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East Tennessee State University
presents the
2008 Appalachian
Student Research Forum
April 2 – 3, 2008
Millennium Centre • Johnson City, TN
Schedule of Events
April 2 – 3, 2008
The Centre at Millennium Park
Johnson City, TN
Wednesday, April 2, 2008
11:00 am – 2:30 pm
Poster Set-Up
Ballroom
3:00 – 5:30 pm
Poster Judging
Ballroom
Refreshments sponsored by:
The Ronald E. McNair Program and
The Center for Community Outreach and Family Services
Thursday, April 3, 2008
8:30 am – 9:00 am
Check-in and Continental Breakfast
9:00 am – 1:00 pm
Poster Viewing
Ballroom
9:00 am – 2:00 pm
Vendor Exhibition
2nd floor
10:30 am – 12:00 pm
Vendor Seminars
Room 230
Shimadzu:
“Fast LC with a Conventional HPLC System”
Promega:
“Maxwell(tm) 16: Automated gDNA, RNA, & HIS-Protein Purification”
Millipore Life Sciences: “Mammalian Cell Culture: Selection and Application of Microporous
Membrane Cell Culture Inserts”
Continental Breakfast and Mid-Morning Break sponsored by:
The Center for Community Outreach and Family Services
2008 Appalachian Student Research Forum Page 1 9:30 am – 12:00 pm
12:00 – 1:00 pm
Oral Presentations
Graduate Students
Residents and
Post-Doctoral Fellows
Room 130
Auditorium
1st floor
BUFFET LUNCH
Lunch sponsored by:
The Office of the Vice President for Health Affairs
The School of Graduate Studies
The Center for Nursing Research
The College of Public Health
1:00 – 2:00 pm
Keynote Address
Auditorium
“Proteinase Regulation in the
Ovarian Tumor Microenvironment”
M. Sharon Stack, Ph.D.
Professor and Vice-Chair for Research
Mulligan Endowed Professor of Cancer Research
Department of Pathology & Anatomical Sciences
University of Missouri, Columbia
2:15 – 3:00 pm
Awards Ceremony
Auditorium
3:00 – 4:30 pm
Poster Viewing
Ballroom
** Posters must be removed by 4:30 pm**
2008 Appalachian Student Research Forum Page 2 Keynote Presentation
“Proteinase Regulation in the Ovarian Tumor Microenvironment”
M. Sharon Stack, Ph.D.
Professor and Vice Chair for Research
Mulligan Endowed Professor of Cancer Research
Department of Pathology & Anatomical Sciences
University of Missouri, Columbia
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•
•
B.S. Clemson University, Biochemistry 1981
Fulbright Scholar, Universitat Bonn, Germany 1982
M.S. Biomedical Science/Biochemistry, East
Tennessee State University 1985
Ph.D. Biochemistry, University of Louisville1989
Post-Doctoral Research Associate, Duke University
1989-1991
Assistant Research Professor, Duke University 19911994
Assistant Professor, Associate Professor with Tenure,
Professor, Northwestern University 1994-2006
Professor and Vice-Chair for Research, University of
Missouri, 2007-present
•
•
•
•
•
ABSTRACT:
The ability to invade host tissues and metastasize is the major cause of cancer-related
death. During tumor invasion, metastasizing cells disrupt normal cell-cell and cell-matrix contacts and
acquire a migratory, invasive phenotype. Subsequent alterations in cellular architecture mediated by
modified extracellular matrix (ECM) attachments induce expression of proteinases that degrade ECM
proteins, facilitating migration through the modified tissue to establish metastatic foci. Epithelial
ovarian carcinoma is the leading cause of death from gynecologic malignancy, as 75% of women with
this disease succumb to complications resulting from disseminated intra-peritoneal metastasis. The
initiating events in the development of ovarian carcinoma are poorly understood. However, ovarian
carcinoma has an unique metastatic mechanism requiring reversible modulation of cell-cell and cellECM contacts that involves shedding of cells from the primary tumor followed by subsequent intraperitoneal adhesion, invasion and proliferation.
Unlike most highly metastatic tumors, the majority of women with advanced intraperitoneal disease
have no clinically apparent lymphatic or hematogenous metastases, implying that a novel mechanism for
metastasis is operative in ovarian cancer. Further, as metastases are largely confined to the peritoneal
cavity, microenvironmental factors that modulate intraperitoneal adhesion, motility and invasion play a
predominant role in ovarian pathobiology. Ongoing research utilizes an integrative approach involving
examination of 2-dimensional (2D) and 3D tissue culture systems complemented by murine tumor
models and analyses of human tumors. Understanding the molecular mechanisms by which tumor cells
orchestrate multiple microenvironmental cues to regulate the expression and activity of these
metastasis-associated proteinases is the major focus of the laboratory.
2008 Appalachian Student Research Forum Page 3 Forum Task Force
Dr. Scott Champney, Chair
Biochemistry & Molecular Biology
Dr. Russ Brown
Dr. Ranjan Chakraborty
Psychology
Health Sciences
Dr. Jay Franklin
Dr. Lee Glenn
Sociology & Anthropology
Professional Roles/Mental Health Nursing
Ms. Margaret Goergen
Dr. Gary Henson
Sponsored Programs
Physics, Astronomy & Geology
Dr. Gary Kukulka
Dr. Dhriendra Kumar
Family Medicine
Biological Sciences
Ms. Amy Leonard
Dr. Foster Levy
Fisher Scientific/Coordinating Vendor
Undergraduate Research/
Biological Sciences
Dr. Cecilia McIntosh
Dr. Louise Nuttle
Graduate Studies/Biological Sciences
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
Dr. Mick Whitelaw
Dr. Michael Woodruff
Physics, Astronomy & Geology
Vice Provost for Research
Ms. Jennifer Vanover
Biomedical Graduate Student/Microbiology
2008 Appalachian Student Research Forum Page 4 2008 Judges
Poster Presentations
Division I – Undergraduates
Division III – Grad Students – 2+ Yrs.
Biomedical Sciences
Biomedical Sciences, Natural Sciences &
Dhirendra Kumar
Mathematics, and Social & Behavioral
Edward Michieka Onyango
Sciences
Natural Sciences and Mathematics
Arpita Nandi
Terry Elisabeth Lancaster
Tricia Metts
Thomas Jones
Gary D. Henson
Division IV – Medical Students
Zulfiqar Ahmad
Yue Zou
Social and Behavioral Sciences
Dennis DeFoe
James Joseph Fox, III
Tiejian Wu
Division V – Residents and Post-Doc Fellows
Judith A. Hammond
Rob Wondergem
Michelle Crimi
Antonio Rusinol
Division II – Graduate Students 1-2 Yrs
Biomedical Sciences
Oral Presentations
Yu-Lin Jiang
Bert C. Lampson
Residents and Post-Doctoral Fellows
Natural Sciences and Math
Jay Mehta
Thomas Laughlin
Doris Hubbs
Padmanabhan Rengasamy
David Linville
Behavioral and Social Sciences
Andrea D. Clements
Graduate Students
Laraine Powers
Beth Bailey
Nathanial Fethke
Chris Newell
2008 Appalachian Student Research Forum Page 5 2008 Forum Sponsors
Philip D. Cooper Memorial Research Trust Fund, Inc.
East Tennessee State University
The Office of Research & Sponsored Programs
The School of Graduate Studies
The Center for Appalachian Studies and Services
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 Computer & Information Services
Department of English
Department of Mathematics
Department of Physics, Astronomy & Geology
The College of Business & Technology
Office of the Dean
The College of Clinical & Rehabilitative Health Sciences
Office of the Dean
Department of Physical Therapy
The Claudius G. Clemmer College of Education
Office of the Dean
Department of Human Development and Learning
2008 Appalachian Student Research Forum Page 6 The James H. Quillen College of Medicine
Office of the Dean
Department of Anatomy & Cell Biology
Department of Biochemistry & Molecular Biology
Department of Obstetrics/Gynecology
Department of Pathology
Department of Pediatrics
Department of Pharmacology
Department of Physiology
Department of Psychiatry & Behavioral Sciences
Department of Surgery
The College of Nursing
Office of the Dean
Center for Nursing Research
The College of Public Health
Office of the Dean
Special Thanks
The 2008 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.
2008 Appalachian Student Research Forum Page 7 Exhibitors
2008 Appalachian Student Research Forum Page 8 Poster Presentations
Division I – Undergraduates
Biomedical Sciences
OXIDATIVE DAMAGE TO THE SUGAR-PHOSPHATE BACKBONE OF DNA CAUSED BY
CARBONATE RADICAL ANIONS
Jeremy Brooks*, Lauren Brooks*, Laura Catron*, Amanda Kelly*, Lindsey McAmis*,
Joe Powell*, Danna Sharp* and Yuriy Razskazovskiy†
*
The Talent Expansion in Quantitative Biology (TEQB) Program, ETSU
†
Dept. of Physics, Astronomy and Geology, ETSU
The carbonate radical anion CO3•- is a biologically relevant oxidizing intermediate generated in seawater
by sunlight and in cells through CO2-catalyzed decomposition of peroxynitrite anions ONOO-. This
species has been shown to induce oxidative damage to guanine bases in DNA that makes it a source of
potentially mutagenic DNA lesions. The focus of this study is the reactivity of CO3•- towards the sugarphosphate backbone of DNA, including its ability of inducing immediate and latent strand breaks through
hydrogen abstraction from the deoxyribose moiety. For the purposes of this project CO3•- was generated
by UV (λ>300 nm) photolysis of inorganic complex [Co(NH3)5CO3]NO3 in phosphate-buffered solution
of calf thymus DNA, and DNA damage products were analyzed by HPLC. Product analysis showed the
formation of heat-labile DNA lesions that released a characteristic low molecular weight product 5methylenefuran-2-one upon spermine-catalyzed thermal decomposition in phosphate-buffered solutions
(pH 6.9). Based on this characteristic behavior we conclude that the carbonate radicals oxidize the sugarphosphate backbone primarily at the C1`-position of the deoxyribose moiety that leads to the formation of
so-called oxidized abasic sites in the DNA structure. Quantitative estimates based on HPLC
measurements show that about 8% of all carbonate radicals generated in the system react through this
pathway.
A COMPARITIVE STUDY OF THE HANDS OF LEMURS
AND OTHER SUSPENSORY SPECIES.
R.A. Deliz
Dept. of Sociology & Anthropology, East Tennessee State University, Johnson City, TN
The hands of prosimian primates play a major role in locomotor activities. The structure of the hands of
extant sifakas exhibits characteristics indicative of leaping and clinging. Their grasping efficacy pales in
comparison to the recently discovered extinct species, whose curved phalanges are convergent with slothlike suspensory positional behaviors or grasping styles of gibbons and siamangs. I have examined
previous studies of the recently extinct species of the subfossil lemurs Paleopropithecus and
Mesopropithecus and explored the morphological heterogeneity between these and modern sifakas, indris,
sloths, gibbons, and siamangs. In addition, I have examined modern sloths and anteaters for comparative
purposes. In each species I measured phalangeal curvature and length and identified skeletal markers of
2008 Appalachian Student Research Forum Page 9 locomotor adaptations, in comparison with the subfossils. Utilizing this data I have drawn inferences
about the relationship between skeletal form and locomotor function in modern lemurs as well as
primates and other suspensory species.
ODD BEAR OUT?
Milan Ganger and Terry Lancaster
Dept. of Health Sciences, East Tennessee State University, Johnson City, TN
Polar Bears are unique bears because they are semi-aquatic. Polar Bears tend to swim with their backs
and neck out of the water. They also have webbed feet for swimming and this layer of waterproof fur over
their black colored skin. This fur is so unique that humans used to hunt polar bears just for their fur to
keep warm in the arctic. Not only do we look at the outer characteristics of the Polar Bear, but the inside
as well. My question is due to their irregular way of swimming, do Polar Bears have some different
distinctiveness along their spine? I will be looking at the transverse and spinous processes to determine if
a Polar Bear is semi-aquatic from a given vertebrae. I will compare the vertebrae of the Polar Bear to
other similar semi-aquatic species (i.e.: Northern Elephant Seal, Hooded Seal, South American Sea Lion,
and the Australian Sea Lion) and two terrestrial bear species (i.e.: American Brown Bear and Black Bear)
as well. Using sliding and spreading calipers, I will measure the length, width, and depth of the transverse
processes and spinous processes of the cervical, thoracic, and lumbar vertebrae. I will compare all
measurements of the Polar Bear with the Northern Elephant Seal, Hooded Seal, South American Sea
Lion, Australian Sea Lion, the American Brown Bear, and the Black Bear. Along with these
measurements, I will measure the length of each region of vertebrae (i.e. cervical, thoracic, and lumbar)
and the curvature of the neck. With these measurements, I will be able to identify some distinct
characteristics that will categorize the Polar Bear as either semi-aquatic or terrestrial by using Principal
Component Analysis (PCA) to identify the semi-aquatic grouping and the terrestrial grouping of
mammals and then attempt to “place” the Polar Bear in one of these groups by its morphology.
SYNTHESIS AND EVALUATION OF GAMMA TOCOPHEROL LYSINATE AS AN ANTI-CANCER
AGENT IN HUMAN PROSTATE AND COLON CANCER CELL LINES
Bassam Lamale, Yu-Lin Jiang and Sharon Campbell
Dept. of Chemistry, East Tennessee State University, Johnson City, TN
Vitamin E is a fat-soluble vitamin represented by eight different forms classified as either tocopherols or
tocotrienols which contain antioxidant properties. Alpha tocopherol is the most bioavailable among the
isoforms in humans and protects cell membranes from oxidation by reacting with lipid radicals produced
in the lipid peroxidation chain reaction. Mediterranean cultures have a lower risk for cancers and
cardiovascular disease due to diets that are rich in vitamin E. Campbell et al. showed that RRR-γtocopherol showed more antiproliferative activity on HCT-116 colon cancer cell lines compared with
RRR-α-tocopherol. Vitamin E isoforms are insoluble in aqueous solution. An inability to absorb a
sufficient amount of fat will result in vitamin E deficiency making the need for a water soluble vitamin E
vital. Altering the chemical structure of RRR-γ-tocopherol (GT) with the addition of polar atoms may
increase the solubility and has been shown to increase the antiproliferative activity in MCF-7 breast
cancer cells. Further, redox silent analogues of Vitamin E that contain polar molecules added to the
hydroxyl antioxidant site by ester or ether link have demonstrated an increase in antiproliferative activity
over the Vitamin E from which it was derived. We began to look for biologically relevant polar
molecules that could be added to the vitamin E and hypothesized that the addition of an amino acid to the
2008 Appalachian Student Research Forum Page 10 antioxidant site may enhance the antiproliferative activity of GT. We selected the amino acid lysine
owing to its three polar groups. The organic synthesis of RRR-γ-tocopherol lysinate (GTL) was carried
with the esterification of GT with protected lysine using dimethylaminopyridine as a catalyst and
dicyclohexylcarbodiimide as a reagent in the first step. The resultant product was then deprotected with
hydrogen chloride in ethyl acetate to the water soluble lysinate of GT. The structure of the product was
validated using NMR. GTL was assayed for its ability to effect cell proliferation in the human cancer cell
lines HCT-116 (colon) and PC-3 (prostate) using the MTT assay. The MTT assay shows that GTL has a
higher percent growth inhibition compared with GT at physiological and pharmacological concentrations
in both cell lines. Western blot analysis for the proliferation and apoptosis proteins PPAR γ, cyclin D1
and Poly (ADP-ribose) polymerase (PARP) was performed. Consistently in both cell lines, western blot
analysis shows that PPARγ is upregulated, cyclin D1 is down regulated and PARP is cleaved to a greater
extent with GTL than GT. These data demonstrate that the addition of the lysine group by an ester
linkage at the antioxidant hydroxyl site improves the ability of the molecule to inhibit proliferation in two
different cancer cell types. Further the genes that are regulated by this molecule are similar to those
previously determined to be important in the GT-mediated cell death and apoptosis. The GTL reaction is
faster than that of GT. Further work needs to be done to determine what makes the kinetics of the
reaction faster with GTL when compared to GT.
EXPRESSION OF HUMAN NEUTROPHIL ELASTASE
USING SUMO FUSION TECHNOLOGY
Jessica Pugh and David Johnson
Dept. of Biochemistry and Molecular Biology, James H. Quillen College of Medicine, East Tennessee
State University, Johnson City, TN
Human polymorphonuclear leukocytes (neutrophils), which play a critical role in innate immunity, have
three active serine proteases (Elastase, Cathepsin G and Protease 3) in their azurophilic granules that aid
in bacterial killing.
Human neutrophil elastase (HNE) has been linked to several diseases including
pulmonary emphysema via degradation of lung elastin, and mutations in the HNE gene (Ela2) result in
low neutrophil counts with susceptibility to bacterial infections. Recombinant HNE has never been
produced and the enzyme isolated from neutrophils is missing 20 amino acids from its C-terminus. A
method of expressing recombinant HNE would allow study of this unusual C-terminal processing and
make site directed mutagenesis studies possible. SUMO (small ubiquitin-related modifier protein)
fusions are known to improve recombinant protein solubility in E. coli. The SUMO fusion vector
pETHSUL, which is also a ligation independent cloning vector (LIC), was obtained from Dr. Patrick Loll
(Drexel University, COM). An image clone of HNE was obtained from American Type Culture
Collection. The coding region of the HNE gene was amplified in a PCR reaction using Stratagene’s
PfuUltra II Fusion HS DNA Polymerase using primers designed to generate LIC compatible products.
The pETHSUL vector was cut with BseRI and then treated with TSAP thermosensitive alkaline
phosphatase to remove the 5’ phosphates. The vector was then gel purified, and treated with T4 DNA
polymerase in the presence of 1 mM dCTP to add a string of Cs. The HNE PCR product was made LIC
ready by removing the 5’ phosphates, followed by adding a string of Gs using T4 DNA polymerase and 1
mM dGTP. A mixture of the LIC ready HNE PCR product and LIC ready SUMO fusion vector
pETHSUL was added to BIOBlue Chemically Competent cells. The vector with its tightly bound HNE
PCR product was taken up by the cells as if they had been ligated together. Transformed cells were
screened on plates with ampicillin. The LIC process is very efficient because all eight colonies screened
by PCR were all positive for the HNE insert and the sequence was correct and in frame from sequencing
data. Rosetta-gami cells will be transformed with pETHSUL-HNE and pSUPER, which contains the
SUMO protease gene which will be used to cut the SUMO off of the SUMO-HNE fusion protein. The
Studier Method for Autoinduction of Protein Expression in E. coli using the T7 promoter will then be
2008 Appalachian Student Research Forum Page 11 used to express SUMO-HNE and the SUMO protease. Stable expression of HNE will be beneficial for
further studies concerning the functions and diseases associated with this protease. Supported by
National Heart, Lung and Blood Institute grant 1R15HL091770.
A COMPARATIVE ANALYSIS OF LONG BONE FORMULAE
TO ESTIMATE LIVING STATURE (HEIGHT)
Michelle Wood, and Terry E. Lancaster
Depts. of Criminal Justice and Health Sciences, East Tennessee State University, Johnson City, TN
Stature estimation from long bones has been a primary focus of anthropological studies since the latter
half of the 1800's. Most stature estimations are done using only a single long bone, such as the humerus,
radius, ulna, femur, tibia, or fibula. The humerus, femur and tibia are the most widely accepted long
bones used for these predictions, and they are rarely used combined in estimation formulae. In this study
we developed our own series of formulae to predict the stature of an individual from the long bones of the
skeleton. Using linear regression models we constructed a series of prediction equations based on length
and midshaft diameter measurements of the humerus, radius, ulna, femur, and tibia using these bones
singly and in combination to produce the best estimator of stature. We then divided our sample by sex and
then by race and recalculated our prediction equations to isolate and identify the effect of these on stature.
Natural Sciences & Mathematics
CONSTRUCTION AND FUNCTIONAL ANALYSIS OF CHIMERIC PROTEINS
FROM PUTATIVE FLAVONOID GLUCOSYLTRANSFERASE CLONES
FROM GRAPEFRUIT
Josephat Asiago, Daniel K. Owens and Cecilia A. McIntosh.
