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2009 Appalachian Student Research Forum April 9, 2009 presented by East Tennessee State University Millennium Centre • Johnson City, TN Table of Contents Schedule of Events ....................................................................... 1 Keynote Presentation .................................................................... 2 ASRF Task Force Members ......................................................... 4 ASRF Judges ................................................................................ 5 ASRF Sponsors............................................................................. 6 Student Artwork on Display ......................................................... 7 Exhibitors...................................................................................... 8 Abstracts (poster presentations) Division I – Undergraduates Arts & Humanities and Biomedical Sciences ............................9 Natural Sciences and Mathematics...........................................14 Social & Behavioral Sciences...................................................21 Division II – Graduate Students, 1 – 2 years Arts & Humanities and Biomedical Sciences ..........................35 Natural Sciences and Mathematics...........................................43 Social & Behavioral Sciences...................................................51 Division II – Graduate Students, more than 2 years ........................56 Division IV – Medical Students .......................................................65 Divisions V and VI – Residents & Post-Doc Fellows .....................69 Abstracts (oral presentations) Division I – Graduate Students ........................................................76 Division II – Residents & Post-Doctoral Fellows............................85 2008 Award Winners.................................................................. 91 TBR No. 170‐105‐08 .15M Schedule of Events April 8 – 9, 2009 The Centre at Millennium Park x Johnson City, TN Wednesday, April 8, 2009 1:00 pm – 4:30 pm Check-in and Poster Set-Up Ballroom Thursday, April 9, 2009 8:00 am – 12:00 pm Poster Judging Ballroom 8:00 am – 4:00 pm Poster Viewing Ballroom 8:00 am – 12:00 pm Vendor Exhibition 2nd floor 9:30 am – 12:00 pm Oral Presentations: Graduate Students Residents & Post-Doctoral Fellows Room 130 Auditorium 12:00 – 1:30 pm BUFFET LUNCH 2nd floor 1:30 – 2:30 pm Keynote Address Auditorium “Novel Aspects of HGF/HGFR Axis in Tissue Homeostasis and Disease” Reza Zarnegar, Ph.D. Tenured Professor, Department of Pathology, University of Pittsburgh 2:30 – 3:00 pm Awards Ceremony Auditorium ** Posters should be removed by 4:30 pm** 2009 Appalachian Student Research Forum Page 1 Keynote Presentation “Novel Aspects of HGF/HGFR Axis in Tissue Homeostasis and Disease” Reza Zarnegar, Ph.D. Tenured Professor Department of Pathology University of Pittsburgh • • • • • • 1996-2000 Associate Professor (with tenure), Department of Pathology, University of Pittsburgh 1991-1995 Assistant Professor, Department of Pathology, University of Pittsburgh 1989-1991 Research Assistant Professor, Department of Pathology, Duke University Medical Center 1987 Ph.D. in Biochemistry, East Tennessee State University M.S. in Microbiology, East Tennessee State University 1978 B.S. in Biology, University of Razi, Iran KEYNOTE ADDRESS ABSTRACT: Dr. Zarnegar is a Tenured Professor of Pathology and a member of the Division of Experimental Pathology. His laboratory is focused on determining the physiological and pathological role of the HGF/Met axis in normal tissue homeostasis and disease processes. Dr. Zarnegar discovered Hepatocyte Growth Factor (HGF) in the 1980’s and since then, has devoted his research efforts to understanding this important signaling factor. HGF and its cell surface tyrosine kinase receptor (Met) play an important role in tissue homeostasis, and the deregulation of HGF/Met axis has pathological consequences such as cancer. An in-depth understanding of the molecular mechanisms of HGF/Met function will not only shed light on normal tissue physiology but also on understanding the basis of its pathogenic effects and thus may provide opportunities for rational drug design to combat various diseases ranging from tissue degeneration to cancer. Through structural and functional studies, Dr. Zarnegar’s laboratory has discovered novel mechanisms by which the Met receptor regulates hepatocyte growth and liver homeostasis. Specifically, his lab found that, in normal liver, the extracellular domain (a.k.a. the ectodomain) of Met directly binds to Fas (a death promoting receptor) and acts as a natural antagonist of FasL thereby prohibiting Fas aggregation and cell killing. They have determined that Fas sequestration by Met is abrogated in human fatty liver disease hence making these livers vulnerable to cell death and development of NASH. The mechanism through which liver cells die is mainly via activation of the Fas axis. Unregulated heapto- 2009 Appalachian Student Research Forum Page 2 KEYNOTE ADDRESS ABSTRACT (cont.): cyte death occurs in a variety of hepatic diseases including hepatitis caused by chemical, viral or metabolic etiologies which may ultimately culminate in end-stage liver disease including liver cancer and hepatic failure requiring liver transplantation. Dr. Zarnegar’s group has mapped the interaction region in Met and Fas to a 12 amino acid region containing a core YLGA motif in Met and show that Met and its synthetic peptide derivatives prohibit FasL binding to Fas, Fas activation and hepatocyte death in vitro and in animal models of NASH. His lab is currently expanding these structure-function studies of Met and he will present his recent findings in this area. In a separate project, Dr. Zarnegar’s lab has discovered that, in human breast cancer, the HGF gene promoter is prone to deletion mutagenesis in a DNA element located 750 bp upstream from the transcription start site. This novel HGF promoter element consists of a mononucleotide repeat of 30As and is referred to as DATE (Deoxy Adenosine Tract Element). Mutation of DATE results in reactivation/lack of proper silencing of HGF gene expression in carcinoma tissues. Normally, the HGF gene is transcriptionally silenced in differentiated breast epithelial cells. Through functional studies, they showed that DATE mutation (shortening) has profound local and global effects on the HGF promoter region by modulating chromatin structure and DNA-protein interactions leading to constitutive activation of the HGF gene promoter in human breast carcinoma cells lines. Interestingly, they found that 51% (19/37) of African Americans and 15% (8/53) of Caucasians with breast cancer harbor the truncated DATE variant (25As or less) in their breast tumors and that the truncated allele significantly associates with cancer incidence and aberrant HGF expression. Dr. Zarnegar will also present these results in his seminar to the ETSU Department of Biochemistry on April 10, 2009. Relevant Recent Publications: 1. Wang, X. DeFrances, M.C., Dai, Y. Bell, A., Michalopoulos G.K. and Zarnegar, R. (2002). A novel mechanism of cell survival: sequestration of the death receptor Fas by direct physical association with the tyrosine kinase receptor Met. Molecular Cell 9: 411422. [Subject of a commentary entitled “A dual purpose anti-death agent” in the Research Roundup section of the Journal of Cell Biology, Volume 156:941, 2002]. 2. Zou, C., Ma, J., Wang, X, Guo, L., Zhu, Z., Stoops, J., Eaker, A.E., Johnson, C., Strom, S., Michalopoulos, G.K., DeFrances, M.C., and Zarnegar, R. Lack of Fas antagonism by Met in human fatty liver disease: The alpha chain of Met is a natural antagonist of FasL via a novel YLGA motif (2007). Nature Medicine. 13 (9): 1078-1085. [Subject of commentaries by Christian Trautwein and Daniela Kroy (2008) entitled “Peptide-Based NASH Therapy, A First Crucial Step Soon Clinical Reality?” Hepatology 47: 757-758] and by Susanne Strand and Peter R. Galle. entitled “Getting the Fat out of Met and Fas”. J. of Hepatology, 49: 479-482 (2008)]. 3. Ma, Jihong, Marie C. DeFrances, Chunbin Zou, Carla Johnson, Robert Ferrell, and Reza Zarnegar. Somatic Mutation and Functional Polymorphism of a Novel Regulatory Element in the HGF Gene Promoter Causes Its Aberrant Expression in Human Breast Cancer (2009). Journal of Clinical Investigations. 119 (3) March issue (Note: available on-line In February 2009 via the JCI site.) 2009 Appalachian Student Research Forum Page 3 ASRF Task Force Dr. Scott Champney, Chair Biochemistry & Molecular Biology Dr. Russ Brown Dr. Ranjan Chakraborty Psychology Health Sciences Ms. Dinah DeFord Dr. William Duncan ETSU McNair Program Vice Provost for Research Ms. Becky Fee Dr. Jay Franklin Office of the Vice Provost for Research Sociology & Anthropology Dr. Wayne Gillespie Dr. Lee Glenn School of Graduate Studies Professional Roles/Mental Health Nursing Dr. Gary Henson Dr. Dhirendra Kumar Physics, Astronomy & Geology Biological Sciences Dr. Foster Levy Dr. Louise Nuttle Undergraduate Research/ Biological Sciences Research and Sponsored Programs Dr. Rebecca Pyles Dr. Mike Ramsey Honors College/Biological Sciences Kinesiology, Leisure& Sport Sciences Dr. Mitch Robinson Ms. Barbara Sucher Biomedical Graduate Program Continuing Medical Education Ms. Carole Thomason Research and Sponsored Programs 2009 Appalachian Student Research Forum Page 4 Judges Poster Presentations Division I – Undergraduates Division III – Grad Students – 2+ Yrs. Arts & Humanities and Biomedical Sciences Biomedical Sciences, Natural Sciences & Yu-Lin Jiang Mathematics, and Social & Behavioral Dhirendra Kumar Sciences Natural Sciences and Mathematics Arpita Nandi Phillip Musich Tricia Metts Douglas Thewke Social and Behavioral Sciences Division IV – Medical Students Gary Henson Mitchell Robinson Thomas Jones Robert Schoborg Leslie McCallister Lev Yampolsky Division V & VI– Residents and Post-Doc Fellows and Case Histories Division II – Graduate Students 1-2 Yrs Sharon Campbell Arts & Humanities and Biomedical Sciences Michelle Duffourc David G. J. Young Ahmad Zulfiqar Natural Sciences and Math James R. Stewart Oral Presentations Terry Elizabeth Lancaster Residents and Post-Doctoral Fellows Behavioral and Social Sciences Doris Hubbs Andrea D. Clements Deborah Mills Thomas Laughlin Ken Olive Graduate Students Beth Bailey David Currie Chris Newell 2009 Appalachian Student Research Forum Page 5 Forum Sponsors Philip D. Cooper Memorial Research Trust Fund, Inc. East Tennessee State University The Office of Research & Sponsored Pgms The School of Graduate Studies The Ronald E. McNair Post Baccalaureate Achievement Program The Office of the Vice President for Health Affairs The College of Arts & Sciences Office of the Dean Department of Biological Sciences Department of Chemistry Department of Mathematics Department of Physics and Astronomy Department of Sociology & Anthropology The College of Business & Technology Office of the Dean The College of Clinical & Rehabilitative Health Sciences Office of the Dean 2009 Appalachian Student Research Forum The Clemmer College of Education Office of the Dean Department of Human Development and Learning The Quillen College of Medicine Office of the Dean Department of Anatomy & Cell Biology Department of Biochemistry & Molecular Biology Department of Microbiology Department of Obstetrics/Gynecology Department of Pathology Department of Pharmacology Department of Physiology Department of Psychiatry & Behavioral Sciences Department of Surgery The College of Nursing Office of the Dean The College of Public Health Office of the Dean Page 6 Special Thanks The 2009 Student Research Forum Task Force would like to recognize and thank The East Tennessee State University Center for Community Outreach and Family Services for their generous support of this year’s Student Research Forum, without which this event would not have been possible. Student Artwork on Display in the Atrium Artist Jessica Augier Kyle Blauw Reese Chamness Brian Godwin Jake Ingram Mikaela Minahan Joseph Reynolds Sarah Olivo Medium Lithograph and Oil Print Bronze Pastel Pastel Textiles Photography Acrylic and Oil 2009 Appalachian Student Research Forum Department Art & Design Art & Design Art & Design Art & Design Art & Design Art & Design Art & Design Women's Studies Advisor Ms. Mia Gerard Ms. Mira Gerard Ms. Catherine Murray Mr. Ralph Slatton Ms. Anita DeAngelis Ms. Deborah Stephens Mr. Michael Smith Dr. Amber Kinser Page 7 Exhibitors 2009 Appalachian Student Research Forum Page 8 Poster Presentations Division I – Undergraduates Arts & Humanities and Biomedical Sciences CELL-ADHESION MOLECULES (CAM’S) AND ASSOCIATED PROTEINS ARE DOWN-REGULATED IN CHLAMYDIA TRACHOMATIS AND CHLAMYDIA MURIDARUM INFECTED HELA CELLS Christopher Daniels, McNair Scholar and Dr. Robert Schoborg, Department of Microbiology, Quillen College of Medicine, East Tennessee State University, Johnson City, TN Chlamydia trachomatis, the most common bacterial STD in the US, can cause chronic infections, making management a major problem. Infection with C. trachomatis, while often asymptomatic, can cause serious sequelae such as ectopic pregnancy, salpingitis, and infertility. Chlamydia infection disrupts multiple host cell functions, including cellcell interactions. Using Western blot analysis, our laboratory has demonstrated that accumulation of host cellular nectin-1, α-catenin, and β-Catenin, proteins involved in forming cell-cell junctions, are significantly decreased in C. muridarum infected genital epithelial cells. We also found significant decrease in nectin-1 and β-catenin in C. trachomatis serovar E infected genital epithelial cells. Thus, we speculate that Chlamydia mediated down-regulation of adhesion molecules and associated proteins may disrupt the junctions between host epithelial cells, aiding chlamydial spread in the human genital tract. THE SYNTHESIS OF TAK-242 (ETHYL 6-[(2-CHLORO-4FLUOROPHENYL)SULFAMOYL]CYCLOHEX-1-ENECARBOXYLATE: A SMALLMOLECULE ANTISEPSIS AGENT James A. Fisher and Michael F. Wempe, Ph.D., Department of Chemistry and Department of Pharmacology, College of Medicine, East Tennessee State University, Johnson City, TN Sepsis may be denoted as a condition where pathogenic organisms, and/or their toxins, are present in the blood and/or tissues. Commonly present in intensive care patients, septic poisoning has been established as a leading cause of death. We sought to prepare TAK-242 ( Ethyl 6-[(2-chloro-4-fluorophenyl)sulfamoyl]cyclohex-1-enecarboxylate), a promising small-molecule antisepsis agent which has been reported to have very potent in 2009 Appalachian Student Research Forum Page 9 vitro and in vivo activity. The goal of our research was to produce an authentic sample of TAK-242, with subsequent studies by a group of colleagues within the Department of Surgery at Quillen College of Medicine. Our sample of TAK-242 possessed analytical data (i.e. 1H NMR, GC/MS, melting point, etc.) consistent with literature values and had biological activity at levels comparable to those reported in the literature. EXPRESSION OF HUMAN NEUTROPHIL ELASTASE IN K. LACTIS Haley Klimecki, Eliot Smith and David Johnson, Department of Biochemistry and Molecular Biology, James H. Quillen College of Medicine, East Tennessee State University,Johnson City, TN Human neutrophils contain the serine proteases Elastase, Cathepsin G, and Protease 3, which are packaged into cytoplasmic granules. Neutrophils are white blood cells that provide the body with a line of defense against foreign microorganisms by attacking and killing bacteria. Mutation of human neutrophil elastase (HNE) can cause neutropenia (a low neutrophil count) that results in susceptibility to infection. HNE is a powerful enzyme that can attack the elastin of the lung; consequently, genetic deficiencies of alpha-1 proteinase inhibitor protein in the blood result in emphysema because HNE released from neutrophils is free to degrade lung tissue. Recombinant HNE is not currently available and the enzyme must be isolated from human blood, which has inherent hazards. Additionally, the lack of recombinant HNE has prevented studies involving site–directed mutagenesis to study the intracellular processing of HNE near its C-terminal end where mutations have been found to result in neutropenia. The HNE DNA sequence was codon optimized for yeast and commercially synthesized. It was then fused with DNA for eGFP (enhanced green fluorescent protein) via a enterokinase cleavage site (D4K). This DNA construct was inserted into the kluyveromyces lactis (K. lactis) pKLAC1 vector, downstream of the alpha mating factor which directs proteins for secretion and sequenced to confirm its correct construction. Protease deficient YAP-3 and YPS-7 strains of K. lactis were transformed with pLAC1-eGFP-D4K-HNE. The gene integrates into the yeast genome upon transformation by electroporation via sequences in pKLAC1 that are homologous with the LAC4 gene that allows galactose utilization. K. lactis is not able to use acetamide as a nitrogen source, but the pKLAC1 vector contains the gene Aspergillus nidulans acetamidase (amdS) that allows only transformants to grow on plates with acetamide as the sole nitrogen source. Selected colonies will be transferred to both liquid and agar-based synthetic media with galactose to induce production of the eGFP-D4K-HNE fusion, and screened via flourescence microscopy for production of eGFP. Cell-free media from eGFP-positive liquid cultures will be assayed by SDS-PAGE and Western blotting with anti-HNE antibody to check for the presence of the HNE portion of the fusion protein. These media samples will also be purified by affinity chromatography and treated with enterokinase, which will remove the eGFP domain and activate the HNE. Eluted fractions will be tested for protein by spectrophotometry measuring absorbance at 280nm. The synthetic substrate Suc-AlaAla-Pro-Val-pNA will be used to assay for enzyme activity. HNE cleavage releases the bright yellow p-nitro-aniline product for calculation of Michaelis-Menten enzyme 2009 Appalachian Student Research Forum Page 10 kinetics. Subsequent studies will examine the effects of C-terminal mutations on processing, trafficking, and activity. Supported by National Heart, Lung and Blood Institute grant 1R15HL091770. DEPRESSION AND THE REGULATION OF BONE-MORPHOGENETIC PROTEIN7 (BMP7) AND OTHER GROWTH FACTORS Erich Lutz, Dr. Greg Ordway, and Attila Szebeni, Department of Pharmacology, College of Medicine, East Tennessee State University, Johnson City, TN During the past 50 years psychopharmacologists have attributed major depression to a decrease in basal levels of the monoamine neurotransmitters serotonin, norepinephrine, and possibly dopamine. This theory is known as the “monoamine hypothesis.” Current antidepressant medications are prescribed to try to restore these monoamine neurotransmitters to normal. These medications are highly effective at rapidly elevating concentrations of brain monoamine, but therapeutic effects are delayed three to six weeks. Due to this latency period researchers are further studying the biochemical mechanisms behind these antidepressant medications. Recently, researchers have focused efforts on understanding how antidepressants alter certain growth factors in the brain such as brain-derived neurotrophic factor (BDNF) and vascular endothelial growth factor (VEGF). These growth factors are proteins that regulate neuronal survival, neuroplasticity, and neurogenesis.2 Recent findings at the East Tennessee State University Quillen College of Medicine have discovered low levels of bonemorphogenetic protein-7 (BMP7) in brains from individuals who had major depression.3 BMP7 is a trophic factor that stimulates growth and development of monoamine neurons, but very little is known about how it is regulated in the brain. HUMAN BMP-7 TRANSCRIPTION START POINT IS DIFFERET IN ASTROCYTES THAN IN KIDNEYS: DETERMINATION VIA 5’ RACE Fritz Prohaska and Michelle M. Duffourc, Department of Pharmacology, College of Medicine, East Tennessee State University, Johnson City, TN Bone morphogenic protein-7 (BMP7 or Osteogenic protein-1) research is of rising importance as it has been shown to improve motor function in rat models of stroke when injected intravenously (Chang et al., 2003) and also shown to be neuroprotective/neuroreparative in rat models of Parkinson’s disease when BMP7 is used to pre-treat the brain (Harvey et al., 2004). We have previously observed that BMP7 is significantly decreased in the locus coeruleus of patients with major depression as compared to matched control patients and that in the mature brain, BMP7 is primarily expressed in astrocytes. In order to use BMP7 as a novel therapy for Parkinson’s, depression, and/or stroke in humans, we must first understand its regulation in astrocytes. 2009 Appalachian Student Research Forum Page 11 Using reverse-transcription PCR we determined that U373 and U138 human astrocytoma cell lines express BMP7. This suggests that the 5’flanking region of the BMP7 gene is not mutated in U373 or U138 cells as compared to primary human astrocytes. This also suggests that the transcriptional machinery for BMP7 expression is present, making these cells appropriate models for expression and study. Putative transcription start points (TSPs) for BMP7 in human kidney and U373 astrocytoma cells, as determined by 5’ RACE, were 433 bp and 309 bp upstream of the translation initiator codon, respectively. The difference between TSPs suggests transcriptional regulation is different in astrocytes than in kidney. The results for human kidney are in agreement with previously published results, thus validating our assay method. The 5’RACE products used for TSP determination will be sequenced to confirm our findings. The DNA sequence around the TSP for each cell type is TATA-less, but GC rich, consistent with a CpG island base promoter. Future studies will establish the minimal promoter region required to drive BMP7 expression and determine transcription factors that interact with the BMP7 promoter. These studies will contribute to our knowledge of how BMP7 expression is physiologically regulated in a mature, normal CNS. BMP7 is a well-documented neuroreparitive and neurotrophic factor; therefore, the aforementioned studies hold great importance for understanding and manipulating glial and neuronal responses to CNS insults. These studies will also aid investigations into the role of BMP7 in neurodegenerative and psychiatric diseases. EXPRESSION AND PURIFICATION OF RECOMBINANT HUMAN ENTEROKINASE LIGHT CHAIN IN KLUYVEROMYCES LACTIS Jessica Pugh, Eliot Smith, and David Johnson, Department of Biochemistry and Molecular Biology, James H. Quillen College of Medicine, East Tennessee State University,Johnson City, TN Human Enterokinase (EK; Enteropeptidase) is a highly specific serine protease produced by cells in the duodenum wall and starts the activation of the pancreatic digestive enzymes. Trypsinogen is activated by EK and active trypsin then activates other pancreatic protease zymogens. Enterokinase cleaves after Lys at sites preceded by four Asp residues (Asp-Asp-Asp-Asp-Lys~X). This property makes it an ideal reagent for cleaving specially constructed fusion proteins resulting in the protein of interest. Recombinant human enterokinase light chain (rhEKLC) was expressed using a yeast system known as Kluyveromyces lactis. Only the light chain was expressed because it has the protease activity. To achieve protein secretion, the gene of interest was cloned into the K. lactis vector pKLAC1 downstream of the α-mating factor domain, which directs the protein through the secretory pathway. The α-MF fusion protein then undergoes processing by signal peptidase in the endoplasmic reticulum, and αMF is removed by the Kex2 protease in the Golgi resulting in the secretion of the native form of the protein. The gene for enterokinase light chain was amplified via a PCR reaction using primers with a XhoI restriction enzyme site and Kex2 cleavage site added to the forward primer and a BglII restriction enzyme site added to the reverse primer. Restriction digests for the purified PCR product and the pKLAC1 vector were performed 2009 Appalachian Student Research Forum Page 12 using XhoI and BglII, and these were ligated to create the pKLAC1-rhEKLC vector. The sequence of pKLAC1-rhEKLC was confirmed by sequencing. pKLAC1-rhEKLC was linearized with SacII and used to transform K. lactis GG799 competent cells. For screening purposes, the transformants were grown up on Yeast Carbon Base media with 5mM acetamide as the only nitrogen source. The pKLAC1 plasmid has an amdS gene for acetamidase which converts acetamide into ammonium for use as a nitrogen source. Only positive transformants containing amdS grow on YCB acetamide plates. For the expression of enterokinase, pKLAC1-rhEKLC transformed K. lactis were grown up in synthetic WM9 media with galactose which induces the LAC4 promoter to produce enterokinase light chain. The yeast secreted active enterokinase light chain into the media as evidenced by increasing activity over five days using the synthetic substrate ZLys-thiobenzyl ester. Enterokinase light chain was purified by affinity chromatography on a Soybean Trypsin Inhibitor Fractogel Matrix. Further purification and characterization will be performed on the expressed enterokinase which will aid in experiments with other fusion proteins. Supported by the National Heart, Lung and Blood Institute via grant 1R15HL091770. QUALITATIVE ANALYSIS OF MOUSE OVARIAN TISSUE EXPOSED TO A MICROGRAVITY ENVIRONMENT Ian M Smith and Allan D Forsman, Ph. D. Department of Health Sciences, East Tennessee State University, Johnson City, TN The effects of space travel are relatively unexplored in regard to the impact on human physiology. An important step in determining any adverse effects is analysis of test animal data. With this in mind, we investigated the ovarian tissue of mice which were flown aboard the space shuttle Endeavour on NASA mission STS-118. The experiment consisted of three groups of animals: two sets of control animals and a single set of flight animals. Each set consisted of twelve individual mice. The flight animals were housed in the Commercial Biomedical Testing Module-2 (CBTM-2) over the twelve day flight conducted 8-21 of August, 2007. One set of control animals (baseline) were housed in standard cages at room temperature. The other set of control animals (ground control) were housed in a ground based CBTM-2, which was environmentally controlled to match the conditions aboard the shuttle Endeavour with a delay of twenty-four hours and subject to normal gravity. The ovarian tissue samples were paraffin embedded, sectioned using a microtome, mounted, stained using standard Hematoxylin and Eosin staining procedures, and cover-slipped. The gross morphology of the tissue was then qualitatively analyzed. The flight animals were compared to the baseline and ground control sets. The study indicates no significant morphological changes occur within ovarian tissue when exposed to a microgravity environment for twelve days or less. 2009 Appalachian Student Research Forum Page 13 BRAIN GLUCOSE UTILIZATION FOLLOWING CHRONIC MANGANESE EXPOSURE IN MALE SPRAGUE-DAWLEY RATS T. Jordan Smith, Department of Physics, King College, Bristol, TN; Noor Tantawy and Todd Peterson, Vanderbilt University Institute of Imaging Science, Vanderbilt University Medical Center, Nashville, TN; Vanessa Fitsanakis, Department of Biology, King College, Bristol, TN Manganese (Mn) is toxic at high concentrations, and prolonged exposure may result in manganism, a neurological disorder similar to Parkinson’s disease. Data suggest Mn accumulates in mitochondria, which may result in toxicity. Unfortunately, few studies have characterized neuronal changes resulting from low-level, chronic exposure. Using non-invasive μPET (micro positron emission tomography), we quantified brain glucose uptake, an indirect measure of cellular respiration, in male Sprague-Dawley rats. Rats were injected (iv) with isotonic saline (CN) or MnCl2 (3 mg/kg/week; TX), and routinely imaged using fluorodeoxyglucose (FDG), a radioactive μPET tracer. Each rat also underwent μCT (micro computed tomography) to aid in identification of brain nuclei in μPET images. Raw data was reconstructed using ordered subsets expectation maximization 2D (OSEM2D), and dose-corrected for each scan. μPET and μCT images were registered the Rubin Rat Brain atlas for analysis of various regions of interest (ROI; striatum, cortex, thalamus, cerebellum, hippocampus and whole brain). ROI were evaluated for standard uptake value (SUV) of FDG. By week 2, treated rats (n = 7) had decreased body mass (p < 0.001) compared to controls (n = 6). Both striatum (p = 0.0074) and hippocampus (p = 0.0072) were also smaller (by mass) in TX compared to CN. Whole brains from TX animals scanned during weeks 4-7 demonstrated a small trend towards increased FDG uptake (11%; p = 0.58), which increased to 17% (p = 0.40) by weeks 8-11. ROI values normalized to whole brain values, to identify areas responsible for this trend, showed increased striatal FDG uptake in weeks 4-7 (4.6%; p = 0.14) that was statistically significant by weeks 8-11 (11.5%, p = 0.00015). These data are consistent with other work suggesting preferential striatal Mn accumulation, with some distribution throughout the entire brain. More importantly, our data suggest that low-level chronic Mn exposure may lead to increased cellular metabolism, perhaps through mitochondrial uncoupling. Natural Sciences & Mathematics PATTERNS OF SPIDER SPACING IN SOCIAL SPIDER WEB WITH ATTENTION TO ONTOGENY AND DIURNAL RHYTHMS Angela E. Cowart, Thomas C. Jones, and Justin T. Peyton, Department of Biological Sciences, East Tennessee State University, Johnson City, TN and Department of Entomology, Ohio State University, Columbus, OH 2009 Appalachian Student Research Forum Page 14 Social spider Anelosimus studiosus is the only known species of social spider living in a temperate climate. It can be found living both socially in colonies containing multiple adult females and solitarily in colonies containing only one adult female and her offspring. This particular species has been shown to be more active at night. The goal of the study was to develop a method which can be used to determine if there is a pattern of spacing between individuals based on ontogeny and circadian rhythms. Both single and multi-female colonies were obtained prior to brood production and were placed in 16.0X1.0 cm round containers. They were maintained on a 12 hour light-dark cycle and photographed each week under light and dark conditions. These photographs were used to mark juveniles in order to determine each individual’s distance to its nearest neighbor. Averages taken for each colony were used to determine tendency towards clustering of randomization using a modified nearest neighbor analysis for confined space. It was expected that spacing of juveniles would increase with ontogeny and would increase during the dark cycle. The study was successful in developing a method to use in future studies describing daily cycles as well as in determining spacing difference based on ontogeny, circadian cycles, and sociality. FORMATION OF FORMALDEHYDE THROUGH HETEROGENEOUS REACTIONS OF OZONE AND D-LIMONENE ON ACTIVATED CARBON IN INDOOR AIR APPLICATIONS Shea Higney and Dr. Tricia Metts, Department of Environmental Health, East Tennessee State University, Johnson City, TN Poor indoor air quality (IAQ) has become a concern in the workplace as it can have a detrimental impact on worker comfort and health. Sick Building Syndrome (SBS) symptoms such as fatigue, sensory irritation, headaches and respiratory problems have been shown by recent studies to be associated with chemical reactions occurring on surfaces or in the air. The objective of this research is to investigate the reactions that lead to the formation of chemical irritants that may contribute to SBS symptoms and complaints. Mechanical ventilation systems containing activated carbon (AC) filters are often used to remove contaminants, but recent research suggests that the formation of strong irritants, like formaldehyde, can occur on the filter itself in the presence of ozone (O3). Formaldehyde is a strong irritant and a class B1 carcinogen; it can cause numerous symptoms in chronic low doses including, but not limited to, itchy eyes, general fatigue, and headaches. This phenomenon of heterogeneous reactions occurring on the surface of AC filters was explored by exposing AC filters spiked with limonene (a chemical frequently used in cleaning supplies) to ozone using conditions typical of indoor air. Effluent air samples were collected on sorbent tubes containing dinitrophenylhydrazine (DNPH)–coated silica gel and analyzed by high performance liquid chromatography (HPLC). In preliminary experiments (N = 4), effluent air of AC loaded with limonene and exposed to O3 (126 ppb, 38 % relative humidity) yielded low levels of formaldehyde (5.0 ± 2.1 ppb). Formaldehyde was not detected in any of the controls. More studies are necessary to investigate other indoor air contaminants, effects of environmental conditions, and other chemical releases from AC that may be enhanced by O3 exposure. 