recognition task, in
which English words and Spanish-like pseudo-words were presented in two prosodic conditions (English vs. Spanish
accent) and two visual conditions (with vs. without fixed abstract pictorial referents) with varying amounts of lag between
repeated presentations (2, 4, 8, and 16). Participants indicated whether they thought a given stimulus item was new or
previously presented; meanwhile response accuracy and RT were recorded. RT was recorded in milliseconds and was
analyzed via Ratcliff diffusion modeling to ascertain drift rate (information quality), boundary separation (speed/accuracy
tradeoff) and non-decision processing. Results: Non-native prosody and phonotactics were associated with lower drift
rate (p=0.0012), higher non-decision time (p=0.0057), and lower boundary separation (p=0.0051). Higher lag values were
also associated with lower drift rates (p=0.0264). Presence of a photographic referent, in contrast, was associated with
significantly higher drift rate (p=0.0042), lower non-decision time (p=0.0123), and higher boundary separation
(p=0.0114). Discussion: As expected, non-native prosody and phonotactics were associated with poorer information
quality. The increase in boundary separation combined with increased drift rate in the presence of a photographic referent
could indicate that information quality precipitates more robust sound memory traces, as evidenced by changes in
processing strategy.
Funding: None
28 Abstract #9
ENDURANCE EXERCISE-DEPENDENT INCREASE IN SKELETAL MUSCLE GLUT4 EXPRESSION IN MICE
OCCURS THROUGH A POST-TRANSCRIPTIONAL MECHANISM
Jami Gurley and Ann Louise Olson Biochemistry and Moleular Biology, University of Oklahoma Health Sciences Center
Peripheral insulin resistance, a hallmark of type 2 diabetes mellitus and obesity, is accompanied by dysregulation of the
insulin-responsive glucose transporter 4 (GLUT4) in adipose (AT) and skeletal muscle (SKM) tissues. Feeding mice fed
a high fat diet (HFD) decreases GLUT4 mRNA and protein in both AT and SKM. In transgenic mice that overexpress
human GLUT4, HFD reduced GLUT4 in AT, but not in SKM. The maintenance of GLUT4 in SKM protected the mice
from hyperglycemia and insulin resistance, which suggests that glucose uptake in SKM is important for maintaining
insulin-independent glucose homeostasis under conditions of nutrient excess. Exposure of mice to voluntary wheel
running (VWR) exercise increases GLUT4 in SKM. In order to determine the mechanism(s) by which HFD and VWR
regulate GLUT4 expression, we assessed GLUT4 mRNA and protein in SKM of lean and obese mice with or without
access to VWR for four weeks. Previous work from our laboratory demonstrated that HFD repressed GLUT4 promoter
activity in AT and that this required the presence of the cis Liver X Receptor element (LXRE). Using transgenic mice
carrying GLUT4 promoter/CAT reporter constructs with either the intact human GLUT4 promoter or a loss-of-function
mutation in the LXRE, we found that HFD also decreased SKM GLUT4 mRNA and that this downregulation was LXREdependent. Immunostaining showed that HFD decreased SKM GLUT4 protein and that VWR returned GLUT4 protein to
regular diet (RD) levels during the exercise recovery period. However, VWR was not sufficient to recover HFD-induced
decreases in GLUT4 mRNA. This suggests that endurance exercise operates via a non-transcriptional mechanism to
upregulate GLUT4 protein synthesis. Interestingly, our data also showed that VWR increased mTOR pathway
activation. Future studies will test the hypothesis that GLUT4 protein synthesis is mTOR-dependent and contributes to
the adaptation that occurs in SKM following prolonged endurance exercise.
Funding:
29 Abstract #10
INFILTRATING IMMUNE CELLS DO NOT CONTRIBUTE TO HSV-1-INDUCED CORNEAL
NEOVASCULARIZATION
Hem R. Gurung1 and Daniel J.J. Carr1, 2 1Department of Microbiology and Immunology and 2Ophthalmology, Dean
McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104
HSV-1 infection of the cornea induces neovascularization of the normally avascular tissue. A robust leukocyte influx
composed primarily of neutrophils is observed by day 7 post infection (pi) and peaks at day 14 pi. This study was
undertaken to investigate the role of infiltrating neutrophils in HSV-1- induced corneal neovascularization. C57BL6/J
mice were infected with 1,000 plaque forming units (PFU) of HSV-1 per scarified cornea. At day 7 pi and every two days
thereafter, the mice were administered 0.5 mg of IgG, anti-Gr-1 (RB6-8C5), or anti-Ly6G (1A8) intraperitoneally (ip).
Leukostasis was performed by cardiac perfusion with FITC-conjugated concanavalin-A. Dexamethasone (DEX)
(10mg/kg) was delivered by ip injection at day 10 pi. Mice were euthanized at day 14 pi, perfused with PBS, and the
corneas excised and processed for confocal microscopy and flow cytometry. Peripheral blood was evaluated for leukocyte
content by flow cytometry. The peripheral blood of anti-Gr-1 or anti-Ly6G antibody-treated mice was leukopenic. In the
cornea, anti-Gr-1 antibody treatment depleted neutrophils (Gr1+F4/80- ), CD3+ T cells, and inflammatory monocytes
(Gr1+F4/80+). In contrast, anti-Ly6G antibody treatment did not deplete any infiltrating cells in the cornea. Unexpectedly,
there was no difference in blood or lymphatic vessel genesis between groups. Upon further interrogation, the majority of
infiltrating cells were found in the luminal side of the blood endothelium at day 14 pi. A single bolus of DEX at day 10 pi
resulted in the significant suppression of blood vessel but not lymphatic vessel genesis at day 14 pi with no apparent
differences in the number of infiltrating cells. Infiltrating cells in the cornea most likely do not contribute to the
development and persistence of HSV-1-induced angiogenesis. We interpret these results to suggest that resident cells
produce pro-angiogenic factors to develop and maintain blood and lymphatic vessels in the cornea.
Funding: R01 EY021238 to D.J.J.C.
30 Abstract #11
IDENTIFICATION OF IMMUNO-REGULATORY CD138HI B CELLS DURING MALARIA INFECTION
Jenna J. Guthmiller, Ryan A. Zander, and Noah S. Butler Department of Microbiology and Immunology University of
Oklahoma Health Sciences Center
Plasmodium infections remain a large public health concern resulting in 225 million malaria cases and approximately one
million deaths each year. Individuals repeatedly infected do not develop severe clinical disease. However, sterilizing
immunity against Plasmodium does not occur, contributing to persistence of the parasite and on-going transmission. It is
not known if Plasmodium infection induces immuno-regulatory cells that limit protective immunity and parasite
clearance. Regulatory B cells (Bregs) are a population of immuno-regulatory cells that limit inflammation and prevent
pathogen clearance in models of autoimmunity and bacterial infection. We hypothesized that Plasmodium infectioninduced Bregs express the regulatory cytokine IL-10, limit the expansion, differentiation, and function of inflammatory
CD4+ T cell responses, and promote parasite persistence. To test this, we utilized IL-10/GFP reporter mice infected with
Plasmodium yoelii. We found that CD138hi B cells are a major producer of IL-10. In an in vitro assay, Plasmodium
infection-induced CD138hi B cells suppressed the proliferation of naïve CD4+ T cells. Adoptive transfer of sort-purified
CD138hi B cells into infection-matched mice reduced parasite-specific antibody responses and delayed parasite clearance.
Collectively, these data show that Plasmodium infection induces the expansion of immuno-regulatory CD138hi B cells that
restrict anti-Plasmodium immunity and parasite clearance.
Funding: This work was supported by grants from the NIH/NIAID (T32AI007633 to R.A.Z.; 1K22AI099070 to N.S.B.)
and the American Heart Association (13BGIA17140002 to N.S.B.). N.S.B. is also an OK-INBRE scholar supported by a
grant from the NIH/NIGMS (8P20GM103447).
31 Abstract #12
LOW-DENSITY LIPOPROTEIN RECEPTOR-RELATED PROTEIN 5 DRIVES RENAL FIBROSIS VIA THE TGFΒ1 SIGNALING PATHWAY
Xuemin He, Rui Cheng, Kyungwon Lee, Yusuke Takahashi and Jian-xing Ma Department of Physiology, The University of
Oklahoma Health Sciences Center
Low-density lipoprotein receptor-related protein 5 (LRP5), a co-receptor of the canonical Wnt/β-catenin pathway, was
recently reported to drive idiopathic pulmonary fibrosis. In our study, we found that LRP5 drives the renal fibrogenic
process, which is independent of its role in the Wnt/β-catenin pathway. In an obstructive nephropathy mouse model,
levels of fibrotic factors were lower in LRP5-deficient mice compared to controls. We then measured the activation of the
Wnt/β-catenin signaling pathway, which was found unaffected by LRP5 deficiency, indicating that attenuated obstructioninduced renal fibrosis in LRP5-deficient mice was not due to mitigated activation of the fibrosis-driven Wnt/β-catenin
pathway. Instead, we detected alleviated TGF-β signaling in LRP5-deficient kidneys in comparison to that in wild-type
kidneys. Interestingly, basal levels of fibrotic factors in LRP5-deficient kidneys were also slightly lower in relative to
controls. Moreover, overexpression of LRP5 in a proximal tubular epithelial cell line resulted in enhanced canonical TGFβ signaling and elevated expression of downstream fibrosis markers in response to TGF-β1 treatment. Consistently,
primary tubular epithelial cells isolated from LRP5-deficient mice displayed attenuated activation of canonical TGF-β
signaling and fibrosis after TGF-β1 treatment, compared to those from wild-type mice. Specific TGF-β1-driven promoter
assay also indicated that overexpression of LRP5 led to enhanced TGF-β signaling activation. In addition, we discovered
potential physical interaction between LRP5 and TGF-β receptor(s), suggesting that LRP5 might act as a co-receptor of
the canonical TGF-β signaling pathway in the regulation of renal fibrogenic process.
Funding: NIH grants EY018659, EY019309 and GM104934.
32 Abstract #13
THE UTILITY OF GENOMIC SEQUENCING IN ASSESSING GAMETE DONORS USED IN DONOR ASSISTED
CONCEPTIONS
Allison L. Hebner1, Susan Hassed1, Karl Hansen2, Andrew F. Wagner2 1Department of Pediatrics, OUHSC 2Department
of Obstetrics and Gynecology, OUHSC
Introduction: For decades, donor assisted conception has been a reproductive option for couples or individuals unable to
conceive. Currently, there are few national genetic screening recommendations regarding requirements to quality as a
gamete donor. With the clinical implementation of whole exome and whole genome sequencing on the rise,
comprehensive genetic screening of gamete donors may offer greater insight into genetic related health risks for families
using these services. Our goal was to assess the utility of this technology to provide more comprehensive patient care for
those who use anonymous gamete donors. Methods: All subjects are U.S. reproductive medicine clinic medical directors
registered with the Society of Assisted Reproductive Technology or North American genetic counselors who are members
of the National Society of Genetic Counselors. Participants were recruited by an email distributed by their respective
professional society. The email directed the participants to an anonymous, web-based survey where both qualitative and
quantitative responses were recorded.
Results: Initial results show that the majority of genetic counselors rate genetic
screening of prospective gamete donors as beneficial or extremely beneficial, while simultaneously rating the use of
whole exome or whole genome sequencing as very infeasible or somewhat infeasible in the scope of their practice. Major
barriers to the use of the technology were listed to be cost, lack of genetic counseling resources, and difficulty with
interpreting the results. There was no significant deviation in opinions when compared with sub-specialty, years in
practice, or work setting. Physician results are unavailable for comparison at this time. Conclusion: These results suggest
that while genetic screening is a valuable tool in assessing gamete donors because of the reduction of the risk of
conceiving a child with a genetic condition, there are barriers to implementing genomic sequencing for donor screening at
this time.
Funding:
33 Abstract #14
GENETIC COUNSELING GRADUATE PROGRAM WEBSITES AND THEIR INFLUENCE ON PROSPECTIVE
STUDENT APPLICATION DECISIONS
Kristina Hermann1, Susan Hassed1, Alix Darden1, Carrie Guy1 1Pediatrics, University of Oklahoma Health Sciences
Center
Genetic counselors are health care professionals trained in both medical genetics and counseling. Currently, 34
universities in the United States and Canada offer a two year Masters-degree program in genetic counseling. The purpose
of this study was to investigate what information was most useful to genetic counseling students when considering which
programs to apply to and how program websites affected their application decisions. Current genetic counseling students
and recent graduates completed an online survey, which asked what information on genetic counseling program websites
was most influential in deciding which programs to apply to. Program/assistant/associate directors were also surveyed to
compare their views. Chi square analysis and t-tests were used to determine significance of results, and a two-sample ttest was used to compare the factors students identified as important on a 5-point Likert scale compared to those identified
by directors. Content analysis identified themes from students’ open-ended responses about program website
improvement. While directors noted limitations to making changes to their program websites, they were interested in how
prospective students use their program website and what information they found most useful. Students indicated there
were specific programs they chose not to apply to due to the difficulty of using that website. Students were significantly
interested in all twelve program factors they were asked about, based on a 5-point Likert scale. A two-sample T-test
showed that students and directors differed significantly in how important they thought information about academic
requirements, application requirements, and course descriptions were in deciding which programs to apply to. Content
analysis revealed three major themes of what students want from individual program websites: easy navigation,
comprehensive information, and impression of the program. This information may help individual genetic counseling
graduate programs to improve the functionality of program websites for prospective students, and improve their applicant
pool.
Funding:
34 Abstract #15
A COMPARISON BETWEEN BAYESIAN AND FREQUENTIST METHODS FOR GEOSTATISTICAL ANALYSIS
OF FINE PARTICULATE MATTER POLLUTION
C. Larry Hill, Jr.1, David M. Thompson1 1Department of Biostatistics and Epidemiology, Oklahoma University Health
Sciences Center
Introduction: Increased levels of fine particulate matter (PM2.5) have been associated with increased incidence of
respiratory illnesses such as asthma, pneumonia, tuberculosis, and chronic obstructive pulmonary disease, and as well as
atherosclerosis. Quantification of air quality is a fundamental issue for identification of exposure to
pollutants. Frequentists methods for predicting the exposure level at unmeasured locations include inverse distance
weighting and kriging methods. Newer approaches emphasize land-use regression techniques as well as Bayesian
statistical methods. Methods: Data from the Environmental Protection Agency’s pollution monitoring stations in an
eight state region of the southern central United States are used. Weekly totals of particulate matter (diameter of 2.5
micrometers or smaller - PM2.5) were collected at 158 locations between January 1 and September 30, 2014. A random
sample of 16 sites is removed from the data, and then Bayesian and frequentist geostatistical methods (inverse distance
weighting, ordinary kriging) were used to predict the PM2.5 levels at the 16 sites. Estimates of the mean square
prediction error were calculated and used to compare the models’ predictive accuracy. Results: Results will be presented
from the model building that is currently ongoing. Results will be analyzed according to the spatial techniques used for
each model. The model with the lowest mean square prediction error will be identified. Conclusion: It is important to
have spatial models which accurately reflect pollution levels at unmeasured locations. By removing 10% of the sample
and comparing the predictions to the actual values, we can better understand each model’s accuracy. More complicated
models can add information on other variables (weather conditions, land-use features, elevations, etc.) to increase the
accuracy of the predictions.
Funding: None
35 Abstract #16
USING BAYESIAN METHODS TO ESTIMATE THE PREVALENCE OF INAPPROPRIATE EMERGENCY
ROOM UTILIZATION IN THE SOONER HAN MEDICAID POPULATION
Juell Homco1, Hélène Carabin2, 1Department of Medical Informatics, University of Oklahoma School of Community
Medicine, 2Department of Biostatistics and Epidemiology, College of Public Health, University of Oklahoma Health
Sciences Center
Introduction: Inappropriate emergency room (ER) utilization is believed to incur a significant burden on the US healthcare
system. Yet, there is no gold standard for classifying these visits as appropriate or not, making it difficult to estimate their
true impact. We use a Bayesian latent-class model to estimate the true prevalence of inappropriate ER visits by combining
information from two imperfect classification systems. Methods: Oklahoma claims data were used to estimate the
inappropriate ER visit prevalence among Sooner Health Access Network Medicaid patients (July 1, 2013—June 30,
2014). Using primary ICD-9 diagnosis codes, ER visit appropriateness was classified using information from Method 1:
the California Emergency Room Coalition and Method 2: the New York University algorithm. A Bayesian latent-class
model was used to combine information from the two methods in WinBugs. Prior knowledge suggests that the prevalence
of inappropriate ER visits is at most 0.50. Based on an expert’s opinion, the prior distribution of the sensitivity and
specificity was assumed to be 0.60 (σ=0.125) and 0.75 (σ=0.125) for Method 1 and 0.40 (σ=0.125) and 0.85 (σ=0.075) for
Method 2, respectively. Results: The unadjusted prevalence of inappropriate ER visits was 0.185 (95% CI: 0.181-0.190)
according to Method 1 and 0.046 (95% CI: 0.044-0.049) according to Method 2. The posterior sensitivity and specificity
were estimated at 0.50 (σ=0.131) and 0.83 (σ=0.016) for Method 1 and 0.25 (σ=0.119) and 0.96 (σ=0.005) for Method 2,
respectively. Based on information from both Methods, the estimated prevalence was 0.054 (95% CI: 0.011-0.182).
Conclusion: The mean misclassification error-adjusted prevalence of inappropriate ER visits fell between the estimates for
each method without adjustment, but was very imprecise. Future work should improve on the uncertainty of this estimate
by increasing the sample size, adding another classification system, and/or eliciting additional expert opinion on priors.
Funding: Sooner Health Access Network
36 Abstract #17
A NOVEL ROLE FOR VERY LONG CHAIN FATTY ACIDS IN BRAIN FUNCTION
Blake R. Hopiavuori 1,3,4, Martin-Paul Agbaga 1,3,4,5, Joseph Wilkerson3,4,5, Richard S. Brush 4, Andria F. Hedrick 2,3,
Debra Saunders7, Faizah Bhatti3,4,8, Nawajes A. Mandal 1,3,4,5, Luke Szweda6, Vibhudutta Awasthi 2,3, Rheal Towner1,7, ,
Robert E. Anderson 1,3,4,5
1
Oklahoma Center for Neuroscience, 2 Department of Pharmaceutical Sciences, 3 University of Oklahoma Health
Sciences Center, Oklahoma City, OK, 4 Department of Ophthalmology, Dean McGee Eye Institute, 5 Department of Cell
Biology, University of Oklahoma Health Sciences Center, 6 Free Radical Biology and Aging Research Program,
Oklahoma Medical Research Foundation, 7 Advanced Magnetic Resonance Center, Oklahoma Medical Research
Foundation, 8 College of Medicine/Peds – Neonatology
Purpose: ELOngation of Very Long chain fatty acids-4 (ELOVL4) is an elongase responsible for biosynthesis of very
long chain (VLC; ≥C28) fatty acids, found as components of complex lipid molecules. ELOVL4 synthesizes the VLC
polyunsaturated fatty acids (VLC-PUFA) in retina and testes, and VLC saturated fatty acids (VLC-FA) in skin and
brain. A 2011 case study reported that homozygous inheritance of the Stargardt’s (STGD3) mutation in ELOVL4 causes
a central nervous system (CNS) phenotype in humans, including seizures, intellectual disability, spastic quadriplegia and
death. We hypothesize that ELOVL4-synthesized VLC-FA play an essential role in neural cell structure and function.
Methods: We generated the first successful animal model for STGD3/STGD3 inheritance. ELOVL4 localization within
the CNS was determined using immunofluorescence. Brain lipids were extracted from hippocampus and separated by
solid phase extraction (SPE) before GC/MS analysis. Positron emission tomography (PET) was used to assess CNS
uptake of fluorodeoxyglucose (FDG) in STGD3/STGD3 mice. HPLC was used to assess intermediary metabolism in
STGD3/STGD3 mice. Membrane fractionation was performed on baboon hippocampus to isolate synaptic membranes for
lipid analysis. Results: Our STGD3/STGD3 mice recapitulate the human phenotype, developing seizures at P19 followed
by death at P21. Immunofluorescence showed the highest enzyme immunoreactivity within the hippocampus, in neurons
of the dentate gyrus. GC/MS confirmed the presence of 28:0 and 30:0 in sphingolipids. PET imaging of STGD3/STGD3
mice revealed a 3-fold increase in the amount of FDG uptake into the CNS. Metabolomic analysis revealed significant
increases in ATP levels in STGD3/STGD3 mice. Membrane fractionation revealed enrichment of 28:0 and 30:0, but not
VLC-PUFA, in synaptic vesicle membranes. Conclusions: This is the first study to demonstrate mutations in Elovl4
causing a CNS phenotype in an animal model, implicating for the first time a potential role of VLC-FA in neural cell
structure and function.
Funding: 1F31NS089358 - NIH grant awarded to Blake R. Hopiavuori 1R21NS090117 - NIH grant awarded to Robert E.
Anderson
37 Abstract #18
FORMULATION OF SHETA2 AS RESPIRABLE MICROPARTICLES FOR TUBERCULOSIS TREATMENT
Mariam Ibrahim1, Doris M. Benbrook2, Lucila Garcia Contreras1 1Department of Pharmaceutical Sciences and
2
Department of Obstetrics and Gynecology. University of Oklahoma Health Science Center.
Introduction: Tuberculosis (TB), an infection caused by Mycobacterium tuberculosis (MTB), is the main cause of death
from a single microorganism. Despite their severe side effects, the efficacy of treatments for drug susceptible MTB is
acceptable but more effective drugs are needed to treat drug-resistant MTB strains. We discovered that SHetA2, an
anticancer drug, is also effective against MTB. Pulmonary delivery achieves high local drug concentration in the lungs,
main site of TB infection, and limits side effects. Thus, we formulated SHetA2 into inhalable microparticles (MPs) for TB
treatment. Methods: Three formulations were manufactured by spray drying (SD): ShetA2 alone, SHetA2+PLGA, and
SHetA2+mannitol. The first two were designed to target alveolar macrophages (where MTB resides) whereas the third
was intended for immediate dissolution and absorption. SD manufacturing conditions (feed concentration, gas flow rate
and temperature) were optimized to yield respirable particles with narrow size distribution (volume diameter, Dv =1-5µm
and geometric standard deviation, GSD<2). The degree of SHetA2 crystallinity in the MPs was determined by differential
scanning calorimtery (DSC) and X-ray diffraction. In vitro dissolution of SHetA2 MPs was evaluated in simulated lung
fluid (mSLF). Results: ShetA2 alone, SHetA2 PLGA and SHetA2 mannitol MPs were small, spherical and hollow
compared to big rod shaped particles of unprocessed SHetA2. The three SHetA2 MPs had optimum properties for
aerosolization with average Dv= 1.72µm±0.26, 1.66µm±0.24 and 3.12µm±0.23, respectively, and a GSD<2. SHetA2 was
chemically stable after SD with no interaction with PLGA or mannitol. ShetA2 changed from crystalline to amorphous
form sfter SD, improving SHetA2 solubility from 3 to 15 µg /ml in mSLF. The dissolution rate of SHetA2 mannitol MPs
was the fastest followed by SHetA2 alone then Sheta2 PLGA MPs. Conclusion: Inhalable SHetA2 MPs suitable for
alveolar macrophage targeting and immediate release have been obtained with optimized spray drying processes.
Funding: PHF seed grant 2014-2015 GSA research award 2014
38 Abstract #19
CHARACTERIZATION OF A PUTATIVE TRANSLOCATION AND ASSEMBLY MODULE (TAM) PROTEIN
FROM BORRELIA BURGDORFERI.
Henna Iqbal, Darrin Akins
Borrelia burgdorferi, the causative agent of Lyme disease, is a dual-membraned spirochete. Similar to Gram-negative
bacteria, B. burgdorferi contains Outer Membrane Proteins (OMPs) that are exported to the surface through a β-Barrel
Assembly Machine (BAM) complex. The central component of the BAM complex in B. burgdorferi is the OMP
BamA. Interestingly, BamA is part of an operon that contains three ORFs; BB0794, BamA (BB0795) and Skp (BB0796),
which is a chaperone protein that escorts OMPs to the BAM complex. Since the BB0794 is a part of an operon that
encodes proteins involved in the BAM complex and OMP transport in general, we were interested in characterizing the
role of this unknown protein in B. burgdorferi. BB0794 has a C-terminal domain that is predicted to be related to a known
protein TamB, which is an inner membrane protein in E. coli that interacts with TamA to help in the efficient secretion
proteins in E. coli. To determine whether BB0794 is associated with the borrelial BAM complex or is involved in protein
secretion, we have performed physicochemical characterization of BB0794 to examine its cellular location, secondary
structure, and topology using various techniques. The combined data suggest that BB0794 is a membrane protein that is
not surface exposed, which is consistent with it being a TamB ortholog. A better understanding of BB0794 could help us
to identify a novel protein secretion/export system related to B. burgdorferi virulence and Lyme disease pathogenesis.
Funding: GRANT AI059373 from NIH/NIAID to DRA.
39 Abstract #20
REVERSAL OF DIET INDUCED-OBESITY BY ALTERING DIET COMPOSITION RAPIDLY RESTORES
INSULIN SENSITIVITY IN MICE
Rob Jackson and Ann Olson Department of Biochemistry and Molecular Biology University of Oklahoma Health Sciences
Center
Obesity is a chronic disease state that is always brought on by over-nutrition and is a significant risk factor for Type II
diabetes, cardiovascular disease, and various cancers. The primary treatment of obesity is reduction of adipose
mass. Because adipose tissue is a major metabolic sensor, we have focused our investigation on the physiologic changes
that accompany the restoration of normal adiposity by altering the diet composition in mice that were previously subjected
to diet-induced expansion of the fat mass. To do this, we ab libitum fed mice for 10 weeks either a High Fat Diet (HFD)
or a Regular chow Diet (RD). Other mice were kept on HFD for the first 5 weeks before swapping to RD for the
remaining 5 weeks. Body weights, fasted blood glucose, and serum insulin levels were collected on a weekly basis, and at
the end of the 10 week period, the visceral fat pads were harvested. All mice were fasted overnight before these
collections, and some mice were either allowed access to food 1h prior or they were given an Insulin injection 30m
prior. Interestingly, we have found that our diet-swapped mice experience massive body weight reduction within the 1st
week and return to control levels by week 2. Insulin signaling molecules, which have diminished responsiveness in HFD
mice, also return to control levels as determined by Western blot detection of phospho-AKT and -AS160. In support of
this, weekly HOMA-IR scores also indicate that diet-swap mice have become more insulin sensitive within this same time
period. All together, we have shown that HFD-induced obesity can be reversed when mice are returned to a standard chow
diet without restricting food intake. Future studies will focus on the metabolic changes that adipose undergoes in order to
accomplish this drastic remodeling during weight loss.
Funding: PHF
40 Abstract #21
BENZENE AND CHILDHOOD ACUTE LEUKEMIA IN OKLAHOMA
Amanda E Janitz, MPH University of Oklahoma, Janis E Campbell, PhD University of Oklahoma, Sheryl Magzamen,
PhD Colorado State University, Anne Pate, PhD Southwestern Oklahoma State University, Julie A Stoner, PhD
University of Oklahoma, Jennifer D. Peck, PhD University of Oklahoma
Background: As a leading cause of childhood mortality, childhood cancer is an important health concern in the US.
However, evidence regarding the etiology is lacking despite numerous studies. One environmental exposure of interest is
benzene as it has been classified as a known carcinogen in adult acute myeloid leukemia (AML). The goal of this study
was to determine if children with acute leukemia have a higher odds of exposure to benzene compared to controls.
Methods: We conducted a case-control study using the Oklahoma Central Cancer Registry as our source for cases
diagnosed between 1997 and 2012 (n=307). Controls were selected from birth certificate records and matched to cases on
week of birth (n=1,013). To evaluate whether exposure to benzene using the National-Scale Air Toxics Assessment
(NATA) was associated with childhood acute leukemia, we used conditional logistic regression. Results: We observed no
differences in benzene exposure between cases and controls in the bivariate analysis or after adjusting for the potentially
confounding factors of urbanization and maternal education. However, in an analysis evaluating benzene stratified by
leukemia type, the estimates for children with AML were stronger than among those with ALL, though none of the
estimates were significant. Discussion: Using the NATA estimates to measure benzene allowed us to assess a specific
pollutant at the census tract level, which provided an advantage over the use of monitor or point source data. Future
studies should consider other markers of traffic, such as NO2 and road density. While we did not observe an association
between benzene and leukemia, it is important to continue evaluating the effects of benzene in areas with higher exposure
concentrations along with other potential health effects of benzene exposure.