Dept. of Biological Sciences, East Tennessee State University; Johnson City, TN
Flavonoids are one of the major groups of plant natural products and many exist in glucosylated form.
They are known for coloration, flavor, plant-plant interaction, plant-microbe interactions, plant-insect
interaction, ultra violet protection and pollen maturation. Plant UDP- glucosyltransferases (UGT’s) are
enzymes that transfer a glucose residue from the activated donor to an acceptor molecule. The transfer of
glucose to the acceptor molecule helps to stabilize, detoxify and solubilize metabolites. Grapefruit is
known for high levels of accumulation of flavanone 7-O-glycosides as well as flavone-7-O-glycosides,
flavonol 7-O and 3-O glycosides. The majority of flavonoid GT’s characterized to date belong to the 3-OGT group (act on flavonols and anthocyanins). There are at least five different flavonoid
glucosyltranferases that have been identified in grapefruit: flavonol 3-O-GT, flavonol 7-O-GT, chalcone
GT, flavone 7-O-GT and flavanone 7-O-GT. This makes grapefruit a good model to look for different
flavonoid GT’s for structure and functional analysis. Eight putative glucosyltranferases have been
identified in our lab. PGT 7 has shown activity with quercetin and characterization is in progress. PGT4
has not shown any activity with flavonoid acceptors. This may be attributed to the low level of soluble
protein or to the fact that this enzyme acts on another group of secondary metabolites. The amount of
soluble protein in PGT4 is currently being optimized by using less rich medium during induction and
collection of samples at different time points. French press instead of sonication will be used to burst
open the cells in an effort to protect the protein from denaturation. The PGT4 protein from the burst cells
will be purified using the HIS TAG-metal affinity column before re-testing for activity. Construction of
2008 Appalachian Student Research Forum Page 12 chimeras will provide important information on the active moieties and their location in PGT 7 and PGT4
and the activity of chimera will be compared to that of the parent proteins. Chimeric protein constructs
were obtained from PGT7 and PGT4 as follows: PGT 4 was digested with restriction enzymes (Sac I
and Xho I). These enzymes are internal to PGT4 and do not cut the vector. The C terminus of PGT7 was
amplified using clone specific primers (CSP 58F and CSP58R) designed to incorporate Sac I and Xho I in
the sense (CSP 58F) and the antisense (CSP 58R) primers respectively. Products were digested with Sac I
and Xho I. The N-terminus section of PGT4 and the C-terminus section of PGT7 were gel purified. These
pieces were ligated and cloned into expression vector. The full structure of chimeric construct will be
confirmed by sequencing the chimeric clone. Once the primary structure is confirmed, the chimeric clone
in expression vector will be transformed into BL21 (DE3), RIL E. coli cells and induced for protein
expression. The expressed protein will be tested for flavonoid GT activity and compared to the parent
proteins.
BIOCHEMICAL IDENTIFICATION AND CHARACTERIZATION OF SABP2-INTERACTING
PROTEINS BY AFFINITY CHROMATOGRAPHY
Raj C. Bant, Kajal C. Patel and Dhirendra Kumar
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Salicylic acid is required by plants to fight pathogen infection and is critical for inducing both local and
long lasting systemic acquired resistance (SAR). SABP2, a 29 kDa plant protein, was identified in a
screen to discover proteins which bind to salicylic acid (SA). In plants, SABP2 catalyzes the conversion
of methyl salicylate (MeSA) into SA, which then activates defense proteins to mediate a resistance
response. Recently, MeSA was identified as the long distance phloem mobile signal required for systemic
acquired resistance (SAR). SABP2 receives this SAR signal in systemic tissue and converts it into SA.
RNAi mediated silencing of SABP2 expression makes the transgenic tobacco plants more susceptible to
TMV. Given the important role of SABP2 in mediating plant resistance it is important to identify cellular
proteins which may interact and regulate its activity. Therefore, we are conducting a screen to identify
and characterize its interacting partners in the cell. As a first step, high amounts of recombinant SABP2
was expressed in E. coli. This recombinant SABP2 was then bound to Ni-NTA agarose column and
utilized as an affinity matrix to bind and purify SABP2 interacting proteins (SBP2IP). Bound proteins
will be analyzed by SDS-PAGE followed by silver staining. Alternatively native SABP2 and its
interacting proteins from tobacco plants could be purified using a affinity column containing anti-SABP2
antibodies linked to protein A/G agarose matrix.
MOLECULAR MECHANISM OF ATP SYNTHASE- ROLE OF CHARGE
IN THE CATALYTIC SITES
Laura E. Brudecki and Zulfiqar Ahmad
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
F1Fo-ATP synthase is the smallest known biological nano motor, found from bacteria to man. It is the
enzyme responsible for ATP synthesis by oxidative or photophosphorylation in membranes of bacteria,
mitochondria, and chloroplasts. ATP synthase is the fundamental means of energy production in animals,
plants, and almost all microorganisms. Synthesis and hydrolysis both are carried out at the catalytic sites
of F1Fo ATP synthase. X-ray structures show the presence of αPhe-291 residue in the phosphate binding
subdomain, in close proximity to the known Pi binding residue, βArg-246. In order to understand the role
of charges in phosphate binding, we mutated αF291 to αF291D and αF291E. The choice of this residue
2008 Appalachian Student Research Forum Page 13 was based on its close proximity to the phosphate analogs AlF3 and SO42- , which are bound in catalytic
sites of x-ray structures of bovine ATP synthase. Using Pi protection from NBD-Cl (7-chloro-4nitrobenzo-2-oxa-1, 3-diazole) inhibition of ATPase in membranes as the assay, we found that these
mutations reduce the ATPase activity while retaining oxidative phosphorylation significantly. This
mutation also led to the abrogation of Pi binding as seen earlier in case of βN243D mutation. We also
found that in contrast to wild-type, ATPase of mutants was not inhibited by MgADP-fluoroaluminate,
MgADP-fluoroscandium, or MgADP-azide.
INVESTIGATING CLIMATE CHANGE IN THE MIDCONTINENT AND SOUTHEASTERN
APPALACHIAN: SPELEOTHEM RECORDS FROM CAVES IN MISSOURI AND TENNESSEE
Jessica Buckles, Yongli Gao, William Hill
Dept. of Physics, Astronomy, and Geology, East Tennessee State University, Johnson City, TN
Two speleothem samples were collected from Power Mill Creek Cave (PMCC), Missouri, for uraniumseries dating at the University of Minnesota and stable isotopic analyses at the University of Kentucky.
The ages of the speleothem samples range between 53,170 and 660 years old. We obtained carbon and
oxygen isotope data on one speleothem for a time period between 53 to 35 ka and reconstructed the
regional climatic and vegetation changes. In general, climatic oscillations and vegetation changes in this
region are consistent with the earlier study from speleothem samples of Crevice Cave, Missouri (Dorale et
al., 1998). The midcontinental climate cooled and forest replaced grassland between 55 and 41 ka. Then,
the temperature suddenly increased at 37 ka. Preliminary analysis of oxygen isotope oscillation shows
that this warming around 37 ka was even more dramatic than that reported in the previous investigation at
Crevice Cave. Recently, we started to process speleothems collected in eastern Tennessee. These
speleothems were dated at different ages. No extended record of continental climate change exists for
eastern North America going back through the past 11 marine isotope stages. The primary objective of
this research project is to assemble a calibrated, highly-resolved composite record of decadal- to
centennial-scale climate change from the mid-latitudes of eastern North America that extends across the
last four interglacial/glacial cycles.
ISOLATION OF THE FULL-LENGTH CLONES OF GRAPEFRUIT 181-1D AND CP1-D10,
PUTATIVE FLAVONOID GLUCOSYLTRANSFERASES
Patricia Campbell, Daniel Owens, and Cecilia McIntosh
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
This research centers within the topic of secondary metabolites produced in the Plant Kingdom, in
particular flavonoids in Citrus paradisi. Flavonoid compounds are found naturally in many plants for
such main purposes as aiding in reproduction and defense. This is accomplished through flavonoids being
responsible for some of a plant's unique odor, taste, and color characteristics as well as having a role in
pollen recognition. Flavonoids also protect leaves from UV light damage, fungal and microbial
pathogens, and act as antifeedants. Flavonoids have been shown to possess antioxidant and possible
anticarcinogenic characteristics in mammals. Our lab is focused specifically on the flavonoid
glucosyltransferases. These are the enzymes whose job as a catalyst is to aid in the transfer of
monosaccarides(sugars) to a receiving molecule, in this case, flavonoids. Glucosyltransferases are key
enzymes in the synthesis of the flavnonoids actually found in plant tissues. There are many putative
sequences for glucosyltransferases in databases, but unfortunately there is little functional characterization
found in the literature. My focus of interest lies within two grapefruit putative GT clones (CP1-D10 and
2008 Appalachian Student Research Forum Page 14 181-1D) that have been found to only differ in a small repeat area so far. This research is designed to test
the hypothesis that these are nearly identical clones with the major difference being confined to the
currently identified repeat area, and try to test their functional similarities. This is being done through a
series of experiments. First, clone specific primers were used in PCR to pull out the desired DNA
fragment. Once amplified, the PCR product was then cloned into a Topo holding vector and transformed
into E.coli competent cells which were then plated for colony growth on LB media plates. The colonies
were tested, such as through PCR screening procedures, to confirm that they contain an insert of the
expected length. DNA sequencing is being performed to determine which clone(s) has been obtained.
Sequences will be analyzed to further assess the degree of identity of the clones. If results confirm the
presence of two distinct putative glucosyltransferase clones with high identity, the clones will be migrated
into a protein expression vector. If time permits, experiments will be conducted to optimize conditions for
inducing protein production followed by assays in the detection of their activity for the structural and
functional characterization to be carried out.
SYSTEMATIC STUDY OF THE LATE NEOGENE NUTS OF CARYA (JUGLANDACEAE) FROM
NORTHEAST TENNESSEE
Graham Cooke, Yusheng Liu
Dept. of Biological Sciences, East Tennessee State University, Johnson City TN
The discovery of the Gray Fossil site, located in Gray Tennessee in 2000 lead to a series of discoveries
lending additional information to the study of the North American-East Asia disjunct, this has laid the
groundwork for comprehensive study of the paleo-climate at the time of the creation of the deposits in the
late Miocene. The site has yielded an unusual amount of fossilized animalia and plantae, notably the
presence of Pristinailurus bristoli, Tapirus polkensis, in addition the presence of Fruit, and or Pollen from
families Juglandaceae, Fagaceae, Pinaceae amongst many other. The fossilized animal remains found in
the deposit indicate the temperate subtropical environment currently found in the locality is far different
to what was present in the late Miocene. Using a combination of techniques we hope to provide
additional information to the reconstruction of the paleo environment, and utilize the currently recognized
phylogeny of Juglandaceae to validate the characteristics used to identify the species of Juglandaceae
present, and in turn their following evolution.
PATTERN OF CALCIUM MOBILIZATION IN A VIVIPAROUS
AUSTRALIAN SKINK, PSEUDEMOIA PAGENSTECHERI
Santiago P. Fregoso, James R. Stewart, and Tom W. Ecay,
Depts. of Biological Sciences and Physiology, College of Arts and Sciences, East Tennessee State
University, Johnson City, TN
In order to obtain calcium during embryonic development, oviparous lizards take advantage of their yolk
and eggshell reserves. However, they do not exploit the eggshell until the last few stages of development.
Viviparous lizards, on the other hand, do not possess an eggshell from which to extract calcium and have
a proportionally smaller yolk. Many species rely heavily on placental provisioning of nutrients during
embryogenesis. Previous studies have shown that highly specialized placental lizards receive most of their
calcium from the placenta. This study aims to expand on that information by ascertaining the mobilization
pattern of calcium provisioning across the placenta during development of a placental scincid lizard,
Pseudemoia pagenstecheri. Samples of yolk and embryos were removed from females at specific
intervals during development and analyzed for calcium using atomic absorption spectrometry. The timing
2008 Appalachian Student Research Forum Page 15 of calcium mobilization to embryos from placental transfer was similar to calcium recovery from the
eggshell of oviparous species. However, oviductal calcium secretion occurs much later in development in
this viviparous species. This difference suggests a timing shift from the ancestral mode and a possible
nutritional advantage to placentotrophy.
EMBRYONIC CALCIUM MOBILIZATION IN AN OVIPAROUS LIZARD, SAPROSCINCUS
MUSTELINUS
Courtney Garland, James R. Stewart, and Tom W. Ecay
Depts. of Biological Sciences and Physiology, East Tennessee State University, Johnson City, TN
Oviparous lizards provide calcium to their embryos through calcium-rich yolk and calcium deposited into
the eggshell. Oviparous skinks receive between 30-50% of their embryonic calcium from the eggshell
with the remainder coming from the yolk. Yolk calcium is deposited during vitellogenesis, while calcium
in the eggshell is secreted during the shelling process in the oviduct. We studied the oviparous scincid
lizard Saproscincus mustelinus. We used atomic absorption spectroscopy to estimate the quantity of
calcium in yolk, eggshell and the embryo at specific time points during incubation. We found that yolk
calcium is mobilized gradually throughout the early stages of embryonic development and uptake begins
to increase during the later stages as embryonic growth accelerates. Calcium from the eggshell is also
mobilized during the later stages while embryonic growth rate is high. We found that Saproscincus
mustelinus receives 54% of its calcium from the eggshell, showing the heavy reliance of the developing
embryo on the eggshell as well as the yolk. The percentage of recovery from the eggshell is higher than
most other oviparous lizards. Therefore, we hypothesize that Saproscincus mustelinus has a more
specialized mechanism for retrieving calcium from the eggshell than most oviparous lizards.
IMROVEMENT OF THE MOTOR FUNCTION OF ESCHERICHIA COLI ATP SYNTHASE BY
SELECTION PRESSURE
Johnny J Grindstaff, Prasanna K Dadi and Zulfiqar Ahmad,
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
ATP synthase is responsible for ATP synthesis by oxidative or photophosphorylation in membranes of
bacteria, mitochondria, and chloroplasts. It is the fundamental means of energy production in animals,
plants, and almost all microorganisms. Escherichia coli ATP synthase in its simplest form contains eight
subunits namely α3ß3γδεab2c10-14. The α/β interface has a total of six sites, three of which are catalytic and
three non catalytic. Synthesis and hydrolysis both are carried out at the catalytic sites which are mainly
contributed by the β subunits in the α/β interface of F1 sector. In earlier studies, our lab has shown that
βR246 is involved in phosphate binding and mutation of βR246A leads to the abrogation of Pi binding.
Phosphate binding is an essential part in the formation of ATP from ADP+Pi. A powerful way to
understand the molecular mechanism of ATP synthase and how ATP is synthesized is the knowledge of
the suppressors (genetic revertants) for mutants which interfere with the Pi binding. The most suitable
mutation appeared to be βR246A. The growth yield of βR246A was tested on minimal glucose and on
non fermentable carbon source succinate. Testing on thousands of succinate plates along with wild-type
as positive control and pUC118 as negative control gave us 23 putative candidates. After further analysis
of the 23 candidates on limiting glucose and 20mM succinate media, we were able to identify six clones
which appeared to be the genetic revertants of β R246A. The 4th colony named GR4 showed maximal
growth on succinate plates which grew better than wild-type, suggesting that the motor function might
2008 Appalachian Student Research Forum Page 16 have improved. This improvement was further confirmed by the growth on minimal succinate media.
Currently, we are in the process of sequencing the plasmids obtained from those six Genetic Revertants.
THE SIERPIŃSKI SQUARE GRAPH AND SOME OF ITS PROPERTIES
Elizabeth Harris, Joshua Berry, Dr. Anant Godbole
Dept. of Mathematics, East Tennessee State University, Johnson City, TN
We generalize the well-studied Sierpiński Triangle and define the Sierpiński Square, a structure obtained
by forming, iteratively, progressive Sierpiński Triangles and an inner square. SQ1 is the unit square, and
SQn, n>1, is obtained from the first n iteration of the above process. Most researchers in graph theory
study a particular property for several families of graphs. Here we take the other, less common, approach,
namely to study a large set of properties for one family of graphs, namely SQn. Properties studied include
the order, size, chromatic number, total edge length, diameter, domination number and Hamiltonicity of
SQn.
SA-METHYL TRANSFERASE INVOLVEMENT
IN DISEASE RESISTANCE SIGNALING
Tazley Hotz and Dhirendra Kumar,
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Salicylic acid (SA) has been shown to play an important role in plant defense response against microbial
pathogens. SA produced is lipid immobile and cannot pass through the chloroplast membrane to enter
into the cytoplasm to mediate the disease resistance signaling. It has been 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 resistant tobacco’s response to tobacco mosaic virus
(TMV). 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 RT-PCR. To investigate the role of SAMT in plant immunity, we are
in the process of making transgenic tobacco by either over expressing SAMT or by knocking out its
expression using RNAi. These transgenic lines will be infected with viral and bacterial pathogens to
determine the role of SAMT in plant defense. Understanding the biochemical pathways involved in plant
defense response against pathogens will help to develop crop plants showing better resistance to everevolving pathogens.
THE EFFECTS OF GALAXY INTERACTIONS
ON STAR FORMATION AND MORPHOLOGY
Sabrina Hurlock, Beverly J. Smith, and Mark Hancock,
Dept. of Physics, Astronomy, and Geology, East Tennessee State University, Johnson City, TN
We have analyzed SARA (Southeastern Association for Research in Astronomy) optical images of
several interacting galaxy systems selected from the Arp Atlas, and compared with GALEX (Galaxy
Evolution Explorer) UV images, Spitzer infrared (3.6 µm and 8.0 µm) images, and Sloan Digital Sky
Survey images. Using SARA, we have observed the galaxies in visible wavelengths including B, V, and
2008 Appalachian Student Research Forum Page 17 R broadband filters and narrowband redshifted H-alpha filters. We used the Image Reduction and
Analysis Facility (IRAF) program to remove the bias and dark current from the images, flat-field the data,
and to align and combine the images. In order to obtain a pure H-alpha image, we first subtracted out the
continuum, or starlight, using the R band image. These continuum subtractions allow us to view areas of
intense star formation that we would otherwise not be able to distinguish due to the light from the older
stars. With this multi-wavelength data set, we are able to determine the effects that interaction has on the
structure of the galaxy including the formation of bridges and tails, and where new star formation is
occurring.
MODELING MICROPARASITIC DISEASES
McKayla Johnson, Tashauna Gilliam, and Istvan Karsai
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Worldwide about 1,415 human pathogens are known, of which over half are microparasites. An infection
begins when a virus or bacterium enters the host and starts to replicate. If the immune system fails, the
host becomes sick and will infect other individuals with the pathogen. Three main properties that control
the transmission and life cycle of the virus are infectiousness, duration of the disease, and the chance of
recovery of the host. Each parameter has the ability to lead to a continuation or an extermination of the
disease within the population. The interplay of the parameters will predict how the disease may affect
populations, and knowing this, we can plan preventive steps that hinder and stop the infection in
populations. Infectiousness, duration, and chance of recovery are used as main parameters in this study.
Each one has an effect on the viability dynamics of the host and the microparasite. The infectiousness
concerns the ease with which a microparasite can infect its host. The chance of recovery concerns how
strong the virulence is in relation to the ability of the host in recovery and the development of immunity
concerning the same strain of disease in the future. The duration of the disease determines how long a
person with the disease may infect others before death or recovery. The three of these parameters
commonly evolve to produce an equilibrium-like situation. If a disease is too aggressive, with little to no
chance of recovery and has a high infectious rate, the hosts will die out, thus eliminating the virus. On the
other hand, if a disease has a long duration and moderate infectiousness, the possibility of the disease
remaining in the population without exterminating itself is higher. The interactions of these three
important parameters were studied via a technique called parameter sweep in the model called Virus
(Wilensky (1998) in the NetLogo 3.1.3 software. In this agent based model, the individuals move
randomly throughout the environment, allowing random contact among the individuals. Our results show
that the more infectious a disease becomes, the more likely it is to extinguish itself. As the disease
becomes more infectious and the duration lengthens, immunity will reach its maximum and then decrease
for each following duration. Regardless of infectiousness with a chance of recovery at 50%, a disease
causes a minimum in healthy individuals if it has the duration of 50 weeks. Results such as these
conclude that several parameters can be combined to model the viability of a disease within a population,
predicting its toll on the population as well as on its own viability.