2009 Appalachian Student Research Forum Page 15 Although the generated formaldehyde concentrations were low, this research provides valuable insight into the origin of chemical irritants in the work place and may lead to mitigation or complete remediation of the SBS problem in the future. FUNCTION OF TOBACCO SALICYLIC ACID METHYL TRANSFERASE IN SALICYLIC ACID MEDIATED DISEASE RESISTANCE SIGNALING Tazley Hotz and Dhirendra Kumar, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Salicylic acid (SA) plays an important role in the defense responses against microbial pathogens in plants. SA is lipid immobile and may not pass through the chloroplast membranes to enter into the cytoplasm where mediation of the disease resistance signaling occurs. It has been recently suggested that SA synthesized in the chloroplast is converted into lipid mobile methyl salicylate (MeSA) by an enzyme, salicylic acid methyl transferase (SAMT). To investigate the role played by SAMT in SA-mediated disease resistance signaling, we examined the expression of SAMT in response to tobacco mosaic virus (TMV) and defense inducing chemical signals like SA, methyl jasmonate (MeJA), benzothiadiazole (BTH), nitric oxide (NO) and ethylene. To compare SAMT expression, we also studied tobacco plants susceptible to TMV expression and a transgenic tobacco plant (NahG) which does not accumulate SA. Expression studies were performed using semi-quantitative Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). To investigate the role of SAMT in plant immunity, we have produced stable transgenic tobacco plants silenced in the expression of SAMT. Effective silencing of endogenous SAMT in these transgenic plants will be determined using semiquantitative RT-PCR. To determine the role of SAMT in disease resistance these transgenic lines will be challenged with the viral pathogen, tobacco mosaic virus (TMV), and/or the avirulent bacterial pathogen, Pseudomonas syringae pv. tomato or virulent Pseudomonas syringae pv. tabaci. Understanding the biochemical pathways involved in plant defense responses against pathogens will help to develop crop plants showing better resistance to ever-evolving pathogens and reduce our dependence on pesticides. IDENTIFICATION OF PHOSPHATE BINDING IN ESCHERICHIA COLI ATP SYNTHASE Wenzong Li, Laura E. Brudecki and Zulfiqar Ahmad, Department of Biological Sciences, East Tennessee State University, Johnson City, TN F1Fo-ATP synthase is the smallest known biological nanomotor, found from bacteria to man. ATP synthase is the fundamental means of cell energy production in animals, plants, and almost all microorganisms. A typical 70 kg human with a relatively sedentary lifestyle will generate around 2.0 million kg of ATP from ADP and Pi (inorganic 2009 Appalachian Student Research Forum Page 16 phosphate) in a 75-year lifespan. ATP synthase works like a nanomotor through gammasubunit rotation, and being the smallest biological nanomotor, it can be used as a base model in developing nanomotors in nanomedicine usage. In order to be able to use it as a base model, it is of paramount importance to know which residues bind Pi to generate ATP. For this purpose we recently focused on determining how Pi binding is achieved to generate ATP through ADP+Pi. So far, we have investigated the relationship between Pi binding and catalysis for eight potential Pi binding residues, namely betaArg-246, betaAsn-243, alphaArg-376, betaLys-155, betaArg-182, alphaPhe-291, alphaSer-347, and alphaGly-351, based on their proximity to the phosphate analogs AlF3 or SO42- in x-ray structures of the catalytic sites. Employing Pi protection against NBD-Cl inhibition of ATPase as an assay, we have found that five residues, namely betaArg-246, alphaArg376, betaLys-155, betaArg-182, and alphaSer-347, are directly involved in Pi binding while three, betaAsn-243, alphaPhe-291, and alphaGly-351, are not. We also found that the highly conserved alpha-subunit VISIT-DG sequence residues appear to play a vital role in the catalysis either directly through Pi binding or indirectly. PLACENTAL CALCIUM PROVISION IN A LIZARD WITH PROLONGED OVIDUCTAL EGG RETENTION Brent J. Linville1, James R. Stewart1, Tom W. Ecay2, Michael B. Thompson3, Jacquie F. Herbert3 and Scott L. Parker3. 1Department of Biological Sciences, East Tennessee State University, Johnson City, TN, 2Department of Physiology, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN, 3Integrative Physiology Research Group and Wildlife Research Institute, School of Biological Sciences, HeydonLaurence Building (A08), The University of Sydney, Australia A prominent scenario for the evolution of viviparity and placentation in reptiles predicts a step-wise pattern with an initial phase of prolonged oviductal egg retention accompanied by: 1) progressive reduction in shell thickness, and 2) placentotrophy. Saiphos equalis is an Australian scincid lizard with a reproductive mode that is uncommon for squamates: 1) eggs are retained in the oviduct until late developmental stages, and 2) the embryonic stage at oviposition varies geographically. We studied calcium mobilization by embryos in two populations with different oviductal egg retention patterns to test the hypothesis that the pattern of nutritional provision of calcium is independent of the embryonic stage at oviposition. Females from one population oviposit eggs containing fully formed embryos that hatch within 2 days, whereas embryos in oviposited eggs of the second population are morphologically less mature and these eggs that hatch in 6 days. Yolk provided the highest proportion of calcium to hatchlings in both populations. Eggs of both populations were enclosed in calcified eggshells but shells of the population with a shorter period of egg retention had twice the calcium content of the second population and embryos recovered calcium from these eggshells. Placental transfer accounted for a substantial amount of calcium in hatchlings in both populations. Hatchling calcium concentration was higher in the population with the shorter egg retention period because these embryos mobilized calcium from yolk, the eggshell and the placenta. This pattern 2009 Appalachian Student Research Forum Page 17 of embryonic calcium provision in which both a calcified eggshell and placentotrophy contribute to embryonic nutrition is novel. The reproductive pattern of S. equalis illustrates that calcified eggshells are not incompatible with prolonged oviductal egg retention and that viviparity is not requisite to calcium placentotrophy. MODELING MICROPARASITIC DISEASE Meng Liu and Istvan Karsai, Department of Biology, College of Art and Science, East Tennessee State University, Johnson City, TN We performed a focused parameter sweep using an agent based model developed in Netlogo. This model simulates the transmission and perpetuation of a virus in a human population. The model begins with 150 people, of which 10 are infected. People who move randomly on a grid will have three states: healthy and susceptible to infection, sick and infectious, and healthy and immune. People may die of infection or old age. When the population dips below the environment's 'carrying capacity' (set at 700 in this model) healthy people may reproduce healthy and susceptible offspring. There are some critical factors in this model: The first is the density of the population. The second one is population turnover, which represents the case that individuals who die of old age or die from virus, will be replaced by those who are born as susceptible. Reproduction rate is constant in this model. The third one is degree of immunity, which represents how immune is a person to the virus after being infected and recovered. In this model, immunity does last forever and is secure. Infectiousness has also been introduced in this model. It shows us how easily the virus spreads. The duration of infectiousness tells us how long a person is infected before they either recover or die. This length of time is essentially the virus's window of opportunity for transmission to new hosts. In this system, we made initial population size and population fitness constant, which are 150 and 75%, respectively. We focused our study how infectiousness, duration of disease and degree of immunity (vaccinated) are affect the population. High ratio of sick people in the population allows virus to thrive and spread. A virus whose infectiousness is very low may spread slowly or die out. Transmittion may occur between the sick and susceptible only through contact with a given probability. Lower degree of immunity allows virus to attack more potential host. However, extremely high infectiousness and very low immunity are not helpful for virus' thrive. High infectiousness or too low immunity may allow the virus spread through population very fast and easily. However, since most individual became infected, most dies or becomes immune as a result, the potential number of hosts is will be often limited. So the virus may be too successful at first, but may not survive in the long term. 2009 Appalachian Student Research Forum Page 18 EXPLOITATION OF FOOD SOURCES BY ANT COLONIES: AN AGENT BASED SIMULATION. Carly Manning and Istvan Karsai, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Ants have an interesting form of communication using pheromones. This is different from most species of insects because ants use pheromone trails to inform colony members of the location of food. The methodology of the species of ants that use these trails has influenced many problem solving methods such as the ant colony optimization algorithm (ACO). Studying how ants are able to solve simple problems is of interest to those studying the behavior of cooperation and self-sustainability. Using an agent based NetLogo computer model “Ants,” the effect of the evaporation and diffusion rates on the ability of the ant to find three different food sources spread around the nest was recorded. A focused parameter sweep was used to study the relationship of food patch exploitation and the physical nature of the chemical. The evaporation and diffusion rates did not have an effect on which food source was exploited first, but they did have an effect on how much time was required to exploit each food source. The data suggests that if the evaporation rate is high, the ants will exploit the food significantly faster; and that the diffusion rate has no effect on the exploitation of the food sources as long as the evaporation rate is high. This study can contribute to the understanding of how ants have optimized the location and exploitation of food sources. INCIDENCE AND PREVALENCE OF FROG VIRUS 3 (FV 3) IN TEMPORARY WETLAND POPULATION OF AMBYSTOMA MACULATUM IN SHADY VALLEY Mehdi Pourmorteza and Dr. Tom Laughlin, Department of Biology, East Tennessee State University, Johnson City, TN Ranaviruses in the family Iridoviridae are a global concern and an emerging infectious disease in amphibians worldwide. Poor water quality caused by cattle use may stress resident populations and hence, increase pathogen prevalence in amphibians. To better understand the dynamics of the ranaviruses, the incidence and prevalence of Frog virus 3 (FV3) was measured in a Ambystoma maculatum population. We compared its prevalence in larval tissue in four different sites in a pond with cattle access in Shady Valley, Tennessee, USA. DNA from spotted salamanders was extracted using Qiagen DNeasy tissue kits. The isolated DNA was used in PCR reactions using hemi-nested primers targeting the major capsid protein gene. 2009 Appalachian Student Research Forum Page 19 OPTIMAL EXTRACTION OF NITROGEN CONTAINING DISINFECTION BYPRODUCTS FROM DRINKING WATER THROUGH SOLID PHASE MICROEXTRACTION Justin W. Quinn and Christine N. Dalton, Department of Chemistry, Carson-Newman College, Jefferson City, TN In most countries, primary sources of drinking water are disinfected prior to human consumption to prevent transmission of waterborne diseases. However, disinfectants react with naturally-occurring species, such as natural organic matter, algae, and bromide, in the water to form organohalogen species known as disinfection by-products (DBPs). Studies have shown that nitrogen-containing DBPs (N-DBPs) are more genotoxic and carcinogenic than other DBPs. The aim of this research is to optimize an extraction method to monitor nitrogen containing disinfection byproducts (N-DBPs) in drinking water. We have developed a reproducible and accurate analytical method to detect two classes of N-DBPs (haloacetonitriles and halonitromethanes) in drinking water using solid phase micro-extraction (SPME) followed by analysis with a gas chromatographmass spectrometer. The method parameters optimized to ensure maximum sensitivity of the analytical method were extraction time, sample temperature, stirring, holding time, and use of an internal standard. The best result was chosen as the optimal setting for use in the final extraction method. This method will be used in conjunction with the EPA to monitor these carcinogenic and genotoxic compounds in drinking water from various municipalities across the United States." STRUCTURAL AND THERMODYNAMICS STUDIES UNFOLDED STATES OF E. COLI ATP SYNTHASE Saurabh Sachan, Steven Sherman, Nitin Sachan, and Zulfiqar Ahmad, Department of Biology, East Tennessee State University, Johnson City, TN ATP synthase is the smallest known biological nanomotor, found from bacteria to man. This enzyme is responsible for ATP synthesis by oxidative phosphorylation in membranes of bacteria, mitochondria, and chloroplasts. The ATP synthase is critical to human health. Malfunction of this complex has been implicated in a wide variety of diseases including Alzheimer’s, Parkinson’s, and the class of severely debilitating diseases known collectively as mitochondrial myopathies. In its simplest form, in Escherichia coli, ATP synthase contains eight different subunits namely α3β3γδεab2c10. Till now, no structural and thermodynamic data is available on the unfolded states of this enzyme. Earlier studies from our laboratory have identified five phosphate biding residues in the α/β interface of the catalytic site. Here, we would like to study the unfolding of this enzyme by known denaturing agents GdnHCl, Urea, LiCl, LiCO4, LiBr, and NaBr. Our initial results show that ATP synthase gets reversibly unfolded in presence of 4M GdnHCl and 6M Urea. 2009 Appalachian Student Research Forum Page 20 SYSTEM DYNAMICS ANALYSIS OF PREDATOR-PREY RELATIONSHIP IN A FRAGMENTED HABITAT Emily Smith and Istvan Karsai, Department of Biology, East Tennessee State University, Johnson City, TN Habitat fragmentation has been a concern for conversationalists for many years. The division of large habitats into smaller patches not only effects the behavior of individuals within a species it also effects the relationship between species such as predator-prey relationships. Some studies have indicated that metapopulations with intermediate dispersal rates between patches are in fact more stable for both species in a predator-prey relationship. This raises the question of whether a fragmented habitat could in fact function as a mechanism to generate a metapopulation in which the patch landscape can provide a more stable habitat than an unfragmented one. Controlled experiments on the habitat landscape level can be difficult to conduct; however, with the use of computer modeling, the possible effects of fragmentation can be analyzed quickly and efficiently. We used systems dynamic modeling to simulate a simple predation system in fragmented habitats that are connected by corridors. In this case each patch caused by fragmentation in our fictional fragmented habitat was considered a stock and the flow of individuals among patches was analyzed as well as the effect of various variables such as density dependent movement from patches, carrying capacity and predator efficiency. Both the “original” and “modified” Lotka-Voltera equation was used to create the model and the effect of the parameters were predicted for the next 100 years. The model showed that fragmentation with corridors could be beneficial for predators. Social & Behavioral Sciences CRUCIFYING ANXIETY: EXAMINING THE RELATIONSHIP BETWEEN ANXIETY AND RELIGIOUSITY Cristi L. Adams, Charity Tipton, Wesley Ramey, and Leslie Griffith, Department of Psychology, East Tennessee State University, Johnson City, TN Anxiety has become the second most commonly studied aspect of personality in comparison to relationship of mental health and religion (Kalkhoran & Karimaollahi, 2007). Derosiers and Miller (2008) cited that some findings have shown that religion has more of an effect on lowering anxiety than spirituality. This may be because of the differences in the operational definitions used in the research. There is a large heterogeneity in the operational definitions of spirituality and religiosity found in the body of research. Miller and Thoresen (2003) highlighted this issue by noting the breadth of differential operational definitions that view spirituality and religiosity as the same thing, although most authors give these two terms different definitions. The majority of the literature defines religion as it relates to organized beliefs or an institution. Definitions of spirituality focus more on internal choices and individual beliefs or 2009 Appalachian Student Research Forum Page 21 perspectives (Miller & Thoresen, 2003). Psychologists have begun to distinguish the role that religion and spirituality can play on the emotional well-being of an individual. Research has shown that some mental health case have been improved by religion and spirituality. Results have been somewhat conflicting in this research due to some have only had significant differences when a person had high levels of anxiety (Kalkhoran & Karimaollahi, 2007; Derosiers & Miller, 2008). Derosiers and Miller (2008) found that being a part of a religious community had more of an effect on lower anxiety, which is consistent with anxiety being lower due to a strong social support system, rather than religious coping found by Hovey and Seligman (2007). Participants were 832 undergraduate students at a southeastern university (542 female and 290 male), which ranged in age from 18 to 55 years (M=21.56, SD=5.86). Data was collected via an online participant pool and were awarded modest extra credit for their participation. The survey included the Inventory of Religiousness, the Inventory on Spirituality, the Beck Anxiety Inventory, and a demographic questionnaire. Results indicate statistically significant negative correlations between: spirituality and anxiety, spirituality and anxiety, and anxiety and social support. Since the sample is only from a southeastern university, the results may not be generalizable. Future research would want to include a much larger sample in a variety of locations. These results should be used to inform research incorporating religious or spiritual beliefs into coping strategies for anxiety. ANXIETY AND DANGEROUS DRIVING IN A RURAL APPALACHIAN SAMPLE Cristi L. Adams and Christopher S. Dula, Department of Psychology,East Tennessee State University, Johnson City, TN Anxiety describes a chronic state of uneasiness and cautiousness, while the focus of worry distinguishes the various anxiety disorders from each other (Craske & Tsao, 1999). It seems that driving anxiety can be caused by a host of different reasons. Linnell and Easton (2004) stated that a person is more likely to develop a mental health problem after having a motor vehicle accident in which they sustained a physical injury. Delahanty, et al., (1997) compared at-fault and not-at-fault drivers involved in a crash and found that not-at-fault drivers were more likely to develop post traumatic stress disorder. They suspected that not-at-fault drivers feel they have lost control of their driving environment and can not stop adverse situations from happening (Delahanty, et al., 1997). As cited in Taylor, Deane, & Podd (2000), Taylor & Deane found that driver anxiety can be caused by driving with a passenger who negatively evaluates their driving behaviors. In the present study, we hypothesized that there would be a positive correlation between anxiety and dangerous driving behaviors. Data was collected via an online participant pool at a southeastern university. Participants were 832 undergraduate students (542 female and 290 male), with ages ranging from 18 to 55 years (M=21.56, SD=5.86). The participants took an online survey and were award modest extra credit for their participation. The survey included the Beck Anxiety Inventory (BAI), the Dula Dangerous Driving Index (Dula & Ballard, 2003), and a demographic questionnaire. The results supported our hypothesis. One limitation of this study is that the measures are self-report, which could impose an effect related to desirable responding. The results of this project can be used to inform therapies for people with driving anxiety. 2009 Appalachian Student Research Forum Page 22 DIFFERENCES IN REASONS FOR LIVING IN COLLEGE METHAMPHETAMINE USERS AND NON-USERS Jenny Barnes and Jon Ellis, Ph.D, Department of Psychology, East Tennessee State University, Johnson City, TN A number of studies suggest an association between substance use and suicidal behavior. However, relatively few studies have examined reasons for living, adaptive beliefs that prevent one from committing suicide, in substance using populations. The current study examines reasons for living and methamphetamine use in college students. Participants are 137 undergraduates at a southeastern university. They were administered a survey to assess demographics and substance use, as well as a suicidal ideation questionnaire and the expanded Reasons for Living Inventory. It was hypothesized that methamphetamine users would reveal themselves to be suicidal ideators more often than non-users, and that methamphetamine users would have a lower total score on the Reasons for Living Inventory. However, few participants admitted to having used methamphetamine, perhaps suggesting that individuals who are successfully attending college are less likely to use methamphetamine. Of the five participants who did report methamphetamine use, one of these indicated having experienced suicidal ideation. Further understanding the risk factors of suicide can lead to the development of better suicide prevention measures. CAUGHT ON CAMERA: IS VIDEO TAPE DATA BETTER THAN IN PERSON Adam Davis, Benjamin Martin, and Dr. Chris Dula, Department of Psychology, College of Arts and Sciences, East Tennessee State Univerisity, Johnson City, TN In the past, data collected on seatbelt use has been recorded by two observers standing in close proximity recording the few variables that can be viewed while the cars pass by (Farrell, Cox, & Geller, 2007; Geller & Hahn, 1984; Weinstein, Grubb, & Vautier, 1986; Zambon, Fedeli, Milan, Brosso, Marchesan, & Cinquetti, 2008). The purpose of this study is to compare inter-rater reliability between two different data collection methods. It was hypothesized that two independent observers coding video taken with a high definition camcorder should be equally or more reliable than the traditional two-observer model. However, there have been no studies conducted with the use of a high definition camcorder. There seems to be little reporting of inter-rater reliability in previous research and it was only monitored in the field. For example, the only instance of inter-rater correlation coefficients we found in seatbelt data collection was in a study by Farrell, Cox, and Geller in 2007. However, this previous literature did not specify whether the observers were allowed to discuss what they had observed in making their determinations (Farrell, Cox, & Geller, 2007). The methodology used in this study has two observers set up a single high definition camcorder and then to independently and without discussion, record seatbelt data and other variables in the traffic which passes while they videotape the same vehicles. The only verbal communication is to announce the color and make of the car being observed, such that two additional independent observers can take data on 2009 Appalachian Student Research Forum Page 23 the same vehicles later. All observers recorded data on car color and vehicle type (the only data spoken of by the observers), the drivers’ seatbelt use, cell phone use, sex, race, estimated age, and the number of passengers in the vehicle. The video recorded data is kept on a computer connected to a high definition monitor to be reviewed later by two different observers. The video observers will record their data independently, but will have the option of running the video in slow-motion or zooming in on aspects of the screen. It is hypothesized that the inter-rater reliability coefficients for the video observers will be similar to or greater than the live observers. FAUNAL ANALYSIS OF LINVILLE CAVE (40SL24) SULLIVAN COUNTY, TENNESSEE Meagan Dennison and Jay Franklin, Department of Sociology and Anthropology, East Tennessee State University, Johnson City, TN A faunal analysis is the interpretation of the animal bones recovered from archaeological sites. In 1990, excavations of Linville Cave yielded more than 2500 animal bones (faunal remains). We conducted a faunal analysis to determine what animal species were consumed by prehistoric Native Americans of the region. This was conducted in the ETSU Archaeology Lab. To begin, remains were separated into bags by excavation unit and level. Bones were identified using comparative skeletons, lab manuals, and discussions with other scholars. Bones were classified according to the Linnaean Taxonomic Classification System. When possible, bones were identified to the species level, but many of the bones fell into higher taxonomic categories such as: Fish, Amphibians, Invertebrates, Mammals, Birds, Reptiles and indeterminate Vertebrates. Where applicable, modifications were recorded which include burned bone, cut bone, and/or gnawed bone. After identifications were made, a database was created in Excel. Quantitative analyses were then conducted. Eleven species were identified. The majority of the identifiable bones belong to game animals, and the majority of the modified bones were burned. After the quantitative analyses were completed, we were able to conclude that Linville Cave was used prehistorically as an intermittent hunting camp. Based on previous botanical analyses, it was also determined that the site was used in the late fall and early winter. Two radiocarbon dates indicate that the site was used most intensely during the Middle to Late Woodland Period (app. 1300 years ago or 700 A.D.) in Southern Appalachia. Once the faunal analysis was completed, the results were compared to two other sites in the region, the Nelson Site and the Eastman Rock Shelter. Both the Eastman Rock Shelter and the Nelson Site date to the Middle and Late Woodland. Like Linville, Eastman is a special purpose rock shelter site; however, the Nelson site is a larger habitation open-air site. When the two are compared to Linville Cave, there is no significant difference in subsistence patterns. This indicates that there is little difference between specialized hunting sites when compared to habitation sites of the region." 2009 Appalachian Student Research Forum Page 24 WHAT LIES BENEATH Ashley Dickson, Ben Martin, Jessica Williamson, and Dr. Chris S. Dula , Department of Psychology, East Tennessee State University, Johnson City, TN The ancient practice of tattooing has been widely accepted in several cultures, but only recently in the western society (Carroll, Riffenburgh, Roberts, & Myher 2002). Tattoo literature currently studies populations of prisoners, military personnel, psychiatric patients, and juvenile delinquents from detention homes. In these captive populations, risk taking and impulsive behaviors may already be prevalent (Laumann & Derick, 2006). According to Forbes, tattooed individuals are categorized as being risky, immoral, nonconforming, and unstable in their behavior patterns (2001). However, when Forbes studied risk-taking and impulsive behaviors in non-confined populations, he found that tattooed individuals conform less to social expectations and perform risk-taking behaviors (2001). Most tattooed individuals reported that they had considerably thought about the tattoo for at least a month before deciding to get it (Forbes, 2001). Participants were 54 faculty members ranging from X to Y years of age (28 women, 26 men, M=45.77 SD=10.44) and a random sample of 175 undergraduate students ranging from18 to 62 years of age (55 men and 120 women, M = 20.5, SD= 4.99). The undergraduates were administered the Barratt Impulsivity Scale (BIS) via Sona Systems, an online participant pool at a regional southeastern university and were granted a modest amount of extra credit. Faculty completed the MSATS and the stigma scores were compared with personality scores from students. Results suggest that people with tattoos are not very different from those without tattoos; however, they are perceived as very different by the non-tattooed population (Forbes, 2001). There were no significant impulsivity differences found between tattooed and non-tattooed individuals. There was not a specified gender difference in impulsivity among the tattooed population; however, there was a lower level of impulsivity among females. This does not support the theory that men with tattoos are more impulsive than women with tattoos. Evidence also does not support the theory that individuals with tattoos are not more impulsive than those without tattoos. Results reveal those with tattoos and particularly females with tattoos, may be unfairly stigmatized as highly impulsive. There were no significant differences found between tattooed and non-tattooed persons in ventursomeness scores. Previous literature is not supported in view of the findings that there is no significant correlation between individuals with tattoos and individuals without tattoos in regards to adventurousness and impulsivity. The results of this study could reduce stigma and may allow tattoos to flourish as a more acceptable art within our culture instead of a taboo." A DOSE-RESPONSE ANALYSIS OF METHYLPHENIDATE SENSITIZATION IN ADOLESCENT RATS: SEX DIFFERENCES IN LOCOMOTOR ACTIVATION Benjamin A. Hughes1, Andrew B. Hughes1, A. Brianna Sheppard1, Brooks B. Pond2, Michael L. Woodruff1, Russell W. Brown1. 1Department of Psychology, East Tennessee 2009 Appalachian Student Research Forum Page 25 State University, Johnson City, TN; 2Division of Pharmaceutical Sciences, College of Pharmacy, East Tennessee State University, Johnson City, TN This study was designed to analyze locomotor sensitization to methylphenidate (MPH, trade name: Ritalin) in adolescent rats. MPH is the most common medication prescribed to adolescents for treatment of attention deficit-hyperactivity disorder (ADHD). Rats were habitutated to a locomotor arena from postnatal days (P)30-32 and began locomotor sensitization testing on P33. Male and female rats were administered one of three doses of methlphenidate (MPH; 1, 3, or 5 mg/kg) or saline every other day through P49, for a total of nine exposures to the drug. Approximately 10 min after each injection, all animals were placed into a locomotor arena and behavioral activity was analyzed. Results showed that adolescent females administered the 5 mg/kg dose of MPH demonstrated an approximate 100% increase in locomotor activity as compared to the 5 mg/kg dose of MPH administered to males. Additionally, females administered the 5 mg/kg dose of MPH demonstrated locomotor sensitization. No other dose of MPH produced locomotor sensitization in males or females. Interestingly, the lowest dose of MPH (1 mg/kg) produced locomotor suppression in females compared to controls, whereas in males, both the 1 and 3 mg/kg of MPH produced locomotor suppression. These results show that there are robust differences in locomotor activation to MPH that is dependent upon dose. Further, it appears that females are more sensitive the locomotor activating effects of MPH than males, and also demonstrate sensitization to the highest dose, suggesting abuse potential of MPH may be higher in adolescent females as compared to adolescent males. FUNCTIONAL IMPAIRMENT AND DEPRESSIVE SYMPTOMS:EXAMINING THE COMPONENTS OF TRAIT HOPE Sherry Kaye1, Jameson K. Hirsch, Ph.D. 1, and Jeffery M. Lyness, Ph.D. 2, 1Department of Psychology, East Tennessee State University, Johnson City, TN; 2Department of Psychiatry, University of Rochester Medical Center, Rochester, NY As older adults age and become subject to illness they may experience a loss of ability to function independently, resulting in increased depressive symptoms. In past research, positive characteristics such as hopefulness (Snyder, 1989; Snyder, Harris et al., 1991; Snyder, Irving, & Anderson, 1991) have been found to buffer against the effects of depressive symptoms. Trait hope has two main components (Agency) refers to one’s belief in his or her ability to effect change and (Pathway) the resources to accomplish change. Snyder determined that both needed to present (1999). Our study focused on the potentially moderating effect of trait hope as a positive, cognitive-motivational characteristic that facilitates psychological and physical adjustment in older adults to functional impairment and the development of depressive symptoms. Participants included 110 older adult (61.8% female; Mean age 74.24 [ SD=5.56], primary care patients. Our sample was predominantly White (91.8%) and well educated (Mean educational level=14.11 years [SD=4.1]. Participants completed the Hamilton Rating Scale of Depression (HRSD), the Mini Mental State Exam (MMSE), and the Goals Scale 2009 Appalachian Student Research Forum Page 26 a measure of Trait Hope. Examiner-rated measures of illness burden (Cumulative Illness Rating scale; CIRS) and functional impairment (Karnofsky Performance Status Scale; KPSS) were completed by a physician after a chart review. Hierarchical multivariate linear regressions were conducted covarying age, gender, and MMSE and CIRS total scores. Patients in this sample had a mean HRSD score of 7.75; a mean Karnofsky Performance Status Scale (KPSS) score of 76.98, indicating that, on average, participants required some assistance to care for personal needs; a mean Cumulative Illness Rating Scale (CIRS) score of 10.16, and a mean Trait Hope score 32.94. In Model I, trait hope moderated the association between functional impairment and depressive symptoms [t = 2.34, p<.05, Un β = .02 (SE = .01)]; In Model II we examined the sub components of trait hope, the agency and pathways subscale, neither of which reached significance. Our results suggest that trait hope moderates the association between functional impairment and depressive symptoms in older, adult primary care patients. Importantly, our study indicates an individual must have both the determination and the resources to accomplish goals to effectively cope with functional impairment. Clinical implications of our findings include the need to promote both self-efficacy (agency) and facilitate environmental support (pathways) for older adult patients with functional impairment, in order to reduce the likelihood of depressive symptoms. GETTING UNDER THE SKIN: THE DEVELOPMENT OF THE MARTIN STIGMA AGAINST TATTOOS SCALE Benjamin Martin and Chris Dula, Department of Psychology, East Tennessee State University, Johnson City, TN Stigma against tattoos and tattooed individuals can lead to self-doubt and shame on the part of the tattooed individual (Dosani, 2005).Although previous studies exist, little is known about the subject. In order to better analyze stigma against tattooed individuals, an instrument was developed to measure this phenomena in a quantitative fashion. This measure, termed the Martin Stigma Against Tattoos Scale (MSAT), was based on literature pertaining to the stigmatization of having tattoos. A 12-item questionnaire consisting of agree or disagree questions was developed. The measure was then distributed through an online survey administration system at a Southeastern regional university. Participants were 175 undergraduate students ranging in age from 18 to 62 years of age (120 women, 55 men, M=20.5, SD=4.99) completed the survey for a modest amount of extra credit. Psychometric analysis was performed and stigma was calculated by summing the responses. In addition to strong face-validity, extraction of one higherorder factor provided evidence of a single overall construct. Confirmatory factor analysis supported the structure of the scale and suggested coefficient reliability. A Cronbach’s alpha was.92 provided evidence of internal consistency reliability. This scale could be applied in future research to gauge whether a stereotype placed on tattooed employees by biased employers has any merit. 