Funding: None
41 Abstract #22
RETBINDIN IS A NOVEL PHOTORECEPTOR-SPECIFIC PROTEIN AND A MEMBER OF THE INTERPHOTORECEPTOR MATRIX
Ryan A. Kelley1, Muayyad R. Al-Ubaidi1, and Muna I. Naash1 1Department of Cell Biology, University of Oklahoma
Health Sciences Center, Oklahoma City, OK 73104
Retbindin is a novel retina-specific protein of unknown function. It has significant sequence homology to the riboflavin
binding protein of chicken oviduct cells. To assess retbindin in the retina, we generated a knockout mouse (Retb-/-), in
which the retbindin coding sequence was replaced with that of eGFP. eGFP in Retb-/- mice was found only in the
photoreceptor cells, which was consistent with the distribution of retbindin in WT animals. Electroretinography revealed
an age- and dose-dependent decline in both rod and cone responses at postnatal days (P)120 and 240. This functional
decline is the result of both rod and cone photoreceptor cell loss. Given the clear importance of this protein for retinal
function, we further explored its properties and localization. Biochemical analysis of retbindin showed that it is secreted
by rod photoreceptors and is maintained within the insoluble interphotoreceptor matrix. Immunofluorescence analysis
localized retbindin to the outer segment/RPE microvilli interface. In vitro binding assays showed that retbindin’s ligand is
riboflavin. Furthermore, riboflavin binding in vitro protected immortalized cone photoreceptors from light induced cell
death. Retbindin’s properties and localization suggest that it may play a role in retinal/RPE metabolite exchange. Future
research will be aimed at investigating its function in vivo to better understand the role of this novel protein in retinal
homeostasis.
Funding: This work was supported by the National Eye Institute R01EY10609 (MIN), R01EY018137 (MRA), and
P30EY021725, and the Foundation Fighting Blindness (MIN and MRA).
42 Abstract #23
CONSEQUENCES OF HSV-1 INFECTION AND LATENCY VARY DEPENDING ON THE TYPE OF TISSUE
INFECTED WITHIN THE NERVOUS SYSTEM.
Chandra Kroll1, Daniel J. Carr1,2. 1Microbiology and Immunology and 2Ophthalmology/Dean McGee Eye Institute,
University of Oklahoma Health Sciences Center
Introduction: Herpes simplex virus 1 (HSV-1) infection can result in a life threatening condition known as herpes simplex
encephalitis (HSE). The direct mechanism(s) responsible for long-term HSE-mediated pathology is unknown. Therefore,
the goal of this study is to characterize viral dissemination pathways in the central nervous system (CNS) and the local
immune responses during acute and latent infection. Methods: Scarified corneas of C57BL/6 or reporter-inducible Rosa
mice were inoculated with HSV-1 and assessed for viral dissemination into the PNS and CNS as determined by RT-PCR
and confocal microscopy. In addition, leukocyte infiltration, T cell function, and resident microglia activation were
analyzed by flow cytometry. Results: It is currently understood that HSV-1 disseminates from the initial site of infection
(the cornea in our model) to the CNS by trafficking through the trigeminal ganglia (TG). Unexpectedly, RFP+ cells,
representing HSV-1-infected cells from Rosa mice, were visualized in olfactory bulb (OB) before other areas of the CNS
were infected (~ 2dpi). Latently infected mice (i.e. >30dpi) express latency-associated transcripts (LAT) in the TG. In
addition, LAT expression was recognized in the OB and ependyma (CNS tissue lining the ventricles also containing the
mid brain). During latency, lytic gene expression was recovered from the ependyma whereas no indication of lytic genes
were expressed in the TG and other CNS regions. At 60dpi, the ependyma and the olfactory bulb maintained elevated
levels of effector T lymphocytes and MHC class II positive resident microglia cells. T cell populations from each CNS
region responded to PMA/ionomycin indicating these cell populations were functional. Conclusion: This is the first study
to evaluate the olfactory bulb as a site of viral dissemination following ocular infection. Results suggest the ependymal
region of the brain acts as a reservoir for lytic infection and maintenance of effector T lymphocyte populations during
latency.
Funding: AI053108-11
43 Abstract #24
HOLDING HOMOLOGS TOGETHER: CENTROMERE-CENTROMERE INTERACTIONS DURING MEIOTIC
PROPHASE DEPEND ON THE N- AND C-TERMINUS OF ZIP1
Emily Kurdzo1,2, Dean Dawson2,1 1Department of Cell Biology, University of Oklahoma Health Sciences Center 2Cell
Cycle and Cancer Biology Program, Oklahoma Medical Research Foundation
Proper chromosome segregation in meiosis is important for maintaining the fidelity of the genome from generation to
generation. In humans, improper chromosome segregation during meiosis is the leading cause of infertility, miscarriage,
and genetic birth defects. During prophase of meiosis in budding yeast, homologous chromosomes become tethered in a
number of ways, but all of these are mediated in part by Zip1. We aim to better understand the contributions made to
meiotic segregation fidelity by Zip1, especially in its roles in mediating centromere interactions. Zip1 “couples” nonhomologous centromeres together in early prophase – the role of coupling is not well understood. In late prophase, just
prior to segregation of the homologous chromosomes, Zip1 aids in the “pairing” of homologous centromeres – the role of
this phenomenon is known to aid in proper segregation in meiosis I. We have created a series of in-frame deletion mutants
of ZIP1 in Saccharomyces cerevisiae that we have used to identify functional domains of Zip1 that are critical for
centromere pairing and coupling. We have identified regions of the N- and C-terminus that are differentially critical for
coupling and pairing. We have concluded that the phenomena of centromere coupling and centromere pairing are distinct
from one another.
Funding: NIH R01-GM-087377
44 Abstract #25
MOLECULAR MECHANISM FOR TBID ACTIVATION OF BAX IN MEMBRANES
Suzanne Lapolla1, Blaine H.M. Mooers1,2, Zhi Zhang1, Justin Kale3, David Andrews3,4 , and Jialing Lin 1,2. 1Department
of Biochemistry and Molecular Biology, 2Peggy and Charles Stephenson Cancer Center, University of Oklahoma Health
Sciences Center, Oklahoma City, Oklahoma 73126, 3Department of Biochemistry and Biomedical Sciences, McMaster
University, Hamilton, Ontario L8N 3Z5, Canada and 4Biological Sciences Sunnybrook Research Institute and Department
of Biochemistry, University of Toronto, Ontario, M4N 3M5, Canada.
Introduction: tBid activates Bax to initiate apoptosis by inducing mitochondrial outer membrane premeabilization
(MOMP). Crystallography predicted an amphipathic a-helix formed by the Bcl-2 homology region 3 (BH3) of tBid binds
to a hydrophobic groove formed by the BH1-3 regions of Bax. However, NMR studies suggested another binding site,
comprised of Bax helices 1 and 6, defined as a trigger pocket. This study’s goal is to determine which interface exists in
the mitochondrial tBid-Bax complex, and contributes to tBid activation of Bax. Methods: Using the BH3-in-groove
crystal structure and a BH3-in-trigger pocket NMR model, tBid and Bax mutants were generated, each with a single
cysteine positioned in the tBid BH3 helix or the Bax groove or trigger pocket. Each tBid/Bax mutant was designed so the
cysteines are in a geometry suitable for disulfide linkage according to their respective model. After in vitro synthesis and
targeting to mitochondria, the mutant pairs were oxidized to induce disulfide formation. Products were analyzed by SDSPAGE and identity confirmed by immunoprecipitation. Mitochondrial cytochrome c release by tBid and Bax mutants,
and others that contain potential interface disruptive mutations, were assayed. Results: Disulfide-linked tBid-Bax
heterodimers were observed with mutant pairs based on the BH3-in-groove structure, confirming this interface in the
membrane-bound complex. No such dimers were detected with most of the mutant pairs based on the BH3-in-trigger
pocket model. A substantial number of mutant pairs with cysteines in tBid’s BH3 region and Bax’s helix 1
formed heterodimers, suggesting a flexible BH3:helix 1 interface. Separate mutations in tBid and Bax that abrogated
heterodimer formation via the BH3-in-groove interface also abolished the MOMP activity. Conclusions: The BH3-ingroove interface mediates a interaction between tBid and Bax required for apoptotic MOMP. Current mutagenesis studies
will help determine the function of tBid BH3:Bax helix 1 interaction.
Funding: Funding: NIH grant R01GM062964 to J.L.
45 Abstract #26
ABSENCE OF THE ANTIOXIDANT TRANSCRIPTION FACTOR NRF2 EXACERBATES OPTIC NEURITIS IN A
MOUSE MODEL OF MULTIPLE SCLEROSIS
Chelsea Larabee1,2, Bob Axtell3, Agnieshka Agasing3, Shruti Desai2, and Scott Plafker1,2 1Oklahoma Center for
Neuroscience, University of Oklahoma Health Sciences Center, 2Free Radical Biology and Aging, Oklahoma Medical
Research Foundation, 3Arthritis and Clinical immunology, Oklahoma Medical Research Foundation
Optic nerve inflammation, or optic neuritis, is experienced by a majority of multiple sclerosis (MS) patients and is
typically characterized by episodes of acute, monocular vision loss. These inflammatory episodes can lead to damage or
degeneration of the retinal ganglion cells (RGCs) that comprise the optic nerve. MOG peptide-induced experimental
autoimmune encephalomyelitis (EAE) is a well-established model of MS in which mice are immunized to produce a
neuro-autoimmunity and present with progressive ascending paralysis. This model recapitulates the cardinal hallmarks of
the human disease, namely, increased oxidative stress, demyelination, and neurodegeneration. Previous studies have
shown that knockout (KO) of the master antioxidant transcription factor Nrf2 increases the susceptibility of mice to EAEassociated motor deficits, but the visual system in these knockouts has not been investigated. Our studies test the
hypothesis that Nrf2 protects RGCs and visual function in EAE. We measured visual acuity by reflexive optokinetic
tracking (OKT) daily and discovered that wildtype (WT) EAE mice exhibit acute, relapsing loss of visual acuity in one
eye reminiscent of that observed in multiple sclerosis patients. We further found that Nrf2 KO mice experience earlier,
more severe visual deficits compared to their WT counterparts. Histological analyses demonstrated more severe EAEinduced loss of RGCs and optic nerve inflammation in Nrf2 KO mice relative to WT. RGC loss was primarily observed in
the central retina surrounding the optic nerve head as opposed to peripheral retinal areas in both WT and KO mice. We
eliminated the caveat that these differences are due to the role of Nrf2 in immune system development by using flow
cytometry to show that baseline immune cell profiles of WT and KO mice are very similar. This is the first study to
report that genetic ablation of Nrf2 exacerbates visual deficits by increasing RGC death and optic nerve inflammatory
infiltrates.
Funding: NIH/NINDS 5R00NS075099-04
46 Abstract #27
SALIVARY GLAND FIBROSIS IS INCREASED IRRESPECTIVE OF AGE IN PRIMARY SJÖGREN’S
SYNDROME AND CORRELATES WITH CLINICAL MEASURES OF DISEASE
K. M. Leehan1, 2, 3, M. Brown2,3, C. Montgomery2, 3, K. L. Sivils1, 2, 3, A. D. Farris1, 2, 3 for the OSSCORT 1Department of
Pathology, University of Oklahoma Health Sciences Center, 2Oklahoma Medical Research Foundation, 3Oklahoma
Sjögren’s Syndrome Center of Research Translation (OSSCORT)
BACKGROUND. Primary Sjögren’s Syndrome (pSS) is an autoimmune disease featuring severe dry eyes and mouth and
inflammation of lacrimal and salivary glands (SG). Because onset of pSS is typically in the fifth decade and SG fibrosis
increases with age, SG fibrosis in pSS has been considered a consequence of aging. As fibrotic damage occurs in other
autoimmune disorders, we hypothesize that SG fibrosis is a pathologic feature of pSS. METHODS. OSSCORT patients
with sicca symptoms and available hematoxylin and eosin stained SG biopsy slides were classified by American European
Consensus Group criteria (exclusions applied) as either pSS or not meeting criteria (DNMC). Both discovery (n= 66; 36
pSS, 30 DNMC) and validation (n=66; 36 pSS, 30 DNMC) cohorts were evaluated. SG cross-sections (4-6 cross-sections
per patient) were imaged and digitally reconstructed. Fibrosis was quantified in a blinded manner using a digital
grid. Sections were scored and reported as average percent area fibrosis per individual. Relationships of degree of fibrosis
with age and clinical measures were evaluated using Spearman correlations. Logistic regression was implemented to
assess the predictive value of fibrosis for disease classification. RESULTS. No differences in age between pSS and
DNMC cohorts were observed. Fibrosis was greater (Wilcox test) in pSS versus DNMC SG in discovery (p<0.0001) and
validation (p = 0.006) groups. In pSS, degree of fibrosis correlated (Spearman, two-tailed) with lymphocytic infiltrates (r=
0.300, p = 0.012), age (r =0.24, p=0.04) and corneal damage (r =0.25, p=0.03). In DNMC, fibrosis correlated with age
alone (r=0.30, p = 0.04). In a multiple regression model including fibrosis and age, SG fibrosis predicted disease
classification irrespective of age in both cohorts and in meta-analysis (68% prediction accuracy; fibrosis meta-p=8.1x10-5,
age meta-p=0.482). CONCLUSION. SG fibrosis is part of pSS pathology and not only a consequence of aging.
Funding: NIAMS 1P50AR060804, OMRF Barrett Pre-Doctoral Fellowship
47 Abstract #28
DISTINCT BIOCHEMICAL CHARACTERISTICS BETWEEN TWO HOMOLOGOUS ENDOSOMAL RABS
M. Caleb Marlin1, Yaoyao Qi2, Zhimin Liang1, and Guangpu Li1 1Department of Biochemistry and Molecular Biology,
University of Oklahoma Health Science Center, Oklahoma City, OK 2Key Laboratory of Biopesticide and Chemical
Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fujian, China
Recent studies from our lab have demonstrated the importance of the endosomal Rab GTPases, Rab5 and Rab22, in the
neuron’s response to nerve growth factor (NGF) via its high affinity receptor, TrkA. Rab5 and Rab22 show striking
similarities in sequence and structure, yet produce contrasting physiological outcomes during neuron differentiation. We
hypothesize that TrkA trafficking and intracellular signaling are differentially governed by Rab5 and Rab22. To better
understand what factors lead to the differences between Rab5’s and Rab22’s functions, we pulled-down Rab5 and Rab22
with a common effector, Rabenosyn-5. Interestingly, wild-type Rab5 did not pull-down with Rabenosyn-5, but the Rab5
constitutively active mutant along with Rab22 wild-type and its mutant did. By repeating this assay with Rab22/Rab5
chimeras, we determined that a small portion of the protein, encompassing six amino acids in length, was in part
responsible for the difference in Rab5 and Rab22 pull-down efficiencies. Substituting one amino acid within this sequence
drastically changed both the pull-down efficiency with Rabenosyn-5 and the rate of GTP hydrolysis of these proteins. The
data suggest that this previously unappreciated segment of Rab5 and Rab22 plays an important role in the differences
found between these two highly homologous proteins. This work, along with future studies, will further clarify the
implications of the minor differences seen in Rab5 and Rab22 upon the complex regulation of receptor–mediated
endocytosis, trafficking, and intracellular signaling in neurons.
Funding: NIH R01GM074692
48 Abstract #29
QUITLINE UTILIZATION AND OUTCOMES AMONG TOBACCO USERS WITH A GENERAL EDUCATIONAL
DEVELOPMENT (GED) DIPLOMA
Sydney Martinez1, Laura Beebe1 1University of Oklahoma Health Sciences Center, Department of Biostatistics and
Epidemiology
Introduction: The inverse association between educational attainment and smoking is well established; however, analyses
with refined categories indicate those with a General Educational Development (GED) certificate do not fit this pattern
and smoke at higher rates than high school (HS) dropouts. Oklahoma Tobacco Helpline registration and 7-month followup data were used to evaluate quitline utilization and effectiveness for individuals reporting GED as the highest level of
education completed. Methods: Adult tobacco users ages 25 or older who enrolled in a single- or multiple-call
intervention from July 1, 2012 to June 30, 2014 were included in analyses. Chi-square tests were used to test differences
in demographics, tobacco use behaviors, and service utilization. Cessation outcomes were evaluated using 30-day
respondent and intention-to-treat quit rates at 7-month follow-up. P-values of <0.05 were considered statistically
significant. Results: During the study period, 8.3% (n=3,823) of registrants reported a GED as their highest education
completed. Adults with a GED (29.4%) were less likely to smoke a pack or more cigarettes per day compared to HS
dropouts (33.3%, p<.0001) and more likely than HS graduates (26.5%, p=.0004). Respondent quit rates were higher
among HS graduates (34.4%) compared to HS dropouts (30.9%) or those with a GED (31.4%), although the differences
were not significant. Intention-to-treat quit rates were significantly lower among GED recipients (9.8%) compared to HS
graduates (13.4%). The quit rates for smokers with a GED were similar to HS dropouts (10.1%). Conclusions: Tobacco
users with a GED who called the Helpline are different from HS graduates in demographic characteristics, tobacco use
behaviors, and quit rates. It is important to evaluate quitline utilization and effectiveness for the GED population
separately from HS graduates. Understanding the differences between groups can help determine whether quitline
promotion and services should be tailored to more effectively serve this unique population.
Funding:
49 Abstract #30
AGE-RELATED COMMONALITIES AND SEXUAL DIMORPHISMS OF DNA METHYLATION IN THE MOUSE
HIPPOCAMPUS
Dustin R. Masser1,2,3, Benjamin C. Wronowski1,3, David R. Stanford1,3 and Willard M. Freeman1,2,3 Departments of
1
Physiology and 2Geriatric Medicine, 3Reynolds Oklahoma Center on Aging, University of Oklahoma Health Sciences
Center, USA
Aging is the major risk factor for diseases including heart disease and diabetes. Normative aging processes also affect the
brain leading to deficits in spatial learning and memory without neurodegeneration. The genome is hypothesized to
undergo hypomethylation with aging, creating a permissive environment for gene expression. Recent evidence suggests
DNA methylation is a dynamic regulator of gene expression allowing for sustained methylation as well as rapid
demethylation. We hypothesize changes in DNA methylation occurring with aging are specifically directed to gene
regulatory regions and that sex-specific changes in methylation with age may underlie differences in the aging process
among males and females. To identify molecular changes accompanying the aging process, we assessed mRNA and DNA
methylation from the hippocampus of Young(3M), Adult(12M), and Aged(24M) male and female C57Bl/6 mice (n=34/group). RNA was subjected to transcriptomic analysis and DNA methylation quantitation was performed through
bisulfite conversion and in-solution oligonucleotide capture of CG islands and gene promoters (~120Mb total sequence)
followed by sequencing (MethylCap-Seq). There was no change in global DNA methylation with age in the assayed
regions at CG (>1,500,000), CHG or CHH sites (>7,500,000), however a number of significant (p<0.001) age
(>13,000CGs) and sex-specific age-related (>3,000CGs) changes in methylation were observed across all autosomes. The
majority of significant age-related changes in DNA methylation occurred in intronic and intergenic regions (80% of CGs),
while promoter and exons contained the rest (20% of CGs) with the majority of these sites within 2kb of a transcriptional
start site. Of these changes, 28% overlapped with CG islands. Inflammation related genes (Cd52) were more highly
expressed in aged females compared to aged males. The combination of mRNA expression profiling with DNA
methylation quantitation provides a comprehensive analysis of epigenetic regulation of gene expression with aging and
identifies changes unique to each sex with age.
Funding:
50 Abstract #31
VISUALIZATION OF RETINOIC ACID SIGNALING IN THE DEVELOPING CHICK EYE
Kathryn Matthews1, Jody Summers2 1OUHSC Department of Pediatric Genetics, 2OUHSC Department of Cell Biology
Introduction: All-trans-Retinoic acid (atRA) is a known primary metabolite of retinol (Vitamin A) that has a key role in
signaling processes during embryonic eye development. Despite numerous studies on the role of atRA in development of
a number of systems, direct visualization of atRA gradients remains ambiguous. Using a specific reporter construct that
allows for the visualization of atRA signaling based on fluorescence resonance energy transfer (FRET) between two
fluorophores, we propose to visualize atRA signaling during pre- and post-natal eye development through the
development of “transgenic” chick embryos and chickens. Methods: A pminiTol2 backbone was used to generate a
genetically encoded probe for atRA (GEPRA). The GEPRA sequence contains a retinoic acid receptor (RAR) ligand
binding domain (LBD), which is flanked by cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP). When
atRA binds to the RAR-LBD, FRET occurs between the CFP/YFP fluorophore pair (Shimozono, S., et al., 2013). 293 T
cells transfected with GEPRA were imaged using a Fluoview 1000 Confocal inverted microscope. The FRET ratio was
then calculated before and after atRA treatment. In parallel, the minitol2 transposon system was used to deliver the
construct to the circulating primordial germ cells (PGCs) in stage 14-16 chicken embryos. Results: GEPRA undergoes
CFP-YFP FRET in response to atRA (500 nM to 10,000 nM) in a dose-dependent manner. Embryos that had been
injected with the GEPRA construct expressed YFP preferentially in the gonads. Co-localization of YFP with the stem cell
antigen, SSEA-1, further confirmed the presence of transgenic PGCs within the gonads of 7 and 14 day incubated
embryos. Conclusion: The ability to directly visualize retinoic acid and/or its signaling pathways will elucidate the
mechanisms by which retinoic acid mediates eye development, as well as the development of many other organ systems.
Funding:
51 Abstract #32
ADDRESSING ADULT PHYSICAL ACTIVITY THROUGH PUBLIC SAFETY POLICIES: A REGIONAL AND
RACIAL/ETHNIC ANALYSIS FROM 2006-2013
Breanca Merritt1 and Roy F. Oman1 1Department of Health Promotion Sciences, OUHSC College of Public Health
Introduction: The number of enacted state policies that intend to create safer environments for physical activity have
increased in the past 10 years. The purpose of this study was to examine associations between aggregated state policies
related to public safety and differences in physical activity behavior of U.S. adults among racial/ethnic groups. Methods:
Individual-level data (N=158,842) were obtained from the National Health Interview Survey (NHIS) from 2006-2013.
The sample consisted of white (n=95,078), black (n=26,422), and Hispanic (n=33,038) adults. Data on public safety
policies (N=116) were obtained from the CDC’s Chronic Disease State Policy Tracking System and merged with NHIS
data. The main outcome variable was minutes of moderate leisure-time physical activity (LTPA). LTPA was categorized
into adults who were inactive or insufficiently active compared to those who were sufficiently or highly active, based on
CDC recommendations. The public safety policy variable was lagged by one year. Logistic regressions estimated
relationships between moderate LTPA and public safety policies, adjusting for potential confounders. Results: The mean
number of public safety policies enacted per year among U.S. Census regions ranged from 0 to 28. Public safety policies
were associated with increases in moderate LTPA for white (OR=1.02, 95% CI 1.01-1.03) and Hispanic adults (OR=1.02,
95% CI 1.00-1.03) living in the Western U.S., and white adults living in the Midwest (OR=1.03, 95% CI 1.00-1.06)
Public safety policies were negatively associated (OR=.98, 95% CI .98-.99) with moderate LTPA among Hispanic adults
living in the South. Public safety policies had no significant relationship with moderate LTPA for black adults in any
region. Conclusion: Policies designed to create safe environments for physical activity may work more effectively for
some regions and for some racial/ethnic groups than others. Understanding the content of these policies may clarify why
these policies vary in effectiveness.
Funding: N/A
52 Abstract #33
UNHEALTHY BEHAVIORS AMONG CANCER SURVIVORS
Dana Mowls, Laura Beebe Department of Biostatistics and Epidemiology, College of Public Health, University of
Oklahoma Health Sciences Center; Oklahoma Tobacco Research Center
The number of cancer survivors in the community is increasing, yet little is known about the prevalence of behavioral risk
factors across all types of cancer diagnoses. This research examines lifestyle behaviors that may impact survivorship and
quality of life among cancer survivors in Oklahoma. Cross-sectional data were obtained from the Oklahoma Behavioral
Risk Factor Surveillance System (2011-2013). Of 24,783 respondents, 2,436 said ‘yes’ to ever having any type of cancer
and were identified as cancer survivors. An additional 1,824 had skin cancer diagnosis only or missing for any type of
cancer and were excluded. Respondents with no prior cancer diagnosis served as the comparison group (n=20,523).
Weighted prevalences and 95% confidence intervals (CI) were computed. Odds ratios (AOR) adjusted for age, sex, race,
and education were computed by logistic regression to assess differences between cancer survivors and the comparison
group. Overall, 7.2% (CI: 6.8, 7.5) of respondents were cancer survivors. Approximately one in four (23.1%) cancer
survivors reported smoking cigarettes, two in five (40.8%) were physically inactive, two in three (67.7%) were
overweight or obese, and the vast majority (86.9%) did not consume the recommended daily servings of five fruits or
vegetables. After controlling for socio-demographic factors, the odds of being a current smoker (AOR: 1.20; CI: 1.03,
1.39) or physically inactive (AOR: 1.20; CI: 1.06, 1.35) were 20% higher in survivors than the comparison group. The
odds of consuming the recommended daily servings of fruits or vegetables (AOR: 1.45; CI: 1.16, 1.81) were significantly
higher in survivors. No difference was observed for overweight or obesity in adjusted analyses. Cancer survivors report
high prevalences of unhealthy behaviors, deviating from the American Cancer Society guidelines. Cancer survivors are
now living longer making it imperative to reduce unhealthy behaviors that may increase risk of re-occurrence and worsen
quality of life.
Funding: Oklahoma Tobacco Settlement Endowment Trust
53 Abstract #34
VISION AND HEARING LOSS ASSOCIATED WITH USHER SYNDROME TYPE 2A
Maggie L. Mwoyosvi1, MichaelAnne Gratton2, and Muna I. Naash1 1Department of Cell Biology, University of Oklahoma
Health Sciences Center, Oklahoma City, OK 73104, USA 2Otolaryngology-Head, Neck Surgery, Saint Louis University,
Saint Louis, MO, USA
Usher syndrome (USH) is the leading cause of combined deafness and blindness; and the mechanisms of sensory loss are
unknown. The usherin c.2299delG mutation is the most common cause for USH2A in patients. To better study the role
of usherin in audiovisual impairment we generated a knockin (KI) mouse model expressing this human mutation. The
c.2299delG mutation causes a frame shift resulting in a premature stop codon. In addition to introducing the c.2299delG
mutation, the KI construct also contains an internal ribosomal entry site followed by GFP (to assess promoter
activity). GFP evaluation identified new tissues expressing usherin not been previously reported and will be explored
further for structural abnormalities. Due to the location of usherin at the connecting cilium of the photoreceptors, proper
translocation of phototransduction machinery upon light exposure was evaluated. Retinal assessments via IHC and ERG
up to postnatal day 180 (P180) appear normal; however there is a delay in recovery of the rod ERG amplitudes at P180 as
well as incomplete translocation of transducin and arrestin upon light exposure at P30. Furthermore, animals at P360
have a significant decrease in maximum scotopic amplitudes (p<0.05), a phenomenon resembling retinitis pigmentosa in
patients. Cochleae scanning electron micrographs (SEM) show outer hair cell abnormalities consistent with hearing loss
at P180. This mouse is the first USH2A model with a human mutation and the first to show retinal dysfunction without
rearing in higher cyclic light. Data are consistent with USH patients having a later onset of visual impairments compared
to hearing loss. GFP expression evaluations will allow for further study of possible systemic effects usherin may have in
conjunction with deafness/blindness. Full characterization of this model will lead to an understanding of the human
disease mechanism and will be valuable in developing targeted therapies for treatment.
Funding: This project was supported by Fight for Sight Summer fellowship, Foundation Fighting blindness, and NEI
(EY10609, EY18656, EY022778)
54 Abstract #35
UNCONSTRAINED KEAP1 RESTRICTS STRESS-INDUCED MITOCHONDRIAL CLUSTERING
Gary B. O'Mealey, William L. Berry, and Scott M. Plafker
Proteasome inhibition initiates a cascade of responses aimed at preventing the accumulation of toxic aggregates that
propagate proteotoxicity. Here, we demonstrate that the mitochondrial network forms microtubule-dependent,
juxtanuclear clusters as an early response to this stress. Clustering facilitates the delivery of a stable, mitochondrial
population of the anti-stress transcription factor Nrf2 to the nucleus to induce chaperone expression and suppress the
impending proteotoxicity. Nrf2 is anchored at the mitochondria in complex with the CUL3 E3 ligase substrate adaptor
KEAP1 and the mitochondrial phosphatase PGAM5. Disrupting this complex reduces clustering and the mitochondrial
delivery of Nrf2 to the nucleus, which in turn delays expression of the Nrf2 target gene, Hsp70, a key chaperone in the
maintenance of proteostasis. PGAM5-KEAP1-Nrf2 complex disruption abrogates clustering by giving rise to a
neomorphic population of KEAP1 on mitochondria that leads to destabilization of Miro2, a mitochondrial GTPase
required for mitochondrial translocation along microtubules. Collectively, our data identify a novel function for Nrf2 at
the mitochondria and show that the mitochondria function as a beacon of proteotoxic stress. These findings could have
pathophysiological implications, as Nrf2 loss is associated with numerous age-dependent diseases.