2008 Appalachian Student Research Forum Page 18 ALLENE-INOC REACTION
Rachel Keith, Dr. David Young
Dept. of Chemistry, East Tennessee State University, Johnson City, TN
The goal of the set of experiments is to synthesize an allene to be used in an intramolecular nitrile oxide
cycloaddition, abbreviated INOC. The reaction has already been published using simple alkenes,
however, we propose to perform the experiment with allenes. Progress on our route to an allene to be
used in this reaction will be reported.
DEVELOPMENTAL EXPRESSION OF CARBONIC ANYHDRASE II
IN THE EXTRAEMBRYONIC MEMBRANES OF THE CORN SNAKE,
PANTHEROPHIS GUTTATUS
Maleka Khambaty1, James R Stewart2, Tom W Ecay1.
Dept. of Physiology, East Tennessee State University, Johnson City, TN
2
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
1
The eggshell provides 28% of the calcium in a hatchling corn snake, with the remainder coming from
yolk. The chorioallantois contributes to calcium homeostasis during embryogenesis by mediating the
transport of shell calcium to the embryo. Our objective is to understand how the embryo regulates
dissolution of mineralized shell calcium and its transport during development. We hypothesize that cells
of the chorioallantois secrete acid onto the eggshell thereby dissolving calcium carbonate crystals. Acid
production is catalyzed by carbonic anhydrase II (CA II). Immunoblotting demonstrates an increase in
CAII expression throughout development in the chorioallantois but no detectable expression in the yolk
sack. Increased CAII expression is coincident with increased calbindin-D28K expression, a marker for
calcium transport. Histochemical staining identifies CA positive cells at the outer surface of the
chorioallantois adjacent to the eggshell. CAII and calbindin-D28K immunohistochemisty are being used
to determine if acid secretion and calcium transport occurs in the same or different cells of the
chorioallantois. (Supported by the NSF and an APS Undergraduate Summer Research Fellowship.)
SYNTHESIS AND CHARACTERIZATION OF DNA BASE RECEPTORS
Bassam Lamale1, Lee M. Daniels2, and Yu Lin Jiang1
1
Dept. of Chemistry, East Tennessee State University, Johnson City, TN
2
Rigaku Americas Corporation, 9009 New Trails Drives, The Woodlands, TX
Two DNA base binding receptors 1 and 2 have been synthesized by three-step reactions, including Suzuki
arylation of dihaloarenes, followed by metallization with n-butyl lithium and carboxylation with carbon
dioxide. Crystals of the receptors were obtained from slow cooling of hot solution of the compounds in
acetonitrile. The crystal structures of the compounds have been determined using X-ray crystallography.
Both receptors form dimers through binding of one carboxylic group to the other. The structure of 1
shows that carboxylic group is in the same plane as the phenyl group; however, the pyrenyl group is
perpendicular to the phenyl group. The structure of 2 shows that both carboxylic and pyrenyl groups are
parallel to each other but perpendicular to the naphthyl group. The molecular recognition of DNA base 9butyl adenine has also been carried out. The binding of 9-butyladenine by 1 results in the decrease of
2008 Appalachian Student Research Forum Page 19 pyrene fluorescence intensity, but the binding of the 2 by 9-butyladenine results in increase of pyrene
fluorescence.
O
OH
OH
O
1
2
ENVIRONMENTAL FACTORS CONTRIBUTING TO COLONY SUCCESS
IN A SOCIAL SPIDER SYSTEM
Brent J. Linville and Thomas C. Jones
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Population density is affected by an organism’s survival and reproductive success in a particular area,
which in turn are affected by environmental factors. Population densities of the northern social spider,
Anelosimus studiosus, vary significantly among seemingly suitable habitats within its range in east
Tennessee. For example, the spider occurs in high densities at Bays Mountain Lake, but not at all at
Watauga Lake. We hypothesize that this variance in density can be attributed to several environmental
factors including prey density, temperature, and relative humidity. To test this, we surveyed populations
at six lake locations in east Tennessee that vary in population density. We also placed experimental
colonies at each of these sites to quantify their survival and reproductive success at each site. Over the
course of the active season (May to October) we monitored temperature, relative humidity and prey
density, as well as colony survival. At the end of the season we collected the nests for a final census of
the number of juveniles. We will present results of nest survival and success with regard to natural
population density and environmental data.
STOICHIOMETRIC DETERMINATION OF HYDROGEN-PEROXIDE
DECOMPOSITION VIA COPPER CATALYSIS
Christopher Moore and Dr. Jeffrey Wardeska
Dept. of Chemistry, College of Arts and Sciences, East Tennessee State University, Johnson City, TN
Hydrogen peroxide has a decomposition reaction that is generally known by the stoichiometric equation
of 2H2O2 Æ 2H2O + O2. Using two imidazole adducts of the tridentate-copper catalyst, [CuSaLAD]2, we
were able to catalyze the decomposition of hydrogen peroxide within minutes. Our research focused on
the stoichiometric decomposition of H2O2 by the copper-imidazole catalysts. To obtain this
stoichiometric decomposition directly, we measured the O2 formed from the catalysis reaction in a closed
system. By injecting the H2O2 into the system, a pressure change due to the formation of O2 from the
catalysis reaction was calculated. Subtracting the pressure measured from a blank run, the pressure of O2
was then applied to the Ideal Gas Law to obtain a molar quantity for O2. The stoichiometric relationship
obtained was not of the general equation mentioned above. In order to further investigate the reaction’s
mechanism we used kinetic studies, cyclic voltammetry, and a chemical test for copper(I) formation. By
allowing the closed system under high pressure to remain closed, a decrease in pressure over time was
obtained. From this data collection, rate constants were calculated for equimolar and excess copper (Cu :
2008 Appalachian Student Research Forum Page 20 H2O2) solutions while H2O2 remained constant. The rate constants were compared for each copperimidazole catalyst resulting in no copper-copper bridging through the imidazoles. The comparison also
displayed that no steric effect was observed between the placements of the methyl group. The cyclic
voltammetry provided evidence of Cu(I) formation as a new peak on the oxidative scan. This peak soon
disappeared, resulting in the possible reversible oxidation of Cu(I). Finally, to assure that the peak from
the voltammetry experiment was from the formation of Cu(I) when H2O2 is catalytically decomposed, a
chemical indicator was used. The 2,2-biquinoline indicator was added to the copper-imidazole catalyst
prior to the addition of H2O2 resulting in a pink color after addition and the evidence of Cu(I) formation.
The data collected from all experiments have been compiled to compare to previous research and
mechanisms in order to develop the best possible mechanism. Once the mechanism has been determined,
the stoichiometric determination of catalytically decomposed H2O2 can be shown.
ROLE OF SABP2 IN THE INDUCTION OF SYSTEMIC ACQUIRED RESISTANCE
IN RESPONSE TO BACTERIAL PATHOGENS IN PLANTS
Cory N. Phillips, Kajal C. Patel, Raj C. Bant, Elizabeth P. Huntley and Dhirendra Kumar
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Plants do not possess an adaptive immune system like animals; however, they can deploy various
defenses, performed and induced, that are used to combat pathogen invasions. One compound that
triggers these defenses against pathogens is a plant hormone called salicylic acid (SA) which is made in
the chloroplast of plant cells via the shikimate pathway. SABP2 is a 29 kDa protein which binds to SA
with very high affinity and is critical for both local and systemic acquired resistance. SABP2 converts the
methyl salicylate (MeSA), synthesized by plants in response to pathogen infection into active SA, which
then activates defense proteins. Recent studies identify methyl salicylate as the long distance phloem
mobile signal required for onset of systemic acquired resistance (SAR). SABP2 is required in the
uninfected systemic leaves to receive and process this SAR signal. All the host resistance response studies
involving SABP2 have been conducted using viral tobacco pathogen, tobacco mosaic virus (TMV). The
role of SABP2 in SAR induced responses to bacterial and fungal pathogens has not been investigated.
Bacterial diseases cause great losses of agricultural crops, which are important for food security.
Therefore, we are investigating the role of SABP2 in SAR and local resistance induced by virulent,
avirulent, and non-host bacterial pathogens. These results will not only have biological importance, but
also have major implications for farming in the future by reducing dependence on harmful pesticides and
insecticides. They may even provide a break away from pesticides to a more sophisticated solution, such
as enhancing plants’ own natural defenses.
THE INCIDENCE AND PREVALENCE OF BATRACHOCHYTRIUM DENDROBATIDIS IN
TEMPORARY WETLAND POPULATIONS OF AMBYSTOMA MACULATUM IN THE
HOLSTON RIVER VALLEY
Mehdi Pourmorteza and Tom Laughlin,
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Chytridiomycosis is an emerging infectious disease affecting amphibians worldwide. It is caused by the
chytrid fungus Batrachochytrium dendrobatidis, a fungus causing sporadic morbidity and mortality in
amphibian populations. The fungus infects the surface layer of the frog’s skin, causing damage to the
keratin layer. Many researchers believe that this infection inhibits the frog’s ability to respire and
osmoregulate. To better understand the dynamics of the chytrid fungus, the incidence and prevalence of
2008 Appalachian Student Research Forum Page 21 Chytridiomycosis will be measured in salamanders in three different populations; one in Shady Valley
where water is polluted with cattle waste, and the other two in the South Holston River floodplain and at
Bays Mtn. Park where water conditions are relatively unpolluted. DNA from salamanders has been
extracted from two different populations thus far using Qiagen DNeasy tissue kits. The isolated DNA
will be used for PCR reactions using primers previously developed for this species.
OSTEOCRANIAL DEVELOPMENT IN THE OVIPAROUS CORN SNAKE, PANTHEROPHIS
GUTTATUS (SERPENTES: COLUBRIDAE)
Elizabeth K. Price1, Rebecca A. Pyles1, James R. Stewart1 and Tom W. Ecay2
Dept. of Biological Sciences and 2Dept. of Physiology, Quillen College of Medicine, East Tennessee
State University, Johnson City, TN
1
Study of osteocranial development in vertebrates serves as an important source of evidence for functional,
systematic, evolutionary, and developmental questions. Osteocranial development has been studied
extensively in fishes, amphibians, birds and mammals; however, such studies are particularly lacking for
squamate reptiles---the lizards and snakes. Most previous studies of squamate skeletal development have
been based on small sample sizes, have not reported variation, and have utilized embryo or postnatal size
to compare specimens. Because size varies considerably among species and with incubation temperature,
results of these studies are not comparable. This study was designed as a baseline study of osteocranial
development in an egg-laying (oviparous) snake, Pantherophis guttatus, commonly called the corn snake.
Data from this study are intended to provide a ‘developmental clock’ describing the uptake and use of
calcium for ossification of the embryonic skeleton. Eighty embryos and young were identified to
developmental stage utilizing a standard table (Zehr, 1962) and size of a skeletal element (Meckel’s
cartilage, with or without investing bones; to nearest 0.05 mm) used as a relative measure of growth.
Specimens were cleared and double-stained (Hanken & Wassersug, 1981; Taylor & van Dyke, 1985) to
reveal both developing cartilage and bone in whole-mount. In the cranium, first evidence of ossification
was a palatal bone (palatine, stage 29—31). During stage 32, ossification of the mandible (prearticular,
surangular) and one jaw suspensory bone (pterygoid) proceed quickly, while the first upper jaw and skull
roofing bones show only initial centers of ossification. Ossification of particular bones show variation
across developmental stages, but a relatively consistent pattern can be identified. By stage 37, just prior
to hatching, most bones of the cranium are ossified, except for the parietals and occipital complex of the
skull roof, and the basisphenoid of the floor of the braincase. Interestingly, the earliest indication of
calcium in the embryonic cranium appears as calcium-containing deposits sequestered in endolymphatic
fluids within the inner ear (otic capsules). These deposits are present in the earliest developmental stages
observed (stage 27), and well before ossification of any skeletal bone begins, and they persist through all
later stages or until superficial ossification of the otic capsules obscure observations. Comparison of our
results, based on our interpretation of comparable developmental stages, to ossification patterns reported
for Python and Naja (cobra) indicate that substantial variation in ossification has been significantly underrepresented in previous studies. However, a generally consistent pattern of ossification is proposed in
which palatal and jaw bones ossify early and braincase ossifies late. The presence of calcium in
endolymphatic fluids very early suggests that mobilization of calcium to the embryo occurs well before
skeletal ossification begins during development. Results of this study provide a model approach to enable
direct comparison of skeletal development among different species.
2008 Appalachian Student Research Forum Page 22 THE INCIDENCE AND PREVALENCE OF RANAVIRUS IN TEMPORARY WETLAND
POPULATIONS OF AMBYSTOMA MACULATUM IN THE HOLSTON RIVER VALLEY
Joel Waddell and Tom Laughlin
Dept. of Biology, East Tennessee State University, Johnson City, Tennessee
Frog virus 4 (FV4), a ranavirus in the family Iridoviridae, is an infectious disease of recent concern
because it has been associated with morbidity and mortality in wild and commercial amphibian
populations on several continents. Recent evidence suggests that environmental degradation can impair
amphibian immune systems. This project will investigate the incidence and prevalence of FV4 in three
Ambystoma maculatum populations in the Holston River Valley. I will screen DNA samples for the
presence of FV4 from approximately 100 individuals from each of three different populations in Sullivan
County, Tennessee characterized by varying degrees of human impact. The extracted DNA will be
screened using PCR to amplify a~500 base pair region of the major capsid protein of FV4. If FV4 in
present, I expect to find a direct correlation between poor water quality and an increased incidence and
prevalence of this ranavirus.
MOLECULAR WIRES: CURRENT RESEARCH AND FUTURE APPLICATIONS
J. Ryan McLain and Jeffery Wardeska
Dept. of Chemistry, East Tennessee State University, Johnson City, TN
Molecular wires are going to revolutionize technology in much the same way the microchip did in the
20th century. They will allow us to create molecular circuits, which are circuits consisting of individual
molecules; this is significant due to the size ratio allowing for so many technological leaps. Currently the
focus is on organic molecules, but this research is shifted towards the inorganic. It is based upon multinuclear mixed-valence iron complexes. Fe(CN)6 and FeHEDTA were used to create different ratio
complexes which could be built upon to chain out and create a full molecular wire. FT-IR, UV-Vis,
Cyclic Voltammetry, and Magnetic Moments were the analysis techniques employed in order to test the
complexes. These showed that using different ratios when mixing the compounds, would in fact, lead to
different ratios within the complexes. This technology is brings us a few steps closer to that of nano-scale
electronic devices.
PICTURE THIS! USING 2D IMAGES AS A MEANS
OF MEASURING SKELETAL ELEMENTS
Chris Swartz
Dept. of Computer and Information Sciences, East Tennessee State University, Johnson City, TN
For several reasons, having a method of measuring bones or other specimens without actually traveling to
the collection is quite beneficial. For researchers this reduces or even eliminates travel costs, especially if
one is traveling just to see one or two specimens. For curators this alleviates the loaning of even one or
two valuable specimens. A database of images and measurements would benefit both of these situations
and in the case of very large bulky items that are difficult to maneuver or small delicate items that should
be handled as little as possible, it could eliminate the headache of handling these specimens all together
for both parties. For my project, I was asked to develop a means of taking measurements from
2008 Appalachian Student Research Forum Page 23 photographs of skeletal elements. I used a digital camera to take the photographs and measured the
specimens from the photos using SigmaScan Pro ™. These measurements were compared against actual
physical measurements taken from the same specimens using calipers and an osteometric board. This was
done to determine if the photographic measurements were a comparable means of data collection. I
wanted a variety of skeletal elements to work with, so the skeletal data collection consists of extinct and
extant mammals such as seals, sea lions, bears, whales, Eocene whales, lemurs, sloths, great apes,
anteaters, walruses, and humans using data collected by my sponsoring professor and fellow students in
her lab. These specimens were collected from the Smithsonian Institution: National Museum of Natural
History. The current results of this project show that my measurements taken from a digital photo are
acceptable within a 95% confidence range when compared to measurements taken manually from the
actual bones. The major errors in measurement from digital photos come from improper placement of
scales and the distance ratio of the element being measured to the camera. The beauty of this method is
threefold in that 1) it works for the skeletal elements I was asked to originally make it work for; 2) it has
the potential to work for more than just bony elements, such as pottery, pelts, or any materials that can be
photographed as long as the guidelines are observed; and 3) despite the simplicity of this method, further
study has shown a potential for 3D imagery and measurements from this method. This third aspect of my
technique is part of my continuing research in this area.
Social & Behavioral Sciences
DRIVING IN APPALACHIA
Breana Chappell, Paul Nease, Robin Leonard, Ben Martin, Dr. Chris Dula
Dept. of Psychology, East Tennessee State University, Johnson City, TN.
Appalachia has historically been referred to as a regionally unique area. Previous research has shown
certain populations, especially low income populations, are at an increased risk for involvement in
collisions. Additionally, research indicates that as personal income increases, individual risk for
involvement in traffic collisions decreases (Keeler 2003). Some research attributes these decreases to
ability to afford safer automobiles and increased traffic safety education. Little research has been
conducted that focuses specifically on low income areas of the United States, such as Appalachia. Safety
belts and their usage may have gone largely ignored by some populations of this area. Also, the number of
lifetime crashes could be related to the non-usage of seatbelts by drivers in Appalachia. The authors
hypothesize that people who identify themselves as ‘Appalachian’ are less likely to use safety belts while
driving than those who do not identify themselves as being Appalachian. The authors additionally
hypothesize that those who identify themselves as Appalachians would report having been involved in
more traffic collisions than non-Appalachian participants. A survey was given using an online participant
pool to a sample of college students at a regional southeastern Appalachian university. The survey was
completed by 225 students, 156 females and 69 males. The participants mean age was 22.26 years with a
standard deviation of 6.13 years. In the survey the participants were asked if they considered themselves
to be Appalachian. Additionally using a 5 point Likert Scale, they were asked how often they used
seatbelts, how many collisions they had been involved in during their entire life, whether or not they were
the driver. A Chi-Square Test was conducted to ascertain any relationship between seat belt usage and
Appalachian self-identification. The analysis showed a relationship but failed to reach significance, p=
.059. An independent t-test was conducted to determine what relationship, if any, exists between lifetime
crashes and Appalachian self-identification. This relationship was found to be statistically significant, t=
2.499, p= .013. Although not statistically significant, a relationship was determined to exist between seat
belt usage and Appalachian self-identification.
2008 Appalachian Student Research Forum Page 24 PERCEPTIONS OF TATTOING ON ONESELF AND OTHERS
Matthew A. Dorton, Benjamin A. Martin., Sonia L. Coney, Chris S. Dula
Dept. of Psychology, East Tennessee State University, Johnson City, TN
In social psychology there are many terms to incorporate the aspects of the self and it’s interactions with
society. Understanding of the concepts regarding self-awareness, social self, and the executive self is an
essential part to the correct perceptions of the self’s identity within its society. Previous research has
found prevalence of tattooed individuals within subpopulations (Edgerton & Dingman, 1963) and
stereotypes about personalities as deviant (Howell, Payne, & Roe, 1971). Discovering the impact that
tattoos have on society may be understood through the analysis of social comparisons, intrinsic and
extrinsic motivations, self-esteem, self-presentation, action identification; as the connection between
principles and actions, self-regulation, and self-defeating behavior (Baumeister &Newman; 1994; Deci,
1971; Ferrari, 2001; Festinger, 1954; Jones & Berglas, 1978; Locke & Kristof, 1996; Shoda, Mischel, &.
Peake, 1990; Tangney, Baumeister, & Boone, 2004; Vallacher & Wegner, 1985, 1987). Previous
literature has made some attempts at categorizing the reasons for getting a tattoo (Garcia, 1987).
However, existing categories lacked a complete definition. Data was collected from two open-ended
demographic questions: “why did you get a tattoo” and “why do you think others get a tattoo.” The
positive reasons and themes are 1. Religious/Beliefs (Garcia, 1987) 2. Remembrance/Commemoration/
Sentimental (Bezaury & Moron, 1983; Garcia, 1987) 3. Art Collection/Decorative/Aesthetic (Garcia,
1987) 4. Social group identification/Popularity (Garcia, 1987; Roberts, Koch, & Armstrong, 2006) and 5.