2009 Appalachian Student Research Forum Page 27 WHAT'S YOUR PLEASURE: THE DEVELOPMENT OF THE SELF-INJURY SELF-REPORT MEASURE Benjamin Martin and Chris Dula, Department of Psychology, East Tennessee State University, Johnson City, TN Despite the amount of research conducted on self-injury there is a lack of empirically validated instruments for which to measure self-injurious behavior. A major problem in classifying self-injury is the lack of agreement on the nomenclature. The terms autoaggression, malingering, Munchausen’s, symbolic wounding, masochism, local selfdestruction, delicate self-cutting, parasuicide, and focal suicide have all been used to denote self-injurious behavior (Clarke & Whittaker, 1998). Understanding self-injury is also problematic due to self-injury having been identified as a symptom of various diagnoses. Despite the range of labels, those most often found in the literature are: selfinjury (e.g., Gratz, 2003), self-harm (e.g., Pattison & Kahan, 1983), and self-mutilation (e.g., Favazza, 1998a), sometimes used interchangeably. The present study details development of the Self-Injury Self-Report Survey (SISRS) Undergraduate students (n = 184) were administered a set of surveys to assess demographics, self-injurious behavior, suicidal ideation, Axis I and Axis II disorders, and impulsivity. A principle components factor analysis indicated the SISRS to be a valid measure and able to assess the extent of self-injury and related features. Subscale items with factor loadings below .50 were deleted. Pearson correlations were performed on SISRS scores in conjunction with scores on the MCMI-III scales. Total scores on the SISRS were calculated by adding subscale scores together. A t-test was performed to determine whether there were significant differences on SISRS scale scores with regard to gender. Such a measure should enable clinicians to better assess self-injury and researchers to more fully examine self-injury and its relationship to demographic factors and pertinent clinical issues. EXAMINING THE ASSOCIATION BETWEEN POSITIVE AND NEGATIVE AFFECT AND SELF-REPORTED HEALTH IN LUNG CANCER PATIENTS H. Mason, J. K. Hirsch, and B. Gibson, Department of Psychology, Laboratory of Rural Psychological and Physical Health, East Tennessee State University, Johnson City, TN; and P.R. Duberstein, Department of Psychiatry, University of Rochester Medical Center, Rochester, NY Lung cancer is the third most common cancer and the leading cause of cancer deaths in the United States with a 15% chance of survival after 5 years. Lung cancer patients may report distress with diagnosis, prognosis or treatment, and may experience low mood and anxiety. Negative affective states are common; research suggests that up to 98% of lung cancer patients experience negative affect, which may contribute to poor physical, psychological and social quality of life. Importantly, most lung cancer patients are able to identify positive or meaningful aspects of their illness; 63% of a sample of older adult lung cancer patients reported positive meaning and 53% describe their illness as a 2009 Appalachian Student Research Forum Page 28 “challenge” to be overcome. We examined the association between positive and negative affect and self-reported health outcomes in 133 lung cancer patients (mean age = 63.68 years old; SD = 9.37). Positive and negative affect were assessed using the NEO-FFI Personality Inventory; facets of positive and negative affect were derived from the broad domains of extraversion and neuroticism. Self-assessed health outcomes were measured using SF-36 subscale scores, including: physical functioning, role limitations due to physical health and emotional problems, social functioning, pain, and general health. We conducted a bivariate correlation analysis, as well as univariate and multivariate linear regression analyses, covarying age, gender, education, and stage of cancer. Our results suggest that both positive and negative affect are independently associated with selfreported health beliefs, behaviors and outcomes. In univariate analyses, negative affect was significantly associated with poorer physical functioning, role limitations due to emotional problems, difficulties with social functioning, bodily pain, and poor general health. In univariate analyses, individuals with greater positive affect had less role limitations due to emotional problems, better social functioning, less bodily pain, and increased general health. A clinical trend existed toward an association of positive affect and reduced role limitations due to emotional problems (p = .09). Importantly, our multivariate model suggests that, despite the presence of negative affect, there are many areas of health functioning that continue to be beneficially influenced by positive affect, including social functioning, perceived general health and, potentially, reduced impairment as a result of emotional problems. Identifying and bolstering such characteristics may serve to improve quality of life and survival of the lung cancer patient, and could be strategically targeted in therapeutic interventions. COMPARATIVE STUDY OF PUBLIC EDUCATION: UNITED STATES AND CHINA Robert Scott and Dr. Weixing Chen, Department of Political Science, East Tennessee State University, Johnson City, TN This is a comparative analysis of the quality of public education in China and the United States. The purpose of this analysis is to understand which areas in education the United States would need to improve upon in order to compete with another industrialized nation in the 21st century. The United States is in an economic recession and has been struggling to compete against rising economic superpowers. There needs to be a change in politics and curriculum in order for Americans to succeed in a global marketplace and workforce. China was chosen as a nation to study because it is a growing economic superpower and is projected to surpass the United States. The areas of education that are studied are curriculum, teacher training, and parental involvement. Some of this is readily available and other is not. The research project will only cover as much information as can be accessed. Test scores will not be used to assess how well one country has placed over another because there is no access to the test scores in China. Curriculum will be studied to differentiate between the subjects that are taught from elementary to high school. The point of this to address the issue that other countries teach subjects, which are considered advanced, at an elementary level. For example, 2009 Appalachian Student Research Forum Page 29 whereas the United States might not introduce a subject until the student has reached the 8th grade level, a student in China may have been introduced to the subject in the 5th grade. Under curriculum, the study will also compare the number of days and hours in which students from each country spend in school and studying at home, along with the workload that is expected to be completed. Teacher training will be studied to understand the amount of teachers per student each country possesses and the quality of training each teacher has undergone in both China and the United States. The level of parental involvement is an important factor on how well a student performs in school. Therefore, the study will identify if the level of parental involvement in China is greater or less than that in the United States. This analysis is meant to be a tool to understand in which ways the public education system in the United States needs to be improved in order to obtain the level of economic prosperity that it once enjoyed. There is a need for students to gain higher levels of education in a technologically advanced global community where the competition is no longer against a handful of college graduates from around country. Instead, students will need to compete for against graduates from all over the world for positions in a global workforce. PERCIEVED CONTROL: A MECHANISM EXPLAINING INTIMATE PARTNER VIOLENCE OUTCOMES Desta A. Taylor and Stacey L. Williams, Ph.D., Department of Psychology, East Tennessee State University, Johnson City, TN Prior research has shown that intimate partner violence (IPV) leads to negative mental health outcomes, and has evidenced increased anxiety and decreased self-esteem among the abused. The factors that explain these links between IPV and negative outcomes have been studied with less frequency. Perceived control, or the sense that one can impact or change the outcome of one’s life, may serve as a mediating mechanism for how partner violence comes to be linked with negative outcomes. Indeed, lower levels of perceived control have been associated with higher levels of depression and anxiety (Clements, Ogle, and Sabourin, 2005). Yet, although previous research has shown that traumatic events can lead to lower levels of perceived control (Brown & Seigel, 1988; Katz & Arias, 1999), no prior work has investigated perceived control as a direct mediating mechanism in the context of IPV. It was hypothesized that IPV would lead to increased anxiety and decreased self-esteem, and that these relations would be mediated by the decreased perceived control experienced following IPV. Participants included 652 (224 male, 424 females, 4 did not specify) undergraduates from a southeastern university. The mean age of participants was 21 years, but ranged from 17-55 years. Participants completed the survey electronically, and received extra credit for their participation. The survey assessed IPV, and was coded as a composite variable comprised of physical, psychological, and sexual violence. These experiences in the past year were assessed using the following scales: CTS-2; Straus et al., 1996; Tolman’s (1999) Psychological Maltreatment of Women Inventory; Marshall’s (1999) Psychological Abuse Scale. Other measures in the study included: Anxiety: 10 items, anxiety symptoms (SCL-90, anxiety subscale; Derogatis, 1994); Self-Esteem: 10 items, self-related beliefs (Rosenberg, 1965); 2009 Appalachian Student Research Forum Page 30 and Perceived Control: 7 items, (mastery; Pearlin, Meneghan, Lieberman, & Mullan, 1981). Hypotheses were tested using a series of multiple regression analyses examining paths to mediation (Baron & Kenny, 1986). Results provide support for study hypotheses. Partner violence was related to increased anxiety symptoms (b= .40, se=.04, p=.000), and decreased self-esteem (b=.08, se=.04, p=.055). In addition, IPV was related to decreased perceived control (b=.24, se=.07, p=.001) which in turn was related to decreased selfesteem (b= .26, se=.02, p=.000) and increased anxiety symptoms (b= .07, se=.03, p=.007). In further support, the Sobel test indicated significant mediation for self-esteem (3.25, p<.01) and partial mediation for anxiety (2.09, p<.05). Thus, results evidence perceived control may help to mediate, or explain, how partner violence links with outcomes that have long been associated with IPV. Findings highlight the need to focus on perceived control as an area for possible intervention to increase self-esteem and decrease psychological symptoms following partner violence experiences. RELIGION BEHIND THE WHEEL: THE RELATIONSHIP BETWEEN RELIGIOSITY AND DANGEROUS DRIVING IN AN UNITED STATES SAMPLE Desta A. Taylor, Cristi L.Adams, and Chris S. Dula, Ph.D., Department of Psychology, East Tennessee State University, Johnson City, TN Prior research has focused on different lifestyle factors associated with dangerous or risky behavior, including dangerous driving (El. Chiliaoutakis, Koukouli, Lajunen, & Tzamalouka, 2005, Jessor, Turbin, and Costa, 1997). Jessor et al. (1997) even suggest that hazardous driving in young adults could indicate other risky behaviors. Dangerous driving is defined as, “dangers from lapses of attention while driving that are inherent in off-task behaviors such as using a cell phone, attending to one's appearance, eating, drinking, smoking, or adjusting a car stereo. Any of these behaviors may occur without the presence of negative emotion or intent to harm” (Willemsen, Dula, Declercq, & Verhaeghe, 2008). Religiosity, or the degree to which one feels connected to a higher power, has been associated to lower levels of dangerous driving (El. Chiliaoutikis et al., 2005, Jessor et al., 1997, Gnardellis, Tzamalouka, Papadakaki, & El. Chliaoutakis, 2008). However, the body of research whose focus is the relationship between dangerous driving and religiosity in the United States is sparse, demonstrating a need for research in this area. For instance, El. Chiliaoutikis et al. (2005) found religiosity to be negatively associated to dangerous driving in a Greek sample. The goal of this study is to examine the relationship between religiosity and dangerous driving in a United States sample. It was hypothesized that religiosity would be negatively correlated to dangerous driving behaviors. Participants included 832 undergraduates from a midsized Southeastern university with an age range of 16-55 years with X= 21.56 and SD= 5.86. Three Measures were used to assess the relationship between dangerous driving and religiosity including: a demographics questionnaire, The Dula Dangerous Driving Index (DDDI), which encompasses 28 items that assess negative emotions while driving, aggressive driving behavior, and risky driving (Dula & Ballard, 2003), and The Inventory on Religiousness consisting of 41 items measuring general religious views (Rayburn, 1997). Participants completed the survey via an online survey system, and were awarded modest 2009 Appalachian Student Research Forum Page 31 extra credit in a psychology course for their participation. Results confirmed prior research and the hypothesis that religiosity would be negatively correlated with dangerous driving (r = -.156, p= <0.01). One limitation of the study is that the sample consisted of only college undergraduates, which may not be representative of the general population. Future research should focus on identifying which variables mediate the relationship between religiosity and dangerous driving to inform interventions aimed at decreasing dangerous driving in nonreligious persons. Future research should also investigate the differing effects of various religious denominations in regards to driving behaviors, and more specifically dangerous driving. PERCEIVED BARRIERS TO INTIMATE PARTNER VIOLENCE SCREENING IN PRIMARY CARE Jamie A. Tedder, Stacey L. Williams, Ph.D., and Peggy J. Cantrell, Ph.D., Department of Psychology, East Tennessee State University, Johnson City, TN Intimate Partner Violence (IPV) is associated with negative physical (e.g., gynecological, stomach, chronic pain problems) and mental (e.g., depression, anxiety) health outcomes (e.g., Coker et al., 2000; McCauley et al. 1995). These negative health effects can be present in medical settings. Women with a history of IPV may be more likely to utilize health care services and visit their primary care physicians (e.g., Rivara et al., 2007). As such, screening for IPV in primary care may be a vital part of women’s health. Previous research has examined the frequency primary care physicians screen for IPV, and how patients feel about being screened. Studies have shown that routine screening rates for IPV are low (1.2-24%) in primary care (Chamberlain et al 2002; Rodriguez et al. 1999), despite that most patients feel screening would be beneficial (86.1%; Gielen et al. 2000), and would disclose IPV (68%; Caralis and Musialowsky 1997). These low rates have prompted examinations of barriers to screening that physicians face (e.g., Limandri et al. 1996). Although some barriers to IPV screening have been outlined previously, the present study assessed a thorough list of potential barriers that might prevent physicians from screening patients for IPV. Participants in this study included 36 physicians and 3 RN/MSNs (25 male, 14 female). The mean age for participants was 48. The majority of respondents practiced in rural areas (66.7%). Physicians were recruited during their attendance of a medical conference in the southeast. Several announcements were made during the conference asking that physicians complete a brief survey. The survey consisted of items about personal- and practice-related demographics, screening practices, and barriers. To determine barriers to IPV screening, physicians were asked to rate 24 potential barriers on how much each was a barrier to IPV screening. The potential barriers represented areas such as limitations of IPV screening measures, beliefs about patient compliance with physician recommendations, and physician-related beliefs about IPV and screening in a medical practice (e.g. the competing demands of other health problems). Findings showed the barriers most commonly reported by physicians included their belief the patient would be noncompliant with recommendations (82%), their personal lack of confidence in dealing with IPV (79.5%), the perceived lack of effective IPV interventions (77%), their belief that health insurance would not adequately 2009 Appalachian Student Research Forum Page 32 reimburse (74.4%), and the perceived inadequacy of IPV screening measures (71.8%). Results suggest that physicians perceive a range of barriers to screening for IPV and highlight a need for more adequate screening measures and interventions for IPV which will reduce barriers to screening. Barrier reduction might be accomplished by integrating into the primary care practice a mental health professional who can directly screen and intervene for IPV. FRUITS OF WAR: TO WHAT EXTENT DID ISRAEL WIN THE SECOND LEBANON WAR? Treston Wheat, Ronald McNair Program, University of Tennessee, Knoxville, TN This study was used to observe the Second Lebanon War in 2006 between the terrorist organization Hezbollah and the state of Israel. The objectives were to determine to what extent did Israel win the war with its own objectives. Israel’s objectives were, broadly speaking, to eliminate Hezbollah by destroying their military capabilities and neutralizing the leadership. In addition, Israel wanted to create an Israeli deterrent, meaning they wanted to present themselves as a “strong man.” This “strong man” would prevent other organizations and states from attacking Israel because of their military power. As this was a qualitative study, the method was to utilize the research of academics looking at the situation to determine how Israel did in the war. Though it is a general consensus that Israel lost, it is important to look at why they lost, to what extent they lost, and how this will affect them in the future. This can be seen most notably in the recent IsraeliPalestinian conflict in Gaza. The researcher first looked at how the Israelis waged war. More importantly, how they waged war after they no longer fought states but organizations like Hamas and Hezbollah. Specifically, the researcher looked at the counter insurgency strategy of Israel and how this affected them when they had to fight a standard war against Hezbollah. From there the researcher read numerous academic articles on how well Israel did. From these the researcher was able to draw conclusions as to what extent Israel had lost the war. As previously stated, it is a general consensus that Israel lost. But few understand that Israel barely hurt Hezbollah, and in fact lost their “strong man” status among the Arab nations. This means that war is more likely for Israel rather than just fighting insurgents in Gaza and the West Bank. Though some might see this as having no influence on America, it does. The United States, for good or ill, is a staunch supporter of Israel, meaning the connection between the two is seen by extremist organizations like Al Qaeda. With Israel’s loss against Hezbollah and Iraq seen as a quagmire for America the two countries appear to be losing it together, giving strength through perception to terrorist organizations and the states that sponsor them. From these results, Israel’s loss in the war has profound consequences for future conflicts between Israel and its neighbors, and it has consequences for American foreign policy in the Middle East. 2009 Appalachian Student Research Forum Page 33 SATISFACTION IN THE WORKPLACE Jessica Williamson, Ben Martin, Robin Leonard, and Chris Dula, Department of Psychology, East Tennessee State University, Johnson City, Tennessee How employees perceived their own organizational support, and its relationship to the extent of the employee’s commitment to the organization is important for a company to function efficiently (Eisenberger, Huntington, & Hutchinson; 1986). For example, employees with higher levels of perceived organizational support had fewer absences (Eisenberger, et. al., 1986). Other studies have also shown that employees feel safer with a higher level of perceived management commitment (Michael, Evans, Jansen, & Haight; 2005). Establishment of operational definitions are needed to further recognize the terms used in the proposed research. The purposes of this study are to investigate the impact of socioeconomic status (SES) on the relationship between job satisfaction and productivity between typical “blue collar” and “white collar” employees. Participants included were university students who had been employed at least some time during the 6 months prior to their participation in this research. Participants were given the Survey of Perceived Organizational Support, Measure of Dutifulness, and Measure of Satisfaction. It was predicted that there would be a significant relationship between socioeconomic status and job satisfaction. Results from this study may help Industrial/Organizational Psychologists solve ongoing problems of minimum wage and attrition. IMPULSIVENESS AND RISKY DRIVING BEHAVIORS Jiaju; Wu, Shufang Sun, Cristi L. Adams, and Rachael Byington, Department of Psychology, East Tennesse State University, Johnson City, TN As cited in Smith, Waterman, & Ward, (2006), impulsive individuals in a college sample have been found to have a willingness to engage in high-risk behaviors, including driving without seatbelts and driving while under the influence of alcohol. It is predicted that impulsiveness will be connected to the type of impulsive aggression that depicts most aggressive driving (Dalen, et al., 2004). People with higher levels of impulsivity are more likely to act on the spur of the moment, become reckless, and be less likely to comply with regulations (Owsley, McGwin, &McNeal, 2003). The purpose of this study was to examine the relationship between impulsivity and the three aspects of dangerous driving behaviors, as defined by the Dula Dangerous Driving Index (DDDI, Dula & Ballard, 2003). The three aspects of dangerous driving are aggressive driving, negative emotional driving, and risky driving. It has been hypothesized that higher scores of impulsivity would predict higher scores on the DDDI total. Data has been collected via an online participant pool at a southeastern university. Participants were 832 undergraduate students (542 female and 290 male). The age of participants ranged from 18 to 55 (M=21.56, SD=5.86). The participants took an online survey and were awarded modest extra credit for their participation. The survey included the Barratt Impulsivity Scale, the Dula Dangerous Driving Index (Dula & Ballard, 2003), and a demographic questionnaire. 2009 Appalachian Student Research Forum Page 34 Results showed that there was a positive correlation between impulsivity scores and the DDDI total scores. Possible practical uses of these results include helping determine automobile insurance companies’ policies and prices, as well as use in driver’s education classes and defensive driving courses. Since only students from a southeastern university were used in the sample, the results might not be generalizable to all populations. Division II – Graduate Students 1-2 Years Arts & Humanities and Biomedical Sciences SOURCE AND LEVEL OF PROTEIN FED TO MICE MAY AFFECT THEIR INTESTINAL MUCIN PRODUCTION Bupe Martha Habiyambere, Edward M Onyango. Biological Sciences, Health Sciences, East Tennessee State University, Johnson City, TN Mucins are large heavily glycosylated proteins which form the major component of mucus that protects epithelial surfaces of ducts in the body. The effect of the source and level of dietary protein on mucin production and intestinal mucosal morphology was investigated in mice. Thirty two male weanling ICR Swiss mice were randomized into 4 groups of 8 mice each. The four groups of mice were randomly assigned to four diets whose sources of protein were either from plant or animal origin and the level of each protein source was either 10% or 20% crude protein. Mice were provided the respective diets for 14 days and euthanized on the fifteenth day. Duodenal and jejunal sections of their intestines were removed, fixed in methacarn, embedded in paraffin, sectioned and stained with alcian blue-periodic acid Schiff (AB-PAS) stain to visualize mucins and quantify the goblet cells. AB-PAS stained duodenal and jejunal sections showed an increase in mucus in mice fed diets containing proteins of animal origin. The number of goblet cells was higher in diets containing protein of animal origin (p<0.05). The increase in goblet cell number was greater for the higher protein level. The length of the villi did not seem to vary among the four different diets. It is concluded that the source and level of protein fed to mice may be influencing their intestinal mucin production. 2009 Appalachian Student Research Forum Page 35 CONVERSATIONAL RECASTS VS IMITATION INTERVENTION FOR SPECIFIC LANGUAGE IMPAIRMENT: A CRITICAL APPRAISAL OF THE EVIDENCE Molly Millard, Sundeepkumar Venkatesan, and Lynn Williams, East Tennessee State University, Communicative Disorders, Johnson City, TN Recasts and imitation are two widely used treatment techniques used to facilitate acquisition of speech language structures in young children with language impairment. There have been several studies evaluating their effectiveness and there is evidence to support both approaches. The debate of which to use is ongoing as research continues within an evidence based practice (EBP) framework that is used to inform clinical decisions. The purpose of this presentation is to evaluate the available experimental evidence to compare the relative effectiveness of conversational recast and imitative treatment for acquiring grammatical structures in preschool children with specific language impairment (SLI). Following the specification of our clinical question, a set of inclusionary and exclusionary criteria was established for selecting articles to include in our review. Our first criteria was that the study featured an experimental design that was a randomized control trial or a quasi-experimental design, as these designs provide the best evidence of casualty. Additional criteria for inclusion of studies were the participants’ age, ranging from 2 to 6 years old, and the type of disability, which was SLI. Only English-language studies published in peer reviewed journals were included in the review. Databases that were searched included PSYCH INFO, ERIC, American Speechlanguage-Hearing Journals (JSLHR, LSHR, LSHSS), PubMed, and JSTOR. A total of 22 studies were retrieved, of which full text reviews were completed on 7. From these, only two studies were found to meet the specified criteria. These two studies involved a total sample size of 35 children: 7 with SLI and 7 language normal (LN) children (Nelson, Camarata, Welsh, Butkovsky, & Camarata, 1996), and 21 SLI children (Camarata, Nelson, & Camarata, 1994). Both studies used similar criteria for choosing their participants, and Nelson et al., (1996) matched the LN group based on relevant language measures. All 35 children received both imitative and conversational recast intervention in a clinic setting using randomization for subject assignments. The number of intervention sessions ranged from 12-18 sessions with each child receiving treatment twice a week. Outcome measures in both studies focused on the number of spontaneous productions produced for each absent or partially mastered target. Both studies had similar criterion for selecting targets as well as measuring spontaneous responses. Converging evidence across the two studies indicated a benefit of conversational recast treatment over imitative treatment for SLI children when addressing the acquisition of grammatical structures. Critical Appraisal of Treatment Evidence (CATE; Dollaghan, 2007) was completed on each study to evaluate the quality and level of evidence. Based on this evaluation and calculation of effect sizes, the findings that conversational recasts are more effective than imitative techniques are “suggestive.” 2009 Appalachian Student Research Forum Page 36 TH-1 SKEWING OF THE NEONATAL IMMUNE RESPONSE UPON EARLY MICROBIAL EXPOSURE Sean Fox1, Subhadra Nandakumar1, Darshan Shah2, Gayatri Balasubramanyam2, Sandeep Chilakala2, Linda Shanks2, Michael DeVoe2 and Uday Kumaraguru1 1Dept. of Microbiology, 2Dept. of Pediatrics, Quillen College of Medicine, East Tennessee State University, Johnson City, TN The CD4 T helper (Th) immune response may be divided into the Th-1 and the Th-2 immune response based on cytokine secretion pattern. The Th-2 immune system is mainly linked with allergies and is the type of response that is predominant in infants and continues to remain so till late infancy/ early childhood. This scenario could continue until the immune system encounters any microbe or microbial components. The microbial components, called PAMPs (pathogen associated molecular patterns) skew the immune system towards the Th-1 response. Th-1 is the type of CD4 T cell response required for infection prevention. Our previous murine model studies support this hypothesis. Thus to further understand this phenomenon in humans, the study was extended to neonates reporting to ETSU clinics in the Pediatrics department. This translational human study was undertaken after Institutional Review Board approval. Two groups were included: Group A included babies born before 32 weeks of gestation and remained in the neonatal intensive care unit (NICU) for a minimum period of 28 days (Pre-term) and Group B consisted of neonates that were born after 32 weeks of gestation and had no NICU stay (Full term). Both the groups of babies were apparently healthy and were between 9-12 months of age. The hypothesis being tested was that, the NICU stay would have exposed the neonates to various microbes including nosocomial pathogens, thereby stimulating the immune system early in life. Blood samples were collected from all the study subjects after obtaining informed consent from the legal guardian. Isolated PBMCs were stimulated with a cocktail of PAMPs and evaluated for the ability of CD4 T cells to produce Th-1 or Th-2 type of cytokines. IFN- &gamma, TNF-&alpha and IL-2 levels were measured to determine the Th-1 type response while IL-10, IL-4 and IL-13 cytokines were used to determine the Th-2 response of the CD4 T cells. As demonstrated in the mouse model study, the human infants that were exposed early in life (pre-term and hospital stay) to microbes had a better Th-1 response. The CD4 T cells of these babies had a significantly higher ability to produce Th-1 cytokines compared to the unexposed group. The study emphasizes the importance of low microbial exposure from the early stage of life, which may be in form of vaccine antigens, symbiotic microflora or normal environmental microbes. Possible mechanisms that skew the immune system towards the Th-1 response will result in better resistance to infection and allergy. 