Funding: NIH (1R01GM092900-03), OCAST, Beckman Initiative for Macular Research
55 Abstract #36
ROLE OF REDUCED SYNAPTOBREVIN 2 LEVELS IN AGE-RELATED COGNITIVE DECLINE
Albert Orock1, 2, Sreemathi Logan2, Ferenc Deak1, 2 1Oklahoma center for neuroscience, 2 Reynolds Oklahoma center on
aging, OUHSC, Oklahoma City, OK 73104
Introduction: One of the major disabilities that affect the aging population is age related cognitive decline. Synaptic
dysfunction is emerging as the major cause of cognitive impairment and dementia. Synaptic plasticity is a central
mechanism in learning and memory. Our laboratory and others have identified synaptobrevin-2, a SNAP Receptor
(SNARE) protein as a key player in synaptic transmission and plasticity. Synaptobrevin-2 (syb2) is the major SNARE
protein of synaptic vesicles (SV) which is highly expressed in the cerebral cortex and hippocampus, an essential brain
center for spatial learning. Syb2 protein levels have also been shown to decrease with age but functional consequences of
this lower expression are still elusive. We hypothesize that reduction of syb2 protein levels with age mediates age related
cognitive decline. Methods: We have been using heterozygous syb2 knock-out mice which express about the same protein
levels at 6 month-of-age as old (24 mo) wild-type animals. This is 50-60% of the protein levels found in young wild-type
littermates (p<0.01, Student t-test). We performed behavioral tests for spatial learning and memory using the radial-arm
water maze (RAWM) and a complex cohabitate environment (Intellicage). Neuronal plasticity was assessed by long-term
potentiation (LTP) assays on hippocampal slices. We used live fluorescence microscopy to measure the SV release rate in
neuronal cultures. Results: The behavioral test with syb2 heterozygous resulted in maintained basic spatial memory
acquisition for simple place finding (RAWM) but impaired learning in complex tasks in Intellicage. This impaired spatial
memory was reflected in reduced CA1 hippocampal LTP and Syb2 heterozygous neurons have reduced SV release rates
(~35% in 15 seconds, p<0.01) compared to wild-type controls. Conclusions: This is the first study to demonstrate that
syb2 levels have a causative role in synaptic failure and dementia and our results suggest syb2 reduction with age
mediates age related cognitive decline.
Funding: COMAA Research Fund to F. D.
56 Abstract #37
THE EFFECT OF “SPEAK-OUT!®” VOICE THERAPY ON PROSODY IN PERSONS WITH PARKINSON’S
DISEASE
Eunsun Park1, Christina Santos2, Justin Dvorak1, Jason Gates2, Frank Boutsen, PhD1 1Motor Speech and Prosody
Research Lab, Department of Communication Sciences and Disorders, College of Allied Health, University of Oklahoma
Health Sciences Center, 2INTEGRIS Jim Thorpe Rehabilitation Center, Oklahoma City, OK
Parkinson’s disease (PD) is a progressive neurodegenerative disorder affecting 1 in 100 adults over the age of 60. A
majority of persons with PD manifest hypokinetic dysarthria, which is characterized by reduced loudness, breathy voice,
mono-pitch, intermittent rapid rushes of speech, and imprecise production of consonants. SPEAK OUT!® is a voice
therapy program to improve functional communicative ability, stressing “speaking with intent” to increase
amplitude. Using readings of the “My Grandfather” passage (GP), vowel prolongation, diadochokinetic tests (DDK), and
conversational speech samples, this study evaluated the efficacy of the 12-session over 4 weeks SPEAK-OUT!® program
in terms of prosody changes such as speech rate, speech intensity and pitch range for persons with idiopathic PD. Data
have been collected on 5 male participants (mean age 71.6±8.0 yrs, mean duration of PD 5.0±1.6 yrs). Data included
participants’ scores on the Voice Handicap Index (VHI) and Voice-Related Quality of Life (V-RQOL) questionnaires,
audio recordings, and demographic data. Participants were asked to read the GP, produce a sustained vowel sound (/ah/),
DDK syllables, and conversational speech. Post-treatment speech intensity was significantly improved vs. pre-treatment
in both conversational (p=0.002) and GP reading (p=0.0046) tasks. Post-therapy pitch range in the GP reading was
significantly greater than pre-treatment (p=0.012). Post-therapy VHI score was significantly lower than pre-treatment
(p=0.004). While participants’ age did not significantly influence change in intensity, greater duration of PD was
associated with reduced change in intensity of the GP reading (p=0.024). Conversational speech rate was marginally
slower (p=0.0741) post-treatment. Participants achieved a significant increase in speech intensity and pitch range after
SPEAK-OUT!® training, consistent with self-reports such as the VHI. Participants read the GP with more varied pitch
range, indicating improvement of their reading fluency. These results suggest that the SPEAK-OUT!® program is a viable
treatment for persons with PD.
Funding: None
57 Abstract #38
NOVEL PROTEIN ADTRP NEGATIVELY REGULATES WNT SIGNALING
Maulin Patel1, 2, Cristina Lupu1, Robert Silasi-Mansat1, Chris Sansam1, 2, Florea Lupu1, 2 1 Cardiovascular Biology
Program, Oklahoma Medical Research Foundation (OMRF), 2 Cell Biology Department, University of Oklahoma Health
Science Center (OUHSC)
Background: We are investigating the expression and function of a novel protein encoded by C6ORF105. The novel
protein is androgen-responsive, as well as predicted to be palmitoylated and multispan. Tissue Factor Pathway Inhibitor
(TFPI) is a serine protease inhibitor and its role in coagulation and angiogenesis is well established. Our initial studies on
human endothelial cell lines have shown that C6ORF105 expression co-regulates with TFPI; therefore, this novel protein
has been named androgen-dependent TFPI-regulating protein (ADTRP). Methods and Results: To characterize the
expression and function(s) of ADTRP in vivo, Zebrafish was used as an animal model. Expression analysis during
embryonic development (up to 7 dpf) and in adult Zebrafish showed that Adtrp is expressed in most of the organs, with
relatively higher expression in the jaws, brain, testis, otic vesicle, heart and intestine. Morpholino based knockdown of
Adtrp expression produced a distinct phenotype of aberrant angiogenesis, jaw/cartilage defects and heart malformation. In
vitro experiments of overexpressing ADTRP in HEK293 cells showed significant reduction in Wnt signaling. Conclusion:
This is the first study to characterize the expression and function of the novel protein ADTRP in vivo. Our studies
revealed that this protein could play important roles both during development and in the adulthood possibly by negatively
regulating the Wnt signaling pathway(s).
Funding: Oklahoma Center for Adult Stem Cell Research (OCASCR), Oklahoma Medical Research Foundation (OMRF)
58 Abstract #39
TARGETING TUMORS LIKE A T CELL: EVALUATION AND TARGETING OF HLA-PRESENTED MIF IN
OVARIAN CANCER
Andrea Patterson1, Saghar Kaabinejadian1, Curtis McMurtry1, Wilfried Bardet1, Ken Jackson1, Rosemary Zuna2, Sanam
Husain2, Harold Ames3, Oriana Hawkins3, Jon Weidanz3, and William Hildebrand1 1Department of Microbiology and
Immunology, OUHSC 2Department of Pathology, OU Medical Center 3Department of Immunotherapeutics, Texas Tech
University Health Sciences Center
Background and Objectives: T cells recognize cancer cells via human leukocyte antigen (HLA)/peptide complexes and,
when disease overtakes these immune mechanisms, immunotherapy can exogenously target these same markers. We
previously identified an HLA-A2-presented peptide derived from macrophage migration inhibitory factor (MIF), a
pleiotropic cytokine with multiple tumor-promoting functions, and showed that an antibody against the MIF/HLA-A2
complex (RL21A) specifically targets invasive breast tissues. The objectives of the current study are to determine whether
the MIF peptide is presented by ovarian cancer cells as well, as MIF is found overexpressed in the majority of ovarian
cancer cases, and to assess whether RL21A represents a candidate immunotherapy agent for this deadly disease. Methods
and Results: Expression of a secreted HLA-A2 molecule (sHLA) in cancerous ovarian cell lines enabled high-sensitivity
HLA peptide retrieval and mass spectrometric analysis, and the MIF peptide was identified in all four lines. By flow
cytometry, RL21A was shown to stain all four cell lines, specifically in the context of HLA-A2. Next, 27 ovarian cancer
and 24 normal fallopian tube tissues underwent immunohistochemical staining with RL21A to assess differential
MIF/HLA-A2 complex expression. Ovarian tumor tissues showed significantly increased RL21A staining (p<0.001)
compared to normal fallopian tube epithelium, with minimal staining of normal stroma and blood vessels (p<0.002
compared to tumor cells) suggesting a therapeutic window. We then tested anti-cancer function when RL21A is bound to
a toxin, and a tetrazolium (MTT) cytotoxicity assay demonstrated dose-dependent killing of ovarian cancer cells.
Conclusions: The MIF-derived peptide is prevalently presented and recognized by RL21A on ovarian cancer cell lines
and patient tumor tissues. Targeting the HLA-A2/MIF complex with RL21A can produce ovarian cancer cell death, and
overall this complex is a promising target for ovarian cancer immunotherapy.
Funding: NIAID Ruth L. Kirschstein Training Grant T32AI007633 Emergent Technologies Incorporated Fund
59 Abstract #40
THE ABCS OF CARDIOVASCULAR DISEASE AND STROKE PREVENTION: PHARMACIST AND PAYER
PERSPECTIVES ON SERVICE PROVISION AND PAYMENT
Timothy Pham1, Lourdes Planas1 1 The University of Oklahoma Health Sciences Center, College of Pharmacy: Clinical
and Administrative Sciences
Objective: To describe perceptions among Oklahoma pharmacists, third-party administrators, and professional
organization leaders about pharmacist involvement in the ABCS of cardiovascular disease and stroke prevention
(appropriate low-dose aspirin use, blood pressure control, cholesterol management, and smoking cessation). Methods:
Eight focus groups with 21 community or ambulatory clinic pharmacists were conducted using a qualitative approach
with a phenomenological perspective. Eight third-party payer administrators or professional organization leaders were
interviewed one-on-one. Focus group and interview questions centered on participants' views regarding pharmacist
involvement in, and payment for, the ABCS of cardiovascular disease and stroke prevention. Discussions were audiotaped and transcribed. Two investigators independently analyzed the transcripts and consulted with one another to
condense and categorize common themes. The study was approved by a university and a state health department
institutional review board. Results: Commonly reported barriers among participants included lack of pharmacy-specific
resources, lack of recognition for pharmacist value, and a fragmented healthcare system. Third-party administrators, in
particular, highlighted the need for objective assurance of pharmacist competency, and evidence-based proposals from
pharmacists containing quantitative data on outcomes and cost-savings of their services. Pharmacist participants
considered meaningful relationships with patients and other healthcare providers vital. Most participants viewed
medication adherence as an appropriate indicator to measure pharmacist service effectiveness. Some pharmacists noted
quality measures, such as Medicare CMS star ratings, as opportunities for pharmacists to address medication use needs in
healthcare systems. Implications/Conclusions: Stakeholder perceptions about Oklahoma pharmacist involvement in, and
payment for, the ABCS of cardiovascular disease and stroke prevention described various barriers and facilitators. These
factors should be considered when developing plans to further integrate pharmacists’ patient care activities into
sustainable programs that address the ABCS.
Funding: Oklahoma State Department of Health’s CDC Heart Disease and Stroke Prevention Program
60 Abstract #41
AMYGDALA-MEDIATED MECHANISMS OF VISCERAL PAIN IN ADULTHOOD FOLLOWING EARLY LIFE
STRESS
Dawn K. Prusator† and Beverley Greenwood-Van Meerveld†,‡ *. Oklahoma Center for Neuroscience†; Department of
Physiology ‡, * VA Medical Center, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA
Early life stress (ELS), such as neglect or abuse, has been indicated as a risk factor for the development of visceral pain in
adulthood. Our lab has developed a model that recapitulates the human situation of ELS and induces visceral pain in adult
rats. However, alterations in neural pathways that regulate pain have not been delineated following ELS exposure. We
postulated that ELS alters the function of the central amygdala (CeA), a key brain region in the processing of stress and
pain. Specifically, we hypothesized that ELS alters glucocorticoid receptor (GR) expression and subsequent regulation of
corticotropin-releasing factor (CRF), a known regulator of visceral pain. In neonates, ELS occurred via unpredictable
odor-shock exposure with odor only exposure serving as a control. In adulthood, CeA punches were collected and
expression of target genes was assessed via qRT-PCR. To knockdown GR expression cannulae were implanted into the
CeA for infusion of antisense or random sense oligodeoxynucleotides (ASO/RSO). In adulthood, visceral pain was
assessed via visceromotor behavioral response (VMR) quantified as the number of abdominal contractions in response to
graded pressures (0-60mmHg) of colorectal distension (CRD). Following neonatal exposure to unpredictable ELS, adult
rats showed significantly increased (P<0.01) GR and CRF expression in the CeA compared to odor only controls.
Knockdown of GR in the CeA via ASO infusion increased the number of pain responses in both groups (P<0.001) as
quantified by VMR to CRD. These findings indicate that exposure to unpredictable ELS disrupts the ability of GR to
negatively regulate CRF expression in the CeA. Furthermore, this data suggests that uncoupling the homeostatic
relationship between GR and CRF plays a pivotal role in the development of ELS induced visceral pain. Future studies
will investigate whether direct knockdown of CRF in the CeA will normalize ELS induced visceral pain in adult rats.
Funding: This work was supported by a department of Veterans Affairs Merit Grant to Dr. Beverley Greenwood-Van
Meerveld and an Oklahoma Center for Neuroscience Seed Grant to Dawn Prusator.
61 Abstract #42
MAMMALIAN GLYCOGENIN-1 RELATED GLYCOSYLTRANSFERASE IS IMPORTANT FOR TOXOPLASMA
PROLIFERATION
Kazi Rahman1, Peng Zhao3, L. Wells3, Ira J Blader4, Christopher M West2 1Department of Microbiology & Immunology,
OUHSC; 2Department of Biochemistry & Molecular Biology, OUHSC; 3Complex Carbohydrate Research Center,
University of Georgia, GA 30602; 4Department of Microbiology & Immunology, University at Buffalo, NY 14214
Toxoplasma gondii (Tg) is a protozoan parasite that causes disease by proliferating intracellularly in target cells in muscle
and the central nervous system. Remarkably, Toxoplasma is able to grow at O2 levels all the way down to 0.5%, which
suggests the existence of a homeostatic mechanism that accommodates to varied O2 levels. Previous studies in our lab
showed that Tg-PhyA mediated prolyl hydroxylation of Skp1, a subunit of SCF-class of E3 ligases, is required for optimal
parasite proliferation especially at low O2. In another protozoan, Dictyostelium, the hydroxyproline is further modified
by 5 sugars, which are also required for optimal O2-sensing. Glycosylation appears to be important for Toxoplasma
proliferation because disruption of two glycosyltransferase genes predicted to form the core trisaccharide on Tg-Skp1
results in decreased parasite growth. Mass spectrometric analysis revealed a pentasaccharide on Tg-Skp1 in wild-type, and
the expected truncated glycans in the glycosyltransferase-KOs. This raises the question of whether the terminal
disaccharide is important and what mediates its addition. Using bioinformatics approaches we predicted a candidate
terminal glycosyltransferase gene, GT8A, and examined the effects of its disruption. GT8A-KO parasites exhibited an
intermediate growth rate between those of wild-type and phyA-KO cells. Mass spectrometric analysis confirmed the
predicted accumulation of the tetrasaccharide, suggesting that GT8A adds the terminal sugar on TgSkp1. GT8A shows
high sequence similarity to glycogenin-1, an enzyme that autoglucosylates itself, and primes metazoan and yeast glycogen
synthesis. As known for glycogenin, bacterially-expressed GT8A exhibits auto-glucosylation activity, but surprisingly
also has promiscuous auto-galactosylation and auto-GlcNAcylation activities. The auto-glycosylation activities raise the
question of whether the growth phenotype of GT8A-KO parasites is due to the defect in Skp1 modification, or another
activity possibly associated with glycogen synthesis. Future studies will focus on how GT8A contributes to Toxoplasma’s
glycogen synthesis and growth at low O2.
Funding: This work was supported by grants from the NIH (R01 GM084383), the Oklahoma Center for Advancement of
Science and Technology (HR10-0181), and the Mizutani Foundation.
62 Abstract #43
TLR4-INTERACTING SPA4 PEPTIDE SUPPRESSES NLRP3-INFLAMMASOME AGAINST LPS AND ATP
STIMULI.
Vijay Ramani and Shanjana Awasthi Departments of Pharmaceutical Sciences University of Oklahoma Health Sciences
Center
Introduction - During tissue injury, the pathogen- (PAMPs) and damage-associated molecular patterns (DAMPs) induce a
crosstalk between Toll-like receptor-4 (TLR4) and inflammasome, eventually causing an exaggerated inflammation. The
TLR4-interacting SPA4 peptide (amino acids GDFRYSDGTPVNYTNWYRGE), discovered earlier in Dr. S. Awasthi’s
lab, binds to TLR4 and suppresses lipopolysaccharide (LPS; PAMP and a TLR4 ligand)-induced inflammatory cytokine
(JPET, 2011) and tissue inflammation (Innate Immun, 2013). In this work, we investigated the anti-inflammatory effect of
SPA4 peptide in cells challenged with LPS and adenosine tri-phosphate (ATP; DAMP, an NLRP3-inflammasome
inducer). Methods - Murine bone marrow-derived dendritic cells (nontransfected or transfected with plasmid DNAs
encoding wild-type and dominant negative forms of TLR4) and primary murine alveolar macrophages were challenged
with LPS and ATP, and treated with SPA4 peptide. The mRNA expression and intracellular protein pools of (pro)-IL-1β
and NLRP3 were quantitated by qRT-PCR and immunoblotting, respectively. The ASC-specks were evaluated as an
indicator of NLRP3-inflammasome assembly by fluorescence microscopy. Caspase activity was assessed by incubating
the cell lysates with fluorogenic Ac-YVAD-AMC. Secreted proteins were precipitated and immunoblotted for active
caspase-1 subunit. The IL-1β levels were measured in cell-free supernatants by ELISA. Results - Our data demonstrated
that SPA4 peptide significantly reduced mRNA and cellular pools of (pro)-IL-1β and NLRP3 against LPS and ATP
stimuli. These changes corroborated with reduced NLRP3-inflammasome assembly, caspase activity and IL-1β.
Furthermore, results from genetically-transfected cells suggested that the SPA4 peptide suppression of NLRP3inflammasome is through its effect on TLR4-priming step. Conclusions – In continuation with previously published
results, these findings suggest that the suppression of LPS-TLR4-NF-κB by SPA4 peptide further reduces the NLRP3inflammasome.
Funding: University Growth Fund (to SA), and NIH-NIGMS (P20GM103648 to SA).
63 Abstract #44
NKT CELLS MEDIATED ENHANCEMENT OF HUMORAL IMMUNITY AGAINST CLOSTRIDIUM DIFFICILE
TOXIN B
Pragya Rampuria1, T. Scott Devera1, Gillian A. Lang1, Mark L. Lang1 1University of Oklahoma Health Sciences Center,
USA
To develop more effective antibody-stimulating vaccines, we must improve our understanding of the mechanisms by
which they stimulate the induction and maintenance of long-lasting and antigen-specific memory B cells plasma cells.
This is particularly important for the development of new vaccines against emerging pathogens such as C. difficile. It is
well-established that Natural Killer T (NKT) cells, activated with the glycolipid, a-galactosylceramide (a-GC) enhance
humoral immune responses, but the mechanisms by which they do so are poorly understood. Here we show that activated
NKT cells enhance the humoral immune response and survival against C. difficile Toxin B (TcdB). We and others have
shown that NKT cells may enhance priming of T cells. We therefore hypothesized that T helper and NKT subsets
cooperatively enhance the humoral immune response against C. difficile. We therefore determined whether NKT cell
activation led to changes in number and function of Th and NKT subsets. Flow cytometry analysis demonstrated that
NKT cell activation with a-GC did not increase numbers of T helper (Th) or follicular T helper (Tfh) cells, but increased
the number of NKTfh cells. ELISPOT assays with NKT-depleted lymphocytes indicated that the enhanced secretion of
IL-21 is NKT derived early in the response (day 3). However at day 7 IL-21 is mainly Th-derived. IL-4 on the other hand
is mainly being secreted by Th cells at both early and relatively late time points. Adoptive transfer experiments show that
a-GC did not alter the ability of Th cells to boost antibody production. This suggests that NKT cells may exert direct and
indirect effects (via Th cells) on humoral immunity. Understanding the mechanism by which NKT cells enhance humoral
immunity will help in designing strategies to appropriately activate NKT cells and boost vaccine efficacy.
Funding: NIH-AI078993 and OCAST-HR12-005
64 Abstract #45
CAVEOLIN-1 PROMOTES RETINAL DAMAGE RESPONSE FOLLOWING ACUTE ISCHEMIA REPERFUSION
Alaina M. Reagan1 and Michael H. Elliott1,2. 1Oklahoma Center for Neuroscience, 2Department of Ophthalmology/Dean
McGee Eye Institute, University of Oklahoma Health Sciences Center, Oklahoma City, OK
Introduction: Caveolin-1, (Cav-1) the scaffolding protein of caveolae membrane domains, promotes retinal inflammatory
responses mediated by Toll-like receptor-4 (TLR4) signaling. TLR4 is activated in the retina as consequence of retinal
ischemia/reperfusion (I/R) injury caused by intraocular pressure (IOP) elevation, the primary risk factor in glaucoma.
Intriguingly, polymorphisms at the CAV1/CAV2 gene locus are associated with increased risk of glaucoma but the
mechanism of this association is unknown. In this study, we tested the hypothesis that Cav-1 deficiency suppresses the
TLR4 response induced by I/R and results in reduced retinal tissue damage. Methods: To mimic elevated IOP, transient
ischemia was induced in one eye of Cav-1 knockout (KO) and control mice by inserting a 33-gauge needle connected to a
raised saline reservoir into the anterior chamber. IOP was raised to 100 mmHg for 1 hour followed by retinal
reperfusion. Contralateral eyes were used as controls. 24h post I/R, retinas were taken to assess IL-6 production using an
ELISA kit. Two weeks post I/R, immunohistochemistry was performed on retinal flatmounts to assess ganglion cell death
Results: Following I/R, protein levels of IL-6 were increased in the WT animals, but blunted in the Cav-1 KO animals.
Because there is evidence that Cav-1 enables TLR pathway activation following an insult, Cav-1 ablation resulting in
reduced IL-6 levels follows this trend. Ganglion cell death occurred in both control and Cav-1 KO animals after I/R.
However, cell death within the control retinas was greater than in the Cav-1 KO retinas, indicating that loss of Cav-1 may
prevent TLR4 activation, providing neuronal protection. Conclusions: These data support a regulatory role for Cav-1 in
the TLR4 signaling pathway. Our results are consistent with the idea that Cav-1 ablation dampens TLR4 activation, and
thus downstream cytokine signaling. Reduced cytokine production may then increase neuronal survival in the
glaucomatous retina.
Funding: NIH grants: EY019494 (MHE), core grant EY021725; and an unrestricted grant from Research to Prevent
Blindness, Inc.
65 Abstract #46
THE ROLE OF RIF1 AND THE REPLICATION TIMING PROGRAM IN VERTEBRATE DEVELOPMENT
Joseph Siefert1, Amnon Koren3, Courtney Sansam2, Emily Clowdus1, Duane Goins2, Chris Sansam1,2 1University of
Oklahoma Health Science Center, Department of Cell Biology, Oklahoma City, OK; 2Oklahoma Medical Research
Foundation, Cell Cycle and Cancer Biology Research Program, Oklahoma City, OK; 3Department of Genetics, Harvard
Medical School, Boston, MA
DNA replication timing is the coordinated manner in which different sequences of DNA are replicated during S-phase of
the cell cycle. The DNA replication timing program is stable and very reproducible among individual cell types, and the
spatiotemporal order of replication is highly conserved among metazoans. In spite of this, the mechanisms and
determinants of replication timing have remained largely elusive. However, replication timing changes have been
observed to occur during embryonic stem cell differentiation in vitro as well as in multiple types of cancer. Furthermore,
late replicating sequences are known to have higher mutation rates in cancer. Using zebrafish as a model of vertebrate
development, we have generated replication timing profiles from various stages of development corresponding to different
transcriptional and differentiation statuses. Our results indicate that lineage-independent replication timing can be
observed at the whole organism level and suggest that changes in replication timing occur in vivo coincident with
differentiation. Ongoing analysis will examine the correlations between replication timing, transcription, and epigenetic
marks, which can reveal conserved mechanisms of replication timing. To date, the only metazoan protein shown to have
global effects on replication timing is Rif1. Our Rif1 knockdown in zebrafish has demonstrated that Rif1 is required for
vertebrate development and has revealed defects in hematopoiesis and vascular development. Using knockdown and
overexpression methods, we are investigating the role Rif1 plays in cell differentiation during vertebrate
development. Additionally, Rif1 has known roles in DNA damage response and collapsed replication fork repair, and
several mutations in Rif1 have been identified in breast cancer. Therefore, we have used TALENs, a precise genome
editing technology, to create an allelic series of Rif1 mutants, including a null and several modified versions of the
protein, in order to generate separation of function mutants and elucidate mechanisms of its biological functions.
Funding: John and Mildred Carson Ph.D. Fund Award, OCASCR
66 Abstract #47
PROCESSING OF FACIAL EMOTIONS IN HEALTHY ADULTS
Kayle Sneed1, Justin Dvorak1, Eunsun Park1, Elliot Ross2, Frank Boutsen1 1Motor Speech and Prosody Research
Laboratory, OUHSC, Oklahoma City, OK 2Veterans Affairs Medical Center, Oklahoma City, OK
Purpose/Hypothesis: Clinical research on facial emotions has emphasized differences in mobility and expression between
the right and left hemi face. Some social psychologists suggest that control of facial expression is organized across the
upper-lower facial axis because of the phenomena of facial blends. It has been demonstrated that upper facial emotions are
processed preferentially by the left visual field, and lower facial emotions are processed preferentially by the right visual
field. However, research has also shown that with advanced age, healthy subjects lose their ability to process upper facial
emotions. The purpose of this study was to determine how visual field, attentional condition, and instruction influence the
processing of facial emotions in healthy adults over the age of 65, as measured by initial saccade movements recorded
during tachistoscopic presentation of facial blends of emotions. Materials and Methods: Eight healthy males (n=1) and
females (n=7) ranging from age 67 to 76 years (mean=71.1 SD=2.8) were asked to look at the face or look and identify
the facial emotion. In half the trials, participants were also asked to attend to the upper face. Stimuli were presented
tachistoscopically and randomized using E-Prime software. An ASL 5000-series eye tracking system measured the
direction and gaze position of the subject's initial saccade relative to the central fixation point. Gaze position was
modeled in repeated-measures analysis as a function of visual field, instruction, and attentional condition. Results: The
attend condition (p=0.0043) and the interaction between attentional condition and task instruction were significant
(p=0.0385). When the participants were asked to attend to the upper face without identifying the emotion, their gaze was
higher than when they were asked to look at the upper face and identify the emotion. Discussions/Conclusions:
Attentional condition and task instruction compete for influence over initial gaze position.
Funding: None.
67 Abstract #48
RDS GLYCOSYLATION; A WINDOW INTO THE DIFFERENTIAL ROLE OF RDS IN RODS AND CONES.
Michael Stuck, Shannon Conley, Muna Naash
The photoreceptor specific glycoprotein retinal degeneration slow (RDS, also called PRPH2) is necessary for the
formation of both rod and cone photoreceptor outer segments, however mutations of RDS can lead to either rod or cone
dominant disease through unclear molecular mechanisms. The conserved RDS N-glycosylation has been shown to be
dispensable in rods leading us to propose the hypothesis that RDS glycosylation is important for RDS’ cone specific
function. To test this we generated a knock-in mouse expressing RDS which lacks the single N-glycosylation site
(N229S) through a point mutation in the native RDS allele. N229S animals exhibited normal rod outer segment structure
and function and normal levels of RDS and the non-glycosylated RDS binding partner rod outer segment membrane
protein-1 (ROM-1). However cone ERG responses were decreased by 40% at 6 months of age. Since cones make up only
3-5% of photoreceptors in the WT background, N229S mice were crossed into the nrl-/- background (in which all rods are
converted to cone-like cells) for biochemical analysis. In N229S/nrl-/- retinas, RDS and ROM-1 levels were decreased by
63% and 70%, respectively. These data suggest that the glycosylation of RDS is specifically required for RDS function or
stability in cones
Funding: This work was supported by the National Institutes of Health (EY010609, EY22778, EY018656-MIN,
T32EY023202-MWS), the Foundation Fighting Blindness (MIN), and the Oklahoma Center for the Advancement of
Science and Technology (SMC).
68 Abstract #49
ARID3A-MEDIATED IFNA EXPRESSION IN SYSTEMIC LUPUS ERYTHEMATOSUS B CELLS
Julie M Ward1,2, Michelle L Ratliff1, Troy Templeton1,3, Mikhail Dozmorov4, Kenneth Smith, Graham Wiley4, Patrick
Gaffney4,5, Judith A James2,3,5, and Carol F Webb1,2,3 1Immunobiology and Cancer Research, and 4Arthritis and Clinical
Immunology, Oklahoma Medical Research Foundation; Departments of 2Microbiology and Immunology, 3Cell Biology,
and 5Medicine, University of Oklahoma Health Sciences Center
Introduction: Systemic lupus erythematosus (SLE) is an autoimmune disease presenting with autoantibody production and
chronic flares of inflammation. We previously showed increased disease severity in lupus patients was correlated
(p=0.0038) with expanded numbers of peripheral blood B cells expressing the DNA-binding protein, ARID3a. It is
unclear why ARID3a+ B cells are increased in SLE. Therefore, we asked what induces ARID3a expression in healthy B
cells, and if the relationship between ARID3a+ B cells and disease activity was via an antibody-dependent or –independent
mechanism. Methods: Plasma and mononuclear cells were isolated from peripheral blood from SLE patients. B cells
were isolated via magnetic bead separation for qRT-PCR or in vitro experiments, and analyzed by flow cytometry. Tolllike receptor (TLR) agonists were evaluated for their ability to induce ARID3a expression, and the inflammatory
cytokine, interferon alpha (IFNA). Plasma-stimulated WISH cells, which have limited TLR expression, were used for
indirect detection of IFNA via qRT-PCR. Results: ARID3a expression in B cells did not correlate with the generation of
autoantibodies. However, our data show for the first time that ARID3a+ B cells produce interferon alpha (IFNA) mRNA
and protein, an inflammatory cytokine associated with pathogenesis in SLE. In vitro ARID3a knockdown experiments
showed loss of both ARID3a and IFNA via qRT-PCR, confirming ARID3a-mediated regulation of IFNA. Moreover,
healthy control B cells stimulated with the TLR 9 agonist, CpG, generated ARID3a+IFNA+ B cells. Conclusions:
Together, these data implicate ARID3a+ B cells as a new innate immune effector B cell, and provide the unique
opportunity to compare functional differences between healthy and SLE ARID3a+ B cells, and their potential roles in
inflammation and SLE.