Self-Statements – including: Self-Knowledge/Concepts, and Self-Agency (Executive Decisions) (Bezaury
et. al., 1983; Millner & Eichold, 2001; Rubio, 2003). For the negative reasons and themes the categories
include: 6. Military/Gang/Prison/Sexuality (Bezaury et. al., 1983; Garcia, 1987) 7. Aggression/AntiSocial/ Death or “Dark” themes/Pain enjoyment (Bezaury et. al., 1983) and 8. Impulsivity/Alcohol
Related Reasons/Peer-Pressure (Roberts et. al., 2006). Through the use of the theory of symbolic
interactionism which describes human relations through language and symbols which have acquired a
conventional significance within the individual’s society; predicts that tattoos in society will continue to
become more acceptable and widespread as their fashion and popularity continue to grow (Atkinson
2003; Irwin, 2001; and Pitts, 2003). Participants were 210 undergraduate students aging from 18 to 62
years of age (142 women, 68 men, M=20.5, SD=4.99) completed the survey for a modest amount of extra
credit. A cross- tabulation was performed to detect differences in answers between the categories and a
chi-square analyzed whether or not having tattoos affected the answers to the demographic questions.
Results 3.9% were identified with impulsivity, aggression, & pain enjoyment as their reason for getting a
tattoo. 96.1% identified with various positive reasons such as: religious beliefs, remembrance/art,
aesthetic, popularity, self statement & other executive decisions. 5.2% participants speculated that people
get tattoos for negative reasons. 94.8% of participants speculated that people get tattoos for positive
reasons. Results from this study may provide strength to the theory that, instead of an impulsive behavior,
tattooed individuals indeed make conscious decisions regarding their tattoos. Future research should focus
on possible relationships between theses results, knowledge of tattoos, and stigma against tattoos.
2008 Appalachian Student Research Forum Page 25 THE ROLE OF MODELING IN RELATION TO SEQUENTIAL TOUCHING
James Anthony Garland
Dept. of Psychology, East Tennessee State University, Johnson City, TN
The sequential touching task is a popular nonlinguistic procedure used to tap into infant and toddler’s
categorical understanding. Although children’s performance on the task demonstrates convergent validity
with linguistic measures of categorical understanding, the task also suffers from low reliability. Although
not usually documented, users of the sequential touching procedure are well aware of the difficulty
involved in scoring infants touching behaviors. The purpose of this study is to investigate whether
modeling targeted actions within the sequential touching procedure would result in a more reliable
assessment of infant and toddler categorical understanding. It is suggested that a simple modification of
the procedure would result in the ability to magnify the distinction between categorizing and noncategorizing infants. Participants include 27 infants with a mean age of 15 months. Specifically, it is
expected that children who are explicitly shown distinctions between two categories presented in a
standard sequential touching task, will be better able to demonstrate their categorical understanding as
compared to their initial baseline performance and as compared to their peers in a non-modeling control
group.
IMPACT OF DRUGS AND ALCOHOL ON MANNER OF DEATH BY SEX AND AGE AMONG
AUTOPSY CASES PERFORMED AT THE UPPER EAST TENNESSEE FORENSIC CENTER IN
2007.
Caleb Harrell1, Paul Benson2, Teresa Campbell2, Emily Lemieux1, David Lisenby2, Laura Parsons2, Kathy
Bryant1, Debbie Bailey1, Kim Hughes1, and Kenneth Ferslew1
1
Toxicology and 2Forensic Pathology , Upper East Tennessee Forensic Center, East Tennessee State
University, Johnson City, Tennessee.
The Upper East Tennessee Forensic Center performs autopsies on the questionable and medicolegal
deaths which occur in the eight counties of the First Tennessee Development District. Toxicological
evaluations of specimens collected at autopsy are used to determine if drugs and/or alcohol are involved
in determining the cause and manner of death. A descriptive database was established defining all
parameters and data pertinent in each case (age, sex, cause/manner of death, and toxicological results).
The purpose of our research was to determine descriptive statistics on the impact of drugs and/or alcohol
by manner/cause of death, age, and sex in the autopsies performed in 2007. Specimens (blood, gastric
contents, urine, vitreous humor, and bile) from the autopsies were analyzed for drugs and alcohol using
multiple analytical toxicological procedures (colorimetric, TLC, immunochemistry, GC, GCMS, and
LCMS). Toxicological results were compiled in an electronic database (Microsoft Access) to allow for
analysis and interpretation. Results indicate that out of 277 total cases 66% were male, 34% were
female, 85% were positive for drugs, 27% were positive for alcohol, 23% were positive for both drugs
and alcohol, and 12% had neither drugs nor alcohol. Analysis of the distribution of cases positive for
drugs, alcohol, and drugs/alcohol revealed that males had a greater percentage of cases involving alcohol
alone as well as cases positive for drugs/alcohol than females. Acute drug overdoses accounted for 34%
of total cases with no substantial sexual differentiation. Of the 94 acute overdose cases, 4 (≈4%) were
intentional (suicides) and 90 (≈96%) were accidental. Autopsies were performed on all age groups
(percentage of cases/ years of age): 5% <14, 4% between 15 and 19, 8% between 20 and 24, 14%
between 25 and 34, 22% between 35 and 44, 16% between 45 and 49, 13% between 50 and 54, 10%
between 55 and 64, 6% between 65 and 74, and 2% ≥75 years of age. The distribution of positive drug
2008 Appalachian Student Research Forum Page 26 cases closely mirrored the distribution of cases by age groups. Manner of death analysis revealed (of
total cases) that 47% were accidental, 27% were natural, 15% were suicides 6% were homicides, and 5%
were undetermined. No appreciable disparity in distribution of manner of death was found between the
sexes. Analysis of the results of the toxicological evaluations revealed there were a large number of
cases in which opiates (100), alcohol (75), benzodiazepines (110), sedatives (18), and/or stimulants (14)
were identified. Review of these results leads to the conclusion that drugs and alcohol have a significant
impact in the questionable and medicolegal deaths occurring in Upper East Tennessee.
CELL PHONE USAGE:
CHANGING THE DYNAMIC OF INTERPERSONAL SPACE
Kelley Marie Hatch
Dept. of Communication, East Tennessee State University, Johnson City, TN
Newspaper and magazine article headlines are the harbingers of public sentiment: “Cell Hell,” “Cell
Phone Follies” and “A Call for Cell Phone Etiquette” are a sampling of recent news items expressing
public outrage over the misuse of cell phones. Cell phone usage in public has struck a dissonant chord.
Most people have a cell phone horror story: a romantic anniversary dinner interrupted by an inconsiderate
cell phone user, overhearing someone’s embarrassing medical problem etc. The purpose of this research
is to ascertain the impact of cell phone usage on the dynamics of interpersonal space. For this study,
“interpersonal space” is defined as the spatial relations between individuals in terms of what is public
space versus what is private space. The portability and accessibility of cell phones changed perceptions
of what was once considered public space into private space for the intent of individual conversations.
No longer does it appear that individuals feel the need to take a phone conversation away from other
people in order to maintain a semblance of privacy. This begs the question, what has occurred in society
as catalyst for this change? At a time when our human experience is inundated with technology: laptop
computers, Mp3 players, DVDs, GPs, electronic billboards, talking cars – fast paced, fast moving
machinery --- our desire for human contact is greater than ever. We are for the first time in a position to
need to daily negotiate our humanness amongst technology. Cell phone technology helps by bridging the
divide between technology we must live with, and human contact which we cannot live without. As we
are renegotiating space and technology, the research question then becomes, has cell phone usage
changed what we talk about in public spaces erasing lines between public and private? To test this
research question 400 respondents from 22 states and with an average age of 33 years, found that people
were always aware of their surroundings when placing a cell phone call, and reported being comfortable
placing a call in a public place. Respondents were asked to divulge not only the topics of their
conversations in public space, but also topics overheard in a public space, and specifically any found to be
shocking. Survey results in conjunction with existing literature justify the conclusion that individuals
making or answering a cell phone call in a public space retreat into a personal cell cocoon or PCC, an
oblivion to their immediate surroundings.
2008 Appalachian Student Research Forum Page 27 DEPRESSION AND ALCOHOL ABUSE: ETHNIC DIFFERENCES
IN THE MODERATING ROLE OF HOPE AND HOPELESSNESS
Angela Jones, Jameson Hirsch, Preston Visser,
Dept. of Psychology, East Tennessee State University, Johnson City, TN
Elizabeth Jeglic, John Jay, College of Criminal Justice, Dept. of Psychology, New York, NY
Depression is often complicated by the co-occurrence of alcohol abuse and dependence; nearly one-third
of individuals with major depressive disorder also have a substance abuse disorder. Ethnic differences in
comorbidity exist. Whites have a higher rate of alcohol abuse than Blacks, Hispanics, and Asians;
however, the rates of alcohol abuse have risen sharply for adolescent Blacks and Hispanics. Although the
relationship between depression and alcohol abuse is bi-directional, a cognitive framework suggests that
depressive symptoms may lead to alcohol use as a maladaptive coping strategy. Not all individuals who
are depressed, however, resort to abuse of alcohol, utilizing more adaptive means of problem resolution
instead. Such differentiation in problem-solving may be the result of an ability (hope) or inability
(hopelessness) to identify future oriented goals and conceptualize pathways toward goal achievement.
Hopeful individuals may be able to find alternative solutions to stressful situations, whereas hopeless
individuals may be unable to do so, possibly resorting to abuse of alcohol instead. We examined the
moderating role of trait hope and hopelessness on the relationship between symptoms of depression and
severity of alcohol abuse, in an ethnically diverse sample. We recruited an ethnically diverse sample
(Black, 25%; Hispanic, 41%; Caucasian, 18%; Asian, 6%) of 385 college students (69% female), with a
mean age of 19.61 years old (SD=3.12), from an urban, Eastern university. Participants provided written,
informed consent and completed measures of depressive symptoms, trait hope, hopelessness, and a
screener for alcohol abuse and dependence. Multivariate, hierarchical regressions and moderator analyses
were conducted, statistically controlling for age and gender. Initial models assessed moderator status of
trait hope; secondary models also examined hopelessness as a moderator. Analyses were conducted for
the entire sample and for ethnically stratified groups. With only trait hope included in the model, hope
moderated the association between depressive symptoms and severity of alcohol abuse for the entire
sample, suggesting that, in the context of depression, individuals who are more hopeful report less severe
symptoms of alcohol abuse. In ethnically stratified analyses, trait hope neared significance as a
moderator only for Whites (p=.06). With hopelessness included in the model, neither was significant for
the overall sample. In stratified analyses, hopelessness was a moderator for Hispanics (p<.05) and neared
significance for Blacks (p=.08), indicating that depressed individuals who are also hopeless may manifest
more severe alcohol abuse, whereas trait hope was a moderator for Whites only (p<.05). Increased abuse
of alcohol in the context of depression is not inevitable; indeed, this relationship appears dependent, in
part, on one’s outlook toward the future. Ethnic differences suggest that sociocultural characteristics may
influence the moderating effect of hope and hopelessness, and that these constructs may not be
operationally equitable across ethnicities. Limitations include unknown generalizability to other
recruitment settings and demographic subgroups, and the potential for biased self report of psychiatric
symptoms. Our findings highlight the need for more comprehensive, future research, and suggest that
strategies designed to help young adults identify meaningful and attainable personal goals could be an
important component of substance abuse programs.
2008 Appalachian Student Research Forum Page 28 DEVELOPMENTAL HISTORY OF INDOOR TANNING
IN HIGH SCHOOL FEMALES
Leslie King and Joel Hillhouse, Ph.D.
Dept. of Public Health, East Tennessee State University, Johnson City, TN
There are over a million new cases of skin cancer diagnosed annually in the US with the incidence of
melanoma, the most lethal form of skin cancer, rising faster than any other cancer. It is important to
develop effective interventions that can possibly reduce the future occurrence of and mortality from
melanoma and non-melanoma skin cancers. High school represents an ideal focus for intervention
implementation. There is evidence that high school indoor tanning initiation is associated with later longterm habitual indoor tanning behavioral patterns which are highly resistant to change. This research study
is using focus groups to look at personal, social and family factors related to initiation and maintenance of
indoor tanning behavior in high school students. Efficacious health promotion interventions depend on
strong theoretical modeling to be successful. The focus group information collected in this study is the
typical first step in developing these essential models. This research could lead to more effective
interventions to substantially reduce the future occurrence of melanoma and non-melanoma skin cancers
in this population. While the relationship of modern tanning devices is still under investigation, a number
of case-controlled studies have reported significant positive associations between indoor tanning and
melanoma morbidity with increased risk associated with early and regular use. A recent report out of the
International Agency for Research on Cancer concluded that exposure to sunbeds before 35 years
significantly increases the risk of melanoma and squamous cell carcinoma. In spite of these facts, indoor
tanning is gaining in popularity, especially among the youth. The research study is being carried out at
ETSU. It will include 40 high school females between the ages of 14 and 16 years old. Eight to ten
students will meet with a facilitator in a private group setting. The discussions will last about 50 minutes
per group. Each student will be asked questions relating to their personal beliefs and attitudes regarding
indoor tanning (e.g. perceived susceptibility to skin aging, tanning alternatives attitudes, etc.). They will
also be guided to elaborate on factors contributing to tanning initiation and personal tanning history (e.g.
special events, social influence, etc.). The participants will also be asked to review a brochure designed
for a previous American Cancer Society grant in hopes of a possible adaptation that this age group can
better identify with. The resulting data will be subjected to qualitative analyses to identify key and
emerging themes of indoor tanning initiation, maintenance as well as reasons for quitting. This data will
be the first step in developing conceptual models of indoor tanning decisions in high school student
populations. Review of the brochure may warrant further revision and have the potential to be
implemented in future interventions. There is currently no published research addressing the indoor
tanning related decision-making in this age group. The models that emerge from the focus group results
will lead to more thorough assessment and intervention approaches toward high school indoor tanning.
This will ultimately lead to substantially reducing indoor tanning occurrences.
NICOTINE-CONDITIONED HYPERACTIVITY IN
ADOLESCENT MALE AND FEMALE D2-PRIMED RATS.
Lehmann J, Sheppard AB, Amine L, Brown RW.
Dept. of Psychology, East Tennessee State University, Johnson City, TN
The aim of this study was to determine the ability of a nicotine-conditioned context to elicit locomotor
hyperactivity in a neonatal quinpirole animal model of psychosis, and whether this conditioned
hyperactivity could be blocked by the D2 antagonist eticlopride. Sprague-Dawley rats were treated with
2008 Appalachian Student Research Forum Page 29 either saline or the dopamine D2 receptor agonist quinpirole from postnatal days (P) 1-21 to create
priming of the dopamine D2receptor, a phenomenon that we have shown persists throughout the animal's
lifetime. Beginning on P33, animals were injected i.p. with either nicotine (0.5 mg/kg), the D2 antagonist
eticlopride followed by nicotine, or saline and placed into the arena 10 min after injection every other day
through P49. A non-paired group was included. Results showed that, D2 priming blocked the typical
initial hypoactivity produced by nicotine, and these animals also sensitized to nicotine more rapidly than
controls. Eticlopride blocked sensitization to nicotine in both D2-primed and non D2-primed rats. On
P50, a portion of these animals were administered a drug-free test in which rats were given saline before
being placed into the locomotor arena. Interestingly, control animals administered nicotine demonstrated
conditioned hyperactivity on the drug free test that was blocked by eticlopride, but eticlopride did not
affect nicotine-conditioned hyperactivity in D2-primed rats. This result indicates priming of the D2
receptor was able to overcome D2 receptor blockade in adolescent rats. Further studies are analyzing the
role of nicotinic receptors in this phenomenon, as collaborators have shown that D2 priming produces
alpha7 nicotinic receptor upregulation.
DURATION OF BREASTFEEDING PREDICTS DIMENSIONS OF INFANT TEMPERAMENT
Kellye L. Lingerfelt and Dr. Andrea Clements
Dept. of Psychology, East Tennessee State University, Johnson City, TN
The benefits of breastfeeding and the duration of breastfeeding required to achieve those benefits have
long been debated. The American Association of Pediatrics (AAP) has noted benefits to the infant in such
areas as increased immunity to infectious disease, improved neurodevelopment, reduced rates of infant
mortality, as well possible benefits much later in life included reduced risk of obesity and diabetes. Other
studies have suggested cognitive benefits of breastfeeding. As of 2001, the reported rate of the initiation
of any breastfeeding at all in the US was 70% (46% breastfeeding exclusively—no formula
supplementation), with 33% (17% exclusively) still breastfeeding at 6 months. The purpose of this study
was to evaluate temperament differences among children who differed in breastfeeding history. To our
knowledge no one to date has investigated the relationship between breastfeeding and dimensions of
infant temperament. The data were collected through two studies being conducting by the Program for the
Study of Infancy at East Tennessee State University. One is a study in which mother-infant pairs are
brought into the lab for a one hour temperament, language, and mother-infant interaction procedure, and
the other is an ongoing collection of temperament, language, and child history from a university
laboratory school childcare program. The participants in these studies included 94 rural Appalachian
children (83% White). When participants are first enrolled in these studies, parents are asked to fill out a
Prenatal Birth Stress Inventory which included pregnancy and birth history variables and a temperament
measure (Infant Behavior Questionnaire [IBQ], Early Childhood Behavior Questionnaire [ECBQ], or
Child Behavior Questionnaire [CBQ]). In our study, 71.6% of mothers reported breastfeeding for 3
months or more and 46.1% reported continuing to breastfeed for 6 months or more. We did not ask about
exclusive versus supplemented breastfeeding. After statistical analysis, it was found that several
dimensions of temperament differed in relation to duration of reported breastfeeding. Focused attention
was unexpectedly lower in children who were breastfed for six months or more (n = 31; mean=3.69,
SD=.88) than in children that were breastfed six months or less (n = 32; mean=4.56, SD=.85;
F(1,61)=6.78, p=.012). Expected differences were found in other temperament dimensions. Frustration
(F(1,53)=6.68, p = .013) and sadness (F(1,61) = 8.97, p = .004) were found to be significantly lower in
infants that were breastfed six months or more. The ability to be soothed was rated significantly higher
(F(1,61) = 5.03, p = .029) for the children who were breastfed six months or more than for children
breastfed less than six months. Differences were found for the same variables in the same directions when
comparing ratings of children breastfed 3 months or more compared to children breastfed less than three
months.
2008 Appalachian Student Research Forum Page 30 A LOOK AT THE DEVELOPMENT OF THE MARTIN STIGMA AGAINST TATTOOS MEASURE
Benjamin A. Martin, Paul Nease, Heidi E. Mason, Ashley D. Dickson, and Christopher S. Dula
Dept. of Psychology, East Tennessee State University, Johnson City, TN,
Tattoos are becoming very popular in today's society, but empirical research lacks a way to measure the
stigma that is held against tattooed individuals. Although previous studies have documented and
discussed views of the tattooed and non-tattooed, little is known about the subject. This prior research,
however useful it may be, primarily consists of qualitative data collection methods (e.g. quasi-interview
methods), subtracting from the overall objectivity and generalizability of the results. In order to better
analyze stigma against tattooed individuals, an instrument was developed to measure this phenomena in a
quantitative fashion. The current study details the development of the Martin Stigma Against Tattoos
Survey Measure (MSATS) to measure the stigma against those with tattoos. Undergraduates (n=241,175
women & 66 men) were administered a set of surveys via an online survey system called Sona Systems to
assess social stigma against tattoos. Psychometric analysis was performed and stigma was calculated by
summing the responses. In addition to strong face-validity, extraction of one higher-order factor provided
evidence of a single overall construct. Exploratory factor analysis exhibited one factor, interpreted as
social stigma against tattoos. Confirmatory factor analysis supported the structure of the scale and
suggested coefficient reliability. Cronbach's alpha was .89 for the original 17-item instrument and was .92
for the modified version, which provided evidence of internal consistency reliability. The MSATS was
found to be reliable and valid with a college age sample of individuals with and without tattoos. The
Martin Stigma Against Tattoos Survey Measure will help reduce negative stigmatizing behavior against
those with tattoos and allow researchers the opportunity to examine stigma against tattooing in various
environments or situations. The MSATS could be useful in helping to both identify and reduce
inappropriate stigmatizing behavior. This scale could be applied in future research to gauge whether a
stereotype placed on tattooed employees by biased employers has any merit. This may be helpful in
breaking current actual and perceived employment barriers and allow previously stigmatized persons to
function at a higher occupational level. Another potential benefit of the MSATS could be to assess
managerial candidates within a company to help screen for unnecessary bias within a company toward
tattooed employees.