2009 Appalachian Student Research Forum Page 37 BLOCKING OF THE PROGRAMMED DEATH-1 PATHWAY IN HEPATITIS C VIRAL INFECTION LEADS TO A DOWN-REGULATION OF SUPPRESSOR OF CYTOKINE SIGNALING-1 Ashley D. Frazier, Zhi Q. Yao, and Jonathan P. Moorman, Department of Internal Medicine, College of Medicine, East Tennessee State University, Johnson City, TN Background and Significance: Hepatitis C is a chronic blood-borne viral infection leading to cirrhosis and liver cancer. Healthy T cells exposed to HCV core protein or T cells from individuals with chronic HCV infection exhibit up-regulation of two negative immunomodulatory proteins, PD-1 and SOCS-1, which may contribute to the T cell exhaustion observed during chronic HCV infection. We hypothesize that these negative immunomodulatory proteins are linked together to inhibit T cell functions during HCV infection. Material and Methods: Using peripheral blood mononuclear cells (PBMCs) and T cells isolated from healthy human blood, we incubated these cells with antibodies to block PD-1 and PD-L1 engagement for 6 hours before adding HCV core protein and incubating at 37oC, 5% CO2 overnight. After the overnight incubation, we isolated RNA from our cells using a QIAGEN RNeasy mini kit and subjected this to RT-PCR specific for SOCS-1. Results: HCV core protein exposure leads to up-regulation of PD-1 as well as SOCS-1 mRNA expression in T cells when compared to control protein exposure. Blocking PD-1 / PD-L1 negative regulatory pathways down-regulated the amount of SOCS-1 mRNA expression in HCV core protein treated T cells. Conclusions: SOCS-1 expression in response to HCV core protein treatment in T cells appears to be dependent upon PD-1 signaling pathway. Reversing the negative immunomodulatory impact of HCV infection has the potential to improve the exhausted T cell functions and facilitate viral clearance. RELATIONSHIP OF TRAINING VOLUME TO BONE MINERAL DENSITY IN NCAA DIVISION I CROSS-COUNTRY RUNNERS A.A. Kavanaugh, M. South, K. Painter, R.C. Hamdy, M.E. Stone, M.M. Byrne, M.H. Stone, and M.W. Ramsey, Center of Excellence for Sport Science and Coach Education, Sport Science Laboratory, East Tennessee State University, Johnson City, TN; and G.G. Haff, Exercise Physiology School of Medicine, West Virginia Univ., Morgantown, WV Bone mineral density (BMD) is the quantity of mineral deposited in a given area of bone. High-impact endurance training has been show to increase BMD. Although there may be a threshold level of training volume that stimulates bone formation, higher volumes of endurance training have shown a reduced or negative effect on BMD in some studies. Therefore, weight-bearing sports (i.e. distance/marathon running, triathlons) that demand athletes perform many hours of low-intensity training for several consecutive days, weeks, and months can actually result in a loss of BMD. The mechanical strength of bone is correlated with BMD, and can be measured a number of ways. This study used dual-energy x-ray absorptiometry (DEXA) to measure BMC (g) and BMD (g/cm²). Sites 2009 Appalachian Student Research Forum Page 38 measured by DEXA included, both hips (total, femoral neck, trochanter, shaft, Ward’s triangle area), the lumbar vertebrae, both distal forearms (radius UD, Ulna UD, radius 33%, ulna 33%, total radius, total ulna), and skull. The purpose of this study was to examine the relationship of training volume to BMD of Division I collegiate crosscountry (XC) runners. The BMD at various sites of thirteen (M=7, F=6) D-1 crosscountry runners from East Tennessee State University (ETSU) were compared to fourteen (M=6, F=8) sedentary controls. In addition potential right and left side and sex differences were compared amongst the groups. Self-reported running volume in miles per week was collected one month prior to DEXA measurements. The sedentary control subjects reported no running and no more than two days a week of exercise for less than 30 minutes a day for the previous three months. Statistically, the control versus XC, as well as male versus female group were compared using a two-tailed t-test. The XC runners showed lower BMD (p≤0.05) of the right radius and right ulna compared to sedentary controls. Male XC runners had lower BMD of the left radius (p=0.04) and right radius (p=0.02) and right ulna (p=0.04) compared to male controls. Female XC runners showed lower BMD of the left and right leg (p=0.02) compared to female controls. Overall there was a moderate correlation (r=0.412) between running volume and total body BMD in the XC runners. In males there was a strong positive correlation (r=0.674) between running volume and total body BMD. In females there was a very strong negative correlation (r=-0.757) between running volume and total body BMD. Research suggests that long distance runners have higher bone turnover, which may lead to reduced bone mass dependent on training volume. Important implications to be derived from this data include the importance of monitoring training volume and bone health to reduce the risk of bone injuries related to lower BMD (e.g. stress fractures) in XC runners. Additionally, a properly designed resistance training program should also be implemented into the training process due to the wealth of evidence it has a positive effect on BMD. MACROPHAGE SCAVENGER RECEPTOR A DEFICIENCY PROTECTS AGAINST FOCAL CEREBRAL ISCHEMIA/REPERFUSION INJURY Chen Lu, Fang Hua, Tuanzhu Ha, John Kalbfleisch, John Schweitzer, Jim Kelley, Race Kao, David Williams, and Chuanfu Li. Departments of Surgery, Academic Affairs, Pathology, and Internal Medicine, East Tennessee State University, Johnson City, TN Stroke is the third leading cause of death and remains the leading cause of long-term disability in the United States. About 750,000 Americans suffer a new or recurrent stroke and 157,000 people are killed each year. Recent evident suggests that innate immune and inflammatory responses are involved in thepathophysiology of cerebral ischemia/reperfusion (I/R) injury (stroke). Indeed, we have previously reported that modulation of cellular signaling pathways that mediate innate immune and inflammatory responses attenuated I/R-induced brain injury. Macrophage scavenger receptor A (SR-A) is primarily known for its role in lipid metabolism, but recent evidence indicates that SRA plays an important role in the induction of innate immune and pro-inflammatory responses. We hypothesized that SR-A mediated cellular response contributes to brain I/R injury. To test our hypothesis, we employed SR-A knockout (SR-A-/-) mice to 2009 Appalachian Student Research Forum Page 39 determine the role of SR-A in focal cerebral I/R injury. SR-A-/- mice (n=10) and agematched wild type (WT) mice (n=9) were subjected to focal cerebral ischemia by occlusion of the middle cerebral artery (MCAO) on the left side for 60 min followed by reperfusion for 24 hrs. The brains were harvested and infarct size was determined by TTC staining. Morphology of neurons in brain sections was examined by Nissl staining. In separate experiments, SR-A-/- (n=6) and age-matched WT mice (n=6) were subjected to focal cerebral ischemia (60 min) followed by reperfusion for 24 hrs. Sham surgically operated mice served as sham controls (n=6 in each group). Brains were harvested and cellular proteins were isolated for Western blot analysis of cellular signaling pathway activation. The data showed that infarct size in SR-A-/- mice was significantly reduced by 63.9% (p<0.01) compared with WT mice (6.9 ± 1.91% vs. 19.1 ± 2.49% of whole brain) after cerebral I/R. In SR-A-/- mice, there was less neuronal damage in the hippocampus compared with WT mice following cerebral I/R. Western blots showed that the levels of FasL, Fas, and FADD were significantly increased in WT mice after cerebral I/R. In contrast, these apoptosis associated molecules were not increased in SR-A-/- mice following cerebral I/R. In addition, cerebral I/R increased brain levels of phospho-IkBalpha and NFkB binding activity in WT mice. In SR-A-/- mice, the NFkB activation pathway did not increase following focal cerebral I/R. These data demonstrate, for the first time to our best knowledge, that SR-A plays a critical role in mediating the pathophysiology of focal cerebral I/R injury. Our data also suggest that SR-A-mediated activation of the FasL-mediated apoptotic signaling pathway may be a mechanism of cerebral pathology in I/R. We conclude that modulation of SR-A activity may be a useful approach for ameliorating brain injury in stroke patients. MAPPING TASTE IN THE REWARD SYSTEM: VENTRAL TEGMENTAL AREA (VTA) NEURONAL ACTIVATION VIA TASTE NERVE STIMULATION Marla Perna and Theresa Harrison, Department of Anatomy and Cell Biology, East Tennessee State University, Quillen College of Medicine, Johnson City, TN The ventral tegmental area (VTA) of the brain is known to play an important role in processing the appetitive properties of natural and drug-induced reward. The lateral hypothalamus (LH) also has been implicated in reward processes, particularly those involving food and feeding. Past research has shown that the VTA receives significant projections from the LH, and that the integrity of this LH-to-VTA circuit is necessary for the increased intake of highly palatable foods over ordinary chow seen in satiated rats. Taste is a major determinant of the palatability of foods, but it is unclear just how taste information reaches these components of the reward system. Further, the idea has been advanced that the anterior and the posterior tongue taste receptor systems subserve different functions, such that only taste information conveyed from the back of the tongue via the lingual branch of the IXth (glossopharyngeal) cranial nerve is used to evaluate hedonic (i.e., rewarding) aspects of taste. The present experiments were designed to clarify the route(s) by which taste information reaches the reward system. Using upregulation of the immediate early gene c-fos as a marker for activated neurons, these experiments will compare the ability of electrical stimulation of taste nerves from anterior 2009 Appalachian Student Research Forum Page 40 and posterior tongue to evoked neural activation within the VTA. They will also identify specific populations of neurons involved in taste reward processing, laying the groundwork for future studies of the interconnections of these neurons with taste circuits, and studies of their neuropharmacological regulation. This research will focus on further understanding how taste regulates feeding behavior, as well as how alterations in the neurons and circuits underlying food reward may contribute to palatability-driven overeating and, ultimately, to the disregulation of food intake that leads to obesity. This initial study compares c-Fos expression in the VTA resulting from lingual IXth nerve stimulation with expression in a sham-operated animal. Neurons expressing c-Fos were plotted in drawings through the VTA and their numbers in specific subdivisions were counted. Boundaries and subdivisions of the VTA were defined based on cresyl violet and tyrosine hydroxylase (TH) staining in control brain sections. EXPRESSION OF HUMAN NEUTROPHIL CATHEPSIN G IN K. LACTIS WITH EGFP FUSION FOR DETECTION Eliot Smith and David Johnson, Department of Biochemistry and Molecular Biology, Quillen College of Medicine, East Tennessee State University, Johnson City, TN Human Cathepsin G (CatG) is a serine protease stored in cytoplasmic granules of mast cells and neutrophils and it participates in bacterial killing and inflammatory responses. CatG has the unusual ability to cleave at hydrophobic amino acids (Phe, Tyr, Leu) and at positively charged residues (Lys & Arg); furthermore, this enzyme undergoes an unusual processing step to remove a sequence of residues from its C-terminus. At present, the only source of this enzyme is from human tissue. Expression of recombinant human CatG (rhCatG) would allow further investigation into these poorly understood phenomena and the potential implications of their malfunction. Initial attempts to express rhCatG in the yeast Pichia pastoris met with limited success due to the highly basic character (pI ≈ 11.0) of the enzyme. Current expression attempts in K. lactis show promise in generating a fusion protein of rhCatG linked to domains facilitating expression, secretion, detection, and activation of the recombinant enzyme. The current construct contains an α-mating factor domain at the N-terminus for secretion followed by an enhanced Green Fluorescent Protein (eGFP) domain linked to the active region of CatG via an enterokinase cleavage site. The eGFP domain enables quick initial fluorescence screening for positive expression while the enterokinase cleavage site allows activation of rhCatG by removing eGFP. A gene encoding rhCatG that has been optimized for yeast codon usage was modified using PCR to include a 5’ sequence coding for the enterokinase cleavage site. The genes encoding eGFP and CatG were both modified during PCR amplification to incorporate recognition sequences for specific restriction endonucleases. The restriction sites facilitate subsequent assembly of the vector pKLAC1-eGFP-rhCatG which includes two genes for transformant selection and the “α-mating factor-eGFP-rhCatG” gene. The pKLAC1 vector uses a homologous recombination strategy to incorporate the genetic construct into the native K. lactis genome. Upon transformation, this strategy places the gene under control of the galactose-inducible promoter LAC4. In media containing galactose, properly 2009 Appalachian Student Research Forum Page 41 transformed yeast secrete the eGFP-CatG fusion protein into the media, facilitating purification. Yeast colonies that show eGFP production are further tested to confirm production of rhCatG by enterokinase activation, affinity purification using bovine pancreatic trypsin inhibitor (BPTI), and activity assays. Activity tests determine the rate of enzymatic cleavage of synthetic peptide-thiobenzyl ester substrates, which produce a measureable color change. Upon successful identification of a colony that produces active rhCatG, further studies will modify the amino acid sequence for the enzyme, specifically to improve understanding of the uncommon C-terminal processing and dualspecificity of this enzyme. Supported by the National Heart, Lung and Blood Institute via grant 1R15HL091770 RELATIONSHIP OF STRENGTH, AND POWER CHARACTERISTICS TO OVERHEAD SHOT THROW PERFORMACE IN NCAA DIVISION I MALE THROWERS A.M. Swisher, M.W. Ramsey, M.E. Stone, C. Nelson, A. Kavanaugh, A. Layne, M. South, and M.H. Stone, Center of Excellence for Sport Science and Coach Education, Sports Science Laboratory, East Tennessee State University, Johnson City, TN Particularly in strength-power-oriented sports, such as the throwing events in track and field, it is known that maximum strength, power output, and rate of force development (RFD) are of primary importance to competitive success. Although 7.26 kilogram overhead backward shot put throws (7.26k OH) are often used by coaches in assessing the “fitness” of throwers, surprisingly little is found in the scientific literature concerning its efficacy as a useful assessment tool. The purpose of this study was to examine the relationships between strength and power characteristics of male collegiate throwers and 7.26k OH performance. Male NCAA Division I track and field throwers (n = 18, 19.1 ± 1.2 yrs old.; 185 ± 6 cm tall, 192 ± 9 cm wingspan; body mass 114.9 ±14.8 kg) from several southeastern universities volunteered for testing. Subjects were assessed for maximum strength (mid-thigh isometric clean pull) and power using countermovement (CMJ) and static jumps (SJ) with loads of 0 kg, 11 kg, and 20 kg. In addition to the 7.26k OH, isometric peak force (IPF), jump peak power (PP), and rate of focre development (RFD) were measured. There were significant positive correlations between 7.26k OH and IPF, instantaneous forces at 50, 90, and 250 ms, RFD, and CMJ height. The data also indicate that the 7.26k OH is moderately related to PP in loaded SJs and CMJs. Correlations ranged from r = 0.48 – 65, p < 0.05. Of particular interest is the strong relationship between 7.26k OH and force produced at 250 ms as a thrower has about the same amount of time to generate force in the final phase of a throw. These data suggest that the 7.26k OH may be a good indicator of strength and explosiveness in throwers. Practically, the 7.26k OH could be used by throws coaches as a reliable, quick, and inexpensive means of assessing explosive strength and power as well as monitoring the training progress of throwers. Future research should examine the relationship between the 7.26k OH and throwing performance. 2009 Appalachian Student Research Forum Page 42 Natural Sciences & Mathematics COMPUTER SIMULATION OF WASPS’ NEST BUILDING PROCESS Fadel Adoe1, Istvan Karsai2 and Chris Wallace1, 1Department of Computer Science; 2 Department of Biological Sciences, East Tennessee State University, Johnson City, TN Self organization is a process in which a pattern emerges at the global level of a system solely from numerous interactions among lower-level components of the system. One example is how wasps individually yet cooperatively build a comb from hexagonal cells. However, the underlying behavior that facilitates the emergence of this global pattern is not generally well understood. This paper focuses on a special random building script and characterizes how nests are built using this simple building algorithm. In this model, the nest is first initiated with a single hexagonal cell and then six neighbors that correspond to the sides of the cell are generated and stored as potential building cells (domain cells) of the emerging nest. The next cell added to the nest is picked from the domain cells. Cell picking process is currently done randomly, but with progress will be based on a probability distribution function that is a function of wasps’ building behavior. Neighbors are generated and also stored for the new added cell. This process of picking, generating and storing neighbor cells is done recursively until a predetermine nest size is reached. With this approach we simulate the random building script, when the wasps have neither global information on the nest nor preference to build at any site in the nest. With this approach, various global structures will emerge of which the centre of mass and compatibility would be calculated. These calculated parameters would be used as feedback indicators to optimize the probability distribution function. This work will help gain better insight and visualization through simulation into wasps building behavior and organization. This acquired knowledge can be applied to collective robots and the business industry. EFFECTS OF CLIMATE CHANGE ON PLETODON WELLERI IN SOUTHERN APALACHIAN MOUNTAIN RANGE. Kirsten Borger and Istvan Karsai, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Climate change has the ability to change individual species as well as the entire ecosystem. Predictive models need to be developed to assess future impacts of recent effects. We developed an agent based model to simulate the effect of climate change to Plethodon welleri. This is an endemic species of the Southern Appalachian Mountain Range and their populations would be affected by even small changes in the climate. Different what-if scenarios are generated to predict the population size and genetic diversity under different climate change regime. Our results show that the population range, size and genetic diversity of P. welleri will substantially change with climate. We 2009 Appalachian Student Research Forum Page 43 also present both beneficial and detrimental scenarios. In case of climate change toward a warmer, less humid direction, the population decreased to a few individuals, and the genetic diversity also decreased. This probably will result in the extinction of the species. In case of climate change toward a colder, more humid direction, the population increase and will spread. Further study of this ecosystem and using modeling on the possible effect of various climate factors seems very efficient way to prepare conservation efforts in time. INHIBITORY EFFECTS OF POLYPHENOLS ON ESCHERCHIA COLI ATP SYNTHASE Prasanna K Dadi, Mubeen Ahmad, and Zulfiqar Ahmad, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Recently published X-ray crystal structures of bovine mitochondrial ATP synthase inhibited by resveratrol, piceatannol, and quercetin, suggest that these compounds bind in a hydrophobic pocket between the gamma-subunit c-terminal tip and the hydrophobic inside of the surrounding annulus in a region critical for rotation of the gamma-subunit. Inhibitory effect of five polyphenols, resveratrol, piceatannol, quercetin, quercetrin, quercetin-3-beta-D glucoside and seventeen modified polyphenols on E. coli ATP synthase is shown here. Polyphenols are naturally occurring plant based phytochemicals which possess antioxidant, chemopreventive, and chemotherapeutic properties. Foods such as apples, berries, cantaloupe, cherries, grapes, pears, plums, broccoli, cabbages, and onions are rich in polyphenols. Physiological relevance of dietary polyphenols can be ascribed to their interaction with mitochondrion in eukaryotic cells. Polyphenol led inhibitory studies on bovine mitochondrial suggest that the beneficial effects of dietary polyphenols are, in part, linked to the blocking of ATP synthesis in tumor cells thereby leading to apoptosis. Furthermore, Polyphenols are also known to have antimicrobial activity through inhibitory actions on ATP synthase. It was shown that polyphenols can inhibit biofilm formation and acid production by S. mutans. One of the pathways through which polyphenols are active against traits of S. mutans is the inhibition of proton– translocating F1-ATPase activity. For the first time, our results show that polyphenols, resveratrol, piceatannol, quercetin, quercetrin, or quercetin-3-beta-D glucoside all inhibit Escherichia coli ATP synthase reversibly. It was found that piceatannol was the most potent inhibitor (IC50 ~14 microM) followed by quercetrin (IC50 ~20 microM), quercetin (IC50 ~33 microM), quercetin-3-beta-D glucoside (IC50 ~71 microM), and resveratrol (IC50 ~94 microM). Inhibitory studies by seventeen modified polyphenol compounds based on repositioning and or addition of –OH, COOH, NH2 group and phenolic rings resulted in potent inhibition of E. coli ATP synthase on a molar scale with very low IC50 values 2009 Appalachian Student Research Forum Page 44 POLLINATION BIOLOGY OF CAMPSIS RADICANS (BIGNONIACEAE): IS IT REALLY BIRD-POLLINATED? Andrea A. Edge, Byron N. Van Nest, Jennifer N. Johnson, and Darrell Moore, Department of Biological Sciences, East Tennessee State University, Johnson City, TN While classifying plants into pollination syndromes can be useful to describe potential pollinators, growing research suggests that plant-pollinator interactions are much more intricate than these broad syndromes imply. Many plants are pollinated by visitors other than what a particular syndrome suggests. Pollinator interactions may be strongly influenced by flower morphology as well as the rewards that the flowers offer. A deeper understanding of floral physiology and morphology, including changes that may occur with time of day or with age, may provide insights into the dynamics of plant-pollinator interactions. Flowers of the trumpet vine Campsis radicans are described as possessing the classical bird-pollination syndrome (ornithophily). These flowers are thought to be pollinated primarily by the ruby-throated hummingbird (Archilochus colubris), although we have seen that many insects (including honey bees, bumble bees and sweat bees) visit the flowers frequently. The primary goals of this study are to determine the major pollinators for the trumpet vine, assess their relative importance to reproductive success of the plant, and ascertain the morphological and physiological factors that influence pollinator visits. If trumpet vines indeed exhibit a true ornithophilous (bird pollination) syndrome, then its flowers should contain large amounts of dilute nectar and hummingbirds should be frequent visitors. Also, in agreement with the ornithophilous hypothesis, we would predict that nectar volume will be highest during the daylight hours when it would be most useful in attracting birds. We measured nectar secretion and concentration patterns of unopened and open trumpet creeper flowers with respect to time of day and age. Our findings thus far show that closed Campsis flowers do not produce a significant amount of nectar and sugar until they are 40-50mm in length. Open flowers produce large amounts of dilute nectar, in which both volume and concentration show no significant changes with time of day (or night) or with age. We currently are designing studies to determine (1) if nectar production changes as volumes are removed by pollinators and if additional production changes based on time of day or with the age of flower, (2) the identity and temporal visitation patterns of the various pollinators, in light of the types of visitors that are naturally occurring in the local environment, and (3) patterns of pollen movement within a population to assess other aspects of reproduction such as low fruit set. ABSENCE OF BEHAVIORAL RHYTHMICITY IN THE BEHAVIOR OF HONEY BEE QUEENS (APIS MELLIFERA) Jennifer N. Johnson, Curtis Gill, Emily Hardgrave, and Darrell Moore, East Tennessee State University, Department of Biological Sciences, Johnson City, Tennessee Apis mellifera (honey bee) workers assess the needs of their colony and determine whether a fertilized egg (a female) will become a queen or a worker. The developmental 2009 Appalachian Student Research Forum Page 45 program through which the female larva will be channeled depends on the type and amount of food she is given. The queen is specialized for reproduction for virtually her entire adult life whereas the workers progress through an age-related series of tasks. Younger workers are behaviorally arrhythmic and perform necessary in-hive tasks both day and night. As they age, the worker bees develop a diurnal circadian rhythm just prior to the last task of their lives (foraging). Does the queen also exhibit behavioral rhythmicity? Our working hypothesis is that, once she has navigated her nuptial flight(s), it is not adaptive for the queen to express any circadian rhythmicity in her behavior and therefore there will be no differences in her activity levels between day and night. We tested for the presence or absence of behavioral rhythmicity by using two separate experimental approaches. Both approaches involved detailed observations of queen behavior, as well as the number of attending workers and the level of queen-feeding and queen-grooming behaviors performed by these workers, within glass-sided observation hives. Observations of active (walking, cell inspections, and egg-laying) and inactive (standing) behaviors performed by a single queen were made at four different 2-hour surveillance periods (two during the day and two during the night), over three consecutive days, once each month for five consecutive months (August 2007-Janury 2008). Additionally, we used a scan-sampling approach utilizing multiple queens (August 2008-October 2008). Observations were made every 30 minutes for three-hour increments over 9, 6, or 5 days. Sampling in this manner allowed us to collect data on multiple queens for all hours of the day and night. No day/night (diel) rhythmicity was observed for any queen behavior or queen attendant behavior under either of the two experimental approaches. Furthermore, partitioning the behaviors into the categories of active and inactive also revealed that mated honey bee queens exhibit no significant changes in activity level throughout the 24-hour day, even in the face of natural changes in temperature and light levels between day and night. CLONING, HETEROLOGOUS EXPRESSION, AND ELUCIDATION OF BIOCHEMICAL FUNCTION OF PUTATIVE FLAVONOID GLUCOSYLTRANSFERASES CLONES (PGT2, 3, AND 9) FROM CITRUS PARADISI Venkata S. Mallampalli1, Daniel K. Owens1 and Cecilia A. McIntosh1 & 2 (1Dept. Biological Sciences, 2School of Graduate Studies, East Tennessee State University, Johnson City, TN Flavonoids are a major group of plant secondary metabolic compounds. Flavonoids play a variety of roles for plants including protection against UV light, pigmentation, and flavor (which can be important for pollination and seed dispersal), antimicrobial and antifungal properties, sexual reproduction and pollen development, and cues for microbial symbiont colonization. Flavonoids are also known to have benefit to human health due to antioxidant and anticancer properties. At every branch point of flavonoid biosynthesis, flavonoids can either be converted into other flavonoid classes via the core biosynthetic pathway or can be derivatized to give the distinct compounds found in plants. Glucosylation is a prominent modification reaction in flavonoid biosynthesis and 2009 Appalachian Student Research Forum Page 46 these reactions are catalyzed by glucosyltransferases (GTs). Citrus paradisi (grapefruit) is an excellent model system for studying flavonoid GTs as it is known to contain GTs capable of producing flavanone, flavone, and flavonol 7-O-glucosides, flavonol 3-Oglucosides, and chalcone glucosides. For example, 40-70% of the dry weight of very young Citrus paradisi fruit and leaves is due to a single bitter flavanone diglycoside, naringin. Current efforts are focused on cloning, expression and characterization of putative secondary product GTs from grapefruit. The research described here is designed to test the hypothesis that grapefruit PGT2, 3 and 9 clones are flavonoid glucosyltransferases. PGT2 was obtained by designing primers based on the PSPG (Plant Secondary Product Glucosyltransferase)- box and a full length clone was obtained by walking out to the extreme ends. PGT3 was obtained by directional cloning using an EST mining approach. PGT2 and PGT3 clones have been cloned into pCD1 vector and transformation into BL21 cell lines has been achieved. Results from experiments designed to optimize heterologous expression of soluble PGT2 and 3 protein by testing media composition, culture temperature, and induction duration are presented. Additionally, the harvEST database, containing ESTs from different citrus varieties, was used as a bioinformatics tool to identify additional PGT clones and primers were designed based on consensus sequences to fish putative flavonoid GT’s out of grapefruit leaf cDNA. We present here the cloning, expression, and purification of the PGT9 identified by this means. Upon heterologous expression of soluble protein, expressed proteins will be screened for flavonoid GT activity using a suite of flavonoids that have been identified in citrus. If flavonoid GT activity is not found, the expressed enzymes will be tested with secondary metabolites including coumarins and other phenolic compounds. If activity is found, the expressed enzymes will be rigorously characterized including: elucidation of the reaction product(s), substrate specificity, optimum pH, activators, inhibitors, isoelectric point, and kinetic parameters. QUANTITATIVE ASSESSMENT OF SEXUAL DIMORPHISM IN MARBLED SALAMANDER AMBYSTOMA OPACUM Lok Raj Pokhrel, Istvan Karsai and Thomas Laughlin, Department of Biological Sciences, East Tennessee State University, Johnson City, TN As little is known about some aspects of sexual dimorphism in Ambystomatid salamanders, we tested the hypothesis that two sexes are different in morphological characters and dorsal body pigmentation in marbled salamanders. We tested differences in many body size measurements and the relative body coverage of white pigmentation from 275 live specimens collected from Osceola Island, Sullivan County in Northeast Tennessee. Our results showed that females are significantly heavier, longer with greater dorsal body area, but significantly lesser white skin pigmentation on the dorsum than males. This study presents evidence of sexual dimorphism in marbled salamanders; however the significance of such dimorphism in this species remains to be tested. 