Funding: FUNDING: This work was supported by a Lupus Foundation of America, AI090343 and AI044215 (CFW), and
T32AI7633-12 (JW) from the National Institutes of Health.
69 Abstract #50
SPHINGOLIPID SIGNALING IN CORNEAL NEOVASCULARIZATION.
Joseph L. Wilkerson1,5, Hui Qi2,5, Hunter Porter5, Megan Stiles2,5, and Nawajes A. Mandal1,2,3,4,5 1Departments of Cell
Biology, 2Ophthalmology, 3Physiology, 4Oklahoma Center for Neuroscience, OUHSC, Oklahoma City, OK 73104, USA;
5
Dean McGee Eye Institute, Oklahoma City, OK 73104, USA
Introduction: Sphingosine-1-phosphate (S1P) is a sphingolipid signaling mediator that activates five separate g-proteincoupled receptors linked to cell proliferation, differentiation, inflammatory responses, and angiogenesis. Sphingosine
kinases (SphK) are responsible for generating S1P, their role in ocular vascularization is unknown. Docosahexaenoic acid
(DHA), an n-3 fatty acid, is known to protect from neovascularization in vitro and in the retina, however, if SphK is
inhibited this protection is diminished. The purpose of this study is to determine the role of S1P in neovascularization
through the role of SphK. We propose that S1P mediates the protective action of DHA for reducing NV. Methods: Sphk1
knockout mice were bred with mice containing the transgene, Fat1. FAT1 is able to convert n-6 fatty acids to n-3 fatty
acids; mice containing the transgene therefore have elevated amounts of DHA when they are put on a diet of 10%
safflower oil containing only n-6 fatty acids. Mice on this diet were used for corneal alkali burns as a NV model. After a
10 day period following the initial alkali burn the corneas were harvested and used for immunohistochemistry to
determine NV. Results: Ten days after the alkali burn wild type cornea exhibit a large amount of dense NV originating in
a disrupted limbus toward the central cornea. Sphk1 knockout mice have dramatically reduced neovascularization
extending from the limbus. In respect to DHA, wild type mice that contain Fat1 show protection from NV, while mice that
lack SphK1 and contain Fat1 show similar patterns of NV as Sphk1 knockout mice alone. Conclusions: We conclude that
SphK1 plays a major role in neovascular development in the cornea. We also show that DHA can protect the cornea from
NV. Furthermore, this protective mechanism is mediated by SphK1, most likely through the generation of S1P.
Funding: NIH grants EY022071, RR17703, and EY021725; Foundation Fighting Blindness Inc. USA, and Research to
Prevent Blindness, USA.
70 Abstract #51
CO-INHIBITORY AND CO-STIMULATORY CROSSTALK REGULATES HELPER T CELL DIFFERENTIATION
AND HUMORAL IMMUNITY DURING PLASMODIUM INFECTION
Ryan Zander, Jenna Guthmiller, Divine Kulu and Noah Butler; Department of Microbiology and Immunology, University
of Oklahoma Health Sciences Center
Pathogen-specific T cells are subject to co-stimulatory and co-inhibitory signals, the balance of which promotes pathogen
control while limiting immunopathology. During blood-stage Plasmodium infection, parasite-specific CD4 T cells exhibit
sustained expression of co-inhibitory receptors that limit CD4 T cell function. Blocking these receptors during chronic
Plasmodium infection enhances CD4 T cell function and accelerates parasite clearance, highlighting the relevance of coinhibitory pathways during malaria. In contrast to co-inhibitory receptors, the role of T cell co-stimulatory receptors
during chronic Plasmodium infection has not been investigated. Our preliminary data show that CD4 T cells also exhibit
sustained expression of the co-stimulatory receptor OX40 during both human and experimental malaria. Therefore, we
hypothesized that co-stimulatory signaling can functionally counteract co-inhibitory pathways to maintain protective
Plasmodium-specific CD4 T responses. To test this hypothesis, Plasmodium yoelii-infected mice were administered
agonistic antibodies that activate OX40 signaling in vivo. At regular intervals, flow cytometric analyses were used to
measure parasite burden and evaluate parasite-specific CD4 T cell activity. Humoral responses were assayed using
ELISA. We found that therapeutically activating OX40 during established experimental malaria enhances CD4 T cell
activity, humoral immunity and parasite control. Our data also show that the effects of OX40 signaling are modified
following the simultaneous blockade of the co-inhibitory receptor PD-1, which markedly impairs parasite-specific T
follicular helper cell responses in an IFN-gamma-dependent manner. Collectively, our results identify that OX40 can be
targeted to limit malaria parasite replication and reveal a previously unrecognized role of IFN-gamma as a negative
regulator of Plasmodium-specific humoral immunity.
Funding: This work was supported by grants from the NIH (T32AI007633 to R.A.Z.; 1K22AI099070 to N.S.B.) and the
American Heart Association (13BGIA17140002 to N.S.B.). N.S.B. is also an OK-INBRE scholar supported by a grant
from the NIH/NIGMS (8P20GM103447).
71 Abstract #52
REPLACEMENT OF 1.4-KB PROMOTER OF MURINE SELP GENE WITH ITS HUMAN COUNTERPART
PARTIALLY RECAPITULATES REGULATION OF HUMAN SELP GENE IN VIVO
Nan Zhang1, 2, 3, Zhenghui Liu2, 3, Rodger P. McEver1, 2 1Department of Biochemistry and Molecular Biology, University of
Oklahoma Health Sciences Center 2Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation
3
These authors contributed equally to this study.
Introduction: Leukocytes roll on P-selectin after its mobilization from secretory granules to the surfaces of platelets and
endothelial cells. Tumor necrosis factor (TNF), IL-1β, and lipopolysaccharide (LPS) increase synthesis of P-selectin in
murine but not in human endothelial cells. We have previously demonstrated that upon stimulation of aforementioned
mediators, the 1.4-kb promoter of murine P-selectin gene (Selp) upregulated a reporter gene whereas the counterpart of
human P-selectin gene (SELP) could not at all. Methods: To explore the physiological significance of this sequence in
gene regulation, we made knock-in mice (SelpKI/KI) replacing the 1.4-kb promoter of Selp with its counterpart of SELP.
They were crossed with P-selectin deficient mice to generate SelpKI/- mice to relatively match constitutive P-selectin
expression with wild-type (WT) mice. Basal and inducible P-selectin expression of SelpKI/- and Selp mice were compared
at mRNA, protein, and functional levels. Results: SelpKI/- mice constitutively expressed relatively comparable P-selectin
as WT mice in various tissue lysates as well as platelets and resident peritoneal macrophages. Leukocytes of SelpKI/- mice
rolled at a similar velocity as WT mice in venules of the cremaster muscle subjected to trauma. SelpKI/- mice recruited a
similar number of neutrophils into the inflamed peritoneum as WT mice. However, TNF and LPS dramatically increased
P-selectin mRNA in WT tissues, whereas they only modestly increased P-selectin mRNA in SelpKI/KI tissues. During
contact hypersensitivity, P-selectin mRNA in WT ears was upregulated significantly more than that in SelpKI/- ears.
Consequently, P-selectin in SelpKI/- mice contributed less to ear inflammation than WT mice with E-selectin blockade.
Conclusion: These findings reveal the significance of the 1.4-kb promoter of human P-selectin gene in pathophysiological
circumstances with different inducible expression patterns from WT murine P-selectin promoter in vivo. However, it
does not fully recapitulate all the differences between human and murine P-selectin gene regulation.
Funding: Funding American Heart Association 14PRE18060013 National Institutes of Health 5P01HL085607-08
72 Abstract #53
ELTD1 AND SLIT3 AS NOVEL ANTIBODY THERAPIES AGAINST GLIOMA BIOMARKERS
Jadith Ziegler1 2, Richard Pody1, Nataliya Smith1, Debra Saunders1, Patricia Coutinho de Souza1, Johnathan Wren1, and
Rheal Towner1 2 3. 1Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, 2 Pathology, The
University of Oklahoma Health Sciences Center, 3 Peggy and Charles Stephenson Cancer Center, Oklahoma City, OK,
United States
Gliomas consist of up to 80% of malignant brain tumors that are invasive and typically resistant to treatment. Finding
biomarkers to high-grade gliomas can enable better diagnosis and intervention for this disease. Through bioinformatics
and microarray experiments, we have identified ELTD1 and SLIT3 as biomarkers for high-grade human gliomas. ELTD1
is found to be associated with angiogenesis and SLIT3 is associated with cell proliferation, migration and angiogenesis.
Here, we report our findings in vivo using anti-ELTD1 and anti-SLIT3 antibodies on mouse GL261 glioma models. Using
MRI we investigate tumor growth and animal survival rate, and as well as report histological findings using mouse tumor
tissue. Mice were implanted with GL261 cells and were either untreated or administered anti-ELTD1, anti-SLIT3, antiVEGF or anti-c-Met antibodies. MRI was used to assess tumor growth, calculate tumor volumes and percent survival was
obtained from time-points when mice are euthanized. Additionally, representative histology slides (H&E) obtained from
GL261 glioma-bearing mice that were either untreated or treated with anti-ELTD1, anti-c-Met, or mouse anti-VEGF
antibodies. Histogram of mitotic index and atypical mitosis were identified. There was a significance increase in animal
survival for anti-ELTD1 and anti-SLIT3 and anti-VEGF antibodies as well as a significance decrease in tumor volumes
for anti-ELTD1 and anti-SLIT3 anti-VEGF and anti-c-Met when compared to untreated animals. Significant decreases in
the mitotic index were also found for anti-ELTD1 and anti-c-Met treatment groups compared to untreated mice. Our in
vivo studies have found that anti-ELTD1 and anti-SLIT3 antibodies to decrease tumor volumes and increase animal
survival in mouse GL261 glioma models. Additionally, anti-ELTD1 therapy was found to decrease the mitotic index in
mouse GL261 glioma model. Future studies will investigate anti-ELTD1 and anti-SLIT3 antibody therapies in human
xenografts. Our results indicate that anti-ELTD1 and anti-SLIT3 antibodies could be potential therapies for high-grade
gliomas in humans.
Funding: Oklahoma Medical Research Foundation, NIH (1P20GM103636, 8P20GM103456-09), and Louis Stokes
Alliance for Minority Participation Bridge to Doctorate Fellowship
73 Section IV. Graduate Student Poster Presentation Abstracts #54 -­‐ #81 74 Abstract #54
INDIRECT INHIBITION OF ORGANIC ANION TRANSPORTING POLYPEPTIDE (OATP) 1B1-MEDIATED
TRANSPORT BY LYSOSOME INHIBITOR CHLOROQUINE
Khondoker Alam1, Abuznait Alaa1, Sonia Pahwa1, Kai Ding2 and Wei Yue1 1Department of Pharmaceutical Sciences,
College of Pharmacy, University of Oklahoma Health Sciences Center, Oklahoma City, OK 2Department of Biostatistics
and Epidemiology, College of Public Health, University of Oklahoma Health Sciences Center, Oklahoma City, OK
Purpose Membrane transporter organic anion transporting polypeptide (OATP) 1B1 mediates hepatic uptake of many
drugs including lipid lowering statins. Chloroquine, an antimalarial drug, is also used for the treatment of other diseases
(e.g. rheumatoid arthritis and lupus) and is currently in clinical trials for cancer therapy. Chloroquine has lysosome
inhibition activity and has been implicated in regulating the surface expression of several membrane proteins, yet the
interaction of chloroquine with OATP1B1 has not been reported. The study was designed to determine the effect of
chloroquine on OATP1B1 degradation, surface expression and transport function. Methods The effect of chloroquine on
OATP1B1 protein levels was determined by immunoblot and that on OATP1B1 transport function was determined using
[3H]estradiol 17 β-D-glucuronide (E217βG) and [3H]pitavastatin as substrates in HEK293-OATP1B1 and human
sandwich-cultured hepatocytes (SCH), respectively. Surface OATP1B1 levels were determined by biotinylation assay,
and cytotoxicity was determined by LDH assay. Co-localization of OATP1B1 and lysosome marker LAMP2 was
determined with confocal microscopy. Results Pretreatment with chloroquine for 2 and 5 h significantly increased
OATP1B1 protein levels to 1.57±0.1 and 1.37±0.1 fold of control, respectively. OATP1B1 is partially co-localized with
LAMP2, consistent with lysosomal degradation of OATP1B1. Surface levels of OATP1B1 was decreased to 0.67 ± 0.07
fold of control in HEK293-OATP1B1 treated with chloroquine (100 µM, 5 h) (mean ± range, n=2). Without pretreatment,
chloroquine concentrations up to 100 µM did not affect OATP1B1-mediated [3H]E217βG uptake. However, pre-treatment
with chloroquine (10 and 100 µM) for as short as 30 min significantly decreased [3H]E217βG uptake (1 µM, 2 min) in
HEK293-OATP1B1. Chloroquine pretreatment also significantly decreased [3H]pitavastatin uptake in human SCH.
Negligible cytotoxicity was detected after chloroquine treatment. Conclusions This is the first report that lysosomal
inhibition is associated with decreased OATP1B1 transport function, providing novel insight into OATP1B1-mediated
DDIs.
Funding: NIH R01 GM094268
75 Abstract #55
EXPOSURE TO ELECTRONIC-CIGARETTE AEROSOL EXTRACT INDUCES SIGNIFICANT DNA DAMAGE
Lacy Brame1, Vengatesh Ganapathy1, Ilangovan Ramachandran1, Theodore Wagener2,3, David A. Rubenstein4, and
Lurdes Queimado1-3, 5,6 Departments of 1Otorhinolaryngology, 3Pediatrics and 5Cell Biology; 2The Oklahoma Tobacco
Research Center and 6The Peggy and Charles Stephenson Cancer Center, The University of Oklahoma Health Sciences
Center, Oklahoma; 4Department of Biomedical Engineering, Stony Brook University, New York, USA.
Introduction: E-cigarettes (ECs) have been marketed as an alternative to smoking cigarettes and their use has been rapidly
increasing. All EC products heat a solution that typically contains nicotine turning it into an aerosolized vapor. Inhaled
and exhaled EC aerosols contain nicotine and low levels of carcinogens. ECs appear to be safer than tobacco cigarettes,
however, the risks of EC use are not completely known. Our aims were to determine whether EC aerosols cause persistent
DNA damage in human cells and to define the strand-specific patterns of DNA damage and repair following exposure to
EC aerosol extracts. Methods: EC aerosol extracts from 3 distinct ECs were prepared as we previously described for
mainstream smoke. Human oral epithelial cells were exposed to escalating doses of EC extracts. DNA damage was
quantified in the p53 gene at 1 and 16 hours post-exposure using a novel and highly sensitive primer-anchored DNA
damage detection assay (PADDA) developed in our laboratory. Cell viability was determined by MTT assay. Data were
analyzed by Student’s t-test. Results: Exposure to EC aerosol extracts for 1 hour resulted in significant increases in DNA
damage in oral epithelial cells. The observed levels of DNA damage were lower than those induced by exposure to
mainstream smoke. Sixteen hours post-exposure, the levels of persistent DNA damage varied between EC extracts,
suggesting the ability to repair the DNA damage induced by EC aerosol extracts is composition dependent. No cell death
was observed. Conclusion: Our study demonstrated that exposure to EC aerosol extracts induces significant DNA
damage. These data suggest that EC use leads to variable levels of persistent DNA damage and potentially increases
cancer risk. Our study emphasizes the need to further investigate the health consequences of exposure to EC aerosols and
the importance of regulating EC and exposure to EC aerosols.
Funding: Funding: This work was supported by the Oklahoma Tobacco Research Center and the National Science
Foundation. Dr. Queimado holds a Presbyterian Health Foundation Endowed Chair in Otorhinolaryngology. Lacy Brame
was funded through the Summer Research Experience for Undergraduates Oklahoma EPSCoR program.
76 Abstract #56
HOW BCL-XL INTERACTS WITH TBID AT MITOCHONDRIA TO REGULATE APOPTOSIS
Christina G. Bruxvoort1, Franklin A. Hays1, 2, Justin Kale3, David W. Andrews3, and Jialing Lin1, 2
1
From The Department of Biochemistry and Molecular Biology, 2Stephenson Cancer Center, University of Oklahoma
Health Sciences Center, Oklahoma City, Oklahoma 73126; 3Biological Sciences, Sunnybrook Research Institute and
Department of Biochemistry, University of Toronto, Toronto, Ontario, M4N 3M5, Canada
Introduction: Apoptosis is a controlled way for unwanted and damaged cells to perish during normal growth and
development in any multicellular organism. Either insufficient or excessive apoptosis can cause diseases like cancer,
autoimmunity, and neurodegeneration. BCL-XL and BID are two BCL-2 family members that play antagonistic roles in
the intrinsic pathway of apoptosis. Principally, t-BID activates BAX at the mitochondrial outer membrane (MOM)
eventually permeabilizing the mitochondria and killing the cell. However, BCL-XL sequesters tBID to prevent it from
activing BAX. How BCL-XL and tBid interact at the MOM is largely unknown. Methods: Molecular modeling
programs were used to generate an in silico structural model to predict key interactions within the BCL-XL/tBID
complex. To test this model, disulfide crosslinking of single-cysteine mutant pairs of BCL-XL and tBID followed by
BCL-XL and tBID-specific immunoprecipitation were used to capture the potential molecular interactions between fulllength BCL-XL and tBID at the MOM. Mutagenesis of the identified interface residues was performed to assess their role
in mediating the BCL-XL/tBID interaction and in the BCL-XL inhibition of tBID. Results: The computational structural
model predicted a BH3-in-groove dimer interface in the BCL-XL/tBID complex. Positive disulfide crosslinking data were
obtained from six single-cysteine BCL-XL/tBID mutant pairs, demonstrating the existence of this interface in the MOMbound BCL-XL/tBID complex and was confirmed with immunoprecipitation. Conclusion: The BH3-in-groove
interaction mediates the formation of BCL-XL/tBID dimer at the MOM. The mutagenesis study is underway to test the
functional significance of this interaction. The knowledge from this study is expected to provide clues for developing
modulators of this potentially critical apoptotic interaction that is commonly dysfunctional in cancer, autoimmune
disorders, and neurodegenerative diseases.
Funding: This work was supported by fellowship from OK-LSAMP Bridge To Doctorate Program-Cohort VI NSF-HRD1249206 to C.G.B, and NIH grant R01GM062964 to J.L.
77 Abstract #57
IMMUNOLOGICAL DIFFERENCE BETWEEN TH1 AND TH2 DOMINANT MOUSE STRAINS IN A MODEL OF
ICD
Calhoun, K.N.1, Luckett-Chastain, J.R.1, Kemp, J.M.1, and Gallucci, R.M.1 1. College of Pharmacy, University of
Oklahoma Health Science Center
Irritant Contact Dermatitis (ICD) is the second most prevalent reported occupational injury associated with workman’s
compensation. ICD is characterized by a non-allergic inflammatory reaction initiated by the skin, which functions as an
immune organ, when irritants cause physical damage and barrier disruption. Since the inflammatory response in humans
can vary, ICD was evaluated through utilization of differing mouse strains to account for some of this
heterogeneity. C57BL/6 and BALB/c strains differ in manifestation of an immune response, specifically leaning toward
Th1 or Th2 dominance respectively. C57 and BALB/c mice were exposed to benzoalkonium chloride (BKC) or acetone
(control) for 3 or 7 days. Multiplex protein analysis showed that pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α,
IFN-γ, and GM-CSF were all significantly up-regulated in C57s at Day 3 as compared to BALB/c. In addition,
inflammatory cell chemoattractants CXCL1, 2, and 10, as well as, CCL3, 4, and 5 displayed higher expression in C57s.
Interestingly, not until Day 7 did BALB/c mice show significantly elevated levels of inflammatory mediators such as IL2, IL-4, IL-31, IL-9, IFN-γ, and CCL2 compared to C57s. Cytokine mRNA expression also varied between the two
strains, but did not necessarily correlate with protein data. Flow cytometry indicated an increased amount of macrophages
within the dermis of both C57s and BALB/c at Day 7 in contrast to Day 3. Overall, these results further confirm previous
findings in which BALB/c and C57 exhibit different immune responses, and may provide insight when predicting an
individual’s response to irritant exposure and potential therapeutic immunomodulation.
Funding: This research was funded by CDC/NIOSH grant R01-OH0 010241.
78 Abstract #58
A DOSE CALCULATION ALGORITHM FOR DIAGNOSTIC IMAGING BEAMS BY EMPIRICAL MODELING
Michael Chacko1, Imad Ali2, Jagadeesh Sonnad1, Sulaiman Aldoohan1 1Department of Radiological Sciences, OU Health
Science Center 2Department of Radiation Oncology, OU Health Science Center
Introduction: The increasing use of cone-beam computed tomography (CBCT) and larger collimation widths in traditional
CT has presented unique challenges for standardized dosimetric indices. This study assesses the feasibility of a new
method for quickly and accurately calculating dose in three dimensions from diagnostic imaging beams using empirical
modeling. Methods and Materials: Dose was modeled with photon attenuation measured using depth dose (DD), scatter
radiation in medium, and off-axis ratio (OAR) using reference dosimetry. Measurements were made with a diode (IBA
SFD) in water phantom and a 2-D diode array (MapCHECK 2) on a Varian kV on-board imager (OBI). All model
parameters were measured at 80, 100, and 125 kVp. Measurements were made with and without bowtie filters at field
sizes 1x1 to 40x40 cm2 and water-equivalent depths 0-20 cm. These parameters were used as lookup tables for dose
calculation using MATLAB. Results: Measured dose decreased with depth because of photon attenuation and increased
with field size due to increased scatter radiation. DD increased with higher energies and beam hardening from half-fan
and full-fan bowtie filters. Scatter radiation increased with larger field sizes and energies. Lateral scatter equilibrium was
approached at 20x20 cm2 for all energies. The OAR, accounting for non-uniformities such as the Heel effect, was within
3% for beam profiles inside plateau regions. The presence of bowtie filters attenuated OAR by as much as 80% at the
largest field size. Conclusion: The data demonstrated that it is feasible to calculate dose in water within 5% in regions
with near charge-particle equilibrium (CPE) conditions, outside buildup and penumbra regions, using this method. It
accurately accounts for scatter in water, which is superior to air-kerma or CTDI measurements typically used for
diagnostic imaging systems. Further considerations for patient-specific dose calculations include non-CPE conditions and
heterogeneity corrections.
Funding: None
79 Abstract #59
MOLECULAR CYTOGENETIC CHARACTERIZATION OF SUBTELOMERIC REARRANGEMENT CASES BY
PERFORMING CGH MICROARRAY
Jiani Chen1, Yue Gu1, Carrie Guy1, Susan Hassed1, Shibo Li1
1
. Department of Pediatrics, OUHSC
Subtelemeres are gene rich areas of the human genome and rearrangements in these areas frequently cause intellectual
disability and a variety of congenital anomalies. The characteristic phenotypes and clinical significance of most of the
rearrangements are yet to be unraveled. Traditional subtelemere screening analysis using Fluorescence In Situ
Hybridization (FISH) in individuals with intellectual disabilities, along with routine karyotype analysis, has gradually
been replaced by chromosomal microarray (CMA) as it has become the first-tier genetic test performed on affected
individuals due to its higher sensitivity and efficiency. In this study, we aimed to re-evaluate patients who had
subtelomeric deletion or duplication by FISH by performing microarray based comparative genomic hybridization (array
CGH) on isolated DNA from peripheral blood. A total of 21 qualified patients were contacted by phone. Ten of them
were consented and had blood drawn. Four declined the study, three were deceased, and the remaining four patients were
lost to follow-up. DNA was isolated from each blood sample and array CGH analysis was carried out following standard
laboratory procedure with an Agilent 2x400K v.1.0 microarray (Agilent Technologies Inc.). Results were analyzed
utilizing standard database resources including UCSC hg 19 human genome, Database of Genomic Variants, ClinVar,
Online Mendelian Inheritance in Man (OMIM), and published research or case reports. Array CGH results not only
provided detailed coordinates of the subtelomeric rearrangement known from FISH analysis, some results but also
revealed additional gains and/or losses in the genome. The additional information provided by array CGH analysis will
provide additional information for clinical correlation. The involvement of genes know to have a dosage effect may also
help predict the risk for certain disorders. Comparisons to similar cases published more recently revealed key
characteristics of the rearrangement and possible hot spots in the genome that are prone to breakage.
Funding: Department of Pediatrics, Genetics Laboratory
80 Abstract #60
THE ROLE OF THE DNA UNWINDING ELEMENT-BINDING PROTEIN (DUE-B) IN VERTEBRATE
DEVELOPMENT
Emily Clowdus1, Courtney Sansam2, Duane Goins2, Joseph Siefert1, Chris Sansam1,2 1University of Oklahoma Health
Science Center, Department of Cell Biology, Oklahoma City, OK; 2Oklahoma Medical Research Foundation, Cell Cycle
and Cancer Biology Research Program, Oklahoma City, OK
In order for a eukaryotic cell to successfully proliferate, DNA replication must be properly initiated and executed. In
vertebrates, hundreds of thousands of origins are licensed, or capable of initiating DNA replication, but only a subset of
these actually fire in any given cell cycle. How specific origins are selected is not well understood, but it involves the
conversion of these origins from licensed to active origins of replication. Our lab is interested in the precise role of
specific components involved in this conversion, as well as the mechanisms governing this process. The DNA unwinding
element-binding protein (DUE-B) has recently been shown to be an important component involved in loading factors
necessary for DNA replication initiation. We have generated due-b null zebrafish using the genome-editing tool
TALENs, and have confirmed DUE-B loss by qRT-PCR and western blot. Our mutant zebrafish do not show any obvious
morphological phenotypes, and FACS analysis of DNA content has not revealed any deficiencies in DNA replication or
cell cycle progression. Furthermore, treatment with replication inhibitors has not revealed any sensitivity as compared to
wild-type clutch mates. While we are still investigating an underlying phenotype, DUE-B does not appear to be either
necessary for DNA replication or an essential gene in vertebrates.
Funding: National Institute of General Medical Sciences of The National Institutes of Health
81 Abstract #61
ESTABLISHMENT OF AN IN VITRO CELL CULTURE MODEL FOR FUNCTIONAL STUDY OF THE
OATP1B1-V174A POLYMORPHISM
Alex Crowe1, Sonia Pahwa1, Khondoker Alam1 and Wei Yue1 1Department of Pharmaceutical Sciences, University of
Oklahoma Health Sciences Center College of Pharmacy, Oklahoma City, OK 73126
Purpose: OATP1B1 is a liver-specific membrane transport protein that mediates hepatic uptake of many drugs (e.g., lipid
lowering statins). The c. 512T>C (V174A) polymorphism of OATP1B1 has a 10-20% frequency in non-African
populations, and is the most robust predictor of myopathy induced by OATP1B1 substrate statin. When expressed in vitro,
the V174A variant has decreased transport activity. To date, the mechanism involved in reduced transport activity is not
known. Currently, the lack of commercial availability of OATP1B1 antibody limits the mechanistic studies. The aim of
this study is to establish stable cell lines in HEK293 cells that over-express FLAG-tagged wild-type or V174A variant of
OATP1B1, and to study the localization of wild-type and V174A variant in these cell lines. Methods: Stable cell line
HEK293-FLAG-OATP1B1 and HEK293-FLAG-OATP1B1-V174A were established and were maintained in DMEM
medium with 600 µg/ml G418. FLAG immunoblotting with FLAG antibody was used determine the protein levels, and
immunofluorescent staining of FLAG and LAMP2, a lysosome marker, was conducted to determine membrane
localization and co-localization of FLAG-OATP1B1 with lysosome. [3H]E217G (1uM, 2min) accumulation was used to
determine transport function of OATP1B1. Results: HEK293-FLAG-V174A expresses the OATP1B1-V174A variant
protein at a similar level to the wild type OATP1B1 in HEK293-FLAG HEK293, however has significantly decreased
transport activity. Interestingly, surface level expression of OATP1B1 is similar among these two stable cell lines.
Minimal wild-type and V174A variant OATP1B1 are co-localized with LAMP2. Conclusion: A valid model is
established to study the function and trafficking of the OATP1B1 wild-type and V174A variant proteins. Future research
will be focused on characterizing the differences in the post-translational modifications between the wild-type and V174A
variant OATP1B1 in order to elucidate mechanisms involved in regulating OATP1B1 function.