NICOTINE SENSITIZATION IN ADOLESCENT BETA ARRESTIN-2
KNOCKOUT MICE: CORRELATIONS WITH BDNF.
Noel DM1, Correll JA1, Thompson KN2, Longacre ID,1 Yin D3, Brown RW1.
Dept. of Psychology , East Tennessee State University Johnson City, TN
2
Dept. of Anatomy and Cell Biology, Quillen College of Medicine, East Tennessee State University
Johnson City, TN
3
Dept. of Internal Medicine, Quillen College of Medicine, East Tennessee State University Johnson City,
TN
1
Beta Arrestin-2 is a protein that regulates hydrolysis of the G-protein and is co-localized with the
dopamine D2 receptor. In this study, 3-4 week old adolescent BA-2 KO and wild type C57/B6 mice were
administered either nicotine tartarate (s.c, 0.5 mg/kg) or saline 10 min before being placed into the
locomotor arena on each of seven (Experiment 1) or 14 (Experiment 2) consecutive days. A drug-free
abstention period of seven days followed nicotine sensitization in each experiment, at the end of which
animals received a nicotine challenge (0.5 mg/kg free base). Experiment 1 results showed that BA-2 KO
2008 Appalachian Student Research Forum Page 31 mice were slightly hypoactive and did not demonstrate sensitization by day 7, whereas wild type controls
did not demonstrate an increase in activity as compared to saline-treated wild types by day 7, but did
show an increase in activity over the 7 days of nicotine treatment. On the nicotine challenge, BA2 KO
blocked expression of nicotine sensitization. In Experiment 2, BA-2 KO mice demonstrated sensitization
although not the levels of the controls. On the challenge, beta arrestin-2 again blocked expression of
sensitization. Brain tissue was taken in both experiments for analysis of Brain-derived neurotrophic factor
(BDNF) in the nucleus accumbens, and showed that nicotine produced a significant increase in BDNF
which was blocked by the knockout of beta arrestin-2. These results show that beta arrestin-2 plays a
more important role in expression as compared to induction of nicotine sensitization, and blocks nicotineinduced increases in BDNF in the nucleus accumbens.
RELIGIOSITY, SPIRITUALITY, FUNDAMENTALISM, AND ACTIVELY CARING BEHAVIORS
Robert B. Russell & Chris S. Dula.
Dept. of Psychology, East Tennessee State University, Johnson City, TN
Current research on the helping behaviors exhibited religious individuals is inconclusive. While religious
individuals overwhelmingly identify themselves as frequently committing altruistic acts (Mattis et al.,
2000), few researchers have found religion to be a mediating factor on helping behaviors in controlled
settings (Jackson & Esses, 1997). Religious fundamentalism--the degree to which one believes their
religion to be unquestionably true--has ironically been shown to negatively correlate with caring
behaviors, particularly those outside one’s own religious group (Jacson & Esses, 1997). Spirituality--a
feeling of purpose, connection to others, and one’s general interest in existential queries (Hodge &
McGrew, 2006), however, has been consistently found to positively correlate with both self-reported and
verifiable altruistic acts (e.g. Greenwood & Harder, 2003). Geller (1991) theorized that several factors
play into one’s propensity to ‘actively care’ for others, including high self-esteem, a sense of
belongingness, high self-efficacy, an optimistic attitude, and a sense of self-control. The purpose of this
study was to determine if an individual’s level of spirituality, religiosity, and religious fundamentalism
relate to those factors which influence actively caring. It was hypothesized that spirituality would show a
positive relationship with the actively caring factors; also, religious fundamentalism was expected to
negatively correlate with these factors. A total of 204 undergraduate psychology students (61 Men, 143
Women) ranging in age from 18 to 55 years of age (Median = 20) completed the Inventory of
Religiousness (IoR; Rayburn, 1997), Religious Fundamentalism Scale (RFS; Altemeyer & Hunsberber,
1992), Inventory of Spirituality (IoS, Rayburn & Richmond, 1999), and the Actively Caring Survey
(ACS; Geller, 1995) as measures of religiosity, religious fundamentalism, spirituality, and actively caring
behaviors, respectively. Because self-reported altruistic behavior differed from those observed in
experimental settings, the Marlowe-Crowne Social Desirability Scale (MCSDS; Crowne & Marlowe,
1960) was included as a measure of desirable responding bias. Data was collected anonymously by use of
an online survey administration software, and all participants received a modest amount of extra credit as
compensation for their time. Data analysis consisted of computing correlation coefficients among the IoR,
RFS, IoS, and all subscales of the ACS, while the MSCDM was held as a constant to lessen the effect of
social desirable responding on the results. Because the number of correlations conducted was significantly
large, the Bonferroni method was used to control for Type I errors, requiring a p value of less than .0023
(.05/21) for significance. Seven of the 21 correlations were significant at this level: Spirituality was
positively correlated with the Actively Caring, Self Efficacy, Optimism, Belonging, and Personal Control
subscales of the ACS, along with the total ACS score (.54, .42, .35, .33, .34, & .48, respectively),
supporting the first hypothesis. Religious fundamentalism was found to negatively correlate with only the
ACS self-esteem subscale (-.29); as such, the second hypothesis was not strongly supported. These
findings may indicate that one’s level of spirituality is more directly related with altruistic and helping
2008 Appalachian Student Research Forum Page 32 behaviors than religious affiliation. Limitations and suggested directions for future research will be
discussed.
AMBIVALENT SEXISM AND SEX ROLE IDENTITY
AS FACTORS IN SEXIST HUMOR APPRECIATION
Jennifer Snyder and Dr. Peggy Cantrell
Dept. of Psychology, East Tennessee State University, Johnson City, TN
Overtly prejudiced remarks in public are often shunned in modern society; however, the reactions to and
appropriateness of humor involving potentially sensitive topics, including gender roles and stereotypes of
women, remain mixed and controversial. The possible relationships, if any, among sexist humor and
other variables as well as any potential consequences of the presence of such humor in public or private
domains have been under empirical investigation within psychology. The current study examined
ambivalent sexism, gender, and gender role identity in a college sample (N = 254) for any possible
relationships with sexist humor against women. Funniness ratings and repeating behaviors of 10 sexist
internet jokes were compared. Results show that men were more likely to rate sexist humor as funny and
more likely to repeat sexist humor than women. Ambivalent sexists significantly rated sexist humor as
funnier and were more likely to repeat sexist humor than benevolent sexists and non-sexists; no
significant differences were found between hostile sexists and any other sexism category. No significant
differences were observed among gender role identity classifications. This study provides additional
evidence for the complementary nature of hostile and benevolent sexism in the justification of prejudice.
Results are discussed in light of Prejudiced Norm Theory (Ford, 2004).
CORRELATIONS BETWEEN THE BIG FIVE PERSONALITY TRAITS
AND SUSCEPTIBILITY TO SOCIAL INFLUENCE
Lolita D. Turner, Robert B. Russell, & Chris S. Dula
Dept. of Psychology, East Tennessee State University, Johnson City, TN
Social influence can be defined as influence exerted by an outside person or group(s) to alter an
individual’s behavior or actions. This influence can be exerted both subtlety and overtly, and plays a role
in everyday interpersonal interactions. Susceptibility to this social influence and its varying forms has
been shown to remain stable over the course of an individual’s life (Dula, Lehman, & Geller, under
review), and may be related to numerous other facets of personality; however, no research has been
conducted which examines a possible relationship between this susceptibility and the Big Five traits of
personality. The aim of this study was to examine this possible relationship. Because of findings in the
current literature, it was hypothesized that an individual’s level of Conscientiousness, Agreeableness, and
Extraversion would positively correlate with susceptibility to social influence. A total of 204
undergraduate psychology students (61 Men, 143 Women) ranging in age from 18 to 55 years of age
(Median = 20) completed the NEO short-form (NEO-FFI; Costa Jr. & McCrae, 1992) and the Social
Influence Scale (SIS; Dula, Lehman, & Geller, under review) as self-reported measures of Big Five
personality characteristics and susceptibility to six types of social influence, respectively. Data was
collected anonymously by use of an online survey administration software, and all participants received a
modest amount of extra credit as compensation for their time. Data analysis consisted of computing
correlation coefficients among all subscales of the NEO-FFI and the SIS. Because the number of
correlations conducted was significantly large, the Bonferroni method was used to control for Type I
errors, requiring a p value of less than .0017 (.05/35) for significance. Eleven of the 35 computed
2008 Appalachian Student Research Forum Page 33 correlations were significant at this level: Extraversion was positively related with Reciprocity,
Ingratiation, Scarcity, and Total SIS scores (.37, .28, .39, & .44, respectively), Openness and Scarcity
were positively correlated (.30), Agreeableness correlated positively with Reciprocity, Ingratiation, and
Total SIS Score (.35, .26, & .26, respectively), and Conscientiousness was significantly positively related
to Reciprocity, Ingratiation, and Total SIS score (.33, .36, & .32, respectively). All three major hypotheses
were confirmed, suggesting that susceptibility to social influence effects/is effected by an individual’s
level of Extraversion, Agreeableness, and Conscientiousness. Unfortunately, no directionality can be
derived from the current results: As the study consisted of correlational statistical procures, it is unknown
how personality and social influence interact with one-another. While personality differences may
moderate one’s level of social-influence susceptibility, susceptibility to social influence may contribute to
defining one’s personality. Because so little research has been conducted on the topic, more research is
needed before any definitive conclusions are reached. Limitations and directions of future research will be
discussed.
ADVENTUROUSNESS AND IMPULSIVE BEHAVIOR: IS THERE A RELATIONSHIP?
Benjamin Martin, Audrey Watkins, Jessica Williamson, Robin Leonard, Chris Dula
Dept. of Psychology, East Tennessee State University, Johnson City, TN
The amount of adventure that a person seeks is often an indicator of how that individual views risk taking
behaviors. As stated by Zuckerman and Neeb in 1980, high sensation seekers are likely to participate in
risky activities such as driving fast and recklessly or acquiring body-art and/or engaging in body
modification. In fact, tattooed persons are considered to regularly engage in behavior that is risky,
immoral, non-conforming, and/or unstable (Forbes, 2001). When these behaviors were examined in non
confined populations, Forbes found that tattooed individuals conform less to social expectations and
perform risk-taking behaviors (2001). Most participants reported that they had thought about their tattoos
for at least a month before going to get it (Forbes, 2001). The evidence provided by Zuckerman and Neeb
suggests a possible positive correlation between an individual’s level of adventure/sensation seeking and
prominence of impulsivity. According to Moore and Rosenthal in 1993 there is a strong correlation
between adventure/sensation seeking and impulsivity. The purpose of this study is to assess whether there
is a relationship between adventurousness and impulsivity within tattooed and non-tattooed populations.
It is hypothesized that individuals that seek adventure will have higher levels of impulsivity. It is also
hypothesized that individuals with tattoos will be more adventurous and impulsive than individuals with
tattoos. Participants were 175 undergraduate students (55 men and 120 women). Their ages ranged from
18 to 62 years of age (M = 20.5, SD = 4.99). Participants completed survey through Sona Systems, an
online participant pool at a regional southeastern university. Extra credit was awarded for participation.
Barratt Impulsivity Scale (BIS). Demographic Questionnaire including questions about the number of
tattoos an individual has, if any. It is hypothesized that individuals that seek adventure will have higher
levels of impulsivity. It is also hypothesized that individuals with tattoos will be more adventurous and
impulsive than individuals with tattoos. Results did not suggest impulsivity varies between tattooed and
non-tattooed populations. Thus it may be that tattooed individuals are unfairly believed to be highly
impulsive. Tattooing is viewed by many as an impulsive and irresponsible behavior (Eksteen &
Jankowski, 2006)
2008 Appalachian Student Research Forum Page 34 AWARENESS OF RACISM AMONG SELF-IDENTIFIED NEGROS
AND MULATOS IN ECUADOR
Meagan Stark, Kenneth Mijeski, and Scott Beck
Depts. of Political Science and Sociology & Anthropology, College of Arts and Sciences, East Tennessee
State University, Johnson City, TN
In the fall of 2004 an interesting and unique survey of over 8,000 adult Ecuadorians was conducted in
regard to racism and related topics. The interview items were developed by a team at SIISE, a semigovernmental research organization, and were attached to the employment and income survey conducted
by the Ecuadorian Census Bureau (INEC). In our research we use a number of measures from this survey
that focus on awareness of, and sensitivity to, issues of racism, prejudice and discrimination in Ecuador.
In Ecuador there are four major culturally defined ethnic groups: White, Mestizo (mixed with Spanish and
Indigenous ethnicity), Indigenous, and Afro- Ecuadorian. Survey items dealing with the respondents’
ethnic self-identification allow us to compare and contrast the level of racial awareness across ethnic
groups. In Ecuador, and much of Latin America, those of African decent are often classified as either
Negro (black) or Mulato (mixed). This study is particularly interested in measuring the degree of racial
awareness, or the awareness of discrimination and stereotypical racist attitudes, among those who selfidentified as Negros and Mulatos. Other independent variables of interest are the level of education
attained and the region in which the respondent resides, as well as whether they live in an urban or rural
area. Also, by utilizing 2001 Census data on the racial composition of the parroquia (parish) in which the
respondents reside, we attempt to determine whether those living in more racially heterogeneous areas are
more aware of prejudice and discrimination. The results will be discussed.
2008 Appalachian Student Research Forum Page 35 Division II – Graduate Students
1-2 Years
Biomedical Sciences
DETERMINING SECONDARY PRODUCT GLUCOSYLTRANSFERASE EXPRESSION DURING
Citrus paradisi GROWTH AND DEVELOPMENT
Jala Daniel and Cecilia McIntosh
Depts. of Biological Sciences and Physiology, East Tennessee State University, Johnson City, TN
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. More than 5000 flavonoids result
from this type of reaction and other modifications. Several different kinds of flavonoid
glucosyltransferases 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 8 putative
flavonoid GT clones. One GT bears 98% homology to a limonoid (terpenoid secondary metabolite) GT
and another has been found to have flavonoid GT activity. The roles of the other six are currently being
studied by other laboratory members. The goal of this investigation is to determine the expression
patterns of the 8 putative GT’s during the grapefruit growth and development by quantifying mRNA
expression levels in the roots, stems, leaves. This research is designed to test the hypothesis that these 8
GT’s are expressed constitutively. Alternatively, one or more could be expressed in a tissue-specific
manner or developmentally regulated. Five growth stages have been defined. The first stage focuses on
emerging roots; stages 2-4 focuses on leaves, stems, and roots; and stage 5 focuses on comparison of
older and younger leaves. RNA was isolated using a Qiagen RNeasy Plant Mini Kit. Unique primers for
each putative GT have been designed that are at least 20 nucleotides in length with similar annealing
temperatures. RT-PCR will be used to semi-quantitate mRNA expression. This will include determining
the linear range of amplification, so that all of the 8 GT’s can be compared in a semi-quantitative analysis.
RNA will be reversed transcribed using oligo dT primers and the cDNA will be amplified with gene
specific primers using a Titanium One-Step RT-PCR Kit. In order to carry out a semi-quantitative
analysis, a limited number of PCR cycles (5-20 cycles) will be run for each sample so that the amount of
cDNA made will be proportional to the amount of each specific mRNA species in the extract. Ribosomal
18s RNA will be used as an internal control to ensure that there are equal starting amounts of RNA. This
method of RT-PCR analysis was chosen because small amounts of RNA can be used (as low as 20 ng) for
each reaction and it is efficient for determining gene expression levels for multiple RNA samples. When
trying to establish the biological function of these putative glucosyltransferases, part of the whole picture
is establishing the reaction that is catalyzed and knowing when and where these GT’s are present. The
results of this study will make a significant contribution to the understanding of plant secondary
metabolism by linking biochemical function and gene expression patterns in order to help elucidate the
metabolic and physiological impact of these gene products.
2008 Appalachian Student Research Forum Page 36 URINARY EXCRETION OF FUNGAL (1-3)-β-D-GLUCAN PHOSPHATE FOLLOWING IV
ADMINISTRATION
Debra K. Head, Janet W. Lightner, Tammy Ozment-Skelton, David L. Williams and Peter J. Rice
Depts. of Pharmacology (DKH, JWL, PJR, DLW) and Surgery (TO-S, DLW), James H. Quillen College
of Medicine, East Tennessee State University, Johnson City, TN
(1-3)-ß-D-Glucans are polysaccharide components of cell walls in various fungi and bacteria. These
fungal and/or bacterial glucans are pathogen associated molecular patterns (PAMPs) that circulate during
infection and interact with pattern recognition receptors, including Dectin-1 and scavenger receptor A, to
modulate innate immunity. Our earlier studies established the pharmacokinetics of intravenously
administered glucans and demonstrated that glucans are absorbed following oral administration. In this
study, we investigated the renal excretion of (1-3)-ß-D-glucans following intravenous administration in
rats. Fluorescently-tagged glucan phosphate was prepared from Saccharomyces cerevisiae and
administered (1 mg/kg, IV) to adult male rats anesthetized with isoflurane. Plasma samples were collected
from the tail vein at 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 5 and 6 h, and urine was collected prior to and for 6
hours after glucan administration. Urine samples were passed through an AKTA HiTrap desalting column
(5K MWCO) for analysis and to separate free fluorescence. The glucan fractions were combined and
lyophilized, and the molecular mass of the excreted glucans was determined by gel permeation
chromatography/multi-angle laser light scattering (GPC/MALLS.) Fluorescent-tagged glucan phosphate
showed a half-life of approximately 3 hours following intravenous administration. Peak urine
concentrations occurred within 3 h. Analysis by AKTA and fluorescence spectroscopy showed 71±3% of
fluorescence remained in the >5K MWCO fraction. GPC/MALLS analysis of this fraction showed a
minor peak with a molecular mass (171±11K) corresponding to injected glucan (~150K). Most glucan
detected by GPC was of lower molecular mass (13±8.5K). Our results indicate that while a small fraction
of glucan phosphate may be excreted unchanged in the urine, most (1-3)-ß-D-glucans are excreted by the
kidneys as smaller polysaccharides. The presence of urinary glucans may be an important indicator of
fungal infection.
THE APPALACHIAN REGION: INCIDENCE RATES OF MELANOMA CASES COMPARED TO
NON-APPALACHIAN REGIONS
Lana McGrady, Shashank Jain, Leslie King, Sunil Swami, James Anderson, Ph.D, Joel Hillhouse, Ph.D.
and Tim Aldrich, Ph.D.,
Dept. of Public Health, East Tennessee State University, Johnson City, TN.
Cancer is the second leading cause of death in The United States. In Appalachian region cancer incidence
and mortality rates have been found to be higher than rates for the rest of the country. The Appalachian
region [410 counties] is divided into three sections: Northern, Central, and Southern. The focus of this
study is the Central and Southern Appalachian regions where research has shown that the incidence rates
of skin cancer are higher compared to the national average (22%, 21.8%, and 20% respectively). This is a
region that is considered to be medically underserved especially in its rural areas. Higher rates of health
disparities and poverty contribute to this problem. The US National Statistics report that the incidence
rate of melanoma has increased by 18% since 1995. Cancer related mortality is also increasing nationally
as a result of this rise for melanoma. Melanoma, the most lethal form of skin cancer has shown to have
strong positive associations with exposure to UV rays through outdoor sun exposure and the use of indoor
tanning beds. This is important as melanoma is the most frequent cancer in women 25 to 29 years of age.