2009 Appalachian Student Research Forum Page 47 PHYLOGEOGRAPHIC STUDY OF THE YONAHLOSSEE SALAMANDER Joshua Rudd and Andy Karsai, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Plethodon yonahlossee is the largest eastern Plethodontid salamander and has been classified as a species of greatest conservation need by the Tennessee Wildlife Resources Agency (TWRA). Found only in mountainous areas along the borders of Tennessee, North Carolina, and Virginia, populations of the yonahlossee are considered to be rare and local throughout their range. Genetic differentiation among populations of any species is usually attributable to long-standing, extrinsic barriers to gene flow. Because of their disjunct populations structure and some observed morphological variation, genetic differentiation among yonahlossee salamander populations is predicted. A phylogeographic study of yonahlossee populations was conducted to identify any genetically differentiated populations or evolutionary significant units (ESU). An ESU is a population of a species which shows genetic differentiation and therefore a potential for local adaption. For this project, one mitochondrial DNA marker as well one nuclear DNA marker were amplified using polymerase chain reaction. The amplifiedmarkers were then sequenced for analysis. After analysis, the mitochondrial marker does not suggest significant genetic differences between populations. The nuclear marker does however show some variation between populations. THE STIMULABILITY TREATMENT APPROACH FOR SPEECH SOUND DISORDERS IN YOUNG CHILDREN: A SYSTEMATIC REVIEW Whitney Boyd, Ashley Rymer, Heather Carpenter, and A. Lynn Williams, Department of Communicative Disorders, East Tennessee State University, Johnson City, TN Although there are a number of promising intervention approaches to treat speech sound disorders in children, it is difficult to determine which approach would be the most effective for a particular child. With the current emphasis on evidence-based practice (EBP), clinicians need to critically evaluate the research to choose the best approach for their client. A relatively new approach for young children with speech disorders is the stimulability approach (Miccio & Elbert, 1996). This approach utilizes hand motions associated with character cards for each consonant sound within a turn-taking format to expand a child’s sound inventory. The purpose of this investigation was to complete a systematic review of empirical studies of the efficacy of the stimulability approach. Following the tenets of evidence-based practice, a clinical question was posed following the PICO framework (Centre for Evidence Based Medicine, 2001; Sackett, Straus, Richardson, Rosenberg, &Haynes, 1996), in which P represents the Patient, Patient Group, Population, or Problem; I represents the Intervention under consideration; C represents the Comparison intervention (i.e., either the standard approach or no intervention); and O represents the desired Outcome. Thus, our EBP question was this: For preschool-aged children with moderate to severe phonological impairments, does the 2009 Appalachian Student Research Forum Page 48 stimulability treatment approach provide greater system-wide generalization compared with no treatment. A set of inclusionary/ exclusionary criteria were developed for identifying studies to be included in this review, including English-speaking preschool children ages 2-4 years old with a phonological impairment. The independent variable was stimulability treatment versus no treatment and the dependent measures were child speech outcomes. Databases searched included PsychINFO; Educational Resources Information Center (ERIC); and American Speech-Language-Hearing Association Journal Archives. The search outcome yielded 1,692 studies. Of these, 913 were screened for inclusion, and from this, 18 were selected for further examination of the full-text articles. Of the 18 articles selected for review, 2 articles met the exclusionary and inclusionary criteria. The quality of evidence of these studies was evaluated using the Critical Appraisal of Treatment Evidence (CATE; Dollaghan, 2007). Systematic review of the outcomes showed that the stimulability approach did result in greater system-wide change as opposed to no treatment. With the EBP framework, clinicians can integrate the evidence presented in this review with other sources of information, such as child and family preferences, their own clinical experiences of different intervention approaches, and the culture in which they work, to make the best decisions concerning the treatment models they use to meet the needs of the children with speech sound disorders. ROLE OF SALICYLIC ACID BINDING PROTEIN 2 IN SYSTEMIC ACQUIRED RESISTANCE INDUCED BY ACIBENZOLAR-S-METHYL Diwaker Tripathi, Arrey Enyong, Yu-Lin Jiang and Dhirendra Kumar, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Plant diseases caused by microbial pathogens result in devastating losses in productivity of food and fiber crops. To fight with these microbial pathogens, plants have evolved resistance (R) proteins which recognize and interact with the avirulence (Avr) proteins of the pathogens. This interaction between Avr molecules and R proteins initiates the resistance response of the host towards the invading pathogens. This Local resistance (LR) overtime leads to the development of a broad spectrum resistance (SAR) in uninfected parts of the plants. It acts by increasing the level of reactive oxygen species (ROS) which results in the plant cell death in infected area (hypersensitive response) Moreover it activates the accumulation of salicylic acid (SA) and expression of a set of nine families of pathogenesis-related (PR) proteins. SA, a plant hormone, is essential for the full development of SAR. Treatment of plants with SA makes them resistant to pathogens. SA-binding protein 2 (SABP2) is a methyl salicylate esterase that catalyzes the conversion of lipid mobile methyl salicylate (MeSA) biosynthesized in plants resisting pathogens into SA. Plants which do not express SABP2 are unable to convert MeSA into SA and fail to develop effective SAR. Various functional analogs of SA have been developed and used to activate plants own natural defenses against microbial pathogens. ASM (Acibenzolar- S-Methyl) is one such functional analog which induces similar responses as pathogens in monocot and dicot plants belonging to various families. ASM is commercially available by the name Actigard. How ASM functions in the plant is mostly unclear? Results from our preliminary experiments using TLC and HPLC 2009 Appalachian Student Research Forum Page 49 analyses suggest that SABP2 catalyzes the conversion of ASM ester into its acid form. This research is designed to test whether this conversion of ASM ester into its acid form by SABP2 is essential for the activation of SAR is the focus of this study. For this study we are using two transgenic tobacco lines, 1) control line containing empty silencing vector and 2) SABP2 silenced tobacco line. Expression of defense proteins e.g. PR-1 in both these lines treated with ASM is being tested and results will be presented. SAR induced by ASM in both these lines is currently being tested and results will indicate if SABP2 is required for the conversion of ASM ester into its acid form for the full induction of SAR. This study will help to understand the action mechanism of ASM and will be useful in developing more efficient resistance inducing chemicals. 'SHOULD I STAY OR SHOULD I GO?' INDIVIDUAL DECISION MAKING IN THE HONEY BEE (APIS MELLIFERA) FORAGER OPTIMIZES GROUP FORAGING EFFORTS Byron N. Van Nest and Darrell Moore, Department of Biological Sciences, East Tennessee State University, Johnson City, TN Classical experiments on honey bee time-memory showed that foragers trained to collect food at a fixed time of day will return the following day with a remarkable degree of time-accuracy. We have found that not all time-trained foragers visit the training station on an unrewarded test day immediately following the last day of time-training. Rather, the trained foraging group comprises two sub-groups: persistent bees and reticent bees. Both sub-groups cluster on the dance floor in anticipation of the training time, and persistent bees then leave the hive to reconnoiter their food source. Reticent bees, however, will not leave the hive until they observe a recruitment dance from another bee confirming the availability of their source. In light of the ability of foragers to recruit hivemates (via the famous round and waggle dances) to a productive food source, it is surprising that so many bees within a foraging group are persistent (40 – 90%, depending on the number of days of experience at the source). What is the benefit to the colony in sending so many foragers to investigate a source that may or may not still be productive when only a few foragers are required to reactivate the entire group? We used an agentbased software model to test the energetics underlying several different ratios of persistent and reticent individuals in the foraging group while varying six different behavioral and ecological factors: (1) forager group size, (2) source distance from the hive, (3) source nectar concentration, (4) number of hours during the day the source is available, (5) number of days the source is known to the colony, and (6) the rate at which new unemployed foragers appear on the dance floor, receptive to recruitment by returning foragers. Energy expenditure and energy capture results are based on previously published values. Our model demonstrates that, under typical foraging conditions (intermediate values of source distance and nectar concentration), the energetically optimal strategy is for the foraging group to contain bees with differing levels of experience and thus differing levels of persistence, in agreement with our empirical findings. 2009 Appalachian Student Research Forum Page 50 COMPUTATIONAL INVESTIGATION OF SPIN TRAPS USING HYBRID SOLVATION MODELS Sai Sriharsha Konda and Scott J. Kirkby, Department of Chemistry, East Tennessee State University, Johnson City, TN The cyclic nitrone 5,5-dimethyl-1-pyrroline-N-oxide (DMPO), and the lesser known linear phenyl-N-tert-butylnitrone (PBN) and its phosphorylated analogues have been used as spin traps for the investigation of free radicals in biological systems. Theoretical work on these molecules suggests that there are important differences in their properties between biological systems and isolated molecules in the gas phase, most likely resulting from intra and intermolecular hydrogen bonding. Most dielectric solvation models such as the polarized continuum model and COSMO are incapable of direct determination of solvent-spin trap chemical interactions. To examine this, hybrid models incorporating COSMO for long range effects and discrete solvent molecules for short range effects, at the DFT/B3LYP/6-31G* level of theory, have been used to study the stabilization and alteration of the spin trap molecules properties in protic and aprotic polar solvents. Social & Behavioral Sciences EVALUATING INTEGRATED PEDIATRIC WELL VISTS IN APPALACHIAN TENNESSEE Angela D. Enlow, M. A.1 and Jodi Polaha, Ph.D.2; 1Fielding Graduate University, School of Psychology Santa Barbara, CA; 2Department of Psychology, East Tennessee State University, Johnson City, TN The aim of this project was to examine key medical and behavioral health issues identified in the context of integrated wellness visits. Specifically, this study sought to examine the role of psychologists in pediatric wellness visits, including the early identification of behavioral, developmental, and learning problems, as well as the delivery of brief, effective interventions. The study was conducted at Cherokee Health Systems, a fully integrated pediatric practice in East Tennessee, which follows a fully integrated model of integrated primary care. Data was obtained from a record review of 100 participants, providing a total of 373 well visits. Outcomes demonstrate that behavioral and developmental concerns are raised at a higher rate than in non-integrated practices. Specifically, visits in which the psychologist was present, 44.42% resulted in some concern being raised. Further, 36.24% of visits resulted in psychosocial concern(s) being raised. This figure is significant given that 22% of well visits typically result in discussion of some psychosocial concern consistently found in the research literature. Additionally, of concerns raised, 81.87% were raised directly with/by the psychologist on staff, who is able to offer immediate interventions when concerns are raised. 2009 Appalachian Student Research Forum Page 51 CARDIAC VAGAL TONE AND EMOTIONAL REACTIVITY IN CHILDREN WHO STUTTER Michelle L. Hogg, Department of Communicative Disorders, College of Clinical and Rehabilitative Health Sciences, East Tennessee State University, Johnson City, TN Stuttering is a speech impairment that affects approximately 5% percent of children between the ages of 2-6 years old. There are numerous factors that have been shown to influence the occurrence and maintenance of dysfluencies including linguistic, motoric, and emotional responses. From self reported measures, it has been found that children who stutter are more emotionally reactive and less able to regulate their emotions (Krass et al., 2006). Although self reports from parents can provide information pertaining to emotional reactions of the child, direct psychophysiological measures such as skin conductance and heart rate may provide valuable data regarding emotional reactivity. The present study was planned to evaluate emotional responses of children who stutter. Respiratory sinus arrhythmia (RSA), an index of emotional reactivity, was used to understand the children’s emotionally reactivity to video stimulus. A series of 5 animated film clips was used to elicit emotional responses in participants. A 5-minute film clip of the video, SPOT, a short story of a playful neighborhood dog, was presented as a baseline sample. Four three-minute animated film clips designed to elicit emotional responses were also used: (a) Positive Affect: 3-minute clip of 'Open Season,' ; (b) Negative Affect: 3-minute clip of 'The Lion King,” ; (c) Positive Affect: 3-minute clip of “Ice Age II,” (d) Negative Affect: 3-minute clip of “Mulan II,” Electrodes were placed on the child to measure their heart rate and heart rate variability. Results indicated that there was a difference in emotional response of children who stutter verse children who do not stutter. This study is greatly valuable in understanding the precursors for stuttering as this is the first study to use physiological measure to evaluate emotional response near the onset of stuttering. Further understanding of emotional reactivity as it relates to stuttering will provide great evidence as to whether children who stutter will spontaneously recover without therapy. TEMPERAMENTAL ASSOCIATIONS OF PRESCHOOL GRAMMATICAL DEVELOPMENT Leslie King, Elizabeth Hay, and Wallace E. Dixon, Jr. Ph.D., Department of Psychology, East Tennessee State University, Johnson City, TN Temperament is generally regarded as a cluster of dimensions reflecting biological predispositions. It is observed at birth and remains fairly stable throughout the lifetime. A growing body of research has linked temperamental constructs in infancy to language development. Temperament dimensions such as attentional focusing, low & high intensity pleasure, perceptual sensitivity, and discomfort have all been associated with vocabulary development in 21-month-olds. However, to date, no study has investigated whether temperament continues to associate with language development in later years, 2009 Appalachian Student Research Forum Page 52 especially with regard to grammatical development among preschoolers. Hence, the present study examined the relationship between temperament and language development in preschoolers. We hypothesized that because temperament is fairly stable over time; preschool temperament would associate with preschool language development in a manner similar to that found in the infant literature. Participants included 20 three- and four-year-old children enrolled in a university-affiliated preschool. Children’s grammatical development was assessed on-site at the center using the Preschool Language Scale (PLS-IV). Rothbart’s Child Behavior Questionnaire (CBQ) was completed by parents as a measure of children’s temperament. Standardized scores from both the PLS and CBQ were analyzed using zero-order correlation. Consistent with the infant research, preschoolers’ attentional focusing, low-intensity pleasure, and perceptual sensitivity were positively associated with their grammatical development (r’s = .55, .55, .49 respectively, all p’s < .05). The direction of the relationships indicated that moreadvanced grammatical development was associated with high attentional focusing, high scores in low-intensity pleasure, and high perceptual sensitivity. Contrary to expectations, children’s discomfort scores were also positively correlated with their grammatical development (r = .57, p < .05), suggesting that children who were regarded by their parents as generally discontented tend to have more advanced grammatical development. Finally, also contrary to expectations, children’s high intensity pleasure scores were negatively correlated with their grammatical development (r = -.47, p < .05), suggesting that children low in high-intensity pleasure were grammatically advantaged relative to their peers. An unexpected negative correlation between activity level and grammatical development (r = -.53, p <. 05) suggests that the most temperamentally active children were the least grammatically advanced. Taken together, these findings suggest that while some dimensions of temperament remain associated with language development throughout the preschool period; others dimensions may change their association over this period. The limited amount of related research involving children in this age range as well as the importance of grammatical development makes these findings noteworthy. PHYSICAL ACTIVITY AND WEIGHT PERCEPTION AMONG RURAL APPALACHIAN MIDDLE SCHOOL GIRLS Lauren E. LaBounty, B.S.* and Karen Schetzina, M.D., MPH***East Tennessee State University, Department of Kinesiology, Leisure, & Sport Science, Johnson City, TN; **East Tennessee State University- Quillen College of Medicine, Department of Pediatrics, Johnson City, TN Background: National surveys suggest that girls are less physically active than boys and that their physical activity disproportionately decreases with age.Objective: To better understand physical activity among adolescent girls in the region.Methods: Sixth grade students from five schools in three different school systems in Northeast Tennessee participated during spring 2008 in a written survey developed by the researchers including items from Pathways and Middle School Youth Risk Behavior Surveys.Results: 230 girls and 198 boys participated in the survey with the mean age of 11.7 years for girls and 12 years for boys. Ninety-four percent of girls and 90 percent of 2009 Appalachian Student Research Forum Page 53 boys were white. Students reported having PE an average of 2 days per week. Sports team participation was equal among girls and boys (29 percent did not participate). Fiftytwo percent of girls and 59 percent of boys reported participating in moderate to vigorous physical activity at least 5 days per week. Twenty-six percent of girls and 31 percent of boys felt that they were either slightly or very overweight. Forty-nine percent of girls and 37 percent of boys reported that they were trying to lose weight.Conclusions: Compared to U.S. middle school students, sixth grade students in this sample from rural Appalachia are receiving PE less often, but participating in sports teams more frequently. A greater proportion of girls in this sample reported being compliant with physical activity recommendations than is reported nationally for middle school students. Nearly twice as many girls reported that they were trying to lose weight than described themselves as overweight.Clinical Implications: Physical inactivity and unhealthy weight loss practices place girls at risk for serious health problems that may persist through adulthood ADOLESCENT AMPHETAMINE SENSITIZATION IN A RODENT MODEL OF PSYCHOSIS: CHANGES IN ACCUMBAL DOPAMINE USING MICRODIALYSIS Meredith L. Smith, Jessica J. Smith, Kimberly N. Huggins, and Russell W. Brown, Department of Psychology, East Tennessee State University. Johnson City TN The study will analyze amphetamine-induced behavioral sensitization and dopamine release in the nucleus accumbens (NAcc) utilizing a rodent model of psychosis that is most relevant to schizophrenia, and both male and female adolescent rats will be used as subjects. The rodent model of schizophrenia used in this proposal is based on long-term supersensitization of the dopamine D2 receptor subtype through neonatal administration of quinpirole, a dopamine D2/D3 agonist. This is a phenomenon also known as dopamine D2 receptor ‘priming.’ In this study, only females were used as subjects because previous work has shown more prominent effects of d-amphetamine in adolescent females as compared to adolescent males. Female rats were administered quinpirole (1mg/kg) or saline from postnatal days (P)1-11 and raised to adolescence (P30). After habituation to the locomotor arena, rats were administered d-amphetamine sulfate (1mg/kg) or saline every other day for 12 days from P35-47, and horizontal activity was recorded. The day after sensitization was complete, all animals were surgically implanted with a guide cannula aimed at the nucleus accumbens core. Six days later, a microdialysis probe was implanted through the cannula and baseline samples were taken every 20 minutes for 2 h. Animals were then given either saline or a cumulative dose curve of d-amphetamine (0.1 – 2.5 mg/kg) and samples were taken every 20 minutes for 3 h. ANOVAs were used as the primary statistic, and Newman-Keuls post hoc tests were used for analysis of significant interactions (p=.05). A 2 x 2 x 3 three-way ANOVA on horizontal activity revealed a significant three-way interaction of neonatal drug treatment x adolescent drug treatment x day of testing F(1,23) = 5.44, p<.02. D2-primed rats treated in adolescence with amphetamine demonstrated a significant increase in horizontal activity compared to all other groups by the end of testing. A 2 x 2 x 9 three-way ANOVA on accumbal dopamine release revealed a significant neonatal drug treatment x adolescent drug treatment two-way interaction F(1,16) = 53.09, p<.001. D2-primed rats administered 2009 Appalachian Student Research Forum Page 54 amphetamine demonstrated a 170% increase in accumbal dopamine release compared to non D2-primed rats administered amphetamine. Consistent with past data in schizophrenics, amphetamine produces a significant increase in accumbal dopamine release in D2-primed versus non D2-primed rats. This suggests that schizophrenics may demonstrate increased positive reinforcing effects of the drug, resulting in an increased propensity for amphetamine abuse. ESZOPICLONE FACILITATION OF THE ANTIDEPRESSANT EFFICACY OF FLUOXETINE USING A SOCIAL DEFEAT STRESS MODEL IN THE MOUSE. Jessica J. Smith, Meredith L. Smith, Daniel M. Noel, A. Brianna Sheppard, and Russell W. Brown, Department of Psychology, East Tennessee State University. Johnson City, TN This study was designed to analyze whether the sleep aid eszopiclone (trade name: Lunesta) may facilitate the efficacy of fluoxetine (trade name: Prozac) on social defeat stress (SDS) in the mouse. The SDS model has been shown to be an effective model for major depressive disorder in humans. Recent work has shown that eszopiclone facilitates the antidepressant action of fluoxetine in patients with major depressive disorder. We tested the hypothesis that eszopiclone would facilitate action of fluoxetine in the alleviation of behavioral stress responses produced by SDS. Subjects were adult male ‘intruder’ mice that that were exposed to a retired ‘resident’ male breeder mouse in his home cage for a 5 min period for 10 consecutive days. During this interaction, the resident would establish physical dominance of the intruder. After confrontation, the intruder was housed in a cage that allowed sensory contact with the resident mouse but would not allow physical interaction. Once the 10 days of interaction were complete, all animals were assigned to one of four drug treatment groups, and treatment was given up to 18 days: saline treatment, fluoxetine (10 mg/kg) only, eszopiclone only (3 mg/kg), or fluoxetine + eszopiclone. A defeated and non-defeated group was included (N=6-8 in all groups). A social interaction test was given on days 1, 5, 10, and 15 of drug treatment in which the aggressor was placed in a Plexiglas box within an arena with the intruder. Animals were able to have sensory contact during this interaction, and time spent in avoidance zones were recorded. A 2x4 ANOVA was used for analysis of behavioral dependent measures, and the Newman-Keuls test was used to analyze significant interactions (p=.05).: Most importantly, a significant two-way interaction of defeat x drug treatment on the amount of avoidance time was revealed at days 1 and 5 (p<.01). The group receiving both eszopcilone and fluoxetine spent significantly less time in the avoidance zones than any other group. At days 10 and 15, this group did not differ from a group receiving fluoxetine only, although both of those groups spent significantly less avoidance time than defeated saline-treated controls. This study demonstrates that eszopiclone facilitates the action of fluoxetine in the SDS paradigm, paralleling clinical findings using this drug combination in depressed patients. 2009 Appalachian Student Research Forum Page 55 Division III – Graduate Students more than 2 Years Biomedical Sciences, Natural Sciences & Mathematics, and Social & Behavioral Sciences PRELAMIN A AFFECTS DIFFERENTIAL GENE EXPRESSION AND REGULATION OF CELL CYCLE Christina Bridges, Michael Sinensky, Lia Lerner, and Antonio Rusiñol, Department of Biochemistry and Molecular Biology, Quillen College of Medicine, East Tennessee State University, Johnson City, TN The proteins comprising the nuclear lamina provide structural support to the nuclear membrane and participate in a variety of cellular functions, including chromatin organization and transcriptional regulation. One of these proteins, Lamin A, is initially synthesized as a 74 kDa precursor protein, Prelamin A, which undergoes an unusual maturation process to yield the mature Lamin A (72 kDa). Previous studies have focused largely on evaluating the function of the mature protein, however, emerging evidence in our laboratory indicate a significant biological function for the precursor form, as well. Using a plasmid DNA construct harboring a mutation in the Lamin A gene that renders the protein product uncleavable by the constituents of the Lamin A maturation pathway, we have overexpressed Prelamin A in mammalian cells. Results of DNA microarray analysis demonstrate a significant effect of this protein on the levels of expression of genes involved in several distinct and important cellular pathways. These pathways include DNA replication and repair (31 genes) cell proliferation and cancer (39 genes), and most notably, cell cycle regulation (52 genes). Pathway specific oligoarrays were used to provide further targeted evaluation of Prelamin A regulation of genes involved in controlling the cell cycle. OVEREXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR 165 (VEGF165) PROTECTS CARDIOMYOCYTES AGAINST DOXORUBICININDUCED APOPTOSIS Tingting Chen, Quan Zhu, Tuanzhu Ha, Jim Kelley, Race Kao, David Williams, and Chuanfu Li. Departments of Surgery and Internal Medicine, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN Doxorubicin (Dox) has been employed in cancer chemotherapy for almost three decades. However its clinical application became restricted because of the dose-dependent cardiomyopathy. Recent studies suggested that Dox-induced cardiomyocyte apoptosis is 2009 Appalachian Student Research Forum Page 56 a primary cause of cardiac damage. Vascular endothelial growth factor (VEGF) is a major and specific factor for endothelial cell survival and angiogenesis. We have previously shown that VEGF165 significantly attenuated oxidative stress-induced cardiomyocyte apoptosis. We hypothesized that VEGF165 will protect the cardiomyocytes from Dox-induced apoptosis. To evaluate our hypothesis, we transfected cardiomyocytes H9c2 with adenovirus expressing VEGF165 (Ad5-VEGF165, 1 x 107 pfu/ml, plaque forming units) 24 hrs before the cells were challenged with Dox at a concentration of 2 µM. Ad5-GFP (green fluorescence protein, 1 x 107 pfu/ml) served as vector control. Untransfected H9c2 cells were treated with Dox (2 µM) for 24 hrs. Activation of the apoptotic signaling pathways were used as apoptotic markers. The experiments were repeated three times. Dox challenge significantly increased caspase-3 activity by 168%. The levels of FADD (Fas associated death domain) and caspase-8 activity were significantly increased by 110% and 136%, respectively, after Dox administration. In Ad5-VEGF165 transfected cells, Dox-increased caspase-3 activity was significantly reduced by 33% compared with Dox-challenged cells. Ad-5 GFP did not affect Dox-increased caspase-3 activity. Ad5-VEGF165 transfection also significantly attenuated Dox-increased the levels of FADD and caspase-8 activity. In addition, Ad5VEGF165 transfection significantly increased the levels of phospho-Akt by 20% and Bcl2 by 35% compared with Dox-treated cells, respectively. Our data suggests that Doxinduced cardiomyocyte apoptosis is mediated, in part, by activation of the death receptormediated apoptotic signaling pathway. VEGF165 protected the cardiomyocytes against Dox-induced apoptosis. The mechanisms involve inhibiting activation of the death receptor mediated apoptotic signaling pathway and activating the PI3K/Akt signaling pathway." DETERMINING PUTATIVE SECONDARY PRODUCT GLUCOSYLTRANSFERASE EXPRESSION DURING CITRUS PARADISI GROWTH AND DEVELOPMENT Jala J. Daniel, Daniel K. Owens, and Cecilia A. McIntosh, Departments of Physiology and Biological Sciences, East Tennessee State University, Johnson City, TN Plants produce secondary metabolites to help protect themselves within their environment. The major classes of secondary products include alkaloids, terpenoids, and flavonoids. Flavonoids are secondary metabolites that are not involved directly in cellular respiration and photosynthesis but have significant roles in plant defense and human nutrition. Glucosyltransferases (GT) transfer sugars from high energy sugar donors to other substrates. Several different kinds of flavonoid GT’s exist in the tissues of grapefruit, making it a model plant for studying their structure and function. The focus of the McIntosh laboratory is to understand how flavonoid GT’s influence metabolism as well as elucidating structure and function. This has led to the isolation of 7 putative secondary product GT clones. The goal of this investigation is to determine the expression patterns of the 7 putative secondary product GT’s during the grapefruit seedling growth and development by quantifying mRNA expression levels in the roots, stems, leaves, and flowers. This research was designed to test the hypothesis that these 7 GT’s are expressed constitutively. Alternatively, one or more could be expressed in a 2009 Appalachian Student Research Forum Page 57 tissue-specific manner or developmentally regulated. Six growth stages were defined. The first stage focused on emerging roots; stages 2-4 focused on leaves, stems, and roots; and stage 5 focused on comparison of older and younger leaves. Stage 6 plants were 4-5 years old when flowers were harvested. Findings show that there were variable degrees of PGT expression between the different tissues and stages of development. Therefore, results were more consistent with the alternative hypothesis that putative secondary product GT expression was tissue specific/and or developmentally regulated. SPINAL CORD STIMULATION BLOCKS ATRIAL FIBRILLATION INDUCED BY MEDIASTINAL NERVE STIMULATION David D. Gibbons1, René Cardinal2, E. Marie Southerland1, J. Andrew Armour2 and Jeffrey L. Ardell1 1 Department of Pharmacology, East Tennessee State Univ, Johnson City, TN, 2 Centre de Recherche, Hôpital du Sacré-Coeur de Montréal and Faculté de Médecine, Université de Montréal, Montréal, Canada Objective: To determine the ability of spinal cord stimulation (SCS) to modulate the extrinsic inputs to the intrinsic cardiac nervous system. Methods: Mediastinal nerves on the superior vena cava were stimulated during the atrial refractory period to induce atrial fibrillation. Nerve activity was recorded in the right atrial ganglionated plexus (RAGP). For SCS, the dorsal aspect of the T1-T3 spinal cord was stimulated electrically (50 Hz; 0.2 ms; 90% of motor threshold). Hexamethonium chloride (5 mg/kg) was administered systemically. Results: Mediastinal nerve stimulation caused a three-fold increase in nerve activity in the RAGP. Mediastinal nerve stimulation also caused a period of atrial fibrillation that self-terminates. SCS reduced the level of neuronal activity in the RAGP to baseline levels and blocked the mediastinal nerve stimulation-induced atrial fibrillation by over 50%. Hexamethonium chloride caused the same reduction of nerve activity and blockade of atrial fibrillation although to a greater degree. Conclusions: SCS acts to reduce the ability of extrinsic inputs to the intrinsic cardiac nervous system to alter normal physiological activity of the heart. SCS may act to modify pre-ganglionic inputs imparting cardioprotection from neuronally dependent stressors. (Supported by HL71830) HOMOGENEITY OF NU NO. 6 WORDS COMPRESSED 45% AND 65% Tiffany Hammonds and Richard H. Wilson, Department of Communicative Disorders, East Tennessee State University, Johnson City, TN and VA Medical Center, Mountain Home, TN Time compressed speech is a degraded speech task that reduces the redundancy cues in the temporal domain of average speech. Wilson et al. (1994) published the original work using the compressed materials used in this study on listeners with normal hearing. The 2009 Appalachian Student Research Forum Page 58 results reflected agradual degradation in recognition performance as the compression ratio increased from 45% to 65% but the homogeneity of the mean performances on each word was not considered. In 1996, Humes et al. evaluated the application of a sample of the Wilson et al. compressed speech materials in a group of elderly listeners with and without hearing loss. The results of this study found elderly listeners with hearing loss had recognition scores of approximately 45% and 20% correct for 45% and 65% compression ratio, respectively. Although four experiments were included in the original Wilson et al (1994) report, the issue of homogeneity of the test words under the compressed conditions was not addressed. As Bilger (1984) and Wilson (2003) independently have argued, equivalency or homogeneity of a set of test materials is most appropriately established on the group of listeners for whom the test materials were intended. Based on this principle, an experiment was designed in which the homogeneity of therecognition performances on the individual words compressed 45% and 65% by Wilson et al. (1994) was evaluated on a group of 72 listeners with sensorineural hearing loss. As a reference or control group, the same measures were made on 24 young adults with normal hearing. For each participant a pure-tone audiogram was obtained and performances established at 70-dB HL on the 200 NU No. 6 words compressed 0% and 45%. A third compression condition (65%) was included but based on the results of Humes et al. (1996) that reflected poor performance on the 65% compressed materials, only 50 words (List 3) were included. For comparison purposes, word recognition in quiet at 80- and 104- dB SPL (NU No. 6) and performance on the Words-In-Noise (WIN) test (Wilson, 2003) were obtained. All of the word materials were the VA (female speaker) version of the NU No. 6 words and all conditions were recorded on CD. The materials were presented monaurally. For the 24 participants with normal hearing, the mean percent correct recognitions were 98.8% (Subject SD = 1.2%; Word SD = 2.8%; range 83-100%) at 0% compression, 98% (Subject SD = 1.6%; Word SD = 4.4%; range 75-100%) at 45% compression, and 90.8% (Subject SD = 7.3%; Word SD = 14.8%; range 17-100%) at 65% compression. For the 72 listeners with sensorineural hearing loss, the mean percent correct recognitions were 88.8% (Subject SD = 8.3%; Word SD = 8.0%; range 56-100%) at 0% compression, 77.2% (Subject SD = 14.5%; Word SD = 12.5%; range 33-98%) at 45% compression, and 41.9% (Subject SD = 28.7%; Word SD = 13.2%; range 21-72%) at 65% compression. The data from the compressed conditions and the WIN also will be discussed. WORD-RECOGNITION PERFORMANCE ON TWO TYPES OF BINAURAL SPEECH TASKS BY LISTENERS WITH NORMAL HEARING AND LISTENERS WITH SENSORINEURAL HEARING LOSS Carolyn Hornberger and Richard H. Wilson, East Tennessee State University, Veterans Affairs Medical Center, Mountain Home, TN and East Tennessee State University, Johnson City, TN This project examined word-recognition performances on two binaural word-recognition tasks by 24 listeners with normal hearing (22.9 years) and 72 older listeners with sensorineural hearing loss (67.2 years). First was temporal integration that involved 2009 Appalachian Student Research Forum Page 59 presentation of the Vowel (segments) In One Ear and the Consonant (segments) In The Other (ear) with the temporal sequence of the word maintained. As a control, the VIOECITO also was administered monaurally. Second was the traditional free-recall dichotic-digit paradigm that involved 1-, 2-, and 3-pairs of digits. The Words-In-Noise (WIN) Test was administered to provide information about speech understanding in background noise. For both subject groups, the materials were presented with inserts from CD at 70 dB HL. As expected the listeners with normal hearing performed at ceiling on the VIOECITO with means of 96.8% and 97.6% for the binaural and monaural VIOECITO conditions. Similarly, performance by the listeners with normal hearing on the dichotic digit materials was near or at ceiling but with a modest right-ear advantage evident. Mean performances 12 of the listeners on the digits presented to the left ear and right ear were (1) 1-pair--97.7% and 99.1%, respectively, (2) 2-pairs--92.6% and 97.2%, respectively, and (3) 3-pairs--88.1% and 92.3%, respectively, for the 3-pair digits. Defining an ear advantage on the dichotic paradigm as 2 tokens (11.1%), a right-ear advantage (REA) was observed in three of the listeners for the 3-digit pairs with one of those listeners also having a REA for the 2-digit pairs. No left-ear advantages (LEA) were observed. Thus, both overall performance and the rightear advantage increased as the stimulus pairs increased. Mean performance on the WIN was 2.6-dB S/N, which is well within the normal range of performances on the WIN (≤6-dB S/N). The listeners with hearing loss performed worse on both listening tasks when compared to the normal hearing listeners. The VIOECITO task revealed a means of 86.1% and 92.5% for the binaural and monaural listening conditions respectively. Mean performances on the dichotic digits when presented to the left and right ears were (1) 1-pair--82.1% and 92.2%, respectively, (2) 2-pairs--70.3% and 82.9%, respectively, and (3) 3-pairs--62.3% and 73.7%, respectively. Of the 72 participants, right-ear advantages (REA) for the 1-, 2-, and 3-digit pairs were observed in 56.9%, 66.7% and 66.7%, whereas left-ear advantages (LEA) were observed in 16.7%, 27.8%, and 30.6%. Those numbers were altered substantially when equal performance was defined as 2 tokens or differences >11.1%. Mean performance on the WIN was 12.0-dB S/N with only one listener with hearing loss in the normal range (≤6-dB S/N). CANNABINOID RECEPTOR (CB2) DEFICIENCY IN HYPERLIPIDEMIC LDLR NULL MICE IS ASSOCIATED WITH DECREASED APOPTOSIS AND INCREASED PLAQUE INSTABILITY Courtney Netherland, Alicia Bales, Theresa Pickle and Douglas P. Thewke, Department of Biochemistry, East Tennessee State University, Johnson City, TN Introduction: Macrophage apoptosis is a vital element of the pathophysiology of atherosclerosis. In early stages, macrophage apoptosis slows lesion formation, while in advanced stages; it contributes to plaque instability. CB2 is the primary macrophage cannabinoid receptor and is found in lesions. A cannabinoid, Δ9tetrahydrocannabinol, induces lesion regression in mice via a CB2-dependent mechanism and, in vitro, CB2 regulates several macrophage process associated with atherosclerosis. 