Funding: Supported by NIH R01 GM094268
82 Abstract #62
ELECTRONIC HEALTH RECORD SYSTEM IMPLEMENTATION AND MEANINGFUL USE CERTIFICATION
AMONG OKLAHOMA HOSPITALS
Ngoc Quyen, Duong1; Aaron, Wendelboe1; Ann, Chou2; Robert, Roswell2 1Department of Biostatistics and Epidemiology,
University of Oklahoma Health Sciences Center 2Department of Health Administration and Policy, University of
Oklahoma Health Sciences Center
Introduction: Hospitals are being incentivized to adopt electronic health record (EHR) systems. The American Hospital
Association (AHA) conducts an annual health information technology survey. The AHA 2012 Annual Survey IT
Database surveyed hospitals across the nation regarding their implementation of an EHR, compliance with federal
Meaningful Use standards, and patient engagement functionalities. We aimed to assess the degree to which hospitals in
Oklahoma have 1) implemented EHR systems, 2) achieved meaningful use certification, and 3) made health records
accessible to their patients. Methods: Frequency statistics were computed for variables in the 2012 AHA Annual Survey
IT dataset using SAS. Comparisons were made between Oklahoma hospitals as a group (n = 85) and hospitals from all
other states and Washington, D.C. (n = 3,334). Chi-square tests of homogeneity were calculated to detect differences in
the overall distribution and chi-square tests of proportions were generated to detect significant differences across various
categories of responses for a given variable. Fisher’s Exact Tests were used when one or more categories had expected
cells less than five. Results: Forty-two percent of Oklahoma hospitals have implemented at least a minimal form of an
EHR system (which did not significantly differ from hospitals in the rest of the U.S., p=0.57). The vast majority of
Oklahoma hospitals (89.9%) indicated they were certified as meeting federal requirements for the hospital objectives of
Meaningful Use. This is consistent with the national proportion (89.9%) of hospitals meeting Meaningful Use
certification. In contrast, a minority of Oklahoma hospitals (≤31%) offers patient engagement functionalities (which is
significantly lower than hospitals in other states, p<0.01).
Conclusion: Although hospitals in Oklahoma are making
progress in implementing EHR systems and are on par with the national average in meeting federal Meaningful Use
criteria, they are behind in providing patients with online access to their health/medical records.
Funding: Oklahoma Health Information Exchange Trust
83 Abstract #63
RAPID DOWN-REGULATION OF ORGANIC ANION TRANSPORTING POLYPEPTIDE (OATP) 1B1 AND 1B3
TRANSPORT FUNCTION BY MAMMALIAN TARGET OF RAPAMYCIN (MTOR) INHIBITOR SIROLIMUS
Taleah Farasyn1, Sonia Pahwa1, Kai Ding2 and Wei Yue1 1Department of Pharmaceutical Sciences, College of Pharmacy,
University of Oklahoma Health Sciences Center, Oklahoma City, OK, 2Department of Biostatistics and Epidemiology,
College of Public Health, University of Oklahoma Health Sciences Center, Oklahoma City, OK
Introduction: Organic anion transporting polypeptide (OATP) 1B1 and 1B3 are hepatic transport proteins that mediate
uptake of many drugs including lipid-lowering statins. Inhibition of OATP1B1 and 1B3 transport activity is an important
determinant for statin-induced myopathy. Unexpected drug-drug interactions (DDIs) have been reported stating that
transplant patients developed severe rhabdomyolysis after concurrent administration of statins and sirolimus. The
mechanism of such DDIs is unknown. The current study was designed to determine the effect of sirolimus on OATP1B1
and 1B3 transport function. Methods: Accumulation of ([3H]estradiol-17β-glucuronide (E217βG) (1 µM, 2 min) and
[3H]CCK-8 (1 µM, 3 min) was determined in OATP1B1- and OATP1B3-overexpressing HEK293 cell lines respectively.
HEK293-OATP1B1 and -1B3 cells were pre-treated with sirolimus (0.1 to 5 µM) for up to 1 h. IC50 values of sirolimus
toward OATP1B1 and OATP1B3 were determined with or without pre-treatment The Cmax /IC50 and R-Values
(R=1+fu × Iin, max/IC50) were calculated based on the FDA draft guidance for OATP-mediated drug interaction studies.
Results: After 1 hour pre-treatment, OATP1B1-mediated-[3H]E217G and OATP1B3-mediated-[3H]CCK-8 accumulation
was significantly decreased at a concentration as low as 0.5 µM to 56.1±1.6% and 62.7±1.4% of control, respectively
(n=3). Co-incubation with sirolimus with substrate yielded IC50 values of 0.997 and 0.981 µM with R Values of 1.35 and
1.36 for OATP1B1 and OATP1B3, respectively (n=3). Pre-incubation significantly reduced the IC50 values to 0.250 µM
for OATP1B1 and 0.282 µM for OATP1B3 with R Values of 2.41 and 2.25, respectively (n=3). Conclusion: Sirolimus
potently inhibited OATP1B1 and OATP1B3 transport function. The R-value for sirolimus was increased after pre+cotreatment to a value greater than the FDA-recommended cutoff of 1.25, suggesting clinical DDIs of sirolimus against
OATP1B1 and 1B3 substrates (e.g. statins) may occur via OATP-inhibition. The current findings elucidate a novel
mechanism for impaired hepatic uptake of OATP substrates and provide new insights into predicting OATP-mediated
drug interactions.
Funding: NIH R01 GM094268
84 Abstract #64
PRIVATIZED HEALTHCARE POSSIBILITIES: A COMPARATIVE HEALTHCARE PERSPECTIVE OF
HEALTHCARE DELIVERY IN THE UNITED STATES UNDER THE AFFORDABLE CARE ACT
Michael Carter1, Regina Greuel2, Cassidy Hamilton1, Alexander Heit1, Kalee Howard1 1Department of Political Science
University of Oklahoma 2Department of Health Promotion Sciences University of Oklahoma Health Sciences Center
Introduction Despite devoting more than 17 percent of GDP to the health sector, the United States compares unfavorably
with other industrialized countries on several key indicators of public health, including infant mortality and life
expectancy. The 2010 Patient Protection and Affordable Care Act (ACA) was enacted to improve health care access,
quality, and affordability. Yet, conservative policy institutes assert that a market-driven health system would have
substantial benefits for patients over the ACA, with expanded federal government role in health care via subsidies,a health
insurance mandate, and increased regulation of insurance companies. Methods To evaluate whether a free market
approach to health care in the U.S. would in all or part be less costly, create more patient accessibility or be higher quality
than the public-private healthcare model reflected in the ACA, we collected quantitative and qualitative data from a
number of sources in the disciplines of public health, social epidemiology, public policy and administration, and global
and domestic health care policy. We included peer-reviewed journals and articles, books, and internet and video sources
as they relate to the effect of market practices and incentives in current health care systems. We analyzed the data from a
comparative health perspective. Using measures of ‘true access,’ we contrasted the performance of global health systems
with the U.S. prior to ACA. We relied on these methods to extrapolate likely effects of an expanded private sector role in
the U.S. health system. Results We found that the free-market exacerbates specific health care problems in the allocation
of health resources, social determinants and health disparities. Conclusions We concluded that there are no significant
advantages of market-oriented health care over a universal, or even public-private health model. Expanding the role of the
private sector in the financing and delivery of health care services would promote rather than alleviate disfunctionality in
the health care system.
Funding:
85 Abstract #65
THE ROLE OF THE GLUCOCORTICOID RECEPTOR IN THE REGULATION OF STRESS-INDUCED
NOCICEPTION
Niran Hadad1, Anthony C. Johnson1, Beverley Greenwood-Van Meerveld1,2,3, 1Oklahoma Center for
Neuroscience, 2Department of Physiology, University of Oklahoma Health Science Center, 3VA Medical Center,
Oklahoma City, OK
Introduction: The amygdala is involved in the stress response and in the enhancement of painful stimuli. Previous studies
in our lab found that water avoidance stress (WAS) increased visceral hypersensitivity and decreased expression of the
glucocorticoid receptor (GR) in the central nucleus of the amygdala (CeA). However, the contribution of amygdaloid GR
in developing pain following stress remains unknown. This study tests the hypothesis that knockdown of GR in the CeA
will mimic stress-induced nociceptive phenotypes. Methods: Rats underwent stereotactic surgery in which bilateral
cannulae were inserted into the CeA. One day following surgery animals were subjected to 1-hour of WAS or sham stress
(SHAM) for 7 days. At days 4 -7, WAS animals were infused with 0.5 µL vehicle (VEH). SHAM animals were infused
with either GR specific antisense or random oligodeoxynucleotides (ASO/RSO), or VEH. At day 5, animals were assessed
for somatic pain using von Frey filaments and on day 7 animals were assessed for visceral hypersensitivity by measuring
their response to colorectal distension (CRD). Results: Knockdown of GR in the amygdala resulted in increased visceral
hypersensitivity in response to CRD at 40 mmHg (p < 0.01) and 60 mmHg (p < 0.0001) compared to controls. No
differences were seen when GR knockdown animals were compared to WAS treated animals. GR knockdown also
resulted in decreased withdrawal threshold (p < 0.01) compared with controls. Withdrawal threshold of WAS animals was
significantly lower than controls and showed no difference when compared to GR knockdown animals. Conclusions: Our
findings show that knockdown of GR in the CeA, in the absence of a stressor, is sufficient to reproduce stress-induced
nociception. This highlights the role of GR in regulating amygdala-mediated pain and suggests that molecular alterations
in the amygdala plays a major role in the development of susceptibility to stress-induced pain.
Funding: VA MERIT to B. G-VM.
86 Abstract #66
COMPARING LIPOSOME-ENCAPSULATED OKN-007 TO NON-FORMULATED OKN-007 IN TREATING
GLIOMAS
Hailey Houson, Natalia Smith, Deborah Saunders, Rheal Towner
OKN-007 is a nitrone therapeutic that has demonstrated anti-glioma properties. One downside of the drug is its short halflife in circulation. Gliomas are common forms of brain tumors that are difficult to treat because of the inability of many
drugs to cross the blood-brain barrier. Liposomes are lipid spheres that are able to cross the blood brain barrier. They are
also circulated in the body and when pegelated, can improve the half-life of their contents in the body. The purpose of this
work is to show the difference between mice with gliomas treated intravenously with OKN-007 and those treated
intravenously with liposome encapsulated OKN-007. My work demonstrated our ability to pack liposomes with OKN-007
and an MRI contrast agent. We were able to see the liposomes get into brain tumors and we saw an improvement in
survival and a reduction in tumor size between the mice treated with liposomes and the untreated mice. Significant
difference was not seen between the survival of the mice treated with OKN and the mice treated with liposomes. Our data
showed a decrease in tumor volumes for the mice treated with OKN over the ones treated with liposomes. Further work
can include binding a targeting moiety like anti-VEGF to the outside of the liposomes to improve delivery, changing the
diameter of the liposomes, and optimizing pegelation to improve release kinetics.
Funding: GPIBS, OMRF Advanced Magnetic Resonance Center
87 Abstract #67
YOGA AS A FORM OF VESTIBULAR REHABILITATION: A SYSTEMATIC REVIEW WITH META ANALYSIS
Anna Jilla1; Kristin Winkler1; Carole Johnson1; J. Connor Sullivan1; Emma Hallab1; Christi Barbee1 1Department of
Communication Sciences and Disorders, OUHSC
Purpose: Each year, 11 million elderly fall due to decreased balance function. Audiologists are part of an interprofessional
team of specialists in balance assessment and rehabilitation. Vestibular rehabilitation therapy (VRT) hastens the brains
natural compensation of this loss. A type of VRT, yoga, incorporates physical and mental exercises that increase
flexibility and balance stability. A systematic review with meta-analysis was conducted to determine if yoga, when used
as a form of VRT, reduces the effects of balance dysfunction and aids in the prevention of falls. The purpose of this
systematic review was to evaluate the evidence for the use of yoga as a viable form of VRT in the elderly. Methods:
Inclusion criteria for studies were Level I-III of evidence, reporting of data for subjects over 65 years of age, and
enrollment of subjects with no co-morbidities. Search strings were submitted to OVID, PubMed, CINAHL, and Cochrane
databases to obtain studies exploring the effectiveness of yoga for increasing the strength and flexibility of the elderly. A
meta-analysis provided effect sizes indicating the overall effectiveness of yoga as a form of VRT for increasing balance
function in the elderly. Results: Several studies showed the yoga intervention groups to have less transfer time from sitting
to standing, as well as improved gait and balance. These results indicate that VRT can increase balance confidence and
decrease fear of falling when compared to the findings from control groups or groups with little to no exercise.
Discussion/Conclusions: Some limitations to the studies included a small testing sample, poor data keeping, and
utilization of varied balance measures. Overall there were marked increases in hip flexion, floor transfer and gait function
in the study groups, suggesting that yoga is a viable form of VRT that can be beneficial for the prevention of falls in the
elderly.
Funding: None to report
88 Abstract #68
DEMONSTRATION ON JOINPOINT: A STATISTICAL METHOD TO ANALYZE TRENDS
Sarah E Johnston¹, David M. Thompson ¹, ¹Biostatistics and Epidemiology The University of Oklahoma Health and
Sciences Center
Introduction: Joinpoint is a free statistical software used to analyze breakpoints in regression lines. While it can analyze
any trend data, Joinpoint is most efficient when given data calculated by SEER*Stat, another free statistical software used
to calculate frequency, rates, and survival statistics. These two programs offer a convenient and cheap opportunity to
research cancer and how it affects society at large. Thus, the goal of this presentation is to educate researchers on how to
access, use, and interpret results in Joinpoint software. Methods: SEER*Stat was used to download data on lung and
bronchus cancer mortality from 1975-2011 and then generate a dataset which contained calculated age-adjusted mortality
rates for both men and women in addition to gender specific mortality rates. The dataset was then pulled into Joinpoint
and mortality rates over time were analyzed. Results: When analyzing data for both men and women Joinpoint detected
four breakpoints in 1977, 1983, 1991, and 2003 in lung and bronchus mortality rates. Mortality rates peaked in 1991 and
then fell until 2011. Breakpoints differed between genders. Joinpoint estimated three breakpoints in 1979, 1988, and
2002 for men, and four breakpoints in 1982, 1991, 1999, and 2007 for women. Joinpoint’s comparison test for
parallelism concluded regression lines for men and women were not parallel (p=0.0002). Conclusion: Joinpoint is easy
to use with limited experience in programing or statistics. Further, generating and interpreting results from Joinpoint is
more straightforward compared with other methods used in SAS or R. It takes advantage of free cancer data, thus
maximizing resources for researchers. With a guided example on how to create a dataset via SEER*Stat, and run a
Joinpoint session, researchers will gain a new method for answering difficult questions about cancer trends over time.
Funding: NA
89 Abstract #69
GENETIC SCREEN FOR GENES INVOLVED IN HIGH-SUGAR DIET-INDUCED DIABETES MELLITUS
Yue Li1,2, Zachary Ishikawa2, Hui-Ying Lim2 1Department of Physiology, University of Oklahoma Health Sciences Center,
2
Free Radical Biology and Aging Program, Oklahoma Medical Research Foundation
Diabetes mellitus (DM) is the fifth leading cause of death in the US. Type 2 DM is a chronic and systemic metabolic
disease characterized by hyperglycemia, insulin resistance, and insulin deficiency. It is well established that flies and
vertebrates share the major metabolic, energy-sensing and endocrine signaling networks that control nutrient uptake,
storage and metabolism. Therefore, Drosophila melanogaster represents an excellent model for the large-scale and rapid
screening for genes involved in obesity and diabetes. We used Drosophila in a forward genetic screen to identify and
study new and novel genes and pathways that regulate carbohydrate metabolism in response to a high sugar diet (HSD;
86.4% sucrose). Wide type (WT) flies fed HSD for 3 days after adult eclosion developed hyperglycemia (whole body
glucose level increased by 1.6-fold) and hyperlipidemia (whole body triglyceride level increased by 1.6-fold) compared to
control (normal food diet-NFD; 63.9% sucrose). Our HSD feeding regimen, therefore, generated a diabetic Drosophila
model that faithfully captured the salient features of human diabetes. We further subjected a collection of Drosophila
deficiency (mutant) lines (n=64) to our HSD feeding platform and identified 6 candidate lines that displayed attenuation
or exacerbation in their development of diet-induced diabetes compared to the WT strain. For 3 lines, HSD-fed flies
showed significantly increased glucose levels (>2-fold, p<0.05) compared to control, whereas for another 3 lines, HSDfed flies exhibited unchanged or even slightly decreased glucose levels. The 6 lines above had similar food intake as WT
flies, indicating that the observed change in glucose levels was not due to alterations in feeding behavior (like appetite).
We are fine mapping these lines to identify their causative genes, using smaller deficiency lines as well as RNAi-mediated
gene knockdown. So far, we have identified 4 genes that could protect against the development of diabetes in Drosophila.
Funding: OMRF Institutional Acount
90 Abstract #70
ASSOCIATIONS OF PARENTAL FEEDING PRACTICES, AND PARENT-CHILD FOOD PURCHASE
INTERACTIONS IN GROCERY STORES: A PROPOSAL
K. Lively 1, B. DeGrace, Ph.D.2, S.B. Sisson, Ph.D.1, M. Anderson, Ph.D.3, K. Lora, Ph.D.1 1Department of Nutritional
Sciences, University of Oklahoma-HSC, Oklahoma City, OK; 2Department of Rehabilitation Sciences, University of
Oklahoma-HSC, Oklahoma City, OK; 3Department of Biostatistics and Epidemiology, University of Oklahoma-HSC,
Oklahoma City, OK.
Background: Food socialization behaviors such as parental feeding practices in the home have been associated with child
health outcomes such as weight status and food intake. Few studies have reported the use of these feeding practices in
environments where food is purchased. Examining parental factors that may influence parent-child purchase interactions
while co-shopping and family dynamics that may be associated with feeding behaviors are essential for understanding
how food gets in the home for consumption. Purpose: 1) Examine the association between parental feeding practices (use
of food to calm the child, use of food as reward, and modeling) with parental behaviors related to granting and refusing
children’s requests for foods and beverages (F/B) during grocery shopping, 2) Examine the association between parental
feeding practices and the type of F/B purchased for the child during grocery shopping. Proposed Methods: Cross-sectional
survey with mothers (n=280) of 2-7 year old children. Data will be collected via an online survey sent to staff and faculty
at the University of Oklahoma-HSC. Participants will answer questions pertaining to demographics, feeding practices,
granting and refusing children’s requests for F/B at the grocery store, and type of F/B purchased. Proposed Analysis: The
relationship between parents’ granting or refusing to children’s requests while shopping and parental feeding practices
will be assessed for linearity, and correlations will be estimated. A study-developed questionnaire will be used to estimate
the type of F/B purchased. Within each type of F/B purchased, ANOVA will be used to compare average parental feeding
practice factors. Significance level will be set at p <.05. Relevance: Elucidating routes through which foods reach the
home has the potential to shape children’s preference of healthy foods in addition to further understanding of how child’s
influence can be utilized as a means for bringing healthier foods into the home.
Funding: Funding: This study is supported by the Department of Nutritional Sciences.
91 Abstract #71
FORMULATION OF A SUBLINGUAL TABLET CONTAINING THE IPAD-IPAB-FUSION PROTEIN, AS A
NOVEL VACCINE CANDIDATE AGAINST SHIGELLOSIS
Sanjida Mahjabeen1, Shyamal P. Choudhari2, Mariam Ibrahim1, Wendy L. Picking2 and Lucila Garcia-Contreras1 1The
University of Oklahoma Health Sciences Center, Oklahoma City, OK; 2The University of Kansas, KS
Aim: Shigellosis causes significant mortality worldwide, especially in children from developing countries. IpaD-IpaBFusion (DBF) protein is a novel vaccine candidate that has shown protective efficacy against Shigella spp. after mucosal
immunization in mice. Our goal was to formulate a sublingual tablet containing the DBF protein and lyophilized double
mutant heat labile enterotoxin (dmLT) as adjuvant that retains the immunogenicity of DBF and is stable in the absence of
refrigeration. Methods: The excipients and their proportion in tablet formulations were optimized using Design-ofExperiments statistical software (DoE). Tablet granulates were prepared using different excipients and proportions with
0.05% LDAO as granulation liquid. The wet granulate was filled into calibrated molds and dried to form tablets
containing 20µg DBF protein and 5µg dmLT. Tablet disintegration time (DT) was determined in simulated saliva and the
uniformity of dosage unit was determined by measuring the contents of 20 tablets. Far-UV circular dichroism (CD)
spectrum was recorded to determine DBF protein stability in the tablet. Results: From the 16 individual formulation
suggested by 2 DoE, the formulation containing 80% mannitol and 20% sucrose had DT and uniformity of dosage unit
that met United States Pharmacopeia USP requirements (DT = 28±2 seconds; 105% uniformity of dosage unit). However,
the high solubility of excipients in granulation liquid resulted in a poor yield of tablet batch. Thus, a new DoE was
designed to include bulking agents (microcrystalline cellulose (MCC) and sodium carboxymethyl cellulose (NaCMC)).
The optimal tablet formulation had a DT of 31±1.5 seconds and 113.06% uniformity. CD spectrum indicated that the
DBF in the tablet retained the same secondary alpha helical structure as the native protein. Conclusion: The 4 optimized
sublingual tablet formulations meet USP requirements and retain the potency of the DBF protein, suggesting that they will
be as effective in vivo as the solution.
Funding: NIH Grant 5 R21 AI105467-02
92 Abstract #72
STREPTOCOCCUS PYOGENES CHROMOSOMAL ISLAND SPYCIM1 AND PROPHAGE SF370.1 BURST SIZE
Kimberly McCullor1, Scott Nguyen1, Catherine King1, and W. Michael McShan1 1Department of Pharmaceutical Sciences,
University of Oklahoma Health Sciences Center, Oklahoma City, OK
Introduction: Streptococcus pyogenes, Group A streptococci (GAS), can cause clinical disease spanning from sore throats
to serious conditions such as rheumatic fever. Most strains of GAS contain lysogenic bacteriophages, such as SF370.1,
that carry genes encoding exotoxins that can enhance virulence capabilities of GAS. GAS genomes frequently contain
phage-like elements such as S. pyogenes Chromosomal Island M1 (SpyCIM1) that lack genes for packaging but enhance
virulence by inducing a mutator phenotype. Currently, we are studying how SpyCIM1 may utilize prophage SF370.1’s
packaging ability to disseminate. Specifically, our aim was to determine when phage infections occur during bacterial
growth and to determine SF370.1 abundance in the presence or absence of SpyCIM1. Methods: SF370.1, tagged with
erythromycin resistance (ermB) via homologous recombination with native phage exotoxin C gene (speC) was induced
with mitomycin C from two SF370 derived strains: SpyCIM1+ strain (OKM77) and SpyCIM1- strain (OKM78). Phage
lysates from each strain were collected and used in separate infection experiments using SF370.1 susceptible host strain
CEM1¿1 at various time points of growth. These infected cultures were then plated to erythromycin plates and resistant
colonies counted after 48 hours incubation at 37° C with 5% CO2. Results: The majority of SF370.1 infections occurred
during early to mid logarithmic growth in CEM1¿1 regardless of SpyCIM1’s presence. Overall, lysate from the
SpyCIM1+ strain resulted in higher amounts of SF370.1 infections compared to the SpyCIM1- strain lysate. Conclusion:
Strain CEM1¿1 is more susceptible to phage SF370.1 infection during early to mid logarithmic growth. The presence of
SpyCIM1 did not decrease the abundance of SF370.1 infectious particles as expected but instead increased SF370.1
infections when compared to the SpyCIM1 knockout strain.
Funding: Oklahoma Center for the Advancement of Science and Technology grant (OCAST) HR11-133 and by NIH
Grant Number R15A1072718 to WMM.
93 Abstract #73
RACIAL/ETHNIC DISPARITIES IN COLORECTAL CANCER DIAGNOSIS AND SURVIVAL: AN ANALYSIS OF
OKLAHOMA CENTRAL CANCER REGISTRY DATA
Kaitlin McGrew, Janis Campbell, PhD, Jennifer Peck, PhD, Sara Vesely, PhD Department of Biostatistics and
Epidemiology, University of Oklahoma College of Public Health
Introduction: Colorectal cancer (CRC) is the second leading cause of death in men and women. Racial disparities in CRC
mortality have been reported to be higher for Oklahoma than the US average. Probability of survival is highly dependent
upon stage at diagnosis, which highlights the importance of screening. Socioeconomic factors affect screening patterns
and therefore the risk of being diagnosed at a later stage. We hypothesized that race/ethnicity is associated with CRC
survival and stage at diagnosis and aimed to adjust for socioeconomic variables that could partially explain this
association. Methods: Cases of CRC captured in the Oklahoma Central Cancer Registry database, diagnosed between
2001 and 2008, and followed up until 2013 were included. Survival probabilities stratified by race/ethnicity were graphed
using Kaplan Meier curves. SEER Summary Staging was referenced to classify cases as early (“in situ” or “localized;”
n=6,456) or late (“distant;” n=2,809). Multivariable log binomial regression was performed to quantify the association
between race/ethnicity and stage at CRC diagnosis adjusting for age, gender, type of primary insurance, marital status, and
census tract-level measures of poverty and education. Results: Survival probability varied by racial/ethnic group, with
American Indian/Alaska Natives (AIAN) and African Americans (AA) having the lowest probabilities of survival.
Compared to non-Hispanic white, identifying as AA, AI, or Hispanic was associated with an increased risk of late-stage
diagnosis. After controlling for selected confounders, identifying as AA or Hispanic was no longer significantly
associated with stage at diagnosis, but identifying as AIAN remained a significant risk factor for late-stage diagnosis
(RR=1.13; 95% CI: 1.01, 1.26). Conclusion: Racial disparities in colorectal cancer diagnosis and survival were partially
explained by socioeconomic differences. Further analyses adjusting for additional socioeconomic measures may increase
our understanding of the disparities in CRC diagnosis and survival for the AIAN population in Oklahoma.
Funding:
94 Abstract #74
HEPATITIS C VIRUS TRANSMISSION AMONG INJECTION DRUG USERS
Dana Mowls, Hélène Carabin Department of Biostatistics and Epidemiology, College of Public Health, University of
Oklahoma Health Sciences Center;
Hepatitis C virus (HCV) is primarily spread via needle sharing among injection drug users (IDUs). Oklahoma has the
second highest incidence of acute HCV, making HCV a significant health problem. Modeling the transmission dynamics
of HCV can provide insight to its prevention and elimination. A compartmental frequency-dependent model was used to
understand HCV’s transmission dynamics among IDUs in England. A susceptible-acute infection-chronic infectionrecovery frequency-dependent model was developed and the rates of infectious contacts were obtained from published
studies on needle sharing among IDUs conducted primarily in England. The probability of transmission by needle, rates of
transition from acute to chronic infections and to recovery were taken from the literature. The basic reproductive number,
R0, was calculated from the number of secondary cases by which an index case in either the low or high-risk group causes
for all possible pairs, based on an equation adapted from Kretzchmar & Wiessing. Interventions were proposed and their
effects on HCV transmission were examined. The rate of needle sharing in the past year among low and high-risk IDUs,
were assumed to be 5 and 25 partners, respectively. The overall prevalence of HCV among IDUs was 53%, while the
prevalence among low and high-risk IDUs was 49% and 82%, respectively. Based on the assumption that 50% share withlike, the number of secondary cases was estimated to be 1.5 (high-high mixing), 2.7 (high-low mixing), 0.3 (low-low
mixing), and 0.2 (low-high mixing). R0 was 1.81; indicating HCV will spread through the population of susceptible IDUs.
Previous studies found similar estimates of the prevalence of HCV among England’s IDU population (~50%) and R0
(~2), confirming the model’s suitability. This research has important implications for understanding the prevention and
elimination of HCV transmission in IDUs. Future work includes improving and applying this model to data from
Oklahoma.
Funding: None
95 Abstract #75
A SYSTEMATIC REVIEW OF RISK FACTOR DIFFERENCES BETWEEN ETIOLOGICAL PATHWAYS IN
NECROTIZING ENTEROCOLITIS IN NEONATES
Whitney Richardson1, Aaron Wendelboe1 1Department of Biostatistics and Epidemiology, University of Oklahoma Health
Sciences Center
Introduction: Necrotizing Enterocolitis (NEC) is the most common and severe gastrointestinal emergency in
neonates. NEC occurs in 12% of very low birth weight infants (<1500 g), with mortality rates ranging from 1050%. NEC is a complex disease with many risk factors and three pathological routes: infectious, ischemic, and enteral
feeding. Few advances have been made in the early detection and control of NEC. This may be due to the failure of
researchers to stratify NEC cases according to etiological pathway. We aim to better understand the differences between
infectious and ischemic pathways of NEC, in hopes that these efforts will lead to novel predictive risk models utilizing
directed acyclic graphs (DAG’s). Methods: We conducted a case-control study of premature infants at the OU Children’s
Hospital neonatal intensive care unit. Case and control infants were ≤ 36 weeks gestational age, and case infants received
a NEC diagnosis according to modified Bell’s criteria. The preliminary findings reported are for cases
only. Results: From 2001-2011, our study enrolled 91 cases. Twenty-one (23%) cases were classified as infectious and
70 (77%) were classified as ischemic. Preliminary descriptive statistics indicate that 24 (26.4%) experienced birth
asphyxia, 76 (83.5%) received exchange transfusions, 62 (68.1%) were exposed to prenatal glucocorticoid steroids, 36
(39.6%) had an umbilical arterial catheter in place during the exposure period, and 49 (53.8%) had an umbilical venous
catheter in place. Regarding the mothers of cases, 14 (15.4%) used illicit drugs, 34 (37.4%) had prolonged rupture of
membranes (PROM), 13 (14.3%) had preeclampsia, and 23 (25.3%) received insufficient prenatal
care. Conclusions: Further statistical analysis is required to determine if risk factor distributions differ between
etiological pathways, and if this information can be used to construct a predictive DAG model for NEC. Funding: No
funding was received for this study.