For men, the incidence rate has increased three-fold among ages 45 to 64. The overall incidence rate for
melanoma is increasing faster than for any other cancer. This is sufficient reason for public health
concern. This study sought to gather more comprehensive information about the incidence and mortality
2008 Appalachian Student Research Forum Page 37 rates resulting from melanoma in the Central and Southern Appalachian region. This includes identifying
regional patterns and specific population sub-groups that have a higher risk of melanoma. The Skin
Cancer Prevention Laboratory at East Tennessee State University began by requesting state-wise crude
and age-adjusted incidence and mortality rates of melanoma for seven states that encompass the majority
of Central and Southern Appalachia. Data was received for each state representing rates from the most
recent five years available. We began analyses with Tennessee data and the bordering states including:
Kentucky, Virginia, North and South Carolina, Georgia and Alabama. Data was recoded to differentiate
between Appalachian and Non-Appalachian counties in each state. Preliminary analysis revealed
significantly higher incidence rate of melanoma within some states. For example, North Carolina’s rate
ratio (RR) = 1.6 and Georgia’s RR=1.24. When the data for all states is aggregated, the age-adjusted
incidence rates will be compared to the national average of 18.5 per 100,000 men and women per year.
These initial findings strongly advocate further research on this subject within the Appalachian region.
The need for efficacious interventions is imperative given that melanoma mortality may be prevented if
treated in the initial stages. Such interventions could include an increase in patient screening and raising
awareness about melanoma. This could ultimately result in a decrease in mortality rates and an
improvement in the quality of life within the region.
CANNABINOID RECEPTOR (CB2) DEFICIENCY IS ASSOCIATED
WITH REDUCED LESIONAL APOPTOSIS
IN LOW-DENSITY LIPOPROTEIN RECEPTOR-NULL MICE
Courtney Netherland, Alicia Bales, Theresa Pickle and Douglas P. Thewke
Dept. of Biochemistry, East Tennessee State University, Johnson City, TN
Introduction: CB2 is the primary macrophage cannabinoid receptor and is found in atherosclerotic
lesions; however, its role in atherosclerosis is poorly understood.
A CB2 agonist, Δ9-/tetrathydrocannabinol, induces lesion regression in ApoE mice and some macrophage processes linked
to atherosclerosis are regulated by CB2 in vitro. Recently, we found that isolated CB2-deficient
peritoneal macrophages are resistant to induction of apoptosis by oxidized low density lipoproteins. As
macrophage apoptosis is a key process in the pathophysiology of atherosclerosis, 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 and lesional apoptosis in Ldlr null mice.
Methods: Ldlr-/- mice genetically deficient in CB2 were generated by crossing CB2-/- and Ldlr-/- mice.
Sex matched groups (n ≥ 8) of 8-week old CB2-/-/Ldlr-/- (CB2-/-) and CB2+/+/Ldlr-/- (WT) mice were fed a
Western 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 was quantified in proximal aorta cross
sections by oil red-O staining and in situ TUNEL analysis, respectively. Results: No differences were
seen in plasma cholesterol and triglyceride levels or in lipoprotein profiles between CB2-/- and WT
groups. No statistical difference was found between the mean lesion area of the CB2-/- and WT groups
after 8 weeks (113,907 ± 32,263 vs 115,377 ± 35,457 μm2, p=0.93, for males) or 12 weeks (185,546 ±
30,098 vs 197,616 ± 40,900 μm2, p=0.47, for males; 215,254 ± 44,016 vs 201,718 ± 39,448 μm2, p=0.49,
for females) on the Western diet. However, a significant decrease (~50%) was found in the mean 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, for males; 55 ± 25.7 vs 113 ± 37.5 apoptotic cells per mm2, p<0.001, for females).
Conclusions: Systemic CB2 deficiency in Ldlr null mice does not affect lesion formation, but does
significantly lessen lesional apoptosis. These results are consistent with our hypothesis and imply that
CB2 may be a novel target for therapies aimed at decreasing plaque vulnerability.
2008 Appalachian Student Research Forum Page 38 RELATIONSHIP OF PEAK ISOMETRIC STRENGTH TO RATE OF FORCE DEVELOPMENT
AMONG COLLEGIATE TRACK AND FIELD ATHLETES.
K. Painter, A. M. Swisher, J.M. Kraska, M.W. Ramsey, C. Nelson, M.E. Stone and M.H. Stone.
Sport Performance Enhancement Consortium, East Tennessee State University, Johnson City, TN
Explosive strength, a function of rate of force development (RFD), is a primary determinant for the
amount of peak force that can be produced in critical sport performance time periods (e.g. foot contact
time). Evidence indicates that maximum strength and RFD are correlated. However, the characteristics of
this relationship are not well established among athletes. This study examined the relationship of peak
isometric force (IPF) and RFD from a mid-thigh pull, among male and female track and field athletes.
Athletes were 12 sprinters (S), 10 jumpers (JP) and 12 cross-country runners (XC). Force-time curve
analysis was conducted for each (2 trials) isometric pull and averaged for analyses. IPF and RFD from 0
– 200 ms were determined from appropriate curves. IPFs were normalized using an allometric (IPFa)
scaling equation: absolute force/ (body mass (kg)0.67). ICCs were previously shown to be > 0.9.
Relationships were established with Pearson’s r; statistical differences with a Bonferroni adjusted t-Test.
IPF, IPFa and RFD were greater in males than females for S and JP but not for XC. IPF, IPFa and RFD
were greater for S and JP compared to XC. Similar relationships of IPF to RFD were noted among groups
S(r = 0.85), JP (r = 0.87), XC (r = 0.88). Among males, n = 19, the relationship of IPF to RFD was r =
0.89 and was somewhat higher compared to females, n = 15, (r = 0.78). Overall (n = 34) relationship
between IPF and RFD was r = 0.90. These data indicate a strong relationship between maximum strength
and “explosiveness” that is independent of sport training method.
TOWARD EXPRESSION OF ACTIVE HUMAN CATHEPSIN G
IN PICHIA PASTORIS
Eliot T. Smith and David A. Johnson
Dept. of Biochemistry and Molecular Biology, James H. Quillen College of Medicine, East Tennessee
State University, Johnson City, TN
Human cathepsin G (CatG) exhibits dual specificity for substrates of both trypsin-like and chymotrypsinlike proteases. Stored in cytoplasmic granules of human mast cells and neutrophils, CatG participates in
bacterial killing and inflammatory responses of the immune system. Given the difficulty and high cost of
purification, generation of a more efficient method for isolating adequate quantities of human CatG would
enable more thorough characterization. Initial attempts to express CatG in P. pastoris met with limited
success due to its highly basic character (pI ≈ 11.0). The modified strategy incorporates the gene for
human serum albumin (HSA), a more acidic protein that P. pastoris successfully secretes, into the design.
Target nucleotide sequences recognized by the restriction endonucleases EcoRI, KpnI, and SacII have
allowed successful generation of a vector encoding a fusion protein of HSA and CatG in pPICzα. This
contains a form of the CatG gene optimized for the codon usage of P. pastoris due to the yeast’s
complement of tRNAs; however, the process does not alter the amino acid sequence. PCR amplification
with extended primers allowed incorporation of EcoRI and KpnI sites onto the 5’ and 3’ ends,
respectively, of the HSA gene for subsequent insertion via restriction digestion followed by ligation. The
next step integrated a KpnI site and an enterokinase cleavage recognition sequence (DDDDK) onto the 5’
end as well as a SacII site onto the 3’ end of a CatG gene by the same methods. Sequencing data analyses
confirm the successful assembly of this construct as well as its proper positioning within the target
plasmid. Work will proceed with transfection of the plasmid construct into P. pastoris followed by
induction of expression and secretion. Purification will use a chromatography column with specific
affinity for HSA and enterokinase cleavage at the DDDDK site should remove the HSA fusion and yield
2008 Appalachian Student Research Forum Page 39 active CatG. Cleavage of synthetic peptide thiobenzyl ester substrates will verify the enzymatic activity
by producing a color change. A yeast expression system for recombinant human CatG will facilitate
further studies of this proteolytic enzyme such as its unusual dual specificity. Supported by the National
Heart, Lung and Blood Institute via grant 1R15HL091770.
THE RELATIONSHIP OF PEAK ISOMETRIC STRENGTH TO PEAK AEROBIC POWER AND 3000
M PERFORMANCE IN CROSS-COUNTRY RUNNERS
A. M. Swisher, J.M. Kraska, M.W. Ramsey, K. Painter, C. Gooden, A. Layne, M.E. Stone and M.H.
Stone.
Sport Performance Enhancement Consortium, East Tennessee State University, Johnson City, TN
Certain variables: peak aerobic power (PAP), running economy, and lactate threshold, act as limiting
factors for endurance running. However, all of the mechanisms underlying high-level endurance running
are not completely clear. Alterations in maximum and explosive strength have been shown to effect
positive changes in endurance performance, likely by altering running economy. If strength related factors
affect running economy, then this should be evident in the running performance of long-distance runners
and perhaps PAP. The purpose of this study was to examine the relationships between peak isometric
strength (IPF), explosive strength (rate of force development, RFD), PAP and 3 k performance (3KT)
among 5 female and 7 male (n = 12) X-country runners. Force-time curve analysis was conducted for
each (2 trials) isometric pull and averaged for analyses. IPF and RFD from 0 – 200 ms were determined
from appropriate curves. IPFs were normalized using an allometric (IPFa) scaling equation: absolute
force/ (body mass (kg)0.67). ICCs were previously shown to be > 0.9. Relationships were established with
Pearson’s r; statistical differences with a t-Test (p < 0.05). IPF, IPFa and RFD were not different between
sexes. PAP was statistically greater in males. Correlations were: PAP vs 3KT (r = -0.92); IPF vs PAP (r
= 0.58), 3KT (r = -0.66); IPFa vs PAP (r = 0.45), 3KT (r = 0.45). RFD vs PAP (r = 0.58), 3KT (r = -0.65).
Results indicate that strength characteristics correlate moderately to strongly with PAP and 3KT.
TOLL-LIKE RECEEPTOR MODULATION OF DENDRITIC CELL:
NATURAL KILLER CELL INTERACTION
Stacie N. Woolard, S. Nandakumar, Evelyn Kurt-Jones*, and U. Kumaraguru
Dept. of Microbiology, Quillen College of Medicine, East Tennessee State University, Johnson City, TN
* Program in Immunol. And Virology, U Mass Medical School, Worcester, MA-01655
Toll-like Receptors (TLRs) are pattern recognition receptors (PRRs) that recognize pathogen associated
molecular patterns (PAMPS) usually associated with bacteria and fungi. This early recognition results in a
rapid initiation of innate immune response that sets the course for subsequent adaptive immune response.
Herpes Simplex Virus (HSV) is one of the very few viruses that can trigger a TLR response. The
induction of Dendritic Cells (DCs) maturation by TLR ligands is considered as a pivotal link between
innate and adaptive immune responses. This renders the DCs capable of directing the immune response.
Natural Killer (NK) cells also express some of the TLRs expressed by DCs. It is our hypothesis that HSV
mediates the interaction and early signaling events of NK cell and DC via TLR. To study TLR induced
initiation between NK and DC, the DCs and NK cells were stimulated with various TLR-ligands; HSV
stimulation and un-stimulated DCs served as positive and negative controls, respectively. The DCs and
NK cells were then allowed to incubate overnight. The impact of TLR ligands on NK cells was analyzed
by the up-regulation of activation markers (CD25 and CD69) and the secretion of cytokines (IFN- ). The
enhancement of the function of DCs was studied by measuring the up-regulation of co-stimulatory
2008 Appalachian Student Research Forum Page 40 markers (CD80, CD86, MHC class I and II, CD40 etc.). To further understand the role of TLRs in
NK:DC interaction, DCs and NK cells were generated from MyD88 knock-out mice. MyD88 is the
adaptor protein used by most TLRs and its absence hampers signal transduction negating the effect of
TLR engagement. Our results indicate that Viral TLR-ligands are able to modulate the NK:DC interaction
and this is one of the many mechanisms used by these two innate immune cells to initiate a potent
acquired immune response.
Natural Sciences & Mathematics
MOLECULAR MECHANISM OF FERRICSIDEROPHORE TRANSPORT VIA OUTER MEMBRANE
RECEPTORS
Jennifer K. Cooke, Ranjan Chakraborty
Dept. of Health Sciences, East Tennessee State University, Johnson City, TN
Iron is an essential nutrient for life and growth in most single-celled and multicellular organisms.
Although it is abundant in the natural environment, iron exists mostly as insoluble iron-oxyhydroxide.
Bacteria are able to overcome this problem by several mechanisms, such as the secretion of small organic
molecules called siderophores. Siderophores chelate iron from the environment with high affinity and
transport it into the cell via complex transport system. In E. coli, an outer membrane receptor protein
known as FhuA transports ferrichrome, a fungal siderophore produced by Ustilago sphaerogena. The
crystal structures of FhuA and several such receptors have been determined; the structure consists of a 22stranded beta barrel and a plug which occludes the pore. Opening of the FhuA channel will either require
dislodgement of the plug domain or a conformational change within the plug domain to form a channel.
Our data indicates that during transport, the plug domain does not leave the barrel; rather it undergoes a
conformational change to form a channel. We used site-directed mutagenesis to change four amino acids
in the plug and barrel to cysteine in order to form disulfide bonds and tether the plug in place, preventing
it from being dislodged during transport. The conformational change may involve partial or complete
unfolding of central β-strands. In order to check whether the plug unfolds, the double-cysteine mutant
(104/149C) was constructed to form a disulfide bond connecting the β2 and β4 strands, preventing the
unfolding of the plug. This mutant shows normal binding with completely diminished transport,
indicating that this region has an important structural role. There is an arginine which is conserved in all
the receptors and in FatA, it has been shown that mutation of this arginine to alanine reduces transport
without affecting binding. In FhuA, we mutated this residue to alanine and glutamine and checked its
effect on in vivo binding and transport using radiolabeled iron. These mutants have been overexpressed in
pET21b plasmid for purification, crystallization, and structural analysis.
THE PATHWAY LEADING TO THE SYNTHESIS OF SALICYLIC ACID
IN PLANTS RESISTING PATHOGEN INFECTION
Alexander Eddo and Dhirendra Kumar
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Salicylic acid (SA) is a well known plant signaling molecule that accumulates upon plant-pathogen
interaction. This accumulation consequently corresponds to the plant being resistant to infection without
which it is susceptible. Pathogenesis-related (PR) gene expression and the synthesis of defensive
compounds associated with both local and systemic acquired resistance in plants require SA. The goal of
this study is to identify molecules that might be directly targeted to cause the SA accumulation. Primers
2008 Appalachian Student Research Forum Page 41 of genes involved in the normal biosynthesis of SA were used to perform semi-quantitative RT-PCR in
order to compare their expression levels in susceptible and resistant tobacco plants challenged with
Tobacco Mosaic Virus. Since SA does not accumulate in susceptible plants, differential gene expression
between the two lines could be responsible for the increased levels of SA. SA synthesis shares a common
substrate, Chorismate with the synthesis of aromatic amino acids. HPLC was used to measure amino acid
levels to see if the increase in SA could be due to a decrease in amino acid levels. Preliminary results
showed that genes involved in amino acid synthesis are not down-regulated although there was a decrease
in amino acid levels in both plant lines. This decrease might correspond to an increase in chorismate
levels made available for SA synthesis. We are determining the Chorismate levels to confirm this
hypothesis. Study in this area can help in the creation of resistant crop plants which can lower our
dependence on harmful herbicides and pesticides.
THE DETECTION OF POSSIBLE OUTER MEMBRANE RECEPTOR, 'FEPA', PROTEIN FROM
RHIZOBIUM LEGUMINOSARUM STRAIN IARI 312
D. Hammond Jr., B. Clarke, E. Storey, A. Windham, R.N. Chakraborty
Dept. of Health Sciences, East Tennessee State University, Johnson City, TN
Iron is an essential element for the majority of cell types, with only few exceptions. While the fourth most
abundant metal on the earth’s crust, in normal physiological conditions (biological pH and aerobic
conditions) iron is primarily found in insoluble oxyhydroxides making it unavailable to microorganisms.
The maximum amount of free unassociated iron concentration is 10-17 µM while the amount needed to
sustain microbial life is 1 µM. To combat the iron limiting environment, many gram-negative organisms
produce siderophores. Siderophores are small molecular weight molecules that scavenge ferric ions and
transport them across the cell membranes into the cell via multicomponent transport systems. Iron is
important because it serves as a cofactor for the components of the electron transport chain and a variety
of oxidoreductases. Siderophore mediated iron transport is one of the ways in which microorganisms
scavenge iron from the environment. Rhizobia form a symbiotic relationship with its host plant and fix
atmospheric nitrogen, making it available to the plant in the form of ammonia. Siderophores have been
described in a variety of Rhizobium species and are thought to play an important role in the nodulation
process. This is because the enzyme, nitrogenase, which fixes atmospheric nitrogen, contains many
atoms of iron. We have already characterized the siderophore by R. leguminosarum IARI 312. Previous
data has shown the presence of both a catechol and hydroxamate siderophore produced by IARI 312. The
Electron Spray Mass Spectroscopy (ESMS) data for the catechol-type siderophore produced by IARI 312
are consistent with the known readings of enterobactin, a siderophore produced by Escherichia coli.
SDS-PAGE analysis of the outer membrane of IARI 312 compared to FepA gene in pET17b expressed by
E. coli strain BL21 (DE3) shows the presence of a protein, expressed only in low iron conditions, with a
molecular weight equivalent to FepA. From this data, the present research is focusing on amplification of
the ‘FepA’ receptor in IARI 312. Once the gene is amplified, cloning, sequencing, and expression of the
DNA will further confirm the presence of ‘FepA’ or a homologue in the genome of IARI 312. It is
important to characterize siderophore mediated iron transport systems in Rhizobia for their agricultural
significance as well as to understand in general the mechanism of iron transport in gram-negative
organisms.
2008 Appalachian Student Research Forum Page 42 EFFECTS OF PERSULFATE OXIDATION ON MICROBIAL POPULATION
Kawalpreet Kaur, Lisa Gallagher and Michelle Crimi
Dept. of Environmental Health Sciences, College of Public Health, East Tennessee State University,
Johnson City, TN
The improper disposal of hazardous wastes has resulted in the contamination of air, water and soil and
poses a threat to human health. In Situ Chemical Oxidation (ISCO) is a rapidly growing technique for the
cleanup of contaminated soils and groundwater involving the injection of a strong chemical oxidant in the
contaminated zone to destroy the targeted contaminants. Sodium persulfate (Na2S2O8) is an emerging
oxidant that is being used. Persulfate when activated by heat, elevated pH and transition metals like Fe,
generates sulfate radicals (SO4-.), which are strong oxidants. Due to heterogeneities and low-permeability
layers common to soil, complete removal of the contaminants is not always achieved, leading to rebound
of contaminant concentrations in groundwater. Both intrinsic and engineered bioremediation applications
have the potential to degrade lower concentrations of contaminants in groundwater following an initial
reduction in contaminant mass. Therefore coupling ISCO with bioremediation can potentially effectively
remediate contaminated sites. However, there are concerns regarding the compatibility of ISCO with
persulfate and bioremediation because effects of persulfate on natural microorganisms that participate in
bioremediation are unknown. Persulfate ISCO effects, like generation of free radicals, low or high
resulting pH, and altered redox potential can affect the microbial population. Microbes like sulfatereducing bacteria (SRB) can use the sulfate produced as a byproduct of persulfate oxidation as an electron
acceptor and can degrade contaminants under anoxic conditions. Therefore batch studies were conducted
to investigate the impact of activated persulfate on soil microbial populations. Six treatment solutions
were used, including groundwater (as a control), tryptic soy broth (TSB-Microbial Supplement), low pH
(≤3 as control for low pH that can result from acidic persulfate) persulfate/Fe-EDTA, persulfate/Fe-CitricAcid and persulfate/alkaline pH (≥10.5). These treatment solutions were selected because they are
commonly used in field to activate persulfate. Soils from a petroleum contaminated site were mixed with
treatment solutions in 50ml centrifuge tubes for 48 hours and were centrifuged. Supernatant was then
removed to perform the analyses described below. In the second set of samples additional volumes of
TSB were added to the supernant and were shaked for 7 days allowing the treatment solutions to reequilibrate. This was done to study if microbial populations that are adversely affected by the persulfate
treatment will rebound when background conditions are reestablished. Three analyses, including Acridine
Orange Direct Count (AODC- which estimates biomass), Biolog® Ecoplates (which assesses the
community level physiological profile based on carbon utilization pattern) and Winogradsky columns
(microcosm analysis for SRB populations) were used on the treated soil. Results show that biomass was
not altered significantly by persulfate treatment. However, the community carbon utilization pattern was
changed, indicating that the community has shifted or that different metabolic pathways are utilized post
ISCO. SRB were present and active indicating the presence of anaerobic conditions. Results of the
second set of samples show that microbial populations rebound upon return to background conditions.