2009 Appalachian Student Research Forum Page 60 Still, the role of CB2 in atherosclerosis is unclear. Recently, we observed that CB2 null macrophages are resistant to apoptosis induced by oxidized low density lipoproteins and oxysterols, two potent apoptotic inducers present in lesions. From this, we hypothesized that CB2 influences atherosclerosis, in part, by modulating lesional apoptosis. To test this hypothesis, we examined the effect of CB2 gene deletion on lesion formation, apoptosis and extracellular matrix (ECM) composition in Ldlr null mice, a murine model of atherosclerosis. Methods: Groups (n ≥ 8) of 8-week old male CB2-/-/Ldlr-/- (CB2-/-) and CB2+/+/Ldlr-/- (WT) mice were fed an atherogenic diet for 8 or 12 weeks. Serum cholesterol and triglyceride levels, as well as lipoprotein profiles, were evaluated by standard methods. Lesion area and apoptosis were quantified in cross sections of the proximal aorta by oil red-O staining and in situ TUNEL analysis, respectively. Smooth muscle cell (SMC) and macrophage content was determined by immunohistochemical staining with α-actin and MOMA2 antibodies. Collagen and elastin composition was evaluated by Mason’s trichrome and Verhoeff-Van Gieson staining, respectively. Results: No differences were seen in plasma cholesterol and triglyceride levels or in the lipoprotein profiles between CB2-/- and WT groups. No statistical difference was observed between the mean lesion area of CB2-/- and WT groups after 8 weeks (113,907 ± 32,263 vs 115,377 ± 35,457 µm2, p=0.93) or 12 weeks (185,546 ± 30,098 vs 197,616 ± 40,900 µm2, p=0.47) on the atherogenic diet. However, a significant decrease (~50%) was observed in the number of apoptotic cells in CB2-/- lesions compared to WT lesions (27.2 ± 9.3 vs 50.5 ± 16.1 apoptotic cells per mm2, p <0.001). In addition, lesions from CB2-/- mice fed an atherogenic diet for 12 weeks had increased SMC content in the cap region and showed decreased collagen content and enhanced elastin fiber damage, evidence of ECM degradation. Consistent with an increase in ECM degradation in the CB2-/- lesions, gelatin zymography detected a 1.9 fold increase in matrix metalloproteinase 9 activity secreted from CB2-/- macrophages compared to WT macrophages. Conclusions: Systemic CB2 deficiency in Ldlr null mice significantly reduces lesional apoptosis and alters the ECM composition of lesions without affecting lesion size. These results imply that CB2 may be a novel target for therapies aimed at reducing plaque instability and rupture. A CANNABINOID (WIN 55,212-2) INDUCES APOPTOSIS IN PANCREATIC TUMOR CELLS Theresa Pickle and Douglas Thewke, Department of Biochemistry and Molecular Biology, College of Medicine, East Tennessee State University, Johnson City, TN Pancreatic cancer is the fourth leading cause of cancer related deaths. The mean survival rate is ~6 months, and only 4% of patients survive five years. Despite extensive testing, only one chemotherapy agent (gemcitabine) has been found to produce any benefit to these patients, unfortunately, the clinical response rate to gemcitabine is less than 10% with life prolongation being only 6 weeks on average. Cannabinoids, such as Δ9tetrahydrocannabinol (THC), the active agent of Cannabis sativa, exhibit some anti-tumor properties via induction of apoptotic mechanisms. Cannabinoids produce their biological effects by engaging specific receptor-mediated signaling pathways. To date, two 2009 Appalachian Student Research Forum Page 61 cannabinoid-specific receptors, designated CB1 and CB2, have been cloned and characterized from mammalian tissue. The current study was conducted to investigate the potential use of cannabinoids as an anti-tumor treatment in pancreatic cancer. Using RTPCR we found evidence for expression of CB1 and CB2 in three human pancreatic cancer tumor cell lines (AsPC1, CaPan1, and Mia-PaCa2). Win55,212-2 treatment induced apoptosis, as determined by caspase-3 activity, in AsPC1, CaPan1 and MiaPaCa2 cells but not in non-neoplastic human pancreatic duct epithelial (HPDE) cells. The induction of caspase-3 activity by Win 55,212-2 in Mia-PaCa2 cells was prevented by a CB1 receptor-specific antagonist, AM251, but not by a CB2 receptor-specific antagonist, SR144528. In contrast, induction of caspase-3 activity in AsPC1 cells was inhibited by SR144528 but not AM251, while induction of apoptosis in CaPan1 cells was not affected by either antagonist. Consistent with induction of apoptosis, Win 55,212-2 treatment reduced expression of Bcl-xL, an anti-apoptotic member of the Bcl family, in Mia-PaCa2 cells but not in HPDE cells. These results demonstrate that several pancreatic tumor cell lines express cannabinoid receptors and that a cannabinoid, Win 55,212-2, potently inhibits the viability of these pancreatic tumor cells, at least in part, by inducing apoptosis via CB1/CB2 receptor dependent and independent mechanisms. IMMUNO-MODULATION OF SENESCENT HSV-SPECIFIC CD8+ T CELLS Stacie N. Woolard, Subhadra Nandakumar, and Uday Kumaraguru, Department of Microbiology, College of Medicine, East Tennessee State University, Johnson City, TN In many chronic inflammatory and infectious diseases, there is a persistent activation of T lymphocytes resulting in accelerated replicative immune cell senescence. Accumulation of senescent lymphocytes could probably be the cause for associated immune abnormalities that follow. Observations that patients with inflammatory syndromes and chronic infections have high frequencies of functionally aberrant, senescent T cells provide compelling evidence for premature immunosenescence in disease pathogenesis. Since T cells have a limited proliferative life span like other cells, we propose that replicative stress imposed by chronic HSV reactivation is a cause of functionally impaired immune memory. It is possible that certain T cell activating conditions including DNA demethylation and production of certain transcription factors may induce inhibitory NK receptor expression. The induction of Killer cell lectin like receptor subfamily G1 (KLRG1) expression on T cells is still largely unexplained. We hypothesized that certain cytokines may also induce such expression. This we base on the finding of cytokine IL-21 inducing expression of inhibitory receptors on NK cells, besides induction of maturation. We have demonstrated that the percentage of antigen specific memory CD8+T cells was influenced by presence of pIL-21 during memory phase. Cells from mice treated with pIL-21 produced around 10 times more CD8+IFN+ cells and enhanced expression of the cytolytic marker, CD107 , than cells from control mice. So it remains to be investigated whether IL-21 acts as a factor that can also promote the KLRG1 expression on CD8+ T cells. KLRG1 positive and negative CD8+ T cells were collected from Naïve and HSV latently infected mice. Cells were treated with various concentration of recombinant cytokine IL-21 in the absence or 2009 Appalachian Student Research Forum Page 62 presence of cognate antigen (HSV-gB498-505 peptide – SSIEFARL). The changes we observed in virus specific CD8+ T cells suggested that IL-21 functions to induce terminal differentiation either directly or indirectly resulting in enhanced immune function. This new information about the pleiotropic effects of IL-21 may be used effectively in combination with other immuno-modulators to achieve heightened immune response. MEN'S COPING SKILLS AND INTIMATE PARTNER VIOLENCE IN SOUTHERN APPALACHIA James Gorniewicz and Michael Floyd, Department of Family Medicine, East Tennessee State University, Johnson City, TN Intimate partner violence (IPV) is a substantial public health concern, affecting nearly 7.7 million Americans annually, with a total related cost estimated to be over $8.3 billion. Previous research involving victims of IPV has demonstrated that approach copings skills tend to be associated with positive health outcomes, whereas avoidance coping skills tend to be associated with negative health outcomes. The use of coping skills after an IPV assault can lead to better psychological adjustment and improved health outcomes. However, little is known about the associations between men’s coping skills and IPV perpetration. The purpose of this study was to determine whether men’s coping skills were associated with specific aspects of IPV perpetration. A sample of men (N = 49) was recruited from batterer intervention programs in Northeast Tennessee and Southwest Virginia. All participants completed a demographic questionnaire, the Conflict Tactics Scales (CTS-2) and the Coping Responses Inventory (CRI). Our analysis showed that batterers’ coping skills (approach and avoidance) were related to levels of psychological aggression and conflict negotiation, but not physical assault. Overall, approach coping skills were related to increased negotiation with an intimate partner. Avoidance coping skills were associated with psychological aggression towards an intimate partner. Coping skills were not associated with physical violence. Unexpectedly, avoidance coping skills were associated with negotiation. Future research should examine whether or not coping skills serve as a mediator between IPV and known risk factors such as life stress, substance abuse, and antisocial personality disorder. Further research could lead to the development of new, more effective, therapeutic interventions for batterers. EVALUATION OF THE LISTENING SELF-EFFICACY QUESTIONNAIRE: PRELIMINARY PSYCHOMETRIC RESULTS Carissa La More, Department of Communicative Disorders, East Tennessee State University, Johnson City, TN Self-efficacy is the belief, or confidence, individuals have in their abilities to accomplish a certain behavior (Bandura, 1989), including behaviors related to manage health 2009 Appalachian Student Research Forum Page 63 conditions. Self-efficacy beliefs are domain-specific in that individuals can have high self-efficacy for abilities related to a certain behavior (e.g., performing an exercise routine regularly) and at the same time lack self-efficacy beliefs in their abilities for another specific behavior (e.g., public speaking). Research has demonstrated that high self-efficacy beliefs for managing health conditions have been related to better health outcomes and improved quality of life (Bandura, 1997; Bradshaw et al., 2004; Brody et al., 1999; Fingeld-Connett, 2004; Focht, Rejeski, Ambrosius, Katula, & Messier, 2005; Horowitz, Reinhardt, & Kennedy, 2005; Lorig, Ritter, & Plant, 2005; Marks, Allegrante, & Lorig, 2005; Rogers et al., 2005). The role of self-efficacy in audiology-related areas is a developing area of study. Self-efficacy investigations are emerging in several different domains of audiology, including hearing protective devices, balance, tinnitus, and hearing aids (Smith & Fagelson, 2008; Smith & West, 2006; West & Smith, 2007; Melamed et al., 1996; Tinetti et al., 1990). Because self-efficacy beliefs are individuals' perceptions of their current abilities to perform certain tasks related to a particular behavior, a questionnaire is used to quantify these beliefs in the domain of interest. A primary goal of listeners with hearing loss is to be able to hear and understand conversation. The purpose of this study was to develop and validate the Listening SelfEfficacy Questionnaire (LSEQ), a questionnaire designed to assess individuals’ listening self-efficacy, or beliefs in their abilities to listen to speech in a variety of real-life conditions. To date, 101 older adults, (mean age= 67.6 years) with bilateral mild to moderate sensorineural hearing loss completed the LSEQ and the Words-In-Noise (WIN) test. A principal component analysis suggested that the LSEQ has three subscales, accounting for 73% of the variance, which corresponded to understanding speech in the following listening conditions: (1) complex and/or noisy environment, (2) car/traffic, and (3) single talker. The internal consistency of each subscale was > 0.90 (Chronbach’s α), suggesting good internal reliability. There were moderate correlations (r = 0.34, r = 0.39) between the WIN threshold and two LSEQ subscales, suggesting that individual's perceptions in their abilities to understand speech are moderately related to their objective speech-in-noise performance. Additional data are being collected to evaluate the LSEQ further; however, preliminary results suggest that the LSEQ will be a valid and reliable questionnaire and shows promise as a clinical tool to assess a patient’s beliefs in his/her ability to understand speech. 2009 Appalachian Student Research Forum Page 64 Division IV – Medical Students ENTEROCOLIC LYMPHOCYTIC PHLEBITIS INITIALLY PRESENTING AS INTUSSUSCEPTION IN AN ADULT: A CASE REPORT Jennifer Chavis and John “Trey” Robertson M.D., James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN and John Lawson Surgical Group, Johnson City, TN Enterocolic Lymphocytic Phlebitis, or ELP, is a rare condition characterized by an immune mediated vasculitis affecting veins exclusively in the wall of the mesentery of the colon and or small bowel in the absence of systemic involvement. There is no arterial involvement in ELP and this further differentiates it from systemic diseases such as SLE or Crohn’s disease that can cause mesenteric ischemia, infarction, and subsequent necrosis of the bowel. According to the literature, patients with ELP often present with an acute abdomen and complain of a history of vomiting, diarrhea, and fevers or chills. Many of these patients are elderly and may have other medical problems, specifically hypertension, further complicating the clinical picture. Surgical therapy which usually involves a diagnostic laparoscopy with resection of the affected bowel is both diagnostic and curative, as most patients make a full recovery after the operation. Definitive diagnosis is confirmed based on the histological findings of mixed T and B cell vasculitis present exclusively in the veins and venules of the wall and mesentery of the colon. We describe a case of ELP in a 47 year old man with no significant past medical history of vascular diseases or immune mediated disease who presents with an eighteen hour history of right lower quadrant pain and multiple grossly bloody bowel movements. Unique to our case is that our patient’s CT scan had a “bull’s eye” pattern indicative of intussusception, which is very rare in adults and is often associated with malignancy. To the best of our knowledge no other case of ELP initially presenting as intussusception has been documented in the literature. ELEVATED CALCITONIN LEVELS IN PATIENT WITH MULTIPLE ENDOCRINE NEOPLASIA 2A (MEN2A) AFTER SURGICAL MANAGEMENT Nadia Sabri and Reena Kuriacose, MD. Department of Internal Medicine. James H. Quillen College of Medicine. East Tennessee State University. Johnson City, TN Learning objectives include: to understand the one of the causes of calcitonin elevation; educate regarding genetics in the cancer syndrome Multiple Endocrine Neoplasia 2A (MEN2A); to present an unusual case of a patient with MEN2A. Case Presentation: 48 year old female with history of MEN2A confirmed by genetic testing, medullary thyroid carcinoma with thyroidectomy, resulting hypothyroidism that is medically managed, depression, recurrent UTIs, osteopenia, lymph node dissections from neck and chest, hypercalcemia, parathyroidectomy and parathyroid transposition in left forearm 2009 Appalachian Student Research Forum Page 65 presents with increasing fatigue , palpitations and elevated calcitonin levels (around 3000) even after surgeries. Octreotide scans reveal increased uptake in liver, which could be explained by hemangioma, gallbladder uptake, or cancer metastases. MRIs of neck and chest, bone scans, annual testing for pheochromocytoma, and PET scans are normal. The puzzling finding therefore is the elevated calcitonin and CEA levels in the absence of positive imaging. Other findings include renal stones, increased calcium, PTH at upper limits of normal (82), high ionized (5.7), total calcium values, and elevated urine calcium (413). However, no hyperactivity seen on parathyroid scan and no residual thyroid tissue on thyroid ultrasound. This unusual presentation of persistent calcitonin and CEA levels leads us to believe this may be evidence of either recurrent medullary cancer or some correlation with the elevated PTH level. Therefore, removal of parathyroid transplant in left arm may be a good option. Further evaluation and workup will be done to elucidate cause of persistent rising levels of calcitonin and CEA in a MEN2A patient who is status post thyroid and parathyroid surgeries. Discussion: Multiple Endocrine Neoplasia type 2 (MEN2) is a rare hereditary cancer syndrome exhibiting autosomal dominant inheritance with mutation of RET gene and associated with medullary thyroid carcinoma. This endocrine cancer syndrome has a prevalence of approximately 1in 30,000. Subtypes of Multiple Endocrine Neoplasia type 2 include familial medullary thyroid carcinoma (FMTC), Multiple Endocrine Neoplasia type 2A, and Multiple Endocrine Neoplasia type 2B (MEN2B). MEN2A is suspected when there are at least two of the three common tumors (medullary thyroid cancer, pheochromocytoma, or parathyroid adenoma/hyperplasia) in one person or close relatives (parents, siblings, and children). Patients with MEN 2 are at risk for developing medullary thyroid cancer (MTC), a neoplasm of the calcitonin-producing parafollicular or C cells within the thyroid gland. Total thyroidectomy has been shown to decrease occurrence or recurrence of metastatic MTC, which is the predominant cause of mortality from MEN2. CIGARETTE SMOKE EXTRACT (CSE) ENHANCES INFLAMMATORY CYTOKINE EXPRESSION IN IL-1 BETA-ACTIVATED MAST CELLS VIA REGULATION OF THE NF-ĸB PATHWAY Thomas Soike1, Kenton Hall1, Jonathan Moorman1, Guha Krishnaswamy1, David S Chi1, Tuanzhu Ha2, and Chuanfu Li2, 1Department of Internal Medicine, East Tennessee State University, Johnson City, TN, 2Department of Surgery, East Tennessee State University, Johnson City, TN Human Mast cells are multifunctional cells capable of inflammatory responses. Mast Cells are associated with allergies, asthma, and atherosclerosis. This study aims to examine the effects and mechanisms of CSE on the expression of inflammatory cytokines in mast cells. The inflammatory cytokines of interest in this experiment are IL-6 and IL8. IL-6 plays a key role in the acute phase response of cells and IL-8 plays a role in chemotaxis and recruitment of neutrophils. NF-ĸB is a transcription factor and plays a key role in the cell activation. In this experiment main-stream (Ms) cigarette smoke and side-stream (Ss) cigarette smoke were collected onto fiber filters using a controlled simulated human inhalation. The cigarette smoke was extracted from the fiber 2009 Appalachian Student Research Forum Page 66 filters using RPMI-1640 medium. Two ml of the human mast cells (HMC-1) at 1 x 106 cells/ml were cultured with Ms and Ss CSE in the presence or absence of IL-1β (10 ng/ml) for 24 hours. The expression of IL-6 and IL-8 was assessed by ELISA and RT-PCR. NF-ĸB activation was measured by electrophoretic mobility shift assay (EMSA) and IĸBα degradation by Western Blot. Both Ms and Ss CSE significantly increased IL-6 and IL-8 production (p<0.0001) in IL-1β activated HMC-1. CSE increased NF-ĸB activation and decreased cytoplasmic IĸBα proteins in IL-1β- activated HMC-1. These results suggest that CSE enhances the expression of inflammatory cytokines through increased NF-ĸB activation and IĸBα degradation, which may partially explain why cigarette smoke contributes to lung and cardiovascular disease (Supported by the Ruth R. Harris Endowment and RDC of ETSU). TARGETING C-REACTIVE PROTEIN TO CAPTURE ATHEROGENIC LDL James A. Thompson1, Sanjay K. Singh1, David J. Hammond1, and Alok Agrawal1, Antonio E. Rusinol2, 1Department of Pharmacology, College of Medicine, East Tennessee State University, Johnson City, TN; 2Department of Biochemistry, College of Medicine, East Tennessee State University, Johnson City, TN C-reactive protein (CRP), a pentameric protein comprised of five identical subunits, is best known for its reactions with substances containing phosphocholine and phosphoethanolamine (PEt). CRP is a plasma protein, but has also been seen deposited with low-density lipoprotein (LDL) in human atherosclerotic lesions. In this study, we investigated the interactions between CRP and oxidized LDL (oxLDL) which is an atherogenic form of LDL. Freshly-purified CRP did not bind to immobilized oxLDL. Interestingly, in the presence of the small molecule compound PEt, freshly-purified CRP bound oxLDL efficiently. On the other hand, when an old preparation of purified CRP was used in the assay, CRP bound oxLDL even in the absence of PEt, although the binding was dramatically enhanced in the presence of PEt. Gel filtration analysis of the old preparation of purified CRP revealed the presence of monomeric CRP generated by dissociation of pentameric CRP. These findings indicated that monomeric CRP was capable of binding to ox-LDL and did not require PEt to do so. Thus, although the exact mechanism of action of PEt in CRP-oxLDL interaction is not known, we hypothesize that PEt may be acting by monomerizing pentameric CRP. Previously, we reported that PEt enhanced the binding of CRP to enzymatically-modified LDL, another atherogenic form of LDL. Combined data raise the possibility that the targeting of CRP by PEt may allow CRP to capture atherogenic LDL in vivo and prevent formation of LDL-loaded macrophage foam cells which is one of the processes that contribute to the development of atherosclerosis. 2009 Appalachian Student Research Forum Page 67 BAICALEIN INHIBITS THE ENHANCING EFFECTS OF CIGARETTE SMOKE EXTRACT ON INFLAMMATORY CYTOKINE PRODUCTION IN IL-1βACTIVATED MAST CELLS Fangbai Wu1, Ta-Chang Lin2, Kenton Hall1, Zong Doa Wu2, Jonathan Moorman1, Guha Krishnaswamy1, and David S.Chi1 1Dept. of Internal Medicine, East Tennessee State University, Johnson City, TN, and 2Dept. of Environmental Engineering, National Cheng Kung University Human mast cells have been known to play a vital role in the pathogenesis of inflammatory airway diseases such as asthma and allergy. Mast cells have been shown to produce inflammatory cytokines, such as IL-6 and IL-8, after activated by IL-1β. Previously, we have reported that cigarette smoke extract (CSE) significantly increased IL-6 and IL-8 production in IL-1β-activated human mast cells (HMC-1) in culture. Baicalein (BAI), isolated from Chinese herbal medicine Huangqin, has been shown to have anti-inflammatory properties. The goal of the present study was to examine the effect of BAI on IL-6 and IL-8 production in IL-1β-activated and CSE-treated HMC-1. Two ml of HMC-1 at 1 × 106 cells/ml were cultured either with or without IL1β (10 ng/ml), and in the presence or absence of mainstream (Ms) or sidestream (Ss) CSE (0.25 mg/ml or 0.125 mg/ml), and BAI (30 μM). HMC-1 with various treatments was then incubated for 24 hours and culture supernatants were collected and analyzed for the production of IL-6 and IL-8 by ELISA. Both concentrations of Ms and Ss CSE significantly increased production of both IL-6 (p < 0.0001) and IL-8 (p < 0.0005) in IL-1β-activated HMC-1 in a dose dependent fashion. BAI (30 μm) decreased IL-6 and IL-8 production in the Ms CSE (0.25 mg/ml) IL-1β treated HMC-1 culture from 2615 + 38 to 251 + 19 pg/ml (p < 0.0000001) and from 583 + 37 to 179 + 7 pg/ml (p < 0.0005), respectively. A similar inhibitory effect of BAI was also observed in the Ss CSE + IL-1β treated HMC-1 culture. Our results show that BAI inhibits the enhancing effects of cigarette smoke extract on inflammatory cytokine production in IL-1β-activated mast cells. It suggests that BAI may be a novel candidate for treatment of smoking related inflammatory airway disease (Supported by the Ruth R. Harris Endowment and RDC of ETSU). 2009 Appalachian Student Research Forum Page 68 Divisions V and VI– Residents / PostDoctoral Fellows and Case Histories PROTECTION AGAINST MYOCARDIAL INFARCTION BY TIR/BB-LOOP MIMETIC AS-1 THROUGH DECREASING THE INTERACTION OF IL-1R WITH MYD88 Zhijuan Cao, Yulong Hu, Tuanzhu Ha, Jim Kelley, Race Kao, David Williams and Chuanfu Li, Departments of Surgery and Internal Medicine, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN It is estimated that 500,000 Americans die of heart attacks each year and ischemic heart disease is responsible for 90% of cardiac mortalities. It is well established that innate immune responses are involved in the pathogenesis of myocardial ischemia/reperfusion (I/R) injury and heart failure. The IL-1 receptor (IL-1R)-mediated MyD88-dependent nuclear factor KappaB (NFB) activation pathway plays an important role in the induction of innate and inflammatory responses. However, the role of the IL-1Rmediated MyD88 pathway in myocardial I/R injury has not been investigated entirely. We hypothesized that inhibition of the interaction of IL-1R with MyD88 will attenuate myocardial I/R injury through reducing innate and inflammatory responses. To evaluate our hypothesis, we induced myocardial infarction in male C57 BL/6 mice (n=8) by ligation of the left anterior descending coronary artery for 45 min (ischemia) followed by reperfusion (4 hrs). In the treatment group, after mice (n=8) were subjected to ischemia (45 min), the TIR/BB-Loop mimetic (AS-1), which inhibits the interaction of IL-1R with MyD88, was administered immediately before reperfusion (50mg/kg body). The hearts were subjected to continuous reperfusion for 4 hrs. Myocardial infarct size was determined by TTC staining. In separate experiments, cardiac function was evaluated by echocardiography after myocardial ischemia (45 min) followed by reperfusion (4 hrs). Sham surgically operated mice that were treated with and without AS-1 served as sham controls (n=6 mice in each group). Hearts were harvested and cellular proteins were isolated for immunoprecipitation and immunoblots. The data showed that AS-1 administration significantly decreased myocardial infarct size by 32.9% compared with the untreated I/R group (32.9 ± 2.17% vs. 49.1 ± 2.26%, P<0.05). Cardiac function indices such as ejection fraction and fractional shortening in AS-1 treated mice were also significantly increased by 18.0% and 25.6%, respectively, compared with the untreated I/R group (53.9 ± 2.07% vs 44.2 ± 4.33%, P<0.05 and 27.5 ± 1.92% vs 20.7 ± 3.09%, P<0.05). AS-1 administration significantly decreased I/R-increased interaction between IL-1R and MyD88, attenuated I/R-increased NFB binding activity by 40.0%, and reduced levels of inflammatory cytokines (IL-1, and IL-6) by 37.0 % and 30.0%, respectively, in the myocardium compared with the untreated I/R group. In addition, AS-1 administration significantly decreased myocardial myeloperoxidase activity by 23.6% and neutrophil infiltration in the myocardium compared with the untreated I/R group. The results demonstrated an important role for the IL-1R-mediated MyD88-dependent signaling pathway in myocardial I/R injury. The data suggests that modulation of the IL-1R/MyD88 interaction could be a strategy for reducing myocardial ischemic injury. 2009 Appalachian Student Research Forum Page 69 ON THE MECHANISM OF ANTI-PNEUMOCOCCAL FUNCTION OF C-REACTIVE PROTEIN David J. Hammond, Sanjay K. Singh, Donald A. Ferguson, Jr., Michael K. Pangburn, and Alok Agrawal, Department of Pharmacology and Department of Microbiology, College of Medicine, East Tennessee State University, Johnson City, TN Human C-reactive protein (CRP) protects mice from lethality following infection with Streptococcus pneumoniae type 3 through undefined mechanisms. Because both CRP and type 3 pneumococci are reported to bind to complement regulatory protein factor H, we propose that the factor H-binding capability of CRP may contribute to anti-pneumococcal function of CRP. In this study, we explored interactions between CRP and factor H. Surprisingly, we found that freshly-purified CRP did not bind human factor H. However, in the presence of the well-known CRP-binding ligand phosphoethanolamine (PEt), freshly-purified CRP bound to human factor H efficiently. In contrast, and as reported previously, when an old preparation of CRP was used in the assay, CRP bound to human factor H even in the absence of PEt, although the binding was dramatically enhanced in the presence of PEt. Identical results were obtained with murine factor H. Gel filtration analysis of the old preparation of CRP revealed the presence of monomeric CRP generated by dissociation of CRP which, in the native form, is a pentameric protein composed of five identical subunits. These findings indicated that, in the absence of PEt, it was monomeric CRP that bound to factor H. Although the exact mechanism of action of PEt in CRP-factor H interaction is not known, we hypothesize that PEt may be acting by monomerizing pentameric CRP. Our findings raise the possibility that CRP, once in contact with a PEt-containing surface, is monomerized and gains factor H-binding capacity which facilitates complement-dependent lysis of otherwise complement attackresistant factor H-bound pneumococci. EFFECTS OF GLUCAN, ZYMOSAN AND CANDIDA ON INFLAMMATORY CYTOKINE EXPRESSION IN IL-1BETA-ACTIVATED MAST CELLS Christopher W T Miller1, David S Chi1, Kenton Hall1, Tammy Ozment-Skelton2, David Williams2, David Johnson3, Guha Krishnaswamy2. Departments of 1Internal Medicine, 2 Surgery, 3Biochemistry and Molecular Biology, East Tennessee State University, Johnson City, TN Not much is known about the function of C-type lectin-like receptors (CLRs) on mast cells or about the recently described Dectin-1 (a pattern-recognition receptor). Our lab has had a long-standing interest in the role of receptor-mediated signaling in mast cells and their role in innate immune responses. Fungi cause many health problems, including severe asthma and allergic disorders. Mast cells (immediate response cells) may be involved in antifungal immunity and fungal mannans may be a mechanism of non-IgEmediated mast cell signaling. This study aims to examine the effects of glucan, zymosan and Candida on inflammatory cytokine production and tryptase content in IL-1beta- 2009 Appalachian Student Research Forum Page 70 activated mast cells, compared with resting mast cells. 