Funding: No Funding was received for this study.
96 Abstract #76
THALIDOMIDE BLOCKS SYMPTOMS OF PTSD AND CO-MORBID PAIN IN AN ANIMAL MODEL OF PTSD
I. SCHALO1, P. DIB1, C. Simpson-Durand1, Y. Zhang1 K. M. STANDIFER1,2 1Department of Pharmaceutical Sciences,
College of Pharmacy and 2Department of Cell Biology and Oklahoma Center for Neuroscience, College of Medicine.
Single-prolonged stress (SPS) is an established animal model for post-traumatic stress disorder (PTSD). We have
previously reported that SPS induces long-lasting (35-42 d) mechanical allodynia and thermal hyperalgesia that is
preceded by increased serum TNF-α levels at day 3 post SPS. The maintenance phase of allodynia/hyperalgesia is
accompanied by elevated serum IL-1β. The aim of this study was to determine the effects of TNF-α blockade on
development of PTSD and co-morbid pain symptoms. To achieve this, male Sprague-Dawley rats were subjected to SPS;
half of the SPS and control group rats were immediately presented with 50 mg/kg thalidomide (THL), a potent TNF-α
blocker, in 10% DMSO vehicle, daily, i.p. for 5 days; non-SPS rats received daily vehicle injections. Rats were assessed
for nociceptive sensitivity periodically from day 3-day 21; anxiety behaviors were determined by exploration of the
elevated plus maze on day 9. Blood samples were taken at day 3 and 21; cytokine levels were determined by ELISA and
multiplex assays, respectively. THL prevented TNF-α increases in serum at day 3 and almost completely prevented
mechanical allodynia and thermal hyperalgesia associated with SPS (p<0.05) as early as day 3 post SPS. THL did not alter
nociceptive sensitivity in non-SPS rats. THL treatment blocked anxiety like behaviors (open entries and time spent in
open arms (p<0.05)) in SPS-treated rats, with no effect in sham-treated animals. Examination of IL-1β levels in isolated
peripheral blood mononuclear cells at day 21 of SPS indicated that acute treated with THL prevented SPS-induced IL-1β
increase in serum at day 21 (p<0.05), consistent with reduced nociceptive sensitivity. Our results suggest that short-term
treatment with immunomodulatory drugs such as THL immediately following traumatic stress may offer a novel
therapeutic method to prevent or reduce symptoms of PTSD and co-morbid pain.
Funding: This work and the personnel who conducted it were supported by grants from the Department of the Army
(DMRDP W81XWH-11-2-0077) and the OUHSC VPR (Bridge Grant Program) to KMS
97 Abstract #77
IDENTIFICATION OF BORRELIA BURGDORFERI NOVEL INTEGRAL OUTER MEMBRANE PROTEINS.
Binu Shrestha and Darrin R. Akins Department of Microbiology and Immunology University of Oklahoma Health
Sciences Center, Oklahoma City, Oklahoma.
Lyme disease, the most prevalent arthropod-borne disease in the US, is caused by the pathogenic spirochete Borrelia
burgdorferi. With no vaccine currently available to prevent this debilitating disease, considerable research has focused on
identification of novel integral outer membrane proteins (OMPs) that could be used as vaccine targets or diseasemodulating therapeutics. Freeze fracture electron microscopy has demonstrated that a large number of OMPs are present
in the outer membrane (OM) of the organism, but only a few OMPs have been identified to date. Identification and
characterization of novel OMPs from B. burgdorferi will enable a better understanding of the physiology of the
spirochete. Furthermore, identifying novel OMPs will also provide unique insight into the strategies used by this
spirochete to cause disease and evade the immune response during mammalian infection. In the current study, analysis of
enriched B. burgdorferi OM fractions revealed two hypothetical proteins, encoded by chromosomal ORFs bb0405 and
bb0406, which are present in an operon and conserved throughout different borrelial species. Cellular localization and
liposome incorporation experiments demonstrated that these proteins are present in the OM, are surface exposed, and
capable of being incorporated into lipid bilayers; all characteristics expected of a OMPs. To further assess the roles of
these proteins in virulence, a BB0405-BB0406 mutant was generated in a virulent borrelial strain, and was complemented
in trans with BB0405, BB0406, or both BB0405 and BB0406. These strains are currently being tested in a mouse model
of infection to examine their role(s) in virulence. Taken together, we have identified and characterized two new OMPs
from B. burgdroferi and our future studies will focus on characterizing their roles in B. burgdorferi physiology, virulence,
and disease pathogenesis.
Funding: AI059373 Grant from NIH/NIAID
98 Abstract #78
A NOVEL MISSENSE MUTATION IN THE EXTRACELLULAR DOMAIN OF THE PDGFRA GENE INDUCES
FUNCTIONAL CONSEQUENCES IN VIVO
Amanda K. Templeton1,2, Imelda T. Sandoval1, Richard Glenn C. Delacruz1, Christeena Satterfield1, Braden Miller1, and
David A. Jones1,2 1Immunobiology and Cancer Research Program, Oklahoma Medical Research Foundation, Oklahoma
City, OK 2 Department of Pathology, University of Oklahoma Health Sciences Center, Oklahoma City, OK
Platelet-derived growth factor receptor-α (PDGFRA) belongs to the class III receptor tyrosine kinase family, which also
includes c-KIT, and is characterized by their structural similarity in immunoglobulin-like extracellular domains, a
transmembrane domain, and intracellular protein kinase domains. Additionally, mutations in PDGFRA and c-KIT have
been described in skin malignancies. Exome sequencing of a patient with basal cell carcinoma revealed PDGFRA E459K
missense mutation. This mutation lies within the Ig5 domain of PDGFRA that may parallel mutations in c-KIT that result
in constitutive receptor activation or altered ligand affinity in skin malignancies. Additionally, bioinformatics analysis of
skin malignancy tumors harboring mutations in PDGFRA, the most common mutation is the single amino acid change at
codon 459. We were, therefore, interested in investigating whether mutations in PDGFRA are also implicated in the
pathogenesis of skin cancer. Assessment of functions of PDGFRA and the novel variants were examined by performing
loss-of-function and gain-of function experiments in zebrafish. Analysis of pdgfra expression in vivo revealed strong
staining in the developing retina, pharyngeal arches, and pectoral fins. Knockdown of zpdgfra resulted in blebbing on the
head and body, which was rescued by injection of human wild-type PDGFRA mRNA. Injection of human PDGFRA
carrying the E459K mutation alone resulted in developmental defects in structures derived from neural crest cells. These
defects included a decrease in epithelial pigmentation, altered migration of melanocytes, neural tube defects, defective jaw
formation, and ectopic development of eyes or secondary body axis. Similar developmental defects were observed in
embryos injected with human PDGFRA carrying a characterized gain-of-function mutation, but not in those injected with
only wild-type human mRNA. Collectively, these findings suggest that this novel PDGFRA variant carries functional
consequences that may contribute to skin tumor development. Therapeutic agents blocking PDGFRA may, therefore,
represent novel approaches to treating skin cancer.
Funding: OMRF institutional funds
99 Abstract #79
IS MARRIAGE A BIG HEADACHE --BEING LESBIAN IN CHINA
Tao We, Health Promotion Sciences , College of Public Health, OUHSC Advisor: J. Neil. Henderson
Disproportionately high rates of mental health problems have been reported among LGBT community in Western
countries, in particular, on depression, anxiety, and suicide. However, relevant studies among lesbians in Mainland China
remain scarce Systematic literature review identifies three articles published in peer-reviewed journals, and four reports
posted on gay and lesbian websites based on small-scale surveys among Chinese lesbians. High rates of depression,
anxiety, and suicide are reported among these women with same-sex attraction, compared with their heterosexual
counterparts. These studies also finds that Chinese lesbians face colossal pressure to enter heterosexual marriage that
serve as social, as social, cultural, and familial obligations, rather than simply a personal choice. It is very likely that
mental health of Chinese lesbians is negatively affected by gender-based discrimination and stigma through marriage
mandate defined by patriarchal heteronormativity dominating Chinese society. Experimental studies using sufficient
sample size are needed to test this hypothesis and inform health promotion programs and policies for this sexually
minority group.
Funding: is applying for the grants of the American Psychological Association
100 Abstract #80
IN VITRO COMPARISON OF TWO NASAL DELIVERY DEVICES TO ADMINISTER DRY POWDERS
Hooman Yari, Rahul Kumar Verma, Lucila Garcia-Contreras College of Pharmacy, University of Oklahoma Health
Sciences Center, Oklahoma City, OK
INTRODUCTION Delivery of drugs and vaccines as powders by the intranasal route is an attractive alternative to
injection of liquids. The size of emitted particles and the reproducibility of the emitted dose upon each device actuation
are key to ensure treatment efficacy. This study evaluated the accuracy, reproducibility and robustness of the Fit-lizer™
(SNBL Ltd., Japan) and UDS powder nasal device (Aptar Pharma, Germany) to determine their suitability for use in
efficacy studies with our West Nile Powder vaccine. METHODS Powders with optimum size for nasal delivery but with
different densities were manufactured and evaluated including micronized lactose, trehalose microparticles, and a blend of
lactose and trehalose microparticles. Powders were loaded in each device and the percent emitted dose (ED%) was
measured by gravimetric methods. The volume median diameter (dv50) of emitted aerosols was measured by laser
diffraction. RESULTS The ED% increased with the decrease in powder density for both devices, but the increase in ED%
with the SNBL device was consistently higher and more uniform for all three test powders. When trehalose microparticles
were used, the variability in the ED% from the Aptar device was 5-fold larger (11.63%) than from the SNBL device
(2.21%). As result from the mechanism of aerosol generation, the particle size emitted from both devices was larger than
their corresponding primary particle size, but particles emitted from the Aptar device were more than two-fold larger
compared to the SNBL device. However, while all particles emitted from the SNBL device were within the optimal size
for nasal deposition (20-45 µm), more than 16% of the dose delivered from Aptar device was evaluated oversized.
CONCLUSION The SNBL device appears to deliver a more accurate and reproducible dose than the Aptar device and
thus is considered more suitable for use in efficacy studies with our powder vaccine.
Funding: ---
101 Abstract #81
MOLECULAR CLONING AND CHARACTERIZATION OF A TYROSINE PHOSPHATASE FROM MONOSIGA
BREVICOLLIS
Benjamin Zhao, Zhizhuang Zhao
Protein tyrosine phosphorylation is thought to be a unique feature of multicellular animals. Interestingly, the genome of
the unicellular protist Monosiga brevicollis reveals a surprisingly high number and diversity of protein tyrosine kinases,
protein tyrosine phosphatases (PTPs), and phosphotyrosine-binding domains. Our study focuses on a hypothetical SH2
domain-containing PTP (SHP), which interestingly has a predicted structure that is distinct from SHPs found in animals.
In this study, we isolated cDNA of the enzyme and discovered that its actual sequence was different from the predicted
sequence as a result of non-consensus RNA splicing. Contrary to the predicted structure with one SH2 domain and a
disrupted phosphatase domain, Monosiga brevicollis SHP (MbSHP) contains two SH2 domains and an intact PTP
domain, closely resembling SHP enzymes found in animals. We further expressed the full-length and SH2 domaintruncated forms of the enzyme in E. coli cells and characterized their enzymatic activities. The double-SH2 domaintruncated form of the enzyme effectively dephosphorylated a common PTP substrate with a specific activity among the
highest in characterized PTPs, while the full-length and the N-terminal SH2 domain-truncated forms of the enzyme
showed much lower activity with altered pH dependency and responses to ionic strength and common PTP inhibitors.
This indicates that SH2 domains suppress the catalytic activity. SHP represents a highly conserved ancient PTP, and
studying MbSHP should provide a better understanding about the evolution of tyrosine phosphorylation.
Funding: OCAST
102 Section V. Oklahoma School of Science and Mathematics (OSSM) Poster Presentation Abstracts #82 -­‐ #93 103 Abstract #82
A CELL SOMA ORGANELLE TRANSPORT FUNCTION FOR CAENORHABDITIS ELEGANS CDK-5, SAD-1,
AND SYD-2 IN MOTOR NEURON AXONS AND DENDRITES AND ITS CORRELATION WITH LOCOMOTION
PATTERNS
Sooraj Boominathan1,2, Stacey L. Edwards2, Logan R. Morrison2, Kenneth G. Miller2 1Oklahoma School of Science and
Math; 2Genetic Models of Disease Laboratory, Oklahoma Medical Research Foundation
Some motor neuron diseases are associated with defects in organelle transport. unc-16 encodes a protein conserved in all
animals that regulates organelle transport by an unknown mechanism. A C. elegans unc-16 null mutation causes cell soma
organelles to accumulate in motor neuron axons. A genetic screen revealed that mutations in cdk-5, sad-1, and syd-2
suppress the accumulation of lysosomes in unc-16 mutant axons and cause lysosomes to accumulate in dendrites. One
goal of this study was to determine if cdk-5, sad-1, and syd-2 mutations also suppress the accumulation of Golgi in unc-16
mutant axons. A fluorescent Golgi body marker was integrated into the C. elegans genome and crossed into cdk-5, sad-1,
and syd-2 mutants in unc-16(+) and unc-16(-) backgrounds. Images collected from these strains under standardized
conditions were used to quantify the levels of Golgi bodies in axons, dendrites, and cell somas of identified motor
neurons. unc-16 cdk-5 double mutants had significantly lower amounts of Golgi membranes in their axons, and possibly
slightly higher amounts in their dendrites, but the suppression phenotype was not as strong as that observed for lysosomes.
This result is the first to indicate that CDK-5 has a role in the transport of Golgi in neurons. Since unc-16 null mutants
also display sluggish locomotion patterns, we sought to determine if the organelle suppressor mutations improved the
locomotion of unc-16 mutants. By conducting standardized locomotion rate assays, we found that unc-16 cdk-5 and unc16; sad-1 double mutants displayed a 2.5-4.5-fold improvement in locomotion compared to the unc-16 null mutants. unc16; syd-2 double mutants did not display a significant improvement, but this may be due to the synthetic phenotypes
present in this strain. These results show a correlation between the suppression of organelle accumulation in the axonal
regions of C. elegans and its resulting locomotion behavior.
Funding: Supported by NIGMS grant GM080765-12 to K.G.M and OCAST grant HR14-003 to K.G.M.
104 Abstract #83
CGMP/PKG SIGNALING REGULATES ENDOPLASMIC RETICULUM CALCIUM CHANNELS IN CONE
PHOTORECEPTORS
Jane Chin1, Hongwei Ma2, Melanie Mason2, and Xi-Qin Ding2 1 The Oklahoma School of Science and Mathematics,
2
Department of Cell Biology, University of Oklahoma Health Sciences Center
Introduction: Photoreceptor cyclic nucleotide-gated (CNG) channels play a major role in the process of phototransduction.
Present in rod and cone photoreceptors, they are responsible for maintaining a steady sodium (Na+) and calcium (Ca2+)
influx. Mutations in the cone CNG channel subunits are associated with human achromatopsia and cone dystrophies. Mice
with cone CNG channel deficiency undergo cone degeneration, accompanied with accumulation of cellular cyclic
guanosine monophosphate (cGMP), the native ligand of CNG channels, cGMP-dependent protein kinase (protein kinase
G, PKG) activation, alterations of the endoplasmic reticulum (ER) calcium channel inositol 1,4,5-trisphosphate receptor 1
(IP3R1), and ER stress. This work examined whether cGMP/PKG signaling regulates ER calcium channels and ER stress
in cone photoreceptors. Methods: In our study, the cone-derived cell lines – the Weri cells and 661W cells – were used.
The cells were treated with cGMP at varying concentrations for 24 hours, and were then harvested at the end of treatment
for analysis of PKG activity using ELISA and expression levels of ER calcium channels and ER stress markers using
Real-Time PCR (qRT-PCR) and Western blotting. Results: We found that cGMP treatment significantly increased PKG
activity, decreased the level of phosphorylated IP3R1, and increased the level of phosphorylated ryanodine receptor 2.
Conclusion: Our results support the view that cGMP/PKG signaling stimulates ER calcium-releasing channels and
regulates ER calcium homeostasis, and that cGMP/PKG signaling may play a role in the ER stress resulting from CNG
channel deficiency.
Funding: This work was supported by grants from the National Eye Institute (grant # R01EY019490).
105 Abstract #84
TARGETED TREATMENT OF CANCER USING NEAR INFRA-RED LIGHT
Kevin Weng1, Moses Bio2, Youngjae You3 1Oklahoma School of Science and Mathematics, 2College of Pharmacy,
University of Oklahoma Health Sciences Center
Introduction: Photodynamic therapy (PDT) is one of the treatment modality for cancer with minimal side effects. It
employs a photosensitizing drug, oxygen and light of specific wavelength. The three individual elements for PDT are nontoxic, but when combined, they generate the reactive oxygen species. PDT can also be used as targeted drug delivery by
coupling an anticancer drug to a photosensitizer through a singlet oxygen cleavable linker. Methods: We developed a
prodrug known as Pc-(L-CA4). The prodrug Pc-(L-CA4) was synthesized with silicon(IV) phthalocyanine dichloride and
linked to anticancer drug combretastatin A-4 (CA4) via aminoacrylate. The prodrug was tested in vitro. The prodrug is
cleaved by far-red light (690 nm) and released CA4. Results: It showed improved toxicity similar to CA4 and displayed
bystander effects in vitro. Conclusion: The prodrug showed enhanced cytotoxicity upon illumination and showed that
targeted drug delivery with PDT is a viable method for treating cancer.
Funding:
106 Abstract #85
EPIGENETIC CONTROL OF DNA REPLICATION THROUGH TICRR-BRD4
Maaz Khan1, Courtney G. Sansam2, Duane Goins2, Christopher L. Sansam2,3
Histone acetylation has been well characterized for its role in affecting transcription, but histone acetylation also affects
DNA replication. Histone deacetylases reduce the efficiency of DNA replication fork progression, shorten S-phase, and
increase DNA damage; however, the specific molecular mechanisms by which acetylation affects replication are not
known. In general, acetylation recruits proteins to chromatin through interaction with acetyl-lysines via bromo-domain
containing proteins. We recently identified a physical interaction between the bromo-domain containing proteins BRD2
and BRD4 and the essential replication initiation protein TICRR (TOPBP1-Interacting Checkpoint and Replication
Regulator). Recent evidence indicates that TOPBP1 and BRD4 are present at viral replication origins. Further, we show
that inhibition of BRD4 with the small molecule inhibitor JQ1 causes severe developmental defects in zebrafish embryos
deficient for TICRR. Our goal is to determine the significance of the BRD2/4-TICRR interaction and how it affects DNA
replication initiation.
Funding: 1Oklahoma School of Science and Mathematics, Oklahoma City, OK 2Oklahoma Medical Research Foundation,
Cell Cycle and Cancer Biology Research Program, Oklahoma City, OK, 3University of Oklahoma Health Sciences Center,
Department of Cell Biology, Oklahoma City, OK
107 Abstract #86
DEVELOPMENT OF A STANDARDIZED HLA-E ASSAY PROCEDURE
Storm Mata1, Saghar-Kaabinejadian3, William H. Hildebrand3, Brent Richards1, Rico Buchli2 1Oklahoma School of
Sciences and Mathematics, Oklahoma City, Oklahoma, 73104 2Pure Protein LLC, Oklahoma City, Oklahoma, 73104
3
Oklahoma University Health Science Center, Oklahoma City, Oklahoma, 73104
Human leukocyte antigen-E (HLA-E) is a nonclassical HLA class I molecule that canonically binds peptides derived from
the leader sequence of classical HLA class I. HLA-E can also bind peptides from stress proteins and pathogens,
illustrating the importance of HLA-E for anti-viral/anti-tumor immunity. Unlike classical HLA class I molecules, HLA-E
is characterized by only a limited sequence variability and two dominant protein forms have been described (HLAE*01:01 and HLA-E*01:03). The immunological and clinical relevance of HLA-E molecules is poorly investigated and
our interest is their potential to act as disease marker, particularly if increased serum levels of soluble HLA-E correlates
with disease activity. We aim to provide a standardized method to quantitatively detect sHLA-E. HLA-E forms a
heterodimer with β2-microglobulin, which presents peptides from proteins after infection, immunization, or
transplantation. A recombinant soluble form was generated by removing the trans-membrane portion of the molecule. The
purified molecule was used as a standard probe to design best assay strategies. Multiple antibody combinations were
investigated and optimized for best ELISA performance. Experiments showed that HLA-E share epitopes with classical
HLA class I molecules by reacting with antibodies like W6/32 or α-β2m commonly used to detect the classical branch.
We demonstrated that the combination of specific HLA-E antibodies like 3D12 with broad classical class I-recognizing
antibodies in a sandwich approach perform more accurately and sensitively than combinations with classical HLA class I
antibodies only. The standard sHLA-E molecule, performed linearly, provide a broad dynamic range. Optimal sandwich
ELISA parameters were established for coating, detection, and signal development. The sandwich ELISA procedure
developed in this study was found to be highly sensitive in detecting low levels of sHLA-E molecules whose
identification in serological samples may have important implications in understanding the pathogenesis of immunemediated vascular diseases and for the diagnosis and monitoring of patients.
Funding: Pure Protein, LLC
108 Abstract #87
THE ROLES OF ESCO1 AND ESCO2 IN MITOSIS
Katie McDonald1, Reem Alomer2, Susannah Rankin3 1Oklahoma School of Science and Mathematics, 2Oklahoma Medical
Research Foundation, 3Oklahoma Medical Research Foundation
Sister chromatids are tethered together by the cohesin complex, which helps them segregate correctly during mitosis. It is
important that cells successfully pass on their genetic material because errors can lead to genetic diseases such as cancer.
After cohesin is loaded onto the chromosome, two enzymes, Esco1 and Esco2, can acetylate the complex. This
acetylation helps to stabilize the interaction with chromatin; in the absence of acetylation, cohesin is readily removed. The
acetylation of cohesin allows for Sororin to bind to the complex, which in turn ensures that the sister chromatids remain
tethered together until prophase of mitosis. We investigated the different roles of Esco1 and Esco2 in promoting proper
mitotic division. We used siRNA to deplete Esco1, Esco2, or both Esco1 and Esco2 from HeLa cells, then examined the
effects on sister chromatid cohesion. We used immunoblotting, flow cytometry, and metaphase chromosome spreads to
analyze Esco enzyme levels and function. We found that Esco1 and Esco2 have different roles in mitosis. Our data
indicate that Esco2 is required for normal levels of mitotic chromosome cohesion. We are still investigating the specific
roles of these enzymes in cohesion establishment and maintenance. It is important to understand the roles of the Esco
enzymes in maintaining sister chromatid cohesion because of the deleterious consequences of errors in chromosome
segregation during mitosis.
Funding: funded by NIH R01GM101250 to S.R.
109 Abstract #88
COMPOUND H INCREASES ANTI-AGING PROTEIN KLOTHO EXPRESSION AND ATTENUATES ARTERIAL
STIFFENING AND HYPERTENSION
Samantha Okere1, Kai Chen2, Zhongjie Sun2 Oklahoma School of Science and Mathematics1, Department of Physiology,
University of Oklahoma Health Sciences 2
Background & Objective - The prevalence of arterial stiffness and hypertension increases with age while the klotho level
decreases with age. Compound H elevated klotho protein expression in opossum kidney and Z310 rat choroid plexus cells,
which express klotho endogenously. The objective of this study is to asses if compound H affects arterial stiffening and
hypertension in aged mice. Methods & Results – Aged mice (24-30 months) were treated with or without compound H
(15 mg/kg, IP daily) for 2 weeks, and adult mice without treatments were used as a control (12 months). Pulse wave
velocity (PWV), a direct measure of arterial stiffness, and blood pressure (BP) were increased significantly in aging mice.
Compound H reversed age-related increases in PWV and BP within 2 weeks of treatment. Compound H effectively
increased secreted klotho levels in both kidney and serum. Aging-related arterial stiffness was associated with increased
collagen and decreased elastin contents in the media of aortas. In addition, the expression of MMP2, MMP9, and
TGFβ1was increased in aortas of aged mice. These changes were attenuated by compound H. Conclusion –Aging-related
arterial stiffening and hypertension are attributed, at least in part, to klotho deficiency. Compound H increases klotho
expression and attenuates arterial stiffening and hypertension in aged mice.
Funding: Funding: NIH R01 HL102074, 105302 and DK 093403
110 Abstract #89
NEURONAL AND MICROGLIAL CELLS DIFFER IN THEIR SENSITIVITY TO GLUTAMATE TOXICITY
Caron Song1, Hibah O. Awwad2 1Oklahoma School of Science and Mathematics, 2Department of Pharmaceutical
Sciences, College of Pharmacy, University of Oklahoma Health Sciences Center
Excessive glutamate in traumatic brain injury and neurodegenerative diseases causes cellular damage known as
excitotoxicity. One of glutamate’s toxic mechanisms on neurons includes its ability to enhance phosphorylation of the tau
protein. Tau protein primarily functions to stabilize cytoskeletal microtubules. Upon tau phosphorylation microtubules
become destabilized, resulting in neurodegeneration and eventually neuronal apoptosis. Microglial cells were recently
found to play a neuroprotective role similar to that of astrocytes by converting glutamate into glutamine. Our goal was to
establish a model to simulate excitotoxicity in vitro and to determine the effects of glutamate toxicity on tauphosphorylation in neuronal and microglial cells. Human neuroblastoma SH-SY5Y and microglial murine BV2 cells were
treated with 2, 5, 10, 15, 25 and 50 mM L-glutamate for 20-24 hours. Cellular morphology was captured using bright-field
microscopy and cell viability was measured using the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium
bromide) assay. Additionally, cell lysates were collected after glutamate treatment and stored for immunoblotting assays
to measure tau-phosphorylation. Toxic concentrations of glutamate treatment significantly decreased cell viability in both
cell lines, however, SH-SY5Y cell lines were more sensitive to glutamate and showed a significant decrease in cell
viability at both 25 and 50 mM glutamate (n=2-4, p<0.0001; student’s t-test) whereas BV2 microglia only showed a
significant decrease at 50 mM glutamate (p=0.0016; student’s t-test) when compared to the respective untreated cells.
Immunoblotting experiments to determine tau phosphorylation in both cell lines are ongoing. This study indicates that
neuronal cells are more sensitive to glutamate toxicity than microglial cells. Our immunoblotting results will determine
whether tau phosphorylation is an underlying biochemical mechanism that contributes to this difference. This study
suggests that microglial cells could survive harsher conditions following a brain injury than neurons and understanding
the underlying neurobiology may shed light on potential neuroprotective mechanisms against glutamate toxicity.
Funding: This work was supported by OU College of Pharmacy funds (H.O.A.)
111 Abstract #90
LA POSITIVE, RO60 NEGATIVE SUBSET OF PRIMARY SJÖGREN’S SYNDROME IS A REALITY
Debashish Danda1, Dat Truong2, Marshall Shaw3, Celia Quang4, Kristi Koelsch5, Biji T. Kurien6, Harini Bhagavant7,
Umesh Deshmukh8, Kathy L. Silvis9, R. Hal Scofield10 1The Arthritis and Immunology Program, Oklahoma Medical
Research Foundation, 2The Department of Medicine, University of Oklahoma Health Sciences Center, 3Department of
Medical Service, Veterans Affairs Medical Center, 4Oklahoma School of Science and Mathematics
Introduction Twenty-nine sera from 348 primary Sjögren’s syndrome patients were identified as anti-Ro60 (anti-SSA)
negative and anti-La (anti-SSB) positive by immunodiffusion, line immunoassays and multiplex bead assays. We
hypothesized that a significant portion of these were falsely negative for anti-Ro60. Methods Twenty-nine sera from
primary Sjögren’s syndrome patients, fulfilling four AECG criteria, were tested for the presence of antibodies directed
against La and Ro60 autoantigen. Anti-La was detected on bovine La treated with or without DNAase and RNAase (to
check for false positivity, since anti-La can bind DNA and RNA). Anti-Ro60 antibodies in the sera were detected using
HEp-2000 substrate (in which cells are transfected with human Ro60) and HEp-2 substrate. Anti-Ro60 and Ro-52 were
also tested by in vitro transcription/translation/immunoprecipitation assay. Results Out of the 29 sera, 25 were
unequivocally negative on HEp-2000 (1:40 dilution). Four samples were clearly found to be Ro60 positive with a
speckled pattern and three of the four continued to be positive up to 1:320 dilution, as against only two positive samples
on HEp-2 at 1:40 dilution. This finding suggests false negativity for Ro60 exists in a small fraction (14 percent) of
primary Sjogren’s syndrome patients. However, all the samples were negative for Ro60 and Ro52 by in vitro
transcription/translation/immunoprecipitation assay. Conclusions Contrary to our hypothesis, we found only a small
fraction of Ro negative, La positive sera to show positive HEp-2000 pattern. This suggests that a subset of primary
Sjogren’s syndrome is probably a true entity with Ro60 negativity and La positivity. The clinical significance of the
subset will be revealed during the follow up of our patients.