This work will be useful in coupling persulfate ISCO with bioremediation processes, which can mitigate
the cost and can be the most efficient method of remediation of contaminated sites.
2008 Appalachian Student Research Forum Page 43 MAPPING THE SKIN PATTERN OF SPOTTED SALAMANDERS,
AMBYSTOMA MACULATUM
Lok Raj Pokhrel, Istvan Karsai, Thomas Laughlin, and Rebecca Pyles
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Animals pigment patterns are one of the most obvious traits of animals and serve a variety of functions.
Pigment patterns of adult salamanders typically are produced by melanophores, xanthophores and
iridophores. In spotted salamanders (Ambystoma maculatum), the dorsal spot pattern development begins
during the transformation of the larvae to metamorphs as they leave the water. Morphological color
patterns, especially in spotted salamander, has been of great interest since the species is visually striking
with bilaterally arranged orange to yellow spots throughout the dorsum. We are studying the spotted
salamanders (A. maculatum) as a model for “spot patterns” and making an effort to understand if the
variations in the dorsal spots pattern reflect the stability of the developmental processes in its biphasic life
history. We collected 23 specimens of spotted salamanders at Osceola Island: recreational area, near south
Holston Dam, in Sullivan County, TN and took their snout-vent-length (SVL) and total length (TL). The
salamanders were photographed digitally to capture all dorsal spots, then the spots were counted and spot
area measured via ImageJ (Image analysis software). Statistical analyses were conducted to test the null
hypothesis that there is no significant relationship between left-right spot patterns; which showed that
there is significant relationship between the left-right spots patterns with P<0.05 (P=0.0001). This result
signifies that there should be some common developmental pathways for the generation of the dorsal spot
pattern which is concordant with the basic developmental mechanism of pigment pattern formation
known today. Secondly, the ratio of variances of left spot area to right spot area is not significantly greater
than 1.0 (ratio=1.74) suggesting developmental stability in the population sampled. This could also mean
that this population of spotted salamanders did not encounter significant levels of stressors during their
metamorphosis, and thus were not affected to show significant variation in spot pattern on their dorsum.
Thus, understanding the dorsal pigment pattern would help us understand developmental stability and the
habitat quality for the sustenance of the amphibian populations.
CHARACTERIZATION OF A MUTANT STRAIN OF R. ERYTHROPOLIS
THAT OVERPRODUCES SIDEROPHORE
Melanie Pratt and Bert C. Lampson
Dept. of Health Sciences, East Tennessee State University, Johnson City TN
Iron is abundant in the Earth’s crust, but in the presence of oxygen and at physiological pH, the iron is in
a form that is not soluble and therefore unavailable to the bacterial cell. Through the use of siderophores,
microorganisms are able to obtain this otherwise unusable form of iron, transport it back into the cell and
reduce it to a useful form. Rhodococcus is an environmentally important genus of the actionobacteria and
is potentially useful in bioremediation, the production of chemicals, and the removal of sulfur from oil. R.
erythropolis is known to produce a mixed ligand siderophore called heterobactin. To determine the uptake
and transport of this siderophore, I have used the plasmid pTNR to induce random mutagenesis in the
chromosome of this bacterium. The plasmid pTNR contains a transposon that can act as an insertional
mutagen when transferred into Rhodococcus cells by electroporation, randomly inserting into the
chromosome. Following mutanagenesis, mutants will be screened on CAS (Chrome Azurol S) plates to
detect the production of siderophore. DNA from non-producing and over-producing mutants will then be
extracted and fragments of chromosomal DNA will then be cloned. To do this, a Southern Blot will be
performed with DNA from over-producing strains. Genes that carry the transposon will then be analyzed
and cloned so that the DNA from the clones can be sequenced for possible transport protein genes. We
hypothesize that over-producing strains will be deficient in uptake and transport of the siderophore. To
2008 Appalachian Student Research Forum Page 44 test this, DNA recovered from the mutant strain will be sequence and analyzed to detect genes that may
code for siderophore transport proteins. Interruption of a transport gene by the transposon presumably
causes the bacteria to over-produce siderophore to compensate for a transport defect as siderophores are
important for iron acquisition and bacterial cell survival.
ANALYSIS OF SIDEROPHORE PRODUCING ABILITY OF CLINICAL AND COMMENSAL
ISOLATES OF STAPHYLOCOCCUS AUREUS
E. Presswood1, A. K. Reynolds1,S. Shukla2, R. Chakraborty1;
1
Dept. of Health Sciences, East Tennessee State University, Johnson City, TN,
2
Marshfield Clinical Research Foundation, Marshfield, WI
Staphylococcus aureus is a common cause of skin infections, pneumonia, sepsis, and other diseases in
humans. S. aureus infections are a growing problem, both in and out of the hospital. With many strains
now resistant to multiple drugs, it is necessary to find new targets and strategies for drug design. One
possible target for drug design is the siderophore mediated iron acquisition systems in bacteria. Iron is a
vital element for the growth of most microorganisms and has been correlated to the virulence of
pathogenic bacteria, including S. aureus. Many bacteria, including pathogens, produce small organic
molecules called siderophores under iron restricted conditions, that specifically chelate ferric ions and
transport them to the cell via a complex transport system. Within the human body, the availability of
soluble iron is restricted to 10-9 to 10-18 M due to the presence of several iron binding proteins, such as
hemoglobin, transferrin, and lactoferrin, used for storing and transporting iron throughout the body. This
concentration is not sufficient for the growth of pathogens. The goal of present research is to analyze a
large number of clinical and commensal isolates of S. aureus for their ability to produce siderophore(s). S.
aureus has been reported to produce at least three different types of siderophores: staphyloferrin A,
staphyloferrin B, and aureochelin. In this study we have analyzed the siderophore mediated iron
acquisition systems in 40 hospital associated, methicillin-resistant strains of S.aureus (MRSA), 30
community associated MRSA strains, and 30 commensal isolates. The majority of the isolates have been
found to produce one or more types of siderophores on Chrome Azurol S (CAS) medium. These isolates
have also been analyzed for their ability to hemolyze and acquire iron from heme. The chemical
characterization of siderophores is also under progress. Our long-term goal is to evaluate the suitability
for using the siderophore mediated iron acquisition system as a target for drug design.
SKELETAL MORPHOLOGY AND SYSTEMATICS OF LATE PLIOCENE ALLIGATOR SP
Jeremy B. Stout
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
The American alligator (Alligator mississippiensis) is one of only two species of alligator in the modern
world. It is only distantly related to the other extant species (A. sinensis), separated geographically by
approximately 11 000 km and up to 25 million years of evolution. Much closer relatives to A.
mississippiensis are known from the geologic past of North America. A disparity exists, though, in the
fossil record between A. mississippiensis and its closest relative, the late Miocene – early Pliocene A.
mefferdi. While A. mississippiensis is well known from the early Pleistocene and later, no Alligator was
known from the middle and late Pliocene until the discovery of Haile 7G, a new late Pliocene (~ 2 Ma)
site from Alachua County, Fl. North American fossil Alligator from this gap in the geologic record will
undoubtedly provide invaluable clues to the evolution and historical biogeography of the modern
American alligator lineage. This research seeks to examine the available Alligator material from this site
2008 Appalachian Student Research Forum Page 45 and apply it to the diagnostic skeletal (primarily cranial) character matrix established by Brochu, 1999
and 2004*. This will afford the specimens species-level identification and confirm one of the three
possible hypotheses: a.) that the Haile 7G alligators represent the modern A. mississippiensis, b.) that they
belong to a known fossil taxon (eg. A. mefferdi), or c.) that the specimens cannot be easily placed within
known taxa, and therefore will either become the holotype of a new species or exhibit a suite of characters
from more than one species. The character data will then be applied to a cladistic program (PAUP 4.0) to
place the fossil alligators into a phylogenetic perspective. At present, one large skull is on loan to the
ETSU Museum of Natural History and is being prepared by the author. Based on the preliminary
findings of this specimen, the available character data suggests A. mississippiensis, though the alternate
hypotheses cannot yet be ruled out.
* Poster and/or oral presentation(s) provide complete citation.
INITIAL INDENTIFICATION OF THE GENES INVOLVED IN THE BIOSYNTHESIS AND
TRANSPORT OF SCHIZOKINEN, A DIHYDROXAMATE TYPE SIDEROPHORE PRODUCED BY
RHIZOBIUM LEGUMINOSARUM IARI 917
W. Wright, D. Hammond, E. Storey, B.C. Lampson, R.N. Chakraborty;
Dept. of Health Sciences, East Tennessee State University, Johnson City, TN
Siderophores are small organic molecules produced by many microorganisms under iron deficient
conditions. They scavenge ferric ions and transport them across the cell membranes into the cell via
multicomponent transport systems. With a few exceptions, iron is a vital element for the survival of most
living cells. This is due to the fact that iron serves as a cofactor for the components of the electron
transport chain and a variety of oxidoreductases. Iron is present in abundance in earth’s crust; however,
under physiological pH and aerobic conditions it is not available to the microorganisms as it forms
insoluble ferric oxyhydroxide polymer. Siderophore mediated iron transport is one of the ways in which
microorganisms scavenge iron from the environment. Rhizobia form a symbiotic relationship with its
host plant and fix atmospheric nitrogen, making it available to the plant in the form of ammonia.
Siderophores have been described in a variety of Rhizobium species and are thought to play an important
role in the nodulation process. This is because the enzyme, nitrogenase, which fixes atmospheric
nitrogen, contains many atoms of iron. We have already characterized the siderophore by R.
leguminosarum IARI 917 to be ‘schizokinen’; in the present study we have characterized the binding and
transport kinetics of ‘schizokinen’ and have also tried to identify the genes involved in its biosynthesis.
Random mutations of strain IARI 917 were performed using mini Tn5 mutagenesis. Siderophore
overproducers and non-producers were detected on CAS (chrom azurol S) plates. DNA sequence
analysis of a non-producer mutant carrying the transposon revealed a gene similar to an alpha keto acid
reductase enzyme, which is already reported to be involved in siderophore biosynthesis besides amino
acid biosynthesis. The sequence analysis of other mutants have shown similarities to amino transferases
(may be involved in biosynthesis of schizokinen as it does contain amino groups). DNA sequence of an
overproducing mutant is presently being analyzed. The hypothesis is that the overproducer may have a
defective repressor mechanism, a faulty siderophore receptor, or a mutation in the internalization system.
It is important to characterize siderophore mediated iron transport systems in Rhizobia for their
agricultural significance as well as to understand in general the mechanism of iron transport in gramnegative organisms.
2008 Appalachian Student Research Forum Page 46 Social & Behavioral Sciences
THE VALIDITY OF SELF-REPORTED WEIGHT AMONG FEMALE COLLEGE STUDENTS
Suzanne Allen, Charlena Lau, Kristen Carlosh, & William T. Dalton, III
Dept. of Psychology, East Tennessee State University, Johnson City, TN
Investigating the validity of the self-reported values of weight allows for the proper assessment of studies
using questionnaire-derived data. The study examined the accuracy of self-reported weight in a sample of
female adults. The effects of age, race and ethnicity, depression, defensiveness, and eating disordered
behavior on the degree of discrepancy (the difference between self-reported weight and measured weight)
were considered. Methods. The analysis used data from fifty-four college age females. Self-reported and
measured weights were abstracted and analyzed according to sex, age, measured weight, self-reported
weight, and body mass index (BMI). Results. Significant differences were found among one’s reported
weight and actual weight. The weight discrepancy was positively associated with age. Eating disordered
behaviors, depression, and defensiveness may play predictive roles in gauging how inaccurate or accurate
one’s self-reported weight actually is. Conclusions. Self-reported weight is often used due to ease in data
collection. However, because of the inaccuracies posed by self-reported weight, researchers should
precede using self-report anthropometric measures with caution. Future research directions should focus
on the inclusion of attitudinal scales and other self-report data in improving prediction of actual weight.
DETECTING DIETARY RESTRAINT: A REVIEW OF THE DIS AND RRS
Suzanne Allen, Kristen Carlosh, Charlena Lau, and William T. Dalton, PhD
Dept. of Psychology, East Tennessee State University, Johnson City, TN
There has been much controversy regarding the validity of the Revised Restraint Scale and the Dietary
Intent Scale with regards to identifying restrained eaters. The current study examines whether these two
scales yield parallel results pertaining to binge eating behaviors and body mass index. Introduction.
Both the Revised Restraint Scale (RRS) and the Dietary Intent Scale (DIS) were created in an effort to
identify a restrained eater, one who attempts to restrict their intake of food in general, and/or of
“fattening” foods in particular. For example, the following questions can be seen on the Revised
Restraint Scale (Herman & Polivy, 1980) “Do you have feelings of guilt after overeating?” and “How
conscious are you of what you are eating?” On the basis of previous research and for the purposes of this
study, those individuals scoring 15 or higher on the Revised Restraint Scale will be classified as
restrained eaters and those scoring lower than 15 will be classified as unrestrained eaters. The Dietary
Intent Scale (Stice, 1998) is a nine-item scale measuring dietary restraint over the last six months. Based
on a five-point scale ranging from “Never” to “Always”, participants answered items coordinating with
the frequency of their behavior. Such questions included on this scale are “I often skip meals in an effort
to control my weight” and “I eat low-calorie foods in an effort to avoid weight gain.” Because both scales
were created to identify dietary restraint among individuals, it is presumed that an individual would yield
parallel scores on both questionnaires. Also, if both scales measure the same construct, similar
correlations should be found on other related scales (e.g., Binge Eating Scale). Thus, the current study
examined whether the Revised Restraint Scale and the Dietary Intent Scale yielded similar results in
defining dietary restraint and correlating with other co-morbid characteristics (i.e., binge eating, body
mass index). Design. Fifty-four female undergraduate students were given both the DIS and RRS, along
with the Binge Eating Scale (BES). Additionally, height and weight measurements were taken for each
participant. Pearson’s correlations were then used to determine the relationship among aforementioned
2008 Appalachian Student Research Forum Page 47 variables. Of the 54 participants measured for height and weight, 8 were in the range of obesity, while
11 were considered overweight, 33 were considered of normal weight, and 2 were considered
underweight for their height. More importantly for this study, 24 of the participants were categorized as
restrained eaters by the RRS, while 20 were categorized by the DIS. Results. Significant differences
were found between the RRS and DIS. The RRS correlated positively with the BES. However, such
relationship was not found with the DIS. Perhaps more intriguingly, when compared to the participants’
BMI, the RRS was positively correlated, while the DIS had a negative correlational distribution. This
may indicate that the two scales measure two different and distinct groups. Further implications will be
discussed.
VOICE ONSET TIME AS A CLINICAL INDICATOR OF HYPOFUNCTIONAL VOICE DISORDERS
Amanda Arnold, Lisa Phillips, Lindsay Pickler, Whitney White, and Amanda McCamey
Dept. of Communicative Disorders, College of Clinical and Rehabilitative Health Sciences, East
Tennessee State University, Johnson City, TN
The purpose of this study was to measure and compare the voice onset times (VOTs) of healthy
individuals using a normal and breathy voice in an effort to determine if VOT can be used as a noninvasive clinical indicator of laryngeal function. Recordings were made of 20 adults between the ages of
20-48 with normal laryngeal function, each using a normal (Group 1) and breathy voice (Group 2). The
participants’ productions were designed and collected in such a manner to control for speaking rate,
vowel context, pitch, and loudness; all of which have been shown to influence VOT. A mixed analyses of
variance showed that hypofunctional productions demonstrated longer VOTs across all stop consonants
when compared to normal productions. Within the stops, a significant difference between the voiced and
voiceless stops was noted, although no gender differences were found. It was concluded that VOT can be
used as an indirect clinical indicator of laryngeal function.
TOUCHING, ORAL, AND PENETRATION:
DOES OFFENSE TYPE MAKE A DIFFERENCE?
Brandon C. Bogle, Chris S. Dula, Robin L. Leonard
Dept. of Psychology, East Tennessee State University, Johnson City, TN.
There is an extensive body of literature in the field of sex offender research. However, there is a lack of
research examining the attitudes of the general public toward sex offenders and their offenses. Current
literature indicates that the attitude of the general public towards sex offenders is highly negative (Nelson,
Herlihy, and Oescher, 2002). Along with these negative attitudes, are misconceptions that have been
generalized about these offenders. Federoff and Moran (1997) identified 9 falsely held myths commonly
generalized to all sex offenders. If the public holds staunch and unwarranted negative attitudes toward sex
offenders, there is little hope for released sex offenders to successfully reintegrate into society and thus,
less informal monitoring of former offenders and little incentive to prevent re-offending. However, there
is little information as to what aspects of sex offenders are most stigmatized or to what degree prejudices
might vary toward sex offender. The current project is a continuation of the research examining the
public’s attitudes toward sex offenders and sex offenses. The primary objective of the current project is to
determine if the type of sex offense has an effect on the attitudes toward sex offenders on three distinct
constructs: punishment, perversity, and severity. Data were subjected to a Wilks’ Lambda repeated
measures MANOVA. F (2, 418)= 31.789, p<.001, ηp2= .288. Least Significant Difference Post Hoc Test
(Pairwise Comparisons) yielded significant interaction effects between Touching, Oral, and Penetration
2008 Appalachian Student Research Forum Page 48 offenses on punishment ratings F(6, 204)= 48.871, p<.001, ηp2= .590. The results of this study indicate
that offense type does indeed make a difference in attitudes toward the offense. The three types of
offense, touching, oral, and penetration are distinct, yet touching seems to differentiate from the other two
offenses. It may be that oral and penetration offenses are intrusive in nature whereas the touching offense
is generally of an external nature and is therefore not viewed as negatively as the other two offenses. The
correlation coefficients between the three aspects of attitude, punishment, perversity, and severity are so
high that they seem to indicate co-linearity, which may indicate that they are not distinct constructs and
couch a single construct. The use of traditional college students may limit generalizability to the rest of
the general population. In addition, the sample was taken from a culturally unique region of the U.S.
Future research will examine the effects of various constructs on attitudes toward sex offenders and sex
offenses. These factors include gender, age, victim consent, offender status, victim status, and offender
attractiveness. The authors also plan to further validate the Bogle Sex Offender Scale (BSOAS).
AMPHETAMINE SENSITIZATION IN A RODENT MODEL OF PSYCHOSIS.
Zackary A. Cope1, A. Brianna Sheppard1, Ian D. Longacre1, Marla K. Perna1, Kimberly N. Thompson2,
David S. Roane3, Russell W. Brown1
1
Dept. of Psychology, College of Arts and Sciences, East Tennessee State University, Johnson City, TN
2
Dept. of Anatomy and Cell Biology, Quillen College of Medicine, East Tennessee State University,
Johnson City, TN
3
Dept. of Pharmaceutical Sciences, College of Pharmacy, East Tennessee State University, Johnson City,
TN
This study was designed to analyze the effects of amphetamine on locomotor sensitization in a rodent
model of psychosis developed in our laboratory. Past studies have shown neonatal quinpirole (dopamine
D2/D3 agonist) produces a significant increase in dopamine D2 receptor sensitivity that persists into
adulthood, a phenomenon known as D2receptor priming. An increase in D2receptor sensitivity is
consistent with several behavioral disorders, including schizophrenia. Rats were administered quinpirole
(1 mg/kg) or saline from postnatal days 1-11 and raised to adulthood (postnatal day 60). Beginning in
adulthood, rats were administered d-amphetamine sulfate (1 mg/kg) or saline every other day for 14 days,
resulting in a total of seven exposures to the drug. Approximately 10 min after drug injection, rats were
placed in a locomotor arena and overall activity was analyzed. Results showed that D2-primed rats
receiving amphetamine demonstrated a significant increase in locomotor activity across all seven days of
testing relative to all other groups. Controls receiving amphetamine also demonstrated a significant
increase in activity over days, demonstrating sensitization. Interestingly, D2-primed rats given saline also
demonstrated sensitization to the locomotor arena, but this is consistent with past observations in our
laboratory and suggests a lack of ability in these animal to habituate to the environment. Seven to fourteen
days after locomotor sensitization testing was complete, cerebrospinal fluid samples were taken via
microdialysis from the nucleus accumbens core to be analyzed for dopamine levels, and these samples are
currently being assayed. Supported by NIH grant 1 R15 DA 020481-01 to RWB
2008 Appalachian Student Research Forum Page 49 THE IMPACT OF GRAMMATICAL COMPLEXITY ON THE PRODUCTION
OF SENTENCE DISRUPTIONS
Tessie Burke, Carol Davidson, Karen Mumpower, Katie Sims, and Kerry Proctor-Williams, PhD. CCCSLP
Dept. of Communicative Disorders, College of Clinical and Rehabilitative Health Sciences, East
Tennessee State University, Johnson City, TN
Research suggests that as grammar develops the frequency and type of sentence disruptions a child
produces will change as a reflection of grammatical complexity (Rispoli, 2003). Types of sentence
disruptions include stalls and revisions. Stalls are sentence disruptions that include repetitions, fillers, and
pauses that reflect glitches in performance. Stalls buy the speaker time to produce the utterances.