2 mL of HMC-1 at a concentration of 1,000,000 cells / mL were cultured with various stimulants, including beta-glucan (10 mcg/mL), zymosan (1 mcg/mL), Candida blastospores (10 mcg/mL), and Candida hyphae (10 mcg/mL), in the presence or absence of IL-1beta (10 ng / mL) for a period of 24 hours. The supernatants were harvested and assayed for tryptase and IL-6, IL-8, and MCP-1 by ELISA. Student’s t-test was used to compare the effects of the individual stimulants on HMC-1, and results were significant with a p < 0.05. Statistical analysis was carried out with the STATISTICA software. 89% of HMC-1 were dectin-1 positive by flow cytometry. IL-1beta decreased tryptase activity from 17.5 (in the resting group) to 10.2 miliA405/min (in the stimulated group), with a p value >0.05. Additional stimulation with beta-glucan or zymosan to the IL-1beta-stimulated mast cells led to a further decrease in tryptase activity to 7.6 and 7.4 mA405/min, respectively (p<0.05 in both cases). Cytokine production in the beta-glucan, zymosan and Candida alone groups was similar to that of the medium alone group (for the latter, values were 1.9, 12.0, and 92.8 pg/mL for production of IL-6, IL-8 and MCP-1, respectively), while that of the IL1beta-activated group was significantly elevated (1617.0, 501.0, and 732.0 pg/mL for the same parameters, in all cases p<0.0001). Addition of beta-glucan and zymosan didn't affect cytokine production in IL-1beta-activated HMC-1 cells. However, both candidal blastospores and candidal hyphae significantly increased production of IL-6 (p<0.0001 in both) and IL-8 (p<0.01 and <0.05, respectively), but not MCP-1, in IL-1beta-activated HMC-1. We showed that mast cells express dectin-1, and that beta-glucan, zymosan, and Candida have various effects on activation of mast cells by IL-1beta. IL-1 and IL-6 are acute phase cytokines and essential to phagocyte recruitment and humoral immunity, while MCP-1 is important in mononuclear chemotaxis and T helper polarization. The pathways by which dectin-1 influences mast cell cytokine production and their response to fungal components need to be elucidated, but our study supports the role of mast cells in antifungal innate immunity. GENETIC-MOLECULAR DEFECTS IN MBL-MASP PATHWAY AND ASSOCIATION WITH SELECTIVE IMMUNOGLOBULIN DEFICIENCIES: IMPLICATIONS FOR HOST DEFENSE Christopher W T Miller, David S Chi, Mini A Michael, Tamar Giorgadze, and Guha Krishnaswamy. Division of Allergy & Immunology, Department of Internal Medicine, Quillen College of Medicine, East Tennessee State University, Johnson City, TN Isolated immunoglobulin (Ig) deficiencies (including IgG4, IgM, and IgE) involve humoral responses and can lead to serious infections. Mannose-binding lectin (MBL) deficiency, the most common immunodeficiency in the general population, is an integral part of host immunity, with deficiency predisposing to infection. Coexistent humoral and MBL deficiency would theoretically lead to graver clinical scenarios, as MBL binds carbohydrates on the surface of immunoglobulins to provide pathways for immune complex clearance and complement activation in response to Ig-covered pathogens. We describe three patients with coincident MBL and selective Ig deficiency. Three patients with recurrent infections were assessed for serum Ig and MBL levels. MBL and MASP-2 2009 Appalachian Student Research Forum Page 71 genotyping was performed in 2 patients (IBT Laboratories, Lenexa, KS). Three patients presented with recurrent chronic rhinosinusitis and bronchitis. All had normal levels of IgG. Two patients, however, had selective IgM deficiency (< 50 mg/dl) and very low MBL levels (<50 ng/mL); genotyping was performed in one patient, revealing a mutated MBL2 promoter (LXPA/LYPB, both haplotypes conferring low serum MBL) and wildtype MASP-2 (A/A). The third patient had severe IgE and IgG4 deficiencies. She had received prior intravenous immunoglobulin therapy for several years for presumed common variable immune deficiency. MBL pathway genotype analysis revealed wild type (A/A) MASP-2 and LXPA/LYPB MBL2. This patient’s IVIG infusion was discontinued. Patients with recurrent infections may have various combinations of primary immune deficiency, including defects in both the complement (MBL-MASP) and the humoral (Immunoglobulin) pathways. The prevalence and clinical implications of such combinations are unknown and require further study. INFLUENCE OF BODY MASS INDEX, HEIGHT, AND AGE ON THE MODE OF DELIVERY IN NULLIPAROUS WOMEN IN APPALACHIA James A. Reed, MD and Howard E. Herrell, MD, Department of Obstetrics and Gynecology, East Tennessee State University, College of Medicine, Johnson City, TN The purpose of this study is to determine a patient’s risk of cesarean section based on her Body Mass Index / BMI at the time of delivery, and to determine whether other variables, such as age or height, have an impact on the mode of delivery. After obtaining IRB approval, a retrospective chart review was conducted at three affiliated hospitals in Northeast Tennessee – two community hospitals and one tertiary care facility. Data was collected for a one year period from July, 2007 to June, 2008. The Fisher’s Exact Test was used to assess the data and a two-tailed p-value was calculated. A p-value of <0.05 was considered to be statistically significant. Control BMI was considered 25-30, control height 61-64 inches, and control age 21-25. Data were collected for a total of 2,490 patients. Parous patients and those with incomplete records were excluded from the study. A total of 612 women were included in the study. A statistically significant increase in the cesarean delivery rate was demonstrated when the BMI at the time of delivery was greater than 30 versus the control group (BMI 25-30). This increase continues with increasing BMIs. Also, being less than 61 inches tall demonstrated a statistically significant increased rate of cesarean delivery. The cesarean section rate was found to increase with increasing age when compared to women aged 21-25, but this was only statistically significant in the group of women 31-35. There were not enough women in the study >35 to show statistical significance for this group (n=26), though the actual cesarean delivery was highest among those women (53.8%). The use of BMI at the time of delivery was unique to this study. Prior studies have focused on BMI at the beginning of pregnancy or total weight gain during pregnancy. Data based on the BMI at the time of delivery may be more useful for practicing clinicians. We also assessed height and found that if the patient were <61 inches tall, therewas a significant increase in the rate of cesarean delivery. We were able to show that those patients 31-35 years old had a significant increase in cesarean delivery. Limitations of this study include the manner in 2009 Appalachian Student Research Forum Page 72 which weight was determined at the time of delivery, which was by patient account. We also did not control for operative indications for cesarean delivery independently. The overall observed primary cesarean delivery rate in the study was 37.2%, higher than the national average of 24.3%. This may be partially accounted for by the higher rate of obesity encountered in the Appalachian region. ETHICS IN THE PHARMACEUTICAL INDUSTRY: A RESIDENT SIMULATION Ross R. Spires1 and Martin J. Eason2, M.D., 1D.O.Resident, Department of Obstetrics and Gynecology, Quillen College of Medicine, East Tennessee State University, Johnson City, TN; 2J.D.Director, Center for Experimental Learning, East Tennessee State University, Johnson City, TN Objective: The purpose of this exercise was to improve understanding of current guidelines for ethical behavior among pharmaceutical representatives and physician responsibilities in interacting with the pharmaceutical industry. We also sought to increase alertness of unethical pharmaceutical industry actions which could have the potential to compromise care.Methods used: Residents were brought into a patient simulation where the patient’s chief complaint was vasomotor symptoms. While en route to see the patient, the learners were met in the hall by a real pharmaceutical representative. They were given literature on a hormone replacement therapy (HRT). They were also given tickets to a concert. Upon entering the exam room, the patient already had a copy of the same HRT literature which had been left in the waiting room. The pharmaceutical representative then entered the exam room during the patient interview and violated patient confidentiality in an attempt to increase sales by offering prescription coupons to the patient.Results summarized: Participants of varying training levels undertook the simulation. Performance was assessed in a debriefing session after the simulation. Current guidelines, provided by the Pharmaceutical Research and Manufacturers of America (PhRMA) Code on Interaction with Healthcare Professionals, Quillen College of Medicine Resident Policy on Ethical Guidelines for Gifts to Physicians from Industry, were discussed. The simulation allowed students and residents to learn about professionalism, communication skills, and systems-based practice during interaction with the pharmaceutical industry.Conclusions reached: This simulation mode allowed residents and medical students to learn and utilize ethical principals while interacting with the pharmaceutical industry in a controlled situation. Competencies achieved included: professionalism, communication skills, systems-based practice, and understanding of current guidelines on ethical behavior in interactions with the pharmaceutical industry set down by various governing bodies. 2009 Appalachian Student Research Forum Page 73 INFLUENCE OF THERMOSTAT ADJUSTMENT IN THE DELIVERY SUITE ON AMBIENT TEMPERATURE TO PREVENT NEONATAL HYPOTHERMIA. Sreedhar Tallapureddy, M.D., Howard E. Herrell, M.D., and Martin E. Olsen, M.D., Department of Obstetrics and Gynecology, College of Medicine, East Tennessee State University, Johnson City, TN The normal body temperature of a newborn infant is 97.7-99.5° F. To prevent neonatal hypothermia, the World Health Organization, the American Academy of Pediatrics, and the American College of Obstetricians and Gynecologists recommend drying the baby thoroughly and immediately after birth, replacing any wet blankets, using pre-warmed radiant warmers if necessary, and maintaing the delivery room ambient temperature at 77° F (25° C) or greater.For the comfort of the surgeon and operating team, the operating rooms where cesarean deliveries are performed are often kept at 70° F or less. It is common practice to turn up the thermostat to 77° F or higher just before deliveries to comply with previously mentioned recommendations. We sought to investigate if this intervention has any meaningful impact on ambient room temperature.A Temp 101 Digital Thermometer was used over a four day period to record ambient room temperatures in the operating rooms used for cesarean deliveries at the Johnson City Medical Center. These measurements were taken at various times of day and night. A total of 29 experiments were conducted. In each experiment, the temperature was recorded initially and then the thermostat was increased from its setting of 68° F to 75° F. Then, temperatures were recorded at 5, 10, and 15 minutes. These data were then analyzed using a paired t-test.A statistically significant increase in the ambient temperature of the operating room was seen at 10 and 15 minutes after turning up the thermostat (p value of 0.028 and 0.0035 respectively). However, the maximum increase at 15 minutes was only 0.56° F. Approximately 52.8 minutes would be required to increase the temperature of the operating room from its baseline to 77° F if this trend continued.While preventing neonatal hypothermia is an important goal, our data would indicate that our common practice of turning up the thermostat just prior to a cesarean delivery is fruitless. In order to accomplish this goal, the room thermostat setting would need to be maintained at 77.0° F for at least an hour. Given the goal to accomplish most emergency cesarean deliveries in less than 30 minutes, then this would practically mean leaving in the setting at 77° F permanently.This would undoubtedly make operating conditions more uncomfortable for the surgical team, increasing perspiration and perhaps lead to surgical errors due to perspiration and broken concentration due to discomfort.Since we routinely use radiant warmers at all deliveries, which emit radiant heat at 99.5° F, then maintaining the ambient temperature of the room at 77° F would only affect the infant during the time that it leaves the operative site until it is placed under the warmer, which is usually less than 30 seconds. 2009 Appalachian Student Research Forum Page 74 SIMULTANEOUS REPAIR OF THORACIC AND INFRARENAL ABDOMINAL AORTIC ANEURYSMS USING ENDOVASCULAR STENT GRAFTS. Hao Duy Pham, Greg Champney, Tony Katras, and Daniel Rush, Division of Vascular Surgery, Department of Surgery, James H. Quillen College of Medicine, East Tennessee State University, Johnson City, TN Endovascular stent graft placement has proved an effective intervention for thoracic as well as abdominal aortic aneurysms. However, a literature search yielded only two previous case reports of simultaneous endovascular stent graft placement for repair of concomitant thoracic and abdominal aortic aneurysms. We report the case of a 63-yearold man with asymptomatic thoracic and abdominal aortic aneurysms. The thoracic aortic aneurysm was approximately 5 centimeters at its maximum diameter and was located in the middle descending portion of the thoracic aorta. The infrarenal abdominal aortic aneurysm was larger and more worrisome, with a maximum diameter of 7.8 centimeters. The patient’s comorbidities included asthma, recent pneumonia, and chronic obstructive pulmonary disease on supplemental oxygen, putting him at high risk for pulmonary complications during open repair. Thus, an endovascular repair was scheduled, and both aneurysms were to be excluded in one procedure using stent grafts. First, a cerebrospinal fluid drain was placed to minimize the risk of perioperative paraplegia. Next, the thoracic aortic aneurysm was excluded using a 40 millimeter by 15 centimeter Gore Tag endograft. Then, the abdominal aortic aneurysm was repaired using an Endologix Powerlink 28 by 16 by 140 millimeter main body device endograft. Finally, a proximal aortic cuff was placed using a 40 millimeter by 10 centimeter Gore Tag endograft. Thoracic and abdominal aortograms showed good flow through the stent grafts, the renal arteries, and the iliac arteries with no evidence of endoleak. The patient did not have any complications, and he was discharged on post operative day 4. He continued to do well at vascular surgery follow-up clinic 4 weeks after the procedure. This case report demonstrates that simultaneous endovascular repair of thoracic and infrarenal aortic aneurysms is a viable option for some high-risk patients. 2009 Appalachian Student Research Forum Page 75 Oral Presentations Graduate Students ESCHERICHIA COLI ATP SYNTHASE ALPHA SERINE-347 OF VISIT-DG SEQUENCE: A NEW MEMBER IN THE LEAGUE OF EXTRAORDINARY PHOSPHATE BINDERS Laura E. Brudecki and Zulfiqar Ahmad, Department of Biological Sciences, East Tennessee State University, Johnson City, TN F1Fo-ATP synthase is the smallest known biological nanomotor, found from bacteria to man. This enzyme is responsible for ATP synthesis by oxidative or photophosphorylation in membranes of bacteria, mitochondria, and chloroplasts. Thus, ATP synthase is the fundamental means of cell energy production in animals, plants, and almost all microorganisms. A typical 70 kg human with a relatively sedentary lifestyle will generate around 2.0 million kg of ATP from ADP and Pi (inorganic phosphate) in a 75-year lifespan. The general structure is the same in almost all organisms and has maintained multiple, highly conserved sequence patches throughout evolution. In its simplest form, in Escherichia coli, ATP synthase contains eight different subunits, namely alpha3, beta3, gamma, delta, epsilon, and ab2c10. Three questions that trouble scientists in developing nanomotors which can be used to repair damaged tissues inside the body are how big, how many, and at what speed should the nanomotors work inside a biological system in order to finish the job. ATP synthase works like a nanomotor through gamma-subunit rotation, and being the smallest biological nanomotor, it can be used as a base model in developing nanomotors in nanomedicine usage. In order to be able to use it as a base model, it is of paramount importance to thoroughly understand the catalytic properties of ATP synthase. For this purpose we recently focused on determining how Pi binding is achieved to generate ATP through ADP+Pi. So far, we have investigated the relationship between Pi binding and catalysis for eight potential Pi binding residues, namely betaArg246, betaAsn-243, alphaArg-376, betaLys-155, betaArg-182, alphaPhe-291, alphaSer347, and alphaGly-351, based on their proximity to the phosphate analogs AlF3 or SO42in x-ray structures of the catalytic sites. Employing Pi protection against NBD-Cl inhibition of ATPase as an assay, we have found that five residues, namely betaArg-246, alphaArg-376, betaLys-155, betaArg-182, and alphaSer-347, are directly involved in Pi binding while three, betaAsn-243, alphaPhe-291, and alphaGly-351, are not. We also found that the highly conserved alpha-subunit VISIT-DG sequence residues appear to play a vital role in the catalysis either directly through Pi binding or indirectly. Our catalytic site modulation studies show that the non-Pi binding residues can be altered through site directed mutagenesis to bind Pi or compensate for the known Pi binding residues thus enhancing the catalytic activity of the enzyme. 2009 Appalachian Student Research Forum Page 76 INHIBITION OF ESCHERCHIA COLI ATP SYNTHASE BY NATURAL AND MODIFIED POLYPHENOLS Prasanna K Dadi, Mubeen Ahmad, and Zulfiqar Ahmad, Department of Biological Sciences, East Tennessee State University, Johnson City, TN We have studied the inhibitory effect of five polyphenols namely, resveratrol, piceatannol, quercetin, quercetrin, and quercetin-3-beta-D glucoside and seventeen modified polyphenols on E. coli ATP synthase. Recently published X-ray crystal structures of bovine mitochondrial ATP synthase inhibited by resveratrol, piceatannol, and quercetin, suggest that these compounds bind in a hydrophobic pocket between the gamma-subunit c-terminal tip and the hydrophobic inside of the surrounding annulus in a region critical for rotation of the gamma-subunit. Polyphenols are naturally occurring plant based phytochemicals which possess antioxidant, chemopreventive, and chemotherapeutic properties. Foods such as apples, berries, cantaloupe, cherries, grapes, pears, plums, broccoli, cabbages, and onions are rich in polyphenols. Physiological relevance of dietary polyphenols can be ascribed to their interaction with mitochondrion in eukaryotic cells. Polyphenol led inhibitory studies on bovine mitochondrial suggest that the beneficial effects of dietary polyphenols are, in part, linked to the blocking of ATP synthesis in tumor cells thereby leading to apoptosis. Furthermore, Polyphenols are also known to have antimicrobial activity through inhibitory actions on ATP synthase. It was shown that polyphenols can inhibit biofilm formation and acid production by S. mutans. One of the pathways through which polyphenols are active against traits of S. mutans is the inhibition of proton–translocating F1-ATPase activity. The role of mycobacterial ATP synthase is of particular interest too as tuberculosis (TB) still claims about 2 million lives worldwide. It is interesting that two mutations, D32A and A63P, in the c-subunit of mycobacterium ATP synthase confer resistance to diarylquinoline, a tuberculosis drug. A wide range of natural and synthetic products are known to bind and inhibit ATP synthase. Thus, the knowledge of inhibitory effects of natural and modified polyphenols on E. coli ATP synthase could provide a basis to contend with pathogenic bacteria. The position of the hydroxyl groups, along with two or more phenolic structures of polyphenols, appears to be critical in exerting the inhibitory effect on ATP synthase. For the first time, our results show that polyphenols, resveratrol, piceatannol, quercetin, quercetrin, or quercetin-3-beta-D glucoside all inhibit Escherichia coli ATP synthase reversibly. Out of the above five polyphenols used to inhibit E. coli, piceatannol was the most potent inhibitor (IC50 ~14 micro M) followed by quercetrin (IC50 ~20 microM), quercetin (IC50 ~33 micro M), quercetin-3-beta-D glucoside (IC50 ~71 micro M), and resveratrol (IC50 ~94 micro M). Inhibitory studies by seventeen modified polyphenol compounds based on repositioning and or addition of –OH, COOH, NH2 group and phenolic rings resulted in potent inhibition of E. coli ATP synthase on a molar scale with very low IC50 values. 2009 Appalachian Student Research Forum Page 77 CANCER SUPPORT IN APPALACHIA: FEMALE CANCER SURVIVORS’ NEED FOR DISCLOSURE TO AND PROTECTION OF SOCIAL SUPPORT SYSTEMS Kathryn Duvall1, Dr. Kelly Dorgan1 and Dr. Sadie Hutson2, 1Department of Communication, 2Department of Internal Medicine, East Tennessee State University, Johnson City, TN This study examined how female Appalachian cancer survivors negotiate the need to disclose to and protect their social support system regarding concerns about their diagnosis, treatment, and fears. The Appalachian region is known for its beautiful mountains, strong heritage, and close-knit communities. Sadly, it is also known for health care disparities including higher rates of cancer and premature mortality. The Appalachian region has elevated lung, colorectal, and cervical cancer incidence and mortality rates. Health and cancer disparities in the region have been attributed to lack of access to health care professionals, lack of preventative care, being un- or under-insured, poverty, and lower levels of educational attainment. In light of these challenges being diagnosed with cancer in the Appalachian region presents unique needs. A convenience sample of 28 Appalachian female cancer survivors from Northeast Tennessee and Southwest Virginia participated in a day-long modified story circle event. Women were divided into two groups during the event and asked to share their stories of cancer survivorship in Appalachia in two 4 hour sessions. Open-ended questions probed for what makes the cancer experience in Appalachia unique. Participation was voluntary and informed consent was obtained from each participant. Transcripts from both story circles were transcribed verbatim and uploaded into NVivo 7 to assist with coding and data organization. Qualitative content analysis was used to identify emergent themes, and data was coded both independently and jointly. An analysis of the data revealed three broad themes regarding social support: social support includes family, friends, and religion; the need for social support during a cancer diagnosis and treatment; and the need to protect the social support system from concerns regarding the cancer diagnosis and treatment. Most survivors expressed the challenges of negotiating the need to disclose to their social support system as well as the need to protect the social support system. They needed support from friends, family, and their religious communities regarding the cancer diagnosis, treatment, and concerns, but also expressed concern over worrying or upsetting members of the social support system. These data demonstrate the need to understand the negotiation of protection and disclosure among Appalachian female cancer survivors. 2009 Appalachian Student Research Forum Page 78 THE IMPACT OF A SUMMER FITNESS INITIATIVE ON PHYSICAL ACTIVITY LEVELS OF RURAL HIGH SCHOOL STUDENTS Michele Gourley*, Seth Brown*, Linsey Neuhaus*, Dr. Kenneth Olive, *Department of Internal Medicine, College of Medicine, East Tennessee State University, Johnson City, TN This study was designed to compare the current fitness habits of students in rural Tennessee who had previously participated in a summer fitness initiative called Pro Strength Pro Speed (PSPS) with those who had not. It was hypothesized that high school students who had participated in PSPS would report higher levels of physical activity than students who had never participated in the program. A physical activity survey based on WHO guidelines was administered to all high school students ages 13-18 years in the county where PSPS is located. The survey asked questions about type and duration of physical activity within the past week as well as whether or not the student had participated in the PSPS program. Data was analyzed using t-test and Chi Square. 758 of 1184 students filled out the survey (64%). 199 males (26.2%) had completed the PSPS program as compared to 77 females (10.2%). Eighty percent of PSPS participants reported doing greater than 30 minutes of vigorous physical activity as compared to 46% of non-participants (p<0.001). Fifty-seven percent of PSPS participants reported completing moderate physical activity within the past week compared to 37% of nonparticipants (p<0.001). PSPS participants reported participating in 1.5 more days per week of vigorous activity and 1 more day per week of moderate activity than nonparticipants (p<0.001). Males spent more days doing vigorous and moderate physical activity than females (p<0.001). Sixty-nine percent of males reported engaging in vigorous physical activity compared to 32% of females. Forty-nine percent of males reported doing moderate physical activity as opposed to only 30% of females. Numerous studies have shown that increasing physical activity is an effective way to prevent weight gain, a key component to preventing or reducing the current epidemic of obesity. This study showed a significant difference between the type and duration of physical activity of previous PSPS participants and non-participants. Important limitations of this study include a possible selection bias in that those who participated in the PSPS program may have been more interested in engaging in physical activity than non-participants since pre-program activity levels were not evaluated. Another limitation is that over twice as many males surveyed reported having completed the PSPS program as compared to females. This could be due to a higher propensity among males for engaging in activity than females. According to the results of this study, a summer fitness program geared toward rural high school students could be a potential means of increasing physical activity thus helping curb the epidemic of obesity. Further research is needed to delineate more clearly the pre and post program activity levels of participants. as well as to explore reasons for the discrepancy between activity levels in males and females. 2009 Appalachian Student Research Forum Page 79 ILLNESS BURDEN AND DEPRESSIVE SYMPTOMS: MODERATING EFFECT OF SOCIAL SUPPORT AND FAMILY CRITICISM IN OLDER ADULTS Joshua Hatfield, B.S. 1, Jameson K. Hirsch, Ph.D.1, and Jeffery M. Lyness, M.D.2, 1 Department of Psychology, East Tennessee State University, Johnson City, TN, 2 Department of Psychiatry, University of Rochester Medical Center, Rochester, NY Older adults with chronic medical problems are at increased risk for depressive symptoms. During times of poor health, older adults may rely more heavily on the support of others, particularly family, than those in other age groups. Social support, including frequency of social interactions, perceived satisfaction with social network, and instrumental support may mitigate some adverse effects of illness, and are related to improved treatment adherence and better physical and mental health. Conversely, lack of familial support and perceived family criticism may contribute to depression. We examined family criticism and social support as potential moderators of the association between medical illness burden and depressive symptoms. Respondents were 745 older adult (63.4% female; Mean age 75.12 [SD=6.86]), primary care patients. Our sample was predominantly White (91.8%) and well educated (mean educational level = 14.11 years [SD = 4.1]). Participants completed the Family Criticism subscale of the Family Emotional Involvement and Criticisms Scale (FEICS), the Duke Social Support scale, the Hamilton Rating Scale of Depression, and a physician-rated measure of illness burden (Cumulative Illness Rating Scale; CIRS). Univariate and multivariate hierarchical, linear regressions were conducted, covarying age, gender, education, MMSE total score, and functional impairment (Karnofsky Performance Status Scale; KPSS). In univariate analyses, social integration and perceived social support significantly moderated the relationship between illness burden and depressive symptoms; family criticism was not a significant moderator. In a multivariate model, social integration was a significant moderator. There was also a clinically significant trend toward significance for perceived social support as a moderator (p=.06). Although not a moderator, family criticism was significantly, independently associated with depressive symptoms. Implications for the treatment of depressive symptoms in individuals with medical illness burden may include promotion of support networks, particularly meaningful community and intergenerational relationships. Although it seems intuitive that reducing family criticism may also be an effective intervention to treat depressive symptoms, perhaps this is a less effective approach when considered in the context of medical illness. Some evidence suggests that older adults, as compared to younger adults, are better able to capitalize on their strengths to promote well-being in times of distress. When confronted with medical illness burden, older adults may respond more positively to promotion of meaningful familial and community interactions, than to efforts toward reduction of family criticism. Treatment of depressive symptoms in patients experiencing chronic medical problems may be enhanced by increasing social integration and the quality of a patients’ interpersonal relationships. 2009 Appalachian Student Research Forum Page 80 POTENTIAL USE OF CANNABINOIDS IN THE TREATMENT OF PANCREATIC CANCER Theresa Pickle and Douglas Thewke, Department of Biochemistry and Molecular Biology, East Tennessee State University, James H. Quillen College of Medicine, Johnson City, TN Pancreatic cancer is the fourth leading cause of cancer related deaths. The mean survival rate is ~6 months, and only 4% of patients survive five years. Despite extensive testing, only one chemotherapy agent (gemcitabine) has been found to produce any benefit to these patients, unfortunately, the clinical response rate to gemcitabine is less than 10% with life prolongation being only 6 weeks on average. Cannabinoids, such as Δ9tetrahydrocannabinol (THC), the active agent of Cannabis sativa, exhibit some anti-tumor properties via induction of apoptotic mechanisms. Cannabinoids produce their biological effects by engaging specific receptor-mediated signaling pathways. To date, two cannabinoid-specific receptors, designated CB1 and CB2, have been cloned and characterized from mammalian tissue. The current study was conducted to investigate the potential use of cannabinoids as an anti-tumor treatment in pancreatic cancer. Using RTPCR we found evidence for expression of CB1 and CB2 in three human pancreatic cancer tumor cell lines (AsPC1, CaPan1, and Mia-PaCa2). Cell viability analysis revealed that treatment of these pancreatic tumor cell lines with Win 55,212-2, a potent synthetic CB1/CB2 agonist, results in a significant reduction in cell viability. When AsPC1 cells were subjected to Win 55,212-2 in combination with gemcitabine or 17AAG, an hsp90 inhibitor, a cumulative decrease in cell viability was observed after 48 hours. Treatment of Mia-PaCa2 and CaPan1 cells with Win 55,212-2 in combination with gemcitabine or 17-AAG did not result in additive growth inhibition. Win55,212-2 treatment induced apoptosis, as determined by caspase-3 activity, in AsPC1, CaPan1 and Mia-PaCa2 cells but not in non-neoplastic human pancreatic duct epithelial (HPDE) cells. The induction of caspase-3 activity by Win 55,212-2 in Mia-PaCa2 cells was prevented by a CB1 receptor-specific antagonist, AM251, but not by a CB2 receptor-specific antagonist, SR144528. In contrast, induction of caspase-3 activity in AsPC1 cells was inhibited by SR144528 but not AM251, while induction of apoptosis in CaPan1 cells was not affected by either antagonist. Consistent with induction of apoptosis, Win 55,212-2 treatment reduced expression of Bcl-xL, an anti-apoptotic member of the Bcl family, in Mia-PaCa2 cells but not in HPDE cells. These results demonstrate that several pancreatic tumor cell lines express cannabinoid receptors and that a cannabinoid, Win 55,212-2, potently inhibits the viability of these pancreatic tumor cells, at least in part, by inducing apoptosis via CB1/CB2 receptor dependent and independent mechanisms. 