Funding:
112 Abstract #91
STAPHYLOCOCCUS AUREUS CONTRIBUTES TO THE PATHOGENESIS OF ENDOGENOUS BACTERIAL
ENDOPHTHALMITIS BY DISRUPTION OF THE TIGHT JUNCTIONS BETWEEN RETINAL PIGMENT
EPITHELIAL CELLS OF THE BLOOD RETINAL BARRIER
Kobby Wiafe1, Roger Astley2, Phillip Coburn1, Michelle Callegan2 1Oklahoma School of Science and Mathematics,
2
Department of Ophthalmology, University of Oklahoma Health Sciences Center
Purpose: Endogenous bacterial endophthalmitis is rare but serious infection of the inside of the eye caused by bacteria
which originate from an infection in one part of the body and travel to the interior of the eye. How this occurs is not
known, but it may involve bacteria in the blood breaking down the blood retinal barrier, which normally separates the
interior of the eye from the components of the blood. Methods: Human retinal pigment epithelial cells (ARPE-19, ATCC
CRL-2302) were grown on glass coverslips which were coated with a basement membrane like material. After growing to
confluence the cell cultures were infected with 10,000 colony forming units per milliliter of Staphylococcus aureus (strain
8325-4). At 4, 6, and 8 hours post infection coverslips were fixed in cold methanol. The ZO-1 protein, which forms part of
the blood retinal barrier, was detected by immunohistochemistry (anti ZO-1, Zymed 33-9100), and visualized using
confocal microscopy. Results: For the first 4 hours post infection the growth of Staphylococcus aureus was very slow,
thus resulting in unaffected tight junctions in the infected retinal epithelial cells. Beginning at 4 hours post infection,
however, disintegrated tight junctions were observed. After4 hours and progressing to 8 hours post infection, tight
junctions between the cells disappear in the infected wells, but not in the mock infected wells. In this time period, the
number of Staphylococcus aureus bacteria grew in an exponential manner. This explains why a higher percentage of
degenerated tight junctions were observed following the 4 hour post infection mark. Discussion: These results indicate
that Staphylococcus aureus is capable of disrupting the tight junctions between cells that form part of the blood retinal
barrier. With this information, closely related studies can be conducted so that the proper treatement option(s) can be
offered to infected patients.
Funding: This study was funded by NIH Grant R21EY022466 (to MCC). Our research is also supported in part by,
R01EY012985 (to MCC), R01EY024140 (to MCC), P30EY12191 (NIH CORE grant to Robert E. Anderson, OUHSC),
P20RR17702 (NCRR COBRE grant to Robert E. Anderson, OUHSC), and an unrestricted grant to the Dean A. McGee
Eye Institute from Research to Prevent Blindness.
113 Abstract #92
EPSIN IS REQUIRED FOR DISHEVELLED STABILITY AND WNT SIGNALING ACTIVATION IN COLON
CANCER DEVELOPMENT
Amanda Zhang1, Kandice L. Tessneer2, Baojun Chang2, John MacMacnus2, Xialei Liu2,3, Scott Hahn2, Satish Pasula2,
Hao Wu2, Hoogeun Song2, Yiyuan Chen2, Xiaofeng Cai2, Yunzhou Don2, Megan L. Brophy2,3, Ruby Rahnman2, Jian-Xing
Ma4, Lijun Xia2,3, & Hong Chen2,3. 1Oklahoma School of Science and Mathematics, Oklahoma City, OK 73104 USA.
2
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma
73104, USA. 3Department of Endocrinology and Diabetes, Harold Hamm Oklahoma Diabetes Center, University of
Oklahoma Health Sciences Venter, Oklahoma City, Oklahoma 73014, USA.
Introduction: Uncontrolled canoncial Wnt signaling supports colon epithelial tumor progression. Understanding these
regulatory mechanisms is crucial for elucidating the pathogenesis and identifying new therapeutic targets to combat colon
cancer. Epsins are ubiquitin-binding adaptor proteins upregulated in several cancers. However, the involvement of epsins
in colon cancer is unknown. Methods: We generated a novel mouse model selectively lacking epsin 1 in intestinal
epithelial cells (IEpCs) on an epsin 2 null background by crossing Epn1fl/fl; Epn2-/- mice with mice expressing tamoxifeninducible Cre recombinase under control of the Villin promoter (Villin-ERT2 Cre). To produce inducible IEpC-iDKO
(double knockout) mice, gender and genetic background-matched 10-week wild-type or Epn1fl/fl; Epn2-/-; Villin-ERT2Cre
mice were intraperitoneally injected with tamoxifen on alternating days for 2 weeks. Standard immunofluorescence,
Western blotting, and subcutaneous tumor implantation were used to examine roles of epsin in the colon cancer
progression. Results: Loss of intestinal epithelial epsins combats colon cancer by significantly reducing stability of the
crucial Wnt signaling effector, disheveled (Dvl2), and impairing Wnt signaling. Consistently, epsins and Dvl2 interact and
are correspondingly upregulated in colon cancer. Conclusion: Our findings reveal an unconventional role for epsins in
stabilizing Dvl2 and potentiating Wnt signaling to encourage robust colon cancer progression. This pro-carcinogenic role
of epsins suggests that they are potential therapeutic targets to combat colon cancer.
Funding: NIH grants R01HL-093242, R01HL-118676, P20 RR018758, a National Scientific Development Grant from the
American Heart Association (AHA) (0835544N), grant from the Oklahoma Center for Advanced Science and Technology
(OCAST) HR09-116, and a grant from the Department of Defense W81XWH-11-1-00226 to H. Chen; AHA Postdoctoral
fellowships 13POST16940008 to K.L. Tessneer and 13POST17270006 to S. Pasula; AHA Predoctoral fellowship
RSRCH016952 to X. Liu.
114 Abstract #93
IMPROVING TIME-EFFICIENCY FOR EVALUATING THE PITUITARY GLAND UNDER SEMI-AUTOMATED
METHOD
Julie Zhu, Jim Zhonning Chen, Mary K. Gumerlock, M.D. Zachary Connor, Dee Wu, Ph.D.
Introduction Evaluation of the pituitary has had a long history in the study and the control of acromegaly and gigantism.
Such conditions are often caused by a benign tumor in the pituitary gland causing excess growth hormone release. We
hypothesize that the growth hormone–producing tumors undergo have a direct impact on the growth of tumors very
quickly in young people and more slowly in adults. The goal of this project is image segmentation of the pituitary. Image
segmentation breaks up an image in order to make it more analyzable. By taking data points from the image, we can then
better map the function tumor. Methods The computation and theory behind the image segmentation is first written into
Python, a programming language. Python is a slow language, however, and if many points are taken from the image, the
processing time could be too long to be reasonable. Therefore, the Python code is translated into C, which deals directly
with memory and is a faster language. Once translated, both versions of the code will be run on the same, small data sets
and the results and plots from both versions will be compared to each other, and then to the theoretical results. Our
laboratory is developing a new image segmentation method based on B-splines. Results We have achieved a reduction in
time using the C code. The C-based codes improved responsiveness of the application as well as we were able to use
other numerical libraries to enhance our application. Finally, the application of C coding will make it possible to
implement this on mobile platforms in the future facilitating the translational research component of this
work. Conclusions Our semi-automated methods for evaluating the pituitary produce greater accuracy in geometric
shape and provide this in a time-efficient manner.
Funding:
115 Section VI. Professional Students Poster Presentation Abstracts #94-­‐ #99
116 Abstract #94
CHARACTERIZING OCULAR SURFACE INFLAMMATION INDUCED BY CERAMIDE SYNTHESIS
INHIBITOR FTY720.
Jeff LaCroix1,2,5, Megan Stiles2,5, Jeremy Tan2,5, Tuan-Phat Huynh2,5, and Nawajes A. Mandal2,3,4,5 1University of
Oklahoma College of Medicine, University of Oklahoma Departments of 2Ophthalmology, 3Physiology, 4Oklahoma Center
for Neuroscience, OUHSC, Oklahoma City, OK; 5Dean McGee Eye Institute, Oklahoma City, OK.
Introduction: Ceramide, a signaling sphingolipid, serves as 2nd messenger to induce inflammation in various tissues and
apoptosis in neural cells. FTY720, a drug currently utilized in the treatment of multiple sclerosis, inhibits ceramide
synthesis. Previous studies in our lab have shown that systemic FTY720 treatment prevented photoreceptor cell death in
rat models of light-induced retinal degeneration by reducing de novo ceramide synthesis. Topical delivery of FTY720 in
rat eyes induced inflammation of the ocular surface. Here, we characterized that inflammation and tested whether
FTY720-induced modification of ceramide composition in ocular surface tissues (tear film, meibomian glands, etc.) is the
cause of this inflammation. Methods: Adult wild type, Sprague Dawley (SD), rats were treated with 1% FTY720 eye
drops at 0hrs, 3hrs, and 6hrs. Tissues were harvested at 24hrs and inflammation was characterized by RT-PCR, histology,
and lipid analysis. SD rats treated with vehicle-only drops served as control. Results: Topical delivery of FTY720 brought
about generalized corneal inflammation characterized by an upregulation of: Icam1, Ccl2, CXCL1, IL-10, IL-1β, and
Timp1 mRNA and a downregulation of: TNF-α, Pyrcd, IL-2, and SK2 mRNA. Preliminary lipid analysis reveals, as
expected, that FTY720 treatment decreased levels of some species of ceramides. Histology reveals dense inflammation of
the limbus and lateral cornea proper in all treated samples and scant, if any, inflammation in untreated rats. Conclusion:
Ceramide is crucial for ocular surface homeostasis. Any topical application for FTY720 must account for this factor and
correct for it. Potentially, addition of short chain ceramides or incorporation of anti-inflammatory steroids may reduce this
inflammation and allow FTY720 enter into the eye for retinal protection.
Funding: NIH grant EY022071; Foundation Fighting Blindness, Inc. USA; Research to Prevent Blindness, USA.
117 Abstract #95
CANCER-TREATMENT ADHERENCE AND SENSITIVITY TO THE EMOTIONAL TONE OF VERBAL
COMMANDS
Sudha Lakhwani,1,3 Blas Espinoza-Varas,1,2 & Kai Ding3 1Communication Sciences & Disorders, 2P&G Stephenson
Cancer Center, 3Biostatistics & Epidemiology, OU Health Sciences Center, Oklahoma
Introduction: Treatment adherence impacts significantly on outcomes and requires complying with verbal commands
issued by health-care professionals to impede or instigate treatment-relevant behaviors (e.g., “quit smoking!” or “go on a
diet!”). Since the commands’ emotional voice tone (VT), lenient or stern, specifies optional or mandatory adherence,
having keen VT sensitivity could increase adherence; however, information or response conflict could decrease the
sensitivity pre or post chemotherapy (CHT), and decrease adherence. The ability to identify the commands VT in
information- or response-conflict conditions that activate executive functions (EF) could reflect how compliant a patient is
with verbal commands. Methods: In gynecological cancers (n=14), the ability to identify the VT of voice commands
was assessed under three EF demands: Inhibitory-control conditions presented the cue word “left” or “right” followed by
impeding commands (“quit!”) in lenient or stern tone, mapped onto a left or right response; the cue and ear side could be
congruent or in conflict with the correct response side. Trials presenting instigating commands (“go!”) mapped lenient or
stern onto a right or left response. Task-switching conditions interleaved impeding and instigating commands within the
same trial block, and required switching the mapping rule depending on the command, impeding or instigating. Workingmemory conditions probed whether the command presented on the current trial was equal to or different from the one
presented two trials back. Results: Both prior to and post CHT, the patients’ VT identification accuracy was
significantly lower than that of healthy controls. The patients’ VT identification errors were small in conditions free of
EF demands, but increased significantly when EF demands were imposed, being largest in task-switching conditions;
individual differences were large. Conclusion: Treatment adherence would decrease if the ability to identify the VT of
verbal commands is impaired because patients have difficulty discerning between optional or mandatory compliance.
Funding: Stephenson Cancer Center, Supportive Care and Outcomes Research
118 Abstract #96
PREVALENCE OF TINNITUS AND NOISE INDUCED HEARING LOSS IN OKLAHOMA DENTISTS
Jamie Myers1, Suzanne Kimball1, Terry Fruits2, Andrew B. John1 1Department of Communication Sciences and Disorders,
College of Allied Health, OUHSC; 2Department of Operative Dentistry, College of Dentistry, OUHSC
Purpose: Noise-induced hearing loss (NIHL) is the result of exposing the auditory system to damaging noise over an
extended period of time. Tinnitus, often characterized by continuous ringing in the ears, is the result of damage to the
inner ear, which often presents itself secondary to NIHL. Many studies have measured dental handpieces and found
potentially hazardous noise levels. We used a survey of Oklahoma dentists and measurements of sound pressure produced
by dental handpieces to evaluate risk of NIHL and tinnitus in this population. Methods: Measures of sound pressure
levels produced by dental handpieces were collected in the OUHSC College of Dentistry. A survey was mailed to
members of the Oklahoma Dental Association. Self-reports of tinnitus, hearing loss, and risk factors were compared to
data reported in previous studies. Survey responses regarding hours per day spent using dental handpieces were combined
with handpiece sound level measurements to calculate a range of approximate noise exposures based on Occupational
Safety and Health Administration and National Institute for Occupational Safety and Health permissible exposure
limits. Results: The survey is in the field and data collection and analysis are ongoing but will be complete prior to
presentation. Sound level measurements made in the dental lab indicated that high-speed dental handpieces in conjunction
with suction have the potential to create hazardous noise levels if used for a sufficient period of time. Preliminary analysis
of survey data suggest a higher self-reported prevalence of NIHL and tinnitus in responding dentists than in the US
population overall. Conclusions: Sound level measurements combined with survey responses regarding perceived
hearing loss and tinnitus as well as hours of exposure to noisy dental handpieces suggest that dentists may be exposed to
hazardous noise levels. Dentists may also be at risk of NIHL and tinnitus as a result of recreational noise exposure.
Funding: Student Research and Creativity Grant, College of Allied Health, OUHSC
119 Abstract #97
ACCEPTABILITY AND FEASIBILITY OF AN AFRICAN-AMERICAN HEALTH & HERITAGE NUTRITION
INTERVENTION AMONG AFRICAN-AMERICANS WITH TYPE 2 DIABETES
Micki Hall1, Tanya Nguyen1, 1College of Pharmacy, University of Oklahoma Health Sciences Center
Introduction: African-Americans are more likely than non-Hispanic Whites to develop diabetes and are more likely to
develop long-term, serious diabetes complications, such as lower-extremity amputations. In 2012, the age-adjusted death
rate from diabetes in Oklahoma was 32.1/100,000 in African-Americans compared to 18.1/100,000 in non-Hispanic
Whites. The purpose of this pilot study was to examine the acceptability and feasibility of an African-American Health &
Heritage nutrition intervention among African-Americans with Type 2 diabetes. Methods: The nutritional intervention
included 6 sessions each with an introduction, cultural history, hands-on cooking lesson, and group discussion. Feasibility
was assessed by examining retention and completion rates. Acceptability was assessed using pre- and post-intervention
questionnaires. The Health Belief Model was used to assess perceived barriers and self-efficacy. Descriptive data was
collected and attendance was recorded. The Health Belief Model responses were analyzed by the Wilcoxon signed-rank
tests to compare participants’ responses before and after the nutritional intervention. A cultural competence survey was
given at session 2 and at the last session to examine how the educator was perceived with participants from a different
culture. Results: Fifteen participants signed consent (IRB #4644). Feasibility was good: 93% completed at least 3 and
53% completed 5 of the 6 sessions. Acceptability responses indicated the program was viewed positively by 14
participants who completed the exit assessment. Low positive response rate (<60%) was identified in the perceived
barriers and self-efficacy domains of the Health Belief Model. Cultural competence survey indicated perception that the
educator used everyday language and respected family beliefs and customs while areas for improvement (<70% positive
response rate) included need to provide suggestions that fit participant’s beliefs. Conclusion: Findings indicate this study
was feasible and acceptable. Results highlight the need for further studies to identify and reduce barriers that matter most
to patients while increasing self-confidence to healthier eating.
Funding: Oldways African Heritage &amp; Health Program
120 Abstract #98
IMPROVING PERCEPTION OF MEANINGFUL ACTIVITY IN A GROUP OF LOW-INCOME, OLDER ADULTS:
A PILOT STUDY
Kimberly Parker1, Carrie Ciro1 and Patsy Smith2 1Department of Rehabilitation Sciences, University of Oklahoma Health
Sciences Center 2College of Nursing
Introduction: Engagement in meaningful activity is critical for older adults, yet little is known of the impact of short-term
group intervention in changing perception of and engagement in meaningful activity particularly for those living in lowincome and underserved residential communities. Methods: Exploratory, sequential mixed methods design where Phase
1(qualitative) informed the development of Phase 2 (quantitative). Focus groups were completed with residents of two
low-income, older adult residential communities to illicit their understanding of meaningful activity and
facilitators/barriers of activity within their community. Phase 2 was a quasi-experimental study examining change in
scores from pre-post measurements of the Meaningful Activity Participation Assessment (MAPA) and the Engagement in
Meaningful Activity Survey (EMAS) after weekly, one hour groups for 7 weeks. In Phase 2, parametric and nonparametric statistics were used to examine differences in pre and post scores. Results: Phase 1 analysis revealed lack of
financial resources and activity availability as barriers to meaningful activity and a high interest in activities that improve
hypertension and diabetes. Of the participants that completed Phase 2 (n=11), mean age =65 years, 64% (7/11) were
female, 54.5 % (6/11) were black, 54.5% (6/11) had a HS education or below, 54.5 % (6/11) were depressed, 72.7%
(8/11) had HTN and 36.4% (4/11) had diabetes. Within group analysis revealed a significant difference in MAPA pre-post
scores (p=.007), but not EMAS (p=.33). Between group analysis revealed differences by educational level (p=.02) where
lower educated benefited more. No differences existed by the presence of hypertension (p=0.12), diabetes (p=0.88), or
depression (p=0.95). Discussion: We found that in a relatively short time period, we could impact perception of what is
considered healthy, meaningful activity, despite lower educational levels. More research is needed to determine methods
for improving older adult engagement in the meaningful activities they identify as necessary for good health.
Funding:
121 Abstract #99
GENOME-WIDE ASSOCIATION STUDY OF CIRCULATING 25(OH) VITAMIN D LEVELS IDENTIFIES A
NOVEL LOCUS ASSOCIATED WITH VITAMIN D DEFICIENCY
Bishwa Sapkota1, Andrew Bjonnes2, Piers Blackett3, Richa Saxena2, Dharambir Sanghera1 Department of Pediatrics,
1
Section of Genetics, 3Section of Endocrinology, College of Medicine, University of Oklahoma Health Sciences Center,
Oklahoma City, OK, USA. 2Broad Institute of Massachusetts Institute of Technology and Harvard, Cambridge, MA, USA
Vitamin D, 25-hydroxy-vitamin D [25(OH)D] deficiency is associated with multiple medical complications, including
musculoskeletal, inflammatory, malignancy and cardiovascular risk. Earlier, we reported that 82% of diabetic patients
were 25(OH)D deficient compared to 64% healthy controls, and high prevalence of 25(OH)D deficiency (<50nm/L) was
associated with a significant increase in cardiovascular risk factor including obesity, hypertension, and type 2 diabetes in
Punjabi Sikhs from the Asian Indian Diabetic Heart Study (AIDHS). In this study, we investigated the relationship of
serum 25(OH)D with cardiometabolic risk and performed the first GWAS and meta-analysis to identify gene variants
influencing 25(OH)D deficiency in AIDHS. Our discovery GWAS of 1,616 individuals (842 cases and 774 controls) was
followed by Stage 1 replication of 67 top signals (P<10-5) in an additional Sikhs (n=2,386). On combined discovery and
Stage 1 meta-analysis (n= 4,002), we identified a novel locus represented by IVL gene in association with serum
25(OHD) levels (β = 0.10, p=3.1 x 10-6) after adjustment with age, sex and type 2 diabetes status. These findings are
currently being replicated in other independent larger datasets. Our results also confirmed a previously reported
association with 25(OH)D represented by rs2282679 at the GC (Vitamin D binding protein) with (β = -0.13, p=3.0 x 10-4)
in Sikhs. The IVL gene (chromosome 1q21.3), encodes involucrin. Both IVL and GC genes are implicated in the
synthesis of vitamin D. Genetic variation in the 25(OH) D pathway may have a significant impact in the observed
deficiency of vitamin D, which could be synergistically contributing to increased cardiometabolic risk in this population.
Taken together, the identification of a new locus in a vitamin D pathway gene for influencing serum 25(OH)D levels may
have important clinical implications and should be replicated in independent sample sets.
Funding: Funding: NIH-R01DK082766 (NIDDK), NOT-HG-11-009 (NHGRI), and VPR Bridge Grant (OUHSC).
122 Section VII. Postdoctoral Fellow Oral Presentation Abstracts #100 -­‐ #116 123 Abstract #100
SECRETED KLOTHO AUGMENTS THE THERAPEUTIC POTENTIAL OF MESENCHYMAL STEM CELLS
FOR MONOCROTALINE INDUCED PULMONARY ARTERIAL HYPERTENSION
Quaisar Ali, Rohan Varshney, Chengxiang Wu, ZhongjieSun Department of Physiology, College of Medicine, University
of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma
Introduction: Klotho is an anti-aging gene. The purpose of this study is to investigate if secreted Klotho in synergy with
mesenchymal stem cells (MSC) improves monocrotaline (MCT)-induced pulmonary vascular dysfunction and
pulmonary arterial hypertension (PAH). Methods: We generated lentivirus carrying SKL-GFP or GFP genes driven by a
CMV promoter. MSCs were transfected with SKL-GFP. Four groups of male Sprague Dawley rats were treated with
MCT (60mg/Kg ,IP) while additional group was given saline (control). Three days later, four MCT-treated groups
received IV delivery of MSCs, MSC-GFP, MSC-SKL-GFP and no treatment respectively. Results: MCT significantly
increased right ventricular (RV) systolic blood pressure. Interestingly, MSC-GFP-SKL significantly attenuated the
MCT-induced increase in RV pressure (Saline: 17.27 ± 4.86, MCT: 31.88 ± 1.39, MCT + MSC-GFP-SKL: 23.35 ± 3.26).
Ex-vivo vascular relaxing responses to acetylcholine were decreased in small pulmonary arteries (PA) in MCT-treated
rats, indicating that MCT caused pulmonary vascular endothelial dysfunction. MSCs overexpressing SKL also
significantly attenuated RV hypertrophy. MSCs alone however, improved MCT-induced PAH, PA endothelial
dysfunction and RV hypertrophy but not to a significant level. Conclusion: MSC s over expressing SKL attenuated MCTinduced PAH, PA endothelial dysfunction, and RV hypertrophy. SKL augments the therapeutic effect of MSCs in
PAH.
Funding: 5R01HL116863-02
124 Abstract #101
SURFACE CHARACTERIZATION OF A NOVEL ANTIBACTERIAL DENTAL ADHESIVE RESIN
Fernando Florez1, Kwai-Sum Chan2, Osmir de Oliveira Júnior3, Edgar O’Rear4, Adam Rondinone5, Sharukh Khajotia1 1
College of Dentistry, University of Oklahoma Health Sciences Center, 2 Oklahoma City Community College, 3 UNESP
School of Dentistry, 4 School of Chemical, Biological and Materials Engineering, University of Oklahoma, and 5 Center
for Nanophase Materials Sciences, Oak Ridge National Laboratory
Objectives: Titanium dioxide nanoparticles possess antibacterial behavior when exposed to UV irradiation. Doping of
nanoparticles shifts the wavelength needed to generate reactive oxygen species to the visible range. The objective of this
study was to characterize the surface properties of a novel adhesive resin containing Nitrogen-doped titanium dioxide
nanoparticles (N_TiO2). Methods: Experimental resins were synthesized by mixing N_TiO2 (35mg/mL, Oak Ridge
National Laboratory) in OptiBond Solo Plus adhesive resin (Kerr Dental Products) in ratios of 1:5, 1:2, 1:1 and 2:1.
N_TiO2 were dispersed in resin (sonication: 20kHz, 30s), then thin-film specimens (n=5/ratio, thickness ≈10µm) of the
experimental resins and unaltered resin (control group) were fabricated on glass coverslips. Drops of ultrapure water
(3µL) were dispensed at two random locations on each thin-film’s surface in a contact angle goniometer (OCA15-Plus,
Future Digital Scientific Corp.). High-resolution digital images of the axisymmetric sessile drops were recorded
(25frames/s, 1min, 37±1oC). Drop profiles were analyzed to determine contact angles at time=0s (θINITIAL) and
time=50s (θFINAL). Data were statistically analyzed using General Linear Models and post hoc Student-Newman-Keuls
(SNK) tests (α=0.05). Specimens were subsequently sputter-coated with gold and characterized using SEM and Energy
Dispersive X-Ray (EDX) spectroscopy. Results: Statistically significant differences were observed among the groups
tested (p<0.01). Mean initial θ values ranged from 49.27 degrees for the control group to 58.88 – 74.12 degrees for the
N_TiO2 groups. Mean final θ values ranged from 44.11 degrees for the control group to 50.85 – 55.64 for the N_TiO2
groups.
Conclusion: Adhesive resins containing higher N_TiO2 concentrations were less wettable than the unaltered
control, thereby confirming the more hydrophobic nature of the antibacterial N_TiO2. EDX mapping for N, Ti and O2
confirmed N_TiO2 incorporation and dispersion. SEM images of thin films' surfaces confirmed the presence of more
nanoparticles in adhesives with higher N_TiO2 concentrations.
Funding: Partial funding by Office of the OUHSC Vice-President for Research Postdoctoral Fellow Award and by Grant
Number 8P20GM103447 of the National Institute of General Medical Sciences (NIH). A portion of this research was
conducted at the Center for Nanophase Materials Sciences, which is a DOE Office of Science User Facility.
125 Abstract #102
A UNIQUE DNA DAMAGE DETECTION ASSAY FOR EARLY DIAGNOSIS OF OROPHARYNGEAL CANCER
Vengatesh Ganapathy1, Majd A. Kabbani1, Clifton Woods1, Dana Mowls2, Ilangovan Ramachandran1, Dini Chissoe1,
Greg Krempl1, Yan D Zhao2, Lurdes Queimado1, 3-6 Departments of 1Otorhinolaryngology, 2Biostatistics, 3Cell Biology
and 4Pediatrics; 5The Oklahoma Tobacco Research Center and 6The Peggy and Charles Stephenson Cancer Center, The
University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Introduction: DNA damage is the main initiator of cancer. Unfortunately, existing assays are limited in the types of DNA
damage they detect, and are not practical for biomonitoring. We developed a unique technique named primer-anchored
DNA damage detection assay (PADDA) that reliably quantifies DNA damage in vivo. PADDA has higher sensitivity than
other available assays, can measure DNA damage in any tissue and is convenient for population-based screening. Here,
we aim (i) to measure the levels of DNA damage in peripheral blood cells (PBC) of patients with oropharyngeal cancer;
(ii) to identify the DNA damage threshold that can optimally discriminate between cancer and non-cancer individuals; (iii)
to correlate the levels of DNA damage with clinical and demographic parameters. Methods: DNA damage was quantified
by PADDA in p53 of the PBC of 50 patients with oropharyngeal cancer and 50 non-cancer controls. To identify the DNA
damage threshold that could optimally distinguish between patients and controls, we constructed a receiver operating
characteristic curve (ROC). Linear regression models were used to investigate the association between DNA damage and
specific individual characteristics. Results: Our data show that oropharyngeal cancer patients have significantly higher
levels of DNA damage in PBC than non-cancer individuals (p<0.01) at time of diagnosis. Using ROC curves, we were
able to define the DNA damage threshold that optimally distinguishes between cancer patients and controls with an
accuracy of 93%. A possible correlation between the in vivo levels of DNA damage and individual or clinical parameters
is currently being analyzed. Conclusion: The ROC curve showed that PADDA is an excellent diagnostic test for
oropharyngeal cancer with an accuracy of 93%. PADDA is an affordable assay for the routine evaluation of DNA damage
and may become a vital tool to evaluate cancer risk and guide prevention strategies.
Funding: This work was supported by the Oklahoma Center for the Advancement of Science and Technology (LQ). Dr.
Queimado holds a Presbyterian Health Foundation Endowed Chair in Otorhinolaryngology.
126 Abstract #103
THE POTENTIAL OF EXOSOME MICRORNAS AS BIOMARKERS FOR BREAST CANCER
Bethany N. Hannafon1, William C. Dooley2, 3, and Wei-Qun Ding1,3 1Department of Pathology and 2Department of
Surgery at the University of Oklahoma Health Sciences Center, 3Peggy and Charles Stephenson Cancer Center
Introduction: Circulating microRNAs are promising candidate biomarkers due to their cancer-specific expression.
However, breast cancer-specific microRNAs are difficult to selectively analyze due to the heterogenous population of
microRNAs in the blood. To overcome this challenge we are developing a molecular profile of microRNAs specifically
secreted from breast cancer cells. The key to identifying breast cancer-derived microRNAs relies on capturing and
analyzing the contents of exosomes, which selectively encapsulate microRNAs indicative of their cell of origin. However,
ways to selectively capture specific circulating exosome populations have not been developed. Furthermore, a complete
molecular profile of breast cancer exosomes or an evaluation of their potential as biomarkers has not been explored.
Methods: Breast ductal fluids and plasma samples were collected under an IRB approved protocol with informed patient
consent. Exosomes were isolated using the Exoquick reagent and verified by electron microscopy. Exosome protein
expression was measured by protein antibody array and western blot analysis. microRNA expression was measured by
qRT-PCR. Select exosome populations were isolated from plasma samples by immunoaffinity isolation utilizing
antibodies against CD63 and MUC1. Results: Exosomes were isolated from ductal fluid and plasma samples and
molecularly characterized. We successfully immunoprecipitated plasma exosomes with magnetic beads-conjugated to
MUC1 and CD63 antibodies. These exosomes were verified by electron microscopy and western blot analysis. More
interestingly, the microRNA expression patterns in the MUC1-precipitated plasma exosomes differed between breast
cancer patients and a control subject, indicating a potential new strategy to selectively analyze plasma breast cancer
microRNA levels. Conclusions: Primary breast exosomes can be successfully isolated from ductal fluid samples. MUC1
is a viable membrane protein candidate for selective capture of circulating breast cancer specific exosomes from the
plasma. These results suggest that selective capture and molecular analysis of breast cancer specific circulating exosomes
is a promising method for breast cancer biomarker development.
Funding: Funding: Oklahoma Clinical and Translational Science Institute and the Peggy and Charles Stephenson Cancer
Center
127 Abstract #104
A NOVEL MONOCLONAL ANTIBODY TO THE HLA OF CISPLATIN RESISTANT OVARIAN CANCER CELLS
Authors: Saghar Kaabinejadian1, Andrea Patterson1, Wilfried Bardet1, Kenneth Jackson1, Curtis McMurtrey1, Timea
Wichner2, Oriana Hawkins2, Jon Weidanz2, William Hildebrand1 Affiliations: 1. Microbiology and Immunology,
University of Oklahoma HSC, Oklahoma City, OK, United States. 2. Immunotherapeutics and Biotechnology, Texas Tech
University HSC, Abilene, TX, United States.
Cisplatin is widely used as a chemotherapeutic drug in the treatment of ovarian cancer. Resistance to cisplatin occurs in
about one-third of women during the primary course of treatment. We hypothesized that the HLA class I of cisplatinresistant ovarian cancer cells presents peptides distinct to these cells as compared to sensitive cells and that HLA/peptide
complexes unique to cisplatin-resistant cells would be valuable targets for immunotherapeutic intervention. To identify
the peptides that are uniquely presented by cisplatin-resistant ovarian cancer cells, the intrinsic cisplatin-resistant cells
(SKOV3) and sensitive cells (A2780, OV90, FHIOSE) were used in comparative mass spectrometry. Peptide sequences
distinct to cisplatin-resistant cells were identified including a peptide (VMF11) derived from thioredoxin interacting
protein (TXNIP) that was present in abundance in SKOV3. Next a T cell receptor mimic monoclonal antibody (RL41A)
against A*02:01/VMF11 complex was generated. The specificity and affinity of RL41A toward VMF11/A*02:01
complex was shown by staining peptide-pulsed T2 cells and surface plasmon resonance respectively. Staining of ovarian
cancer cells by flowcytometry also showed that RL41A was able to only stain cisplatin-resistant cells and not the sensitive
ones. We therefore report the successful development of a monoclonal antibody that represents an attractive candidate for
further validation using cisplatin-resistant and sensitive primary ovary tissues.
Funding:
128 Abstract #105
THE POLYCYSTIN COMPLEX MEDIATES WNT/CA2+ SIGNALING
Seokho Kim1,6, Hongguang Nie1,4,6, Vasyl Nesin1, Uyen Tran2, Patricia Outeda3, Chang-Xi-Bai1,5, Jacob Keeling1, Dipak
Maskey1, Terry Watnick3, Oliver Wessely2 & Leonidas Tsiokas1,7 1Department of Cell Biology, University of Oklahoma
Health Sciences Center, 975 NE 10th Street, Oklahoma City, OK 73104, USA 2Department of Cellular and Molecular
Medicine, Cleveland Clinic, 9500 Euclid Avenue/NC10, Cleveland, OH 44195, USA 3Division of Nephrology, Baltimore
PKD Research and Clinical Core Center, University of Maryland School of Medicine, 655 West Baltimore Street,
Baltimore, MD 21201, USA 4Institute of Metabolic Disease Research and Drug Development, China Medical University,
Liaoning Shenyang, 110001 China (H.N) 5Department of Advanced Research on Mongolian Medicine, Research Institute
for Mongolian Medicine, Inner Mongolia Medical University, Hohhot 010110, Inner Mongolia, China (CB) 6These
authors contributed equally to this work.
WNT ligands induce Ca2+ signaling on target cells. PKD1 (Polycystin 1) is considered an orphan, atypical G protein
coupled receptor complexed with TRPP2 (Polycystin 2 or PKD2), a Ca2+-permeable ion channel. Inactivating mutations in
their genes cause autosomal dominant polycystic kidney disease (ADPKD), one of the most common genetic diseases.
Here, we show that secreted WNTs bind to the extracellular domain of PKD1 and induce large whole cell currents and
Ca2+ influx dependent on TRPP2, but independent of Frizzled (FZD) receptors. Pathogenic PKD1 or PKD2 mutations that
abrogate complex formation, compromise cell surface expression of PKD1, or diminish TRPP2 channel activity suppress
activation by a WNT protein. Pkd2-/- fibroblasts lack WNT-induced Ca2+ currents and are unable to polarize during
directed cell migration. In Xenopus embryos, PKD1 acts independently of FZD8, but within the same pathway with
Disheveled 2 to preserve normal kidney tubulogenesis. These data define PKD1 as a new class of WNT (co)receptors and
implicate defective WNT/Ca2+ signaling as one of the causes of ADPKD.
Funding: This work was supported by grant number 81270098 from NSFC (HN), DK080745 from NIH (OW), DK59599
from NIH (LT), Oklahoma Center for the Advancement of Science and Technology (LT), and the John S. Gammill
Endowed Chair in Polycystic Kidney Disease (LT).
129 Abstract #106
CRITICAL INTERACTIONS OF PEPTIDE LIGANDS WITH THE EXTRACELLULAR DOMAIN OF
CALCITONIN AND AMYLIN RECEPTORS: IMPLICATIONS FOR PEPTIDE RECOGNITION MECHANISMS
OF CLASS B GPCRS
Sang-Min Lee and Augen A. Pioszak Department of Biochemistry and Molecular Biology, University of Oklahoma Health
Sciences Center
Introduction: An endogenous peptide calcitonin regulates calcium homeostasis by activating the calcitonin receptor
(CTR). Receptor activity modifying proteins (RAMPs) interact with CTR and form the selective receptor for an
endogenous peptide amylin. Amylin is co-secreted with insulin and has shown anti-diabetic effects. Although an amylin
derivative pramlintide is used to treat diabetes patients, lack of structural understanding on peptide recognition has
hindered rational drug design that leads to more desirable selectivity. Here, we investigate the structural mechanisms of
selective peptide recognition at CTR and the amylin receptor. Methods: Since the extracellular domains (ECD) of RAMPs
and CTR are reported critical for peptide interaction, we established DNA constructs for human CTR ECD alone and
RAMP-CTR ECD complex. These DNA constructs were transiently transfected into mammalian cells for large-scale
protein production, and the expressed proteins were purified. Molecular interactions were investigated with salmon
calcitonin (sCT) and an amylin receptor antagonist peptide AC413 that show high affinity to CTR and the amylin
receptor, respectively. Multiple truncated peptides and alanine substitution mutants were also tested in binding assays.
Results: AC413 showed binding affinity to the RAMP-CTR ECD complex significantly stronger than to CTR ECD alone,
whereas sCT affinity to CTR ECD alone was higher than to the RAMP-CTR ECD complex. This suggested that the
purified proteins recapitulated the peptide selectivity of the amylin receptor and CTR. We also identified the minimal
fragments of sCT and AC413 needed for the receptor ECD binding as well as specific peptide residues that contribute to
the receptor ECD interactions. Conclusion: Our results provide insights into peptide recognition mechanisms of CTR and
the amylin receptor. The structural basis for selective peptide binding will be further investigated using X-ray
crystallography in future studies.
Funding: NIH grant R01GM104251 and a training grant from Harold Hamm Diabetes Center.
130 Abstract #107
AT2R AUTOANTIBODIES BLOCK ANGIOTENSIN II AND AT1R AUTOANTIBODY-INDUCED
VASOCONSTRICTION
Hongliang Li, Campbell Liles, Vineet Veitla, Xichun Yu,† David C. Kem† Endocrinology & Heart Rhythm Institute,
Department of Medicine (C.L., H. L., V.V., X.Y., D.C.K.); University of Oklahoma Health Sciences Center and Veterans
Affairs Medical Center, Oklahoma City, OK † X. Yu and D. Kem served as co- mentors.
Activating autoantibodies to the angiotensin type 1 receptor (AT1R) second extracellular loop (ECL2) are associated with
hypertensive disorders. The angiotensin type 2 receptor (AT2R) is known to counter-regulate the actions of AT1R. Till
now, there have been no reports of activating autoantibodies directed toward the AT2R-ECL2. We investigated whether
AT2R autoantibodies produced in immunized rabbits will activate AT2R and suppress the vasopressor responses to
angiotensin II (Ang II) and AT1R-activating autoantibodies. Five rabbits were immunized with a peptide corresponding to
the AT2R-ECL2. After immunization, all animals developed high AT2R-ECL2 antibody titers. Rabbit anti-AT2R sera did
not produce any direct effect on arteriolar diameter in a rat cremaster arteriole assay; however, when co-perfused with
Ang II or AT1R-activating autoantibodies, the anti-AT2R sera completely blocked their contractile effects. Rabbit antiAT2R sera recognized a predominant sequence near the N-terminus of the second extracellular loop of AT2R. A decoy
peptide based on this sequence effectively reversed the opposing effect of the anti-AT2R sera on Ang II-induced
contraction of rat cremaster arterioles. A similar blockade of the anti-AT2R sera effect was observed with the AT2R
antagonist PD 123319. Rabbit anti-AT2R sera reacted specifically with AT2R. No cross-reactivity with AT1R was
observed. Blood pressure did not change in immunized animals. However, the pressor responses to incremental Ang II
infusions were blunted in immunized animals. In conclusion, we are the first to demonstrate AT2R-ECL2 autoantibodies
produced in immunized rabbits have the ability to activate AT2R and counteract the AT1R-mediated vasoconstriction.
These autoantibodies provide useful and selective tools for study of their roles in blood pressure regulation and possible
therapeutic intervention.
Funding: AHA Postdoctoral Fellowship (HL), VAMC Merit Review, R01 HL056267, NIH grant
GM092238/U26IHS300412, and Oklahoma NARCH Student.
131 Abstract #108
IGF-1 REGULATES LIPID BIOSYNTHETIC PATHWAYS IN HIPPOCAMPUS.
Sreemathi Logan1,3, Jessica Sanders1,3, Nicole Ashpole1,3, Daniel Owen1,3, Julie Farley1,3, Richard Brush2, Robert E.
Anderson2, William E. Sonntag1,3 Depts of Geriatric Medicine1 and Opthalmology2, Reynolds Oklahoma Center on
Aging3, University of Oklahoma Health Sciences Center, Oklahoma City, OK.
Cognitive decline is a critical issue as individuals age. Several aspects of cognitive function are known to be impaired
with increasing age including (but not limited to) spatial learning and memory. For aging subjects who experience
cognitive deficits, the reduction in function can be disabling and decrease health-span and independence. Studies suggest
that age-related cognitive decline is reversible but the etiology for the decline remains unknown, thereby limiting the
development of effective therapeutics. Lipids have been shown to mediate critical aspects of cellular function. Changes in
fatty acid metabolism and subsequent alterations in the lipid milieu have been reported in aging and age-related
neurodegenerative diseases including Alzheimer’s and Parkinson’s disease. Modulation of lipid biosynthesis has also been
implicated in cellular stress resistance and longevity. Insulin-like growth factor-1 (IGF-1) is a major neurotrophic
hormone levels of which decline with age and are associated with impairments in learning and memory. Recent data
indicate that levels of short chain monounsaturated fatty acids (scMUFA) are positively correlated with longevity. Our
hypothesis is that scMUFA synthesis is neuroprotective in the aging brain and retards cognitive impairment; factors, such
as IGF-1, that reverse the age-related decline in cognitive function act, at least in part, by altering lipid metabolism and
synthesis of scMUFAs. We have shown in primary neuronal cultures that IGF-1 induces the expression of enzymes that
mediate the synthesis of scMUFA, namely ELOVL6 elongase and sterol CoA desaturase-1 (SCD-1). Pharmacological
inhibition of IGF-1 signalling attenuates the expression of these enzymes in cultured neurons. Mice deficient in circulating
IGF-1 show significant changes in the expression of these enzymes and synthesis of these scMUFA in the hippocampus.
By understanding the specific role of these fatty acids in brain function and its regulation by IGF-1, we expect to develop
metabolic targets of therapeutic potential to avert or delay age-related cognitive impairment.
Funding: NIA AG37847
132 Abstract #109
INHIBITION OF THYROID HORMONE RECEPTOR PROTECTS CONE PHOTORECEPTORS IN RETINAL
DEGENERATION
Hongwei Ma1, Michael Butler1, Joshua Belcher1, T. Michael Redmond2, Thomas S. Scanlan3 and Xi-Qin Ding1
1
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK; 2Laboratory of
Retinal Cell and Molecular Biology, National Eye Institute, Bethesda, MD; 3Department of Physiology and
Pharmacology, Oregon Health & Science University, Portland, OR
Introduction: Thyroid hormone (TH) signaling regulates cell proliferation, differentiation, and apoptosis. In the retina, TH
signaling plays a central role in cone opsin expression and patterning. We previously showed that suppressing TH
signaling by anti-thyroid treatment preserves cones in mouse models of retinal degeneration. This work investigates
whether inhibition of TH receptors (TRs) affects cone viability in retinal degeneration. Methods: The cone precursor Weri
RB-1 cell line was used to determine the antagonistic activity of the TR antagonists, NH3 and 1-850, on T3-induced Mopsin expression. Rpe65-/- mice were treated with NH3 and 1-850 via intravitreal injection to determine whether treatment
with TR antagonists protects cones. Cone survival was evaluated by examining cone density on retinal whole mounts and
retinal sections using immunohistochemical approaches. Results: Treatment with T3 increased M-opsin expression in
Weri RB-1 cells. The effects of T3 treatment were inhibited by NH3 and 1-850 in a dose-dependent pattern. Cone density
was increased by about 35% in Rpe65-/- mice treated with NH3 or 1-850. Conclusion: We show that treatment with TR
antagonists improved cone survival in cone degeneration model mice. Our findings suggest that suppression of TR
signaling locally in the retina may represent a novel strategy for retinal degeneration management.
Funding: This work was supported by grants from the National Eye Institute (P30EY12190, R01EY019490,
R21EY024583, and T32EY023202) and the Foundation Fighting Blindness.
133 Abstract #110
CELL CYCLE-DEPENDENT UBIQUITYLATION AND DESTRUCTION OF NDE1 BY CDK5-FBW7 REGULATES
CILIARY LENGTH
Dipak Maskey1, Matthew C. Marlin2, Seok-Ho Kim1, Sehyun Kim1, 3, Guangpu Li2, and Leonidas Tsiokas1 1Department
of Cell Biology, University of Oklahoma Health Sciences Center, 975 NE 10th Street, Oklahoma city, OK, 73104, USA
2
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, 975 NE 10th Street,
Oklahoma City, OK, 73104
Primary cilia start forming within the G1 phase of the cell cycle and continue to grow as cells exit the cell cycle (G0).
They start resorbing when cells re-enter the cell cycle (S phase) and are practically invisible in mitosis. The mechanisms
by which cilium biogenesis and disassembly are coupled to the cell cycle are complex and not well understood. We
previously identified the centrosomal phosphoprotein NDE1, as an inhibitor of ciliogenesis and showed that its levels
inversely correlate with ciliogenesis. Here, we identify the tumor suppressor FBW7 as the E3 ligase that mediates the
destruction of NDE1 upon entry into G1. CDK5, a kinase active in G1/G0 primes NDE1 for FBW7-mediated proteolysis.
Cells depleted of FBW7 or CDK5 show enhanced levels of NDE1 and a reduction in ciliogenesis, which is corrected in
cells depleted of both FBW7 or CDK5 and NDE1. These data show that cell cycle-dependent mechanisms can control
ciliogenesis through a CDK5-FBW7-NDE1 pathway.
Funding: This work was supported by PBBEP3-141439 from the Swiss National Science Foundation (DM); GM074692
from NIH (GL); DK59599 from NIH, Oklahoma Center for the Advancement of Science and Technology, and the John S.
Gammill Endowed Chair in Polycystic Kidney Disease (LT).
134 Abstract #111
A NOVEL ONCOGENIC ENZYME, RCL, PROMOTES BREAST TUMORIGENESIS
Sangphil Oh, Sook Shin, Ralf Janknecht Department of Cell Biology, University of Oklahoma Health Sciences Center
RCL is an enzyme that cleaves the N-glycosidic bond of dNMP yielding deoxyribose 5’-monophosphate and a free base.
Conceivably, free bases may increase dNTP levels through the salvage pathway, while deoxyribose 5’-monophosphate
may stimulate angiogenesis. However, these predictions have remained untested and the physiological roles of RCL
unexplored. Here we found that RCL is overexpressed in multiple cancer cell lines and human tumors, suggesting that
RCL has oncogenic potential. To investigate its role in vivo, we generated RCL-deficient mice and crossed them with
MMTV-HER2/Neu transgenic mice, which are known to develop breast tumors within one year. First, RCL expression
was highly upregulated in tumor but not normal mammary glands of HER2/Neu mice. Second, mice lacking RCL showed
delayed HER2/Neu-induced breast tumor onset, reduced tumor multiplicity and metastasis. Intriguingly, while murine
Lewis lung cancer cells expressing enzymatically inactive mutant RCL showed no gross defects in cell proliferation and
long term survival in vitro, their growth was suppressed and tumor vessel density decreased in mouse xenografts. This
suggests that RCL may affect tumorigenesis possibly by regulating tumor angiogenesis through the secretion of
deoxyribose, a conversion product of deoxyribose 5’-monophosphate. Altogether, our findings and its enzymatic function
suggest that RCL is an oncogenic enzyme and, therefore, could be exploited as a new drug target in the treatment of
especially breast cancer patients.
Funding: Startup funds to Dr. Ralf Janknecht
135 Abstract #112
PRE-INCUBATION POTENTIATES THE INHIBITION POTENCY OF DASATINIB AND VEMURAFENIB
TOWARD OATP1B1-MEDIATED TRANSPORT
Sonia Pahwa1, Khondoker Alam1, Sukyung Woo1 and Wei Yue1 1Department of Pharmaceutical Sciences, University of
Oklahoma Health Sciences Center College of Pharmacy, Oklahoma City, OK 73117
Introduction: Organic anion transporting polypeptides (OATP) 1B1 mediate hepatic uptake of many drugs (e.g. statins),
and is involved in clinically significant drug-drug interactions (DDIs). Dasatinib and vemurafenib are tyrosine kinase
inhibitors (TKI) used for targeted cancer therapy. Increased simvastatin exposure has been reported when co-administered
with dasatinib. Current study was designed to determine the time-dependent effects of dasatinib and vemurafenib on
OATP1B1-mediated transport and to elucidate potential mechanism(s) underlying such effects. Methods: OATP1B1mediated transport of [3H]estradiol-17β-glucuronide (E217G) (1 µM, 2 min) was determined in HEK293 cell line overexpressing OATP1B1 or FLAG-tagged OATP1B1 in the presence of vehicle control or inhibitors (dasatinib, vemurafenib
and positive control rifampicin) with or without 1-hour pre-incubation with inhibitors. The half-maximal inhibitory
concentration (IC50) were estimated with Phoenix WinNonlin and R-values calculated. Surface biotinylation assay was
conducted to compare surface levels of OATP1B1 after 1 h pre-incubation with vehicle control, dasatinib and
vemurafenib in HEK293-FLAG-OATP1B1 cells. Results: In HEK293-OATP1B1 cells, pre-incubation step resulted in a
2.7- and 1.6-fold decrease in IC50 values for dasatinib and rifampicin, respectively. Pre-incubation did not affect IC50 of
vemurafenib towards OATP1B1 inhibition, however, resulted in a 1.7- fold increase in Imax . The R-values for dasatinib
and vemurafenib are close to or greater than the FDA recommended cut-off of 1.25. In HEK293-FLAG-OATP1B1 cells,
pre-treatment with dasatinib and vemurafenib (10 µM, 1 h) significantly decreased [3H]E217G accumulation to 47.54 ±
2.04% and 34.39 ± 3.02% of the vehicle control, respectively, and reduced surface levels of FLAG-OATP1B1 to 0.61 ±
0.2 and 0.13 ± 0.08 fold of control, respectively (mean ± range, n=2). Conclusions: Pre-incubation step potentiates the
inhibition potency of dasatinib, vemurafenib and rifampicin toward OATP1B1. OATP-mediated DDIs may occur when
dasatinib or vemurafenib is co-administered with OATP substrates (e.g. statins). Decreased surface levels of OATP1B1 is
involved in the time-dependent inhibition of OATP1B1 by dasatinib and vemurafenib.
Funding: Supported by NIH R01 GM094268
136 Abstract #113
IL-24 REGULATES AKT BY INHIBITING THE HIGH MOBILITY GROUP (HMG) A1/MIR222 SIGNALING
NODE IN LUNG CANCER CELLS
Janani Panneerselvam1,3, Akhil Srivastava 1,3, Ranganayaki Muralidharan1,3, Qi Wang 1,3, Anupama Munshi 2,3, and
Rajagopal Ramesh1,3,4* Department of 1Pathology, 2Radiation Oncology, 3Stephenson Cancer Center, and 4Graduate
Program in Biomedical Sciences, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104,
USA.
High mobility group A1 (HMGA1), a member of non-histone chromosomal proteins commonly referred to as
architectural transcription factor, is involved in the transcription of various genes involved in cell growth and survival.
Overexpression of HMGA1 in several cancer types including human lung cancer has been shown to be associated with
tumor progression and metastasis. Further, inhibition of HMGA1 expression has been shown to suppress tumor cell
proliferation and growth suggesting HMGA1 is molecular target for cancer therapy. A recent study showed HMGA1
activated AKT function by reducing the activity of the protein phosphatase, PPPR2A via the oncogenic micro (mi) RNA
222. Recently we demonstrated that interleukin (IL)-24, a novel tumor suppressor/cytokine inhibited AKT in lung cancer
cells. However, the molecular mechanism of AKT inhibition by IL-24 is not known. In the present study we hypothesized
that IL-24-mediated AKT inhibition involved the HMGA1/miR222 axis. To test our hypothesis we conducted in vitro
studies using human H1299 lung tumor cell line that was stably transfected with a tetracycline-inducible plasmid vector
carrying the human IL-24 cDNA, hereafter referred to as H1299-IL24. Upon addition of doxycycline (Dox; 1µg/ml),
H1299-IL24 cells were induced to express IL-24 protein. Induction of IL-24 expression resulted in a marked reduction in
HMGA1 protein and mRNA levels compared to control cells. Further, IL-24 reduced miR-222-3p and -5p levels as
determined by qRT-PCR. Associated with HMGA1 and miR-222 inhibition was a marked increase in PPP2R2A with
concomitant decrease in phosphorylated AKT T308/S473 expression. Further, siRNA-mediated knockdown of HMGA1 in
combination with IL-24 significantly reduced AKT T308/S473 protein expression and greatly reduced cell migration and
invasion when compared to individual treatments alone. In conclusion our study results demonstrate that IL-24 inhibits
AKT via the HMGA1/miR222 signaling node in human lung cancer cells.
Funding:
137 Abstract #114
ANTHRAX LETHAL TOXIN SUPPRESSES IL-22 PRODUCTION IN THE 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 bioweapon. B.
anthracis requires secreted virulence factors called lethal toxin to gain entry in the host by disrupting the barrier at the
mucosal surfaces such as skin, gut and the lungs. Lethal toxin impairs host innate and adaptive immune systems by
disrupting MAP kinase pathways. 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
protective antigen, a component of lethal toxin required for toxin entry into the host cell, bound to ILC3s as detected by
flow cytometry analysis. Importantly, lethal toxin decreased basal and IL-23 stimulated IL-22 production in splenocytes
from Rag-1 knock out animals (which have increased ILC3s). The decrease in ILC3-IL-22 production by lethal toxin was
dose and time dependent. Lethal toxin decreased IL-22 production in an ILC3 cell line and in human tonsillar
lymphocytes, suggesting conserved actions in mice and humans. Intracellular cytokine staining confirmed IL-22
production was indeed by ILC3s. Our data suggest that B. anthracis can modulate the function of ILC3s to favor its
pathogenesis and thus understanding the mechanism of how B. anthracis modulates ILC3 function may lead to therapeutic
interventions for deadly anthrax infections.
Funding: Pilot project funding, Subaward No. A195F under U19 AI062629-10 to K. Mark Coggeshall, Ph.D. (OMRF),
National Institutes of Health
138 Abstract #115
INTRACELLULAR SURVIVAL OF E. FAECALIS IN MACROPHAGES
Jun Zou, Nathan Shankar Department of Pharmaceutical Sciences, University of Oklahoma Health Sciences Center
Enterococcus spp. are commensal bacteria residing in the gastrointestinal tract of mammals but can cause serious
antibiotic-resistant opportunistic infections through bacterial translocation from the intestinal lumen to extraintestinal
sites. Many strains of E. faecalis have been reported to be capable of surviving within macrophages for extended periods
of time, which may increase the possibility of leading to systemic spread of the infection; however the exact mechanisms
involved are unknown. Here we analyzed the intracellular trafficking of enterococcus, and found that enterococci
containing vacuoles could resist fusion with lysosome after phagocytosis by macrophages. Characterization of
enterococcus containing vacuole during infection of macrophage by TEM revealed that enterococci were surrounded by a
single membrane vacuole, which may preclude a role for autophagy in elimination of intracellular enterococci. By
employing cells transfected with RFP-LC3 plasmid and infected with GFP labeled E. faecalis, we observed there was a
decrease of RFP-LC3 positive punctate dots in macrophages infected with E. faecalis compared with uninfected cells and
no colocalization of RFP-LC3 with E. faecalis during enterococcus infection. Examination of the conversion from LC3-I
to LC3-II by Western blot showed that E. faecalis could trigger inhibition of the production of LC3-II during infection,
indicating that enterococci could decrease the autophagy level in the infected macrophages. Furthermore, we found that
during enterococcal infection, macrophages could produce ROS and RNS, which may play an important role for killing
and elimination of enterococci. Further studies are in progress to understand the molecular mechanisms involved in the
inhibition of autophagy. The results from these studies may offer a significant new approach to therapeutic intervention
for the treatment of serious enterococcal infections.
Funding: This work was supported, in part, by a Presbyterian Health Foundation seed grant (NS)
139 Abstract #116
SNARES INTERACT WITH RDS/ROM-1 DURING CONVENTIONAL AND UNCONVENTIONAL OUTER
SEGMENT TARGETING
Rahel Zulliger1, Shannon M. Conley1, Maggie L. Mwoyosvi1, Michael W. Stuck1, Seifollah Azadi1,2, Muna I. Naash1
1
Department of Cell Biology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, 73104, United
States of America. 2Oklahoma Medical Research Foundation, University of Oklahoma Health Sciences Center, Oklahoma
City
Purpose: Peripherin-2, also known as RDS (retinal degeneration slow), is an important structural protein in the
photoreceptor outer segment (OS), and mutations in it cause several severe forms of retinal degeneration. Together with
its non-glycosylated homolog ROM-1 (rod outer segment protein 1), RDS is synthesized in the inner segment and then
trafficked into the outer segment where it functions in non-covalently linked tetrameric complexes as well as covalently
linked larger complexes. However, in spite of the fact that many RDS mutations interrupt OS targeting, little is known
about the processes which govern RDS OS targeting. Materials and methods: To investigate possible trafficking partners
of RDS and ROM-1, we analyzed the bound fraction of RDS and ROM-1 by mass spectrometry. Potential candidates
were confirmed with binding assays. Immunohistochemistry (IHC) was performed to show co-localization in the retina
and proximity ligation assay (PLA) was conducted to show interactions in situ. Results: We used biochemical and cell
biological techniques to show that RDS/ROM-1 interact with two proteins known to be involved in OS targeting of other
proteins (such as rhodopsin), namely Syntaxin 3B and SNAP-25 from the SNARE family. The interaction was initially
seen with mass spectrometry and was confirmed by co-immunoprecipitation in in vivo and in vitro. The IHC showed a
possible site of interaction at the apical edge of the inner segment and binding between Syn3B and RDS in this region was
confirmed in situ using the PLA. We further show that Syn3B interacts with both covalently linked and non-covalently
linked RDS complexes. Conclusion: Our data suggest that the RDS/ROM-1 complexes are trafficked in vesicles that
utilize Syn3B and SNAP-25 for fusion with the inner segment membrane during OS targeting. This OS trafficking
process also involves other proteins which will be the focus of future research.
Funding: Funding: R01EY10609
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