2009 Appalachian Student Research Forum Page 81 DISPOSITIONAL OPTIMISM AS A MEDIATOR OF THE RELATIONSHIP BETWEEN ETHNIC IDENTITY AND DEPRESSIVE SYMPTOMS Preston Visser1, Jameson Hirsch1, and Elizabeth Jeglic2, 1Department of Psychology, East Tennessee State University, Johnson City, TN 2Department of Psychology, John Jay College of Criminal Justice, New York, NY Ethnic identity refers to the portion of a person’s self-concept composed of knowledge, beliefs, and attitudes about affiliation, belonging and commitment to a social group of which he or she is a member. Ethnic identity has been associated with decreased depression in several ethnic groups; however, this relationship is not clearly understood. Greater involvement in ethnically oriented activities or stronger adherence to cultural values may directly protect against depression, but they may also be indirectly related via optimism, or individuals’ general expectation of experiencing limited discrepancy between actual and ideal life situations. Although dispositional optimism consistently predicts lower levels of depression, it has not been investigated in relation to ethnic identity and depression. We examined the potential association between ethnic identity and depression in several ethnic groups and, additionally, assessed the role of optimism as a mediator of these associations. Participants included 386 undergraduate students [69.2% female; Mean age = 19.6 (SD = 3.1)] from an urban, Northeastern university, consisting of 41.5% (n= 160) Hispanic, 25.4% (n= 98) Black, 18.4% (n= 71) White, and 5.7% (n= 22) Asian students. Ethnic identity was assessed with the Multigroup Ethnic Identity Measure, dispositional optimism with the Life Orientation Test-Revised, and depressive symptoms with the Beck Depression Inventory-Second Edition. We used hierarchical, multivariate regressions, controlling for age and gender, to test for mediation. A main effect of ethnic identity on depression was found in Blacks, Hispanics, and Whites, but not Asians. In stratified analyses, the effect of ethnic identity was reduced to non-significant with the inclusion of optimism in the model, and indirect effects testing further supported the model in all three groups. Ethnic identity was related to decreased depression in each ethnicity except Asians, but small sample size (n = 23) limits findings in this group. Optimism mediated ethnic identity and depression in Blacks, Hispanics and Whites. Stronger ethnic identity may impact an individual’s selfperception or what he or she considers to be an ideal life situation, such that overall discrepancy and, subsequently, psychological distress decrease. For example, expressing desire to reflect one’s own cultural values and norms may be more predictive of success, and less discrepancy, than attempting to adhere to those outside one’s culture. Clinical implications include selective reinforcement of ethnic identifications that will reduce discrepancy between one’s perception of current and ideal life circumstances. Limitations include use of cross-sectional data and a college sample, which limit generalizability. Clinical and community research investigating the role of optimism as a mechanism of influence in the relationship between ethnic identity and psychological distress is needed. 2009 Appalachian Student Research Forum Page 82 REPROGRAMMING OF IMMUNE RESPONSE IN LATENT HSV INFECTION Stacie N. Woolard, Subhadra Nandakumar, Akaike1 and Uday Kumaraguru, Department of Microbiology, College of Medicine, East Tennessee State University, Johnson City, TN; 1Dept. of Biomolecular Engineering, Institute of Technology, Yokoyama, Japan Herpes Simplex Virus type 1 (HSV-1) is a chronic viral infection that remains latent in sensory neurons. The virus persists for life but a potent T cell response is induced each time the virus is reactivated from latency. The stress and the negative impact of repeated and intermittent immune stimulation on responding T cell impairs immune memory. NK cell share with CD8+ CTL subsets several phenotypic and functional features. However, so far only NK inhibitory receptors have been detected on CD8+ T cells. All of the inhibitory NK receptors are characterized by the presence of a tyrosine-based inhibitory motif (ITIM) that is necessary and sufficient for their inhibitory function. Killer cell lectin like receptor subfamily G1 (KLRG1) is one such receptor. We chose to investigate KLRG1 because of the recent description of its ligand –the cadherins. Cadherins have been shown to be modulated by some chronic virus infection but its importance in the context of HSV infection is still unexplored. These proteins are also expressed on immature DCs that first encounter the antigen and help in its processing and presentation to the T cells. Cadherin levels are down-regulated in persistent viral infection, resulting in an aberrant maturation of the DCs and migration to the secondary lymphoid organs. This fact prompted us to investigate the immunological outcome of KLRG1 and cadherin interaction during persistent viral infections. Using a mouse model of latent HSV infection we show that periodic reactivation resulted in the development of discordant CD8+ T cells. One of feature being the expression of an NK receptor, KLRG-1 on CD8+ T cells that increased with duration of latency. Interestingly, at the same time the ligand for KLRG-1, E-Cadherin level went down and was more pronounced on DCs. The Ecadherin expression on the DC is essential for the DC-DC transfer of the antigen and the subsequent presentation to the T cells. The absence of the E-cadherin resulted in the improper maturation of the DCs. Our experiments used DCs isolated from naïve mice and from acute phase and chronic phase of HSV infected mice. DCs were infected with GFP labeled HSV and the functional ability was evaluated. The DCs were analyzed for Ecadherin and GFP levels to correlate the viral antigen processing ability of the DCs to the level of E-cadherin expression. Our results indicate that the lower E-cadherin levels resulted in discordant behavior of anti-HSV CD8+ T cells presumably as a result of inferior antigen processing and presentation by the DCs and also by other regulatory mechanisms. Thus, strategies to manipulate the E-cadherin levels could aid in a better immune response in chronic infections and cancer. Our exploratory study should impact on the future design of vaccines against HSV, particularly those which could be used therapeutically to improve immunity in persons subject to recurrent lesions.Acknowledgements: Department of Microbiol. ETSU- Start up Funds 2009 Appalachian Student Research Forum Page 83 THE ROLE OF DIETARY DIVERSITY IN THE NUTRITIONAL STATUS OF SCHOOL-AGED CHILDREN IN RURAL HONDURAS Michele Gourley, Greg Winters, Dr. Roger Smalligan*, Dr. James Anderson, College of Public Health, East Tennessee State University, Johnson City, TN *Dept. of Pediatrics, Quillen College of Medicine, East Tennessee State University, Johnson City, TN This study was undertaken at the request of a non-governmental organization that was concerned about the rate of malnutrition among school-aged children in their catchment area in rural Honduras. Studies have consistently shown that in developing countries, an increased diversity in diet increases the likelihood that adequate amounts of macro and micronutrients critical for the growth of a child will be consumed. The objective of the study was to determine the prevalence of malnutrition and to evaluate whether or not a correlation exists between the dietary diversity in 8 and 9-year-old children and their nutritional status as measured by body mass index (BMI) (kg/m2). A random sample of 8 and 9-year-olds was chosen from 14 schools in three communities in rural Honduras. A survey with questions about food groups consumed within a 24 hour time period as well as other questions thought to be possible contributing factors to dietary diversity and/or the nutritional status of the children was developed based on similar surveys administered by USAID. Questions aimed at determining the socioeconomic status of the children were also included. Results were calculated using Chi-Square and t-test. 132 of 181 eligible eight and nine-year-old students from 2007 records were surveyed (72.9%) and 127 surveys were included in the study. Sixty-two males participated and 65 females. Height and weight measurements were used to calculate each child’s BMI, and these were compared to current WHO growth parameters. Only 3 percent of children were classified as malnourished and 4 percent were classified as having stunted growth. Individual dietary diversity scores (IDDS) ranged from 3-8 with a mean score of 5.5 and a standard deviation of 1.2. No correlation was found between IDDS and BMI in either gender or age group. However, a significant positive correlation (p=0.04) was shown between those who reported eating beans, lentils, or legumes and BMI. Often a high prevalence of malnourished children exists in developing countries in Central America; however, this study showed few malnourished children in the communities studied in Honduras. Some studies in developing countries have shown an IDDS of 4 to be an indicator of an adequate diet, and the mean IDDS of 5.5 of this study may indicate a low prevalence of malnutrition in this population. The lack of correlation between BMI and IDDS was inconsistent with prior studies. However the overall low prevalence of malnutrition in the study group may have reduced the power of this study to detect a difference in IDDS and BMI. Few studies exist showing the correlation between socioeconomic status and dietary diversity, and this study demonstrated no correlation. The significant correlation between legumes and BMI suggests that the frequency or quantity of legume consumption could serve as a future indicator of adequate nutritional intake by children. 2009 Appalachian Student Research Forum Page 84 Medical Residents and Post-Doctoral Fellows ORGANIZING PNEUMONIA WITH MYELODYSPLASTIC SYNDROME: A RARE ASSOCIATION Venkataramanan Gangadharan M.D., Department of Internal Medicine, East Tennessee State University, Johnson City, TN; and Mayur Patel M.D., Division of Pulmonary/Critical Care, Department of Internal Medicine, East Tennssee State University,Johnson City, TN Mr. R is a 56 year old smoker, who presented to the hospital with progressive shortness of breath. His illness began 2 months earlier with generalized fatigue, productive cough and weight loss. Shortly after the onset of his symptoms, he was treated with a course of antibiotics without improvement. He had no previous medical illness. On presentation he was pale and cachectic. He was normotensive with mild tachycardia. He was in moderate respiratory distress while breathing oxygen by nasal cannula at 4 liters per minute. Coarse breath sounds could be heard bilaterally as well as mild bibasilar crackles. The remainder of the examination was unremarkable. His hemoglobin was 5.1 gm/dl, Hct 16.1 with an MCV of 102.8. The complete blood count and basic metabolic profile were otherwise normal. The patient’s D-dimer was elevated at 3.95 and his arterial blood gas analysis reflected a respiratory alkalosis. His chest radiograph documented a pattern suggestive of loculated pleural fluid as well as an underlying pulmonary alveolar interstitial pattern. Subsequent computerized tomography of the chest confirmed bilateral extensive pleural effusions with extensive consolidation involving the upper and middle lung zones. An enlarged spleen was also identified, which when put together with his acute anemia suggested an associated hematologic process. A subsequent bone marrow biopsy had features consistent with myelodysplastic syndrome (MDS). Recognizing the potential of organizing pneumonia with associated MDS, antibiotic therapy was discontinued and the patient was treated with glucocorticoids with rapid resolution of his pulmonary pathology. His myelodysplastic syndrome was then treated supportively. Cryptogenic organizing pneumonia (COP), also known as Bronchiolitis Obliterans Organizing Pneumonia (BOOP) is a clinical entity characterized by a flu-like illness, non productive cough, and ensuing progressive dyspnea. The diagnosis is often suggested by thoracic imaging and, sometimes, characteristic pathologic specimens obtained at biopsy. Imaging modalities such as HRCT Scan typically show areas of air space consolidation often with lower lung predominance. At times there may be a subpleural or peribronchial distribution of disease, small nodules along bronchovascular bundles, and ground-glass attenuation. The underlying etiology is often indeterminate and ranges from viral infection, connective tissue disease, to drug abuse or medication use. A rare association with underlying hematologic malignancy has also been documented, however, without a specific causal relationship. Malignancies previously documented include the acute leukemias and lymphomas as well as myelodysplastic syndrome. This patient’s presentation should remind the clinician that cryptogenic organizing pneumonia may be a manifestation of an underlying chronic hematologic malignancy and that appropriate therapy will provide early symptomatic benefit. 2009 Appalachian Student Research Forum Page 85 THE HIDDEN DIARRHEA CULPRIT: LYMPHOCYTIC COLITIS Umbar Ghaffar MD, East Tennessee State Uinversity, Department of Internal Medicine, Johnson City, TN; and Roger D Smalligan MD, MPH, East Tennessee State Uinversity, Department of Internal Medicine, Johnson City, TN The objective is to recognize a relatively uncommon but easily treatable cause of diarrhea.A 69-year-old Caucasian man presented with a history of 5 weeks of watery diarrhea associated with nausea and occasional abdominal discomfort. He denied any blood in his stools, fever, sweats, chills or weight loss.He had started taking over the counter weight loss medication 5 weeks prior. He took bismuth subsalicylate for the diarrhea without relief. His past medical history was significant for hypertension, osteoarthritis, obstructive sleep apnea, erectile dysfunction and depression. His medications included multivitamins, aspirin and tidalifil. Physical exam was normal including a soft abdomen without organomegaly .Labs were unremarkable. Colonoscopy with biopsy showed Lymphocytic colitis .The patient was treated with mesalamine and a prednisone taper with complete resolution of symptoms within 2 weeks.IMPLICATIONS/DISCUSSION: Chronic diarrhea is a common complaint of patients presenting to internists and GI specialists alike. The most common etiologies are irritable bowel syndrome, medication or radiation side effects, inflammatory bowel disease or some infectious cause including certain viruses and parasites. Our patient illustrates an uncommon but often treatable cause of chronic diarrhea: lymphocytic colitis (LC). Lymphocytic colitis is one of two types of microscopic colitis, the other being collagenous colitis (CC). Both lymphocytic and collagenous colitis are more prevalent in older adults, affect men and women equally and the cause is still unknown. Since there is an association of microscopic colitis with celiac disease, diabetes, rheumatoid arthritis, thyroid disorders, pernicious anemia and scleroderma, an autoimmune etiology has been proposed. There are also reports of LC and CC occurring in families suggesting a possible genetic component as well. Medications associated with an increased risk of microscopic colitis include NSAIDS, lansoprazole, sertraline, ticlopidine, ranitidine and acarbose. The diagnosis of lymphocytic colitis is typically reached when a patient with chronic non-bloody diarrhea has an endoscopically or radiologically normal colon, but biopsies show unique inflammatory changes including an increased number of CD4+ T lymphocytes in the colonic epithelium. Management of mild cases include initiation of a low fat diet, avoidance of alcohol, caffeine, spicy and lactose containing foods. Carbonated beverages and certain vegetables like beans, cauliflower, broccoli and cabbage may also exacerbate the symptoms. More severe cases may be treated with antidiarrheals, cholestyramine, steroids, or other anti-inflammatory medications like mesalamine or sulfasalazine. This case reminds physicians to keep lymphocytic colitis in the differential diagnosis of chronic diarrhea and shows the important role of colonoscopy with biopsy when the etiology is not readily identifiable by usual historical, exam, and laboratory means. 2009 Appalachian Student Research Forum Page 86 HEPATOCELLULAR CANCER WITH BONE METASTASES: UNDER DETECTION OR UNUSUAL METASTASES. B. Jenigiri, A. Hamati, R. E. Enck, Department of Oncology, East Tennessee State University, Johnson City, TN Bone as a site of initial metastases is considered to be uncommon in patients with hepatocellular carcinoma (HCC), and is a site that is often overlooked during initial work up. Previous case studies and autopsy series have shown that it is the third most common site of metastases after lung and the adrenals. In the past two years three patients presented to our clinic with HCC and were subsequently noted to have bone as the only site of metastases.We discuss one of the cases and review the literature to emphasize the need for a bone scan as part of the initial evaluation of HCC.A 47 year old male with history of hepatitis C for more than 20 years presented to the primary care clinic complaining of increasing abdominal pain for one month. Initial workup showed multiple hypoechoic lesions in the liver. Subsequent CT scan of the abdomen showed a suspicious rib lesion and a bone scan revealed multiple bone metastases. Metastatic workup was otherwise negative. In view of the advanced (stage IV) disease and the inoperability of the primary lesion the patient was treated with sorefenib. Follow up CT scan four months later showed partial remission of the multiple lesions in the liver. Three months later, this patient was readmitted with seizures and an MRI revealed brain metastases,subsequently the patient’s condition deteriorated and the family elected for hospice.This case shows a common presentation for HCC but in the usual work up the bone metastases would have been overlooked. Most compendia guidelines do not include bone scan as a part of initial work up for HCC. There is literature to suggest that HCC should always be considered in the differential diagnoses in patients presenting with bone metastasis, with a few reports suggesting this to be the first manifestation of HCC. The incidence of bone metastasis is 3-20% and recently has shown an upward trend.Bone metastases are unique since they occur early in the course prior to clinical manifestations of HCC become apparent, and majority are asymptomatic. Metastasis to the bones occur likely via portal vein-vertebral vein plexuses due to either portal thrombus and/or portal hypertension which allows bypass through plexus.The bone involvement in patients with HCC is rising and one of the main reasons for this is early detection and better survival of HCC patients due to recent progress made in both the diagnosis and treatment of the disease. The importance of the recognition of this entity is enhanced by reports that patients with HCC and bone metastases do well and show long-term survival after hepatectomy and radiotherapy to the bone lesions. However the use of bisphosphonates in this condition is not widely studied.In conclusion there is a need to change treatment guidelines to include bone scan as a part of initial evaluation of hepatocellular carcinoma and also emphasize the need for further studies in evaluating the role of bisphosphonate therapy in management of bone metastases in HCC. 2009 Appalachian Student Research Forum Page 87 BEWARE OF A BROKEN HEART ! Sumit Kalra, MD, Hassan Ismail, MD, Ransford Brenya, MD, Roger D Smalligan, MD, Department of Internal Medicine, East Tennessee State University, Johnson City TN A 64-year-old postmenopausal woman presented with 1 day of chest pain and shortness of breath. She was in a motor vehicle accident (MVA) two days prior, evaluated and sent home. Her past medical history, family and social history were unremarkable. On exam she had normal vitals, mild sternal tenderness, and normal cardiac, respiratory, abdominal and neurological examinations. Her CBC, lipid and metabolic panels were normal while she was found to have a suppressed TSH, elevated free T3, T4 and positive thyrid peroxidase antibodies. Her chest and rib radiographs and CT scans did not reveal any fractures, lung contusions, or aortic dissection. Her EKG showed 3-4 mm ST-segment elevation in the precordial leads along with a troponin level of 1.99 (<0.5 normal). The patient was taken to emergent left heart catheterization but was found to have normal coronaries. The echocardiogram revealed anterior wall hypokinesis with an ejection fraction of 30%. She was managed medically and had a normal LV function on follow up echocardiogram a few weeks later. The most common diagnoses considered following an MVA in a patient with chest pain are myocardial contusion, aortic dissection, pericardial tamponade and acute coronary syndrome. This case reminds physicians of another important condition which can occur following such a stressful event: Takotsubo or stress cardiomyopathy (Broken heart syndrome). This is a recently recognized syndrome which was first described in detail in 1991. The typical presentation is of a postmenopausal woman with a recent stressful life event that presents with acute chest pain and EKG changes consistent with myocardial ischemia or infarction (ST-segment elevation:56%, T wave inversions:17%), acute (and transient) left ventricular systolic dysfunction on echocardiogram and a left heart catheterization demonstrating normal coronary arteries. The transient LV dysfunction typically reverses within days to weeks. The cardiac enzymes may be elevated above normal although there is usually less elevation than that seen in STEMI. The diagnosis is thus usually established after left heart catheterization in the appropriate clinical setting. Stressful events reported as preceding the condition range from an argument, death of a loved one, illness, heavy exercise, or rarely, following an MVA as in our case. Most cardiologists agree that the mechanism of Takotsubo cardiomyopathy involves the effects of excess catecholamines on the heart. Thyroid hormones have a facilitatory role on the adrenergic system and are known to enhance the myocardial sensitivity to sympathetic stimulation. We believe that the combination hyperthyroidism and sympathetic stimulation from the recent stressful event (MVA) was responsible for the presentation in our case. It is important for internists to be aware of the typical presentation of stress cardiomyopathy and be watchful for the presence of thyrotoxicosis in these patients. 2009 Appalachian Student Research Forum Page 88 A RARE CASE OF A WEAK HEART Venkataramanan Gangadharan M.D.1, Benjamin Holland M.S. M.D.2, and Gretchen Wells M.D. M.P.H. 2, 1Department of Internal Medicine, East Tennessee State University, Johnson City, TN; 2Department of Internal Medicine, Wake Forest University Baptist Medical Center, Winston-Salem, NC A 40-year-old Caucasian female presented to the emergency department with worsening shortness of breath over the past one month. Past medical history was significant for asthma since childhood, hypertension, and diabetes mellitus. Family history revealed premature heart disease but of unknown etiology. Subsequent physical examination revealed an obese woman, in no distress, normotensive but tachycardic, saturating 93% on 2L/min of O2 with an elevated jugular venous pulse. Systemic exam revealed decreased breath sounds with rare inspiratory rhonchi and distant heart sounds but no other abnormalities. CXR demonstrated cardiomegaly with some increased pulmonary vascular congestion. CBC and BMP were essentially normal except for a fasting blood sugar in the 300’s. TSH and D-dimer were within normal limits; however, BNP was elevated at 1000 much higher than her baseline as recorded on previous hospital admissions. With new-onset heart failure the primary diagnosis, the patient underwent extensive testing including a transthoracic echocardiogram which revealed severe systolic dysfunction but no overt valvular abnormalities. A left and subsequent right heart catheterization with endomyocardial biopsy ruled out ischemic, infiltrative, and infectious etiologies. A Cardiac MRI showed prominent biventricular apical trabeculation which appeared hyperintense on T2-weighted images. Left ventricle end-diastolic trabeculae to myocardium maximum thickness ratio was approximately 2-3 where a diastolic ratio >2.3 suggests LV non-compaction. The patient was then placed on appropriate initial therapy including an ACE inhibitor, beta-blocker, and diuretic with improvement in her symptoms. Heart Failure is a common and disabling condition especially in the elderly, where 6-10 percent of those above 65 years of age are affected. Over 60% of cases are secondary to ischemic injury; however, rarely non-ischemic etiologies are diagnosed. Possibilities include infectious or infiltrative disease vs. toxin and drug induced cardiac injury. One such rare cause of non-ischemic dysfunction of the heart is “isolated left ventricular non-compaction”. It is an entity characterized anatomically by numerous prominent trabeculations and deep intertrabecular recesses in the ventricular wall which occurs when normal endomyocardial embryogenesis around 5 to 8 weeks of fetal life is arrested. These patients often go undiagnosed for many years and can present with symptoms of overt heart failure, arrhythmias and/or evidence of thrombo-embolic events in their 40’s. Treatment involves routine medical management for systolic/diastolic heart failure, but emphasis should be placed on anticoagulation, necessity for ICD placement and screening of first degree relatives. This case illustrates that idiopathic cardiomyopathy albeit an uncommon diagnosis should be further investigated to afford early screening measures amongst family members while providing aggressive and appropriate medical therapy. 2009 Appalachian Student Research Forum Page 89 PUTTING CANCER TO SLEEP AT NIGHT. B. Jenigiri, S. Koppisetti, and K. Krishnan, Department of Oncology, College of Medicine, East Tennessee State University, Johnson City, TN Objective: To review vasculogenic mimicry (VM), its role in cancer metastases and emphasize on the inhibitory effect of melatonin on various extracellular matrix components(ECM) and growth factors involved in the synthesis of these capillary like structures. Also discuss the significant anti-cancer effects of melatonin and its additive effect in decreasing chemotoxicity.Methods: A review of pertinent literature on pub med.Results: The ability of tumor cells to metamorphose into vessels that connect to host vessels and carry blood for the tumor growth was observed in several aggressive tumors and was termed Vasculogenic mimicry (VM). Review of the literature on VM reveals several signal transduction factors and ECM components like vascular endothelial growth factor (VEGF), and hypoxia inducible factors-1 (HIF-1) that can be significantly inhibited by melatonin. The efficacy of melatonin in inhibiting these factors is comparable to drugs that are currently used for treating tumors but devoid of the associated adverse effects. The use of melatonin as an anti-cancer agent has been proven by several studies both in vivo and in vitro, but literature suggests its use specifically in tumors metastasizing by VM where, the conventional therapeutic agents that are currently being used have a limited role.Conclusion: Several aggressive tumors with VM like breast cancer and melanomas when treated with the conventional angiogenesis inhibitors still derive blood supply by the alternate channels and survive. This limits the role of the available treatment modalities and calls for a tumor targeted therapy based on the mechanism of metastases.Hypoxia the significant stimulant of angiogenesis, induces the expression of several hypoxia responsive genes like HIF-1 alpha, VEGF, etc. Hypoxia induced gene expression increases the vascular permeability and induces the plasticity of the tumor cells enabling their transformation to pleuripotent cells. VEGF inhibitors like bevacizumab and sorefenib are the latest drugs at the cutting edge of cancer therapeutics. Melatonin significantly decreases the expression of all the hypoxia inducible genes including VEGF. Cyclo-oxygenase-2(COX-2) inhibitors suppress vasculogenic mimicry in breast cancer and other cell lines and are currently being investigated in clinical trials. However, the use of COX-2 inhibitors like celecoxib is associated with several adverse cardiac effects. Melatonin in addition to being a COX-2 inhibitor is a potent antioxidant decreasing the free radical induced damage by chemotherapy.Population studies have shown an increased incidence of breast cancer among night shift workers. Disturbance of the circadian rhythm mediated via the melatonin synthesis and clock genes, increases the risk of hormone-related diseases, including breast cancer. Our review indicates that melatonin at physiological and pharmacological blood concentrations inhibits tumorigenesis putting cancer to sleep at night. 2009 Appalachian Student Research Forum Page 90 2008 Award Winners Poster Presentations Division I – Undergraduates Biomedical Sciences First Place: Bassam Lamale, ETSU. Faculty Sponsor – Sharon Campbell, Dept. of Internal Medicine. Synthesis and Evaluation of Gamma Tocopherol Lysinate as an AntiCancer Agent in Human Prostate and Colon Cancer Cell Lines. Second Place: Jessica Pugh, ETSU. Faculty Sponsor – David Johnson, Dept. of Biochemistry and Molecular Biology. Expression of Human Neutrophil Elastase Using Sumo Fusion Technology. Natural Sciences & Mathematics First Place: Elizabeth Harris, ETSU. Faculty Sponsor – Michelle Hoard, Dept. of Mathematics. The Sierpiński Square Graph and Some of its Properties. Second Place: Brent Linville, ETSU. Faculty Sponsor – Charles Thomas Jones, Dept. of Biological Sciences. Environmental Factors Contributing to Colony Success in a Social Spider System. Third Place: Maleka Khambaty, ETSU. Faculty Sponsor – Dr. Tom Ecay and Dr. James Stewart, Depts. of Biology & Physiology. Developmental Expression of Carbonic Anyhdrase II in the Extraembryonic Membranes of the Corn Snake, Pantherophis Guttatus. Fourth Place: Tazley Hotz, ETSU. Faculty Sponsor – Dr. Dhirendra Kumar, Dept. of Biological Sciences. SA-Methyl Transferase Involvement in Disease Resistance Signaling. Social & Behavioral Sciences First Place: Daniel Noel, ETSU. Faculty Sponsor – Russell Brown, Dept. of Psychology. Nicotine Sensitization in Adolescent Beta Arrestin-2 knockout Mice: Correlations with BDNF. Second Place: Benjamin Martin, ETSU. Faculty Sponsor – Chris Dula, Dept. of Psychology. A Look at the Development of the Martin Stigma Against Tattoos Measure. Third Place: Julia Lehmann, ETSU. Faculty Sponsor – Russ Brown, Dept. of Psychology. Nicotine-Conditioned Hyperactivity in Adolescent Male and Female D2Primed Rats. 2009 Appalachian Student Research Forum Page 91 Division II – Graduate Students, 1-2 Years Biomedical Sciences First Place: Courtney Netherland, ETSU. Faculty Sponsor – Douglas Thewke, Dept. of Biochemistry and Molecular Biology. Cannabinoid Receptor (CB2) Deficiency is Associated with Reduced Lesional Apoptosis in Low-Density Lipoprotein Receptor-Null Mice. Second Place: Stacie Woolard, ETSU. Faculty Sponsor – Uday Kumaraguru, Dept. of Microbiology. Toll-Like Receeptor Modulation of Dendritic Cell: Natural Killer Cell Interaction. Natural Sciences & Mathematics First Place: Lok Raj Pokhrel, ETSU. Faculty Sponsor – Istvan Karsai, Dept. of Biological Sciences. Mapping the Skin Pattern of Spotted Salamanders, Ambystoma Maculatum. Second Place: David Hammond, ETSU. Faculty Sponsor – Ranjan Chakraborty, Dept. of Health Sciences. The Detection of Possible Outer Membrane Receptor, 'FepA', Protein From Rhizobium Leguminosarum Strain IARI 312. Social & Behavioral Sciences (combined) First Place: Marie Fillers, ETSU. Faculty Sponsor – Nancy Scherer, Dept. of Communicative Disorders. Normative Speech Accuracy Measures of Children 24-30 Months of Age. Second Place: Zackary Cope, ETSU. Faculty Sponsor – Russ Brown, Dept. of Psychology. Amphetamine Sensitization in a Rodent Model of Psychosis. Division III – Graduate Students, 2+ Years First Place: Sam Boyd, ETSU. Faculty Sponsor – Darrell Moore, Dept. of Biological Sciences. Investigating Nectar Rhythms in Squash (Cucurbit Pepo): Effects on Honeybee (Apis Melliffera) Foraging Behavior. Second Place: Christina Bridges, ETSU. Faculty Sponsor – Antonio Rusinol, Dept. of Biochemistry and Molecular Biology. Comparison of the Activity and Expression of the ZMPSTE24 Endoprotease in Cellular Quiescence. Division IV – Medical Students First Place: Heather Wright, ETSU. Faculty Sponsor – Beth Bailey, Dept. of Family Medicine. Access to Obstetric Care in Rural Northeast Tennessee: Association with Birth Outcomes. 2009 Appalachian Student Research Forum Page 92 Second Place: Thomas Soike, ETSU. Faculty Sponsor – David Chi, Dept. of Internal Medicine. Effects of Cigarette Smoke Extract on Cytokine Production from IL-1BetaActivated Mast Cells. Division V – Medical Residents and Post-Doctoral Fellows First Place: Daniel Owens, ETSU. Faculty Sponsor – Cecilia McIntosh, Dept. of Biological Sciences. Identification, Recombinant Expression, And Biochemical Characterization Of A Flavonol 3-O-Glucosyltransferase From Citrus Paradisi. Second Place: Subhadra Nandakumar, ETSU. Faculty Sponsor – Uday Kumaraguru, Dept. of Microbiology. Post-Priming Manipulation of Discordant Antiviral T Cells. Third Place: Zia Ur Rahman, ETSU. Faculty Sponsor – Fereshteh Gerayli, Dept. of Family Medicine. What is the Best Treatment of Chronic Fecal Retention?. Oral Presentations Graduate Students First Place: Jeremy Stout, ETSU. Faculty Sponsor – Blaine Schubert, Dept. of Biological Sciences. Skeletal Morphology and Systematics of Late Pliocene Alligator SP. Second Place: Preston Visser, ETSU. Faculty Sponsor – Jameson Hirsch, Dept. of Psychology. Ethnic Differences in Risk and Protective Factors for Depression. Third Place: Stacie Woolard, ETSU. Faculty Sponsor – Uday Kumaraguru, Dept. of Microbiology. Up-Regulating Death Receptor 5 Via Tunicamycin Administration Increases Anti-HSV Immune Response. Medical Residents and Post-Doctoral Fellows First Place: Subhadra Nandakumar, ETSU. Faculty Sponsor – Uday Kumaraguru, Dept. of Microbiology. Molecular Link Between Toll Like Receptors, Obesity and Risk of Cancer. Second Place: Sumit Kalra, ETSU. Faculty Sponsor – Terry Forrest, Dept. of Internal Medicine. A Case of Ventricular Tachycardia Terminated by Carotid Massage. Third Place: Georges El Khoury, An Exceptional Variant of Single Coronary Artery Anomaly, Presenting with Acute St Segment Elevation Myocardial Infarction. 2009 Appalachian Student Research Forum Page 93