Revisions are sentence disruptions that reflect self-correction due to increased language complexity.
Revisions are corrections of utterances produced that do not match the speaker’s intention. The rate of
stalls should remain stable or slightly increase, and the rate of revisions should increase as sentence length
increases for the child in the midst of grammatical development. Our study investigates the relationship
between utterance length and complexity and sentence disruption among preschool children. It examines
the spontaneous language samples collected during story re-tell and story generation tasks from 10
typically developing children ages 3;2 to 4;11 (years; months). The analyzed samples were controlled for
length to include only utterances of 4, 5, 7, and 8 words in length. The results show that, overall, stalls
occurred more often than revisions regardless of utterance length. It was also found that there was no
main effect for length; stalls and revisions were not more likely to occur in longer utterances. However,
there was a trend in this direction. Further, there were no observed correlations between measures of
language development and rate of disruptions. These results supported previous findings that children
produce sentence disruptions while grammatical complexity develops, in contrast to earlier studies,
however, their level of language development did not predict their disruption rate. This reveals that the
relationship between production of disruptions and complexity is intricate. The current findings suggest
that the frequency of disruptions produced during grammatical development peaks and then begins to
level off as language development reaches a certain point. Once the child has reached a comfort level with
longer and more complex sentences, disruptions even out and the relationship is less predictable.
RURAL ELEMENTARY AND MIDDLE SCHOOL PERSONNEL PERCEPTIONS
OF STUDENT NUTRITION, PHYSICAL ACTIVITY, AND OBESITY
Lauren LaBounty and Karen Schetzina M.D., MPH,
Dept. of Pediatrics, College of Medicine, East Tennessee State University, Johnson City, TN
Background: The percentage of children with obesity and type II diabetes in the United States has
increased over the past two decades, particularly in the rural Southeast. Schools are a proposed setting for
prevention initiatives. Purpose: The purpose of this study was to gain a better understanding of school
personnel perceptions of child obesity and diabetes prevention. This understanding will facilitate a unique
collaboration between a regional university, a hospital system, community organizations, and schools in
making healthy changes in policy, curricula, and environments in elementary and middle schools. The
approach will be modeled after the Coordinated School Health Program (CSHP). Methods: The study
was approved by the university Institutional Review Board. Key personnel from six school systems in
Northeast Tennessee were invited to participate in a written survey that included questions about
nutrition, physical activity, and obesity. Results: All school systems participated, with a total of 32
administrators and teachers completing the survey. Sixty-five percent of respondents believed that the
amount of student physical activity at their school was too little. All respondents felt that to be healthy,
2008 Appalachian Student Research Forum Page 50 students needed a total of 30 minutes of physical activity at school each day in every grade level. Eightyfour percent of respondents thought that the nutritional quality of foods and drinks consumed by students
at school was fair, good, or very good. Most teachers did agreed that schools should not sell unhealthy
snacks as fundraisers and that classroom parties should include mostly healthy foods and drinks. Eightytwo percent of personnel were either extremely or very concerned about child obesity in their school
community. Over ninety percent agreed that schools should do more to promote physical activity and
healthy nutrition among both children and teachers. All respondents felt that parents needed to be more
involved in this effort with their children. Conclusion: School personnel from this Northeast Tennessee
sample were concerned about child obesity and seemed willing to implement changes in their schools to
promote physical activity and healthy nutrition. The next phase of this study will test whether a program
modeled after and implemented in conjunction with Tennessee CSHP is effective in producing changes in
school policies, curricula, and environments that will promote physical activity and healthier nutrition.
FAF RESEARCH
Dr. Vijayakumar Guntupalli, Jenny Gilbert, B.A., Lauren Laethem, B.A.A., Lashondra Phillips, B.A.,
Cody Robinson, B.S., Angie Travers, B.A.
Dept. of Communicative Disorders, East Tennessee State University, Johnson City, TN
The purpose of this study was to determine the effect of frequency-altered auditory feedback (FAF) on the
frequency of stuttering events during conversation. Two male children (ages 11 and 10 years) who stutter
participated in this study. An ABAB time-series design was used. Each participant held spontaneous
conversation under both non-altered auditory feedback (NAF) and FAF conditions. During FAF
condition, participant’s speech was routed through a digital signal processor that shifted the frequency of
speech signal either up or down by half octave before being presented to participant ears via insert ear
phones. Frequency of stuttered syllables and number of syllables produced per minute were recorded
during baseline and treatment conditions. Both participants exhibited significantly more disfluencies
under NAF conditions when compared to FAF. Participant 2 showed a significant decrease in stuttering
under the second interval of NAF conditions compared to 1. Other research and theories about the
relationships between FAF and NAF on stuttering related to this treatment are discussed.
2008 Appalachian Student Research Forum Page 51 Division III – Graduate Students
2+ Years
Biomedical Sciences, Natural Sciences & Mathematics,
and Social & Behavioral Sciences
COMPARISON OF THE ACTIVITY AND EXPRESSION OF THE ZMPSTE24 ENDOPROTEASE IN
CELLULAR QUIESCENCE
Christina Bridges and Antonio Rusiñol
Dept. of Biochemistry and Molecular Biology, College of Medicine, East Tennessee State University,
Johnson City, TN
The nuclear lamins are a family of intermediate filament proteins that form a thin, fibrous scaffold lining
the inner nuclear membrane, thereby providing structural rigidity to the nucleus (Goldman RD 2002).
One member of this family, the 72 kDa Lamin A, is initially synthesized as a 74kDa precursor, prelamin
A.
This precursor undergoes an unusual maturation pathway in which two endoproteolytic cleavages
occur (Beck 1990). The first cleavage can be carried out by the endoproteases Zmpste24 or Rce1. The
second cleavage is unique to lamin A in higher vertebrates, and is specifically carried out by ZMPSTE24
(Corrigan DP 2005). Two human genetic diseases, Hutchinson-Gilford Progeria Syndrome (HGPS) and
Restrictive Dermopathy (RD), arise from defects in this endoproteolytic processing pathway. Though
previous work from our laboratory has provided the insight into the processing pathway of prelamin A, as
described above, the functional significance of the pathway remains puzzling. Related literature
(Ukekawa R 2007, Gruber J 2005) indicates a role for accumulation of Prelamin A in quiescence, where
ZMPSTE24 is down-regulated (Ukekawa R 2007, Gruber J 2005), and thus not available to process the
Prelamin A to maturation. Additionally, cells that do not express A/C lamins (Mattia E 1992, Stewart C
1987, M. B. Broers JL 1997) lack a quiescent state. Cells that do express the A/C lamins are typically
able to enter quiescence under conditions of serum starvation or contact inhibition. To compare the
activity of Zmpste24 with cell cycle stage, we induced quiescence in HeLa cells by serum starvation and
measured the rate of Zmpste24 activity compared to the rate of DNA synthesis. For this comparison, we
labeled cells with [3]H-Thymidine and monitored incorporation as an indicator of the rate of DNA
synthesis over a serum starvation time course of 30-32 hours, and measured the rate of Zmpste24
cleavage of its Prelamin A substrate over the same time course. The results demonstrate a rapid posttranslational down-regulation of activity of Zmpste24 in quiescence that mimics the pattern of
downregulation of DNA synthesis as cells enter quiescence, as well as a rapid increase in activity as
serum is returned and DNA synthesis levels increase. We also measured the expression levels of
Zmpste24 by Western Blot and Northern Blot over a 5 day serum starvation time course, and found levels
to be relatively unaltered until the fifth day. These results together suggest that there is a rapid posttranslational down-regulation of activity of Zmpste24 in quiescence that is independent of the slower
regulation of its synthesis. To further investigate, we overexpressed Zmpste24 by transfection into
fibroblast cells, plated at high density to induce quiescence, and monitored BrdU incorporation as an
indicator of DNA synthesis. The results demonstrate a likely ability to bypass the quiescent state when
ZMPSTE24 is overexpressed. This implies the potential that down-regulation of Zmpste24 is, thereby,
required to enter or maintain quiescence.
2008 Appalachian Student Research Forum Page 52 GENETIC SUSCEPTIBILITY TESTING FOR BERYLLIUM:
WORKER KNOWLEDGE, BELIEFS, AND ATTITUDES
James Gorniewicz, Dr. Ken Silver, and Dr. Gary Kukulka.
Depts. of Psychology, Environmental Health, and Family Medicine, East Tennessee State University,
Johnson City, TN
Since the 1940s, thousands of nuclear energy workers have been exposed to beryllium, a metal which
causes a potentially fatal autoimmune mediated disorder known as chronic beryllium disease (CBD).
Genetic testing may provide exposed employees with better information about their individual chances of
developing CBD and thus assist them in making informed choices regarding current employment and
future medical treatment. Collecting genetic information in the workplace raises a variety of legal,
ethical, and social issues. Many individuals fear genetic testing due to concerns over confidentiality,
employability, and insurability. Therefore, we wanted to gain a better understanding of what nuclear
energy workers thought regarding these important issues and genetic testing. Specifically, we wanted to
know more about workers’ beliefs and attitudes concerning ideal ways of collecting genetic information
in the workplace so as to ensure confidentiality and avoid the possibility of genetic discrimination from
employers or insurers. We conducted three focus groups with former and current energy workers in
northern New Mexico. Each group was comprised of five to eleven individuals who were exposed to
beryllium, diagnosed with CBD, or tested positive for beryllium sensitivity. Open-ended questions were
asked on a variety of topics. Focus group sessions were audio taped, transcribed, and then independently
analyzed using grounded theory methodology and NVivo7 qualitative data analysis software. Results of
this work identified: 1) specific domains where increased worker education may be beneficial (genetic
testing, genetic susceptibility, etc.), 2) numerous worker concerns relating to confidentiality, and 3)
worker recommendations for the successful implementation of future genetic testing programs.
Understanding the unique insights provided by energy workers could lead to improved employee
education as well as improved health outcomes for workers and their families. Findings from this study
will be used to suggest priority issues in the development of policies regarding genetic testing and will be
presented to organizations that represent the interests of energy employees. Findings from this study
could be used to better educate primary care clinicians regarding the recognition and management of
CBD and patient concerns - particularly on complex issues related to the advisability and interpretation of
genetic tests. Results of this study may also be used to develop an in-depth questionnaire for a larger
subsequent study of genetic testing.
VITAMIN E (TOCOTRIENOLS) AND PROSTATE CANCER:
A PROTEOMICS APPROACH
Christian Mbangha Muenyi
Dept. of Chemistry, East Tennessee State University, Johnson City, TN
Proteomics is the large scale study of proteins in cells or organisms. The purpose of this study was to
characterize the proteomic alterations occurring in a prostate cancer (LNCaP) cell line after treatment
with delta-tocotrienol (a form of vitamin E not very prevalent from most dietary sources). We found that
both gamma- and delta-tocotrienols induced time and concentration dependent growth inhibition and
programmed cell dead (apoptosis) in LNCaP cells. Secondly, we used two-dimensional gel
electrophoresis (2-DE) to characterize changes in protein expression levels associated with this treatment.
Our results show that a specific set of proteins are regulated at both early and late times following
treatment with delta-tocotrienol and these proteins have been characterized by their apparent molecular
weights and isoelectric points. The alteration observed at early time points are particularly interesting
2008 Appalachian Student Research Forum Page 53 because these changes are likely to reflect the underlying molecular mechanisms for triggering cancer cell
death.
A CANNABINOID (WIN 55,212-2) INDUCES APOPTOSIS
IN PANCREATIC TUMOR CELLS VIA CB1/CB2-DEPENDENT
AND INDEPENDENT MECHANISMS
Theresa Pickle and Douglas Thewke
Dept. of Biochemistry and Molecular Biology, East Tennessee State University, Johnson City, TN
Pancreatic cancer is the fourth leading cause of cancer related deaths. In 2008, an estimated 37,170
Americans were diagnosed with cancer of the pancreas and an estimated 33,370 Americans died of
pancreatic cancer. 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 Δ9-tetrahydrocannabinol (THC), the
active agent of Cannabis sativa, exhibit anti-tumor properties. 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 RT-PCR 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 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 17-AAG, 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. Caspase-3 activity assays revealed
that the growth inhibitory effects of Win 55,212-2 involve induction of apoptotic mechanisms. The
induction of caspase-3 activity by Win 55,212-2 in Mia-PaCa2 cells was prevented by a CB1 receptorspecific antagonist, AM251, but not by a CB2 receptor-specific antagonist, SR144528, suggesting the
mechanism of apoptotic induction in Mia-PaCa2 cells is mediated by CB1. However, AM251 had no
effect on the induction of caspase-3 activity in AsPC1, but there was a reduction in caspase-3 activity in
the presence of SR 144528, suggesting the mechanism of apoptotic induction in AsPC1 cells is mediated
by CB2. In CaPan1, Am 251 and SR144528 had no effect on the induction of caspase-3 activity. In
conclusion, the results presented here 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.
THE HOST ADHERENS JUNCTION MOLECULE NECTIN-1 IS DEGRADED BY CHLAMYDIAL
PROTEASE/PROTEASOME-LIKE ACTIVITY FACTOR (CPAF) IN CHLAMYDIA TRACHOMATISINFECTED GENITAL EPITHELIAL CELLS
Jingru Sun, Robert V. Schoborg
Dept. of Microbiology, College of Medicine, East Tennessee State University, Johnson City, TN
Chlamydia trachomatis, an obligate intracellular human pathogen, is one of the leading causes of sexually
transmitted disease and the most common cause of preventable blindness worldwide. Previous studies in
2008 Appalachian Student Research Forum Page 54 our laboratory have demonstrated that accumulation of the host cell adhesion molecule nectin-1 is
significantly decreased in C. trachomatis serovar E-infected cervical epithelial cells. This downregulation requires active C. trachomatis protein synthesis and replication. RT-PCR analyses indicate that
chlamydial infection does not alter accumulation of any nectin-1 transcripts, demonstrating that nectin-1
is reduced at a post-transcriptional level. In the present study, Western blot analyses show that nectin-1
down-regulation is C. trachomatis concentration-dependent. In addition, the half life of nectin-1 is much
shorter in C. trachomatis-infected epithelial cells than that observed in mock-infected cells, indicating
nectin-1 is likely down-regulated post-translationally. The chlamydia-secreted protease CPAF degrades
several important host proteins, such as USF-1, RFX5, Keratin 8 and pro-apoptotic BH3-only proteins.
Our data have demonstrated that the timing of CPAF expression within infected cells correlates with the
infection time-dependent cleavage of nectin-1. Previous studies by Zhong et al. (2000) have shown that
lactacystin inhibits the proteolytic activity of CPAF while MG132 does not. Our in vivo inhibition
experiments using these two inhibitors have demonstrated that down-regulation of nectin-1 is inhibited by
lactacystin but not by MG132. Finally, a cell-free cleavage assay has shown that nectin-1 is degraded by
the functional recombinant GST-CPAFwt protein but not by the inactive CPAF mutant GST-CPAFD502A.
Degradation of nectin-1 in vitro by recombinant GST-CPAFwt is also inhibited by lactacystin, as was
previously observed in vivo. Taken together, our results suggest that the host adherens junction molecule
nectin-1 is degraded by CPAF in C. trachomatis-infected cells. Since nectin-1 is crucial for formation of
adherens junctions, tight junctions and cell adhesions, nectin-1 down-regulation by CPAF could weaken
contacts between an infected genital epithelial cell and adjacent, uninfected cells. This would allow
release of the intact, infected cell into genital tract lumen, facilitating chlamydial dissemination and
causing chronic, ascending genital tract infections.
INVESTIGATING NECTAR RHYTHMS IN SQUASH (CUCURBITA PEPO): EFFECTS ON
HONEYBEE (APIS MELLIFERA) FORAGING BEHAVIOR
Sam Boyd, Jennifer N. Johnson, Byron N. Van Nest, Darrell Moore,
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
It has been shown that honeybees are able to learn and anticipate the time in which flowers present nectar.
Foragers are able to anticipate the time and location of the sucrose reward after only one day of training.
We have discovered that when the food reward is removed, this time-memory for individual foragers
shows an apparent extinction over several days. However, following a rain event, a greater than expected
number of foragers return to the feeding station, showing that extinction is delayed. The present study
investigates whether this apparent delay in extinction is due to an adaptive response of the bees to an
increase in nectar quality (concentration and/or volume) following a rain event. To test this hypothesis,
we used squash plants (Cucurbita pepo) grown in a greenhouse at ETSU (Johnson City, TN) under
controlled watering conditions. The plants show a remarkably consistent daily rhythm in nectar secretion:
the sugar concentration is highest early in the day, typically 37% (6:00 AM), and diminishes to about
18% just before the flowers close (12:00 PM). Nectar volume, on the other hand, increases during this
period. Volume starts at less than 2ml (6:00 AM) but reaches 22ml by noon. Based on these findings, the
overall objective of this work is to test the effect an increase (mimicking a rain event) or decrease
(mimicking drought) in water availability will have on the nectar rhythm for volume and concentration.
We hypothesize that (1) a simulated rain event will cause an increase in the nectar volume produced, and
(2) the rain event will cause an increase in nectar concentration. Both improvements in nectar quality
increase the profitability of the nectar for the honeybees, therefore increasing foraging intensity. Two
greenhouse experiments (GH1 and GH2) were conducted. GH1 consisted of 3 days of controlled
watering (baseline) before a simulated rain event (2x baseline), followed by three additional days of
baseline watering. GH2 included an 8-day simulated drought immediately preceding the rain event. In
both experiments, nectar was collected daily, in 90-minute intervals, while the flowers were open. At
2008 Appalachian Student Research Forum Page 55 each sample time, nectar volume and concentration for 6-8 randomly chosen flowers were recorded.
These values, as well as time of day and experimental day were used to perform a GLM statistical
analysis (using experimental day to represent watering amount). The results of GH1 show that an
increase in water did not cause a change in volume but did cause a decrease in the concentration on the
day following the simulated rain day with respect to the first three days. The only significant changes for
GH2 were between the volume of the first day and the last three days, possibly due to the deteriorating
health of the plants. Interestingly, both the volume and concentration rhythms were maintained
throughout both experiments, despite the drastic changes in water, indicating the robust nature of the daily
nectar rhythm. However, these results do not explain the delay in honeybee time-memory extinction
following a rain event. Further studies using other plants are needed.
APPLICATION OF MULTIVARIATE STATISTICAL ANALYSES
TO MICROBIAL WATER QUALITY PARPAMETERS IN FOUR GEOGRAPHICALLY SIMILAR
CREEKS IN NORTHEAST TENNESSEE
TO IDENTIFY PATTERNS ASSOCIATION LAND USE TO FECAL POLLUTION
K.K. Hall, B.G. Evanshen, K.J. Maier and P.R. Scheuerman,
Dept. of Environmental Health, College of Public Health, East Tennessee State University, Johnson City,
TN
The addition of surface water bodies to impaired waters (303D) lists for pathogen impairment and the
need to address these through the Total Maximum Daily Load (TMDL) process has resulted in increased
research to find methods that effectively and universally identify fecal pollution sources. A fundamental
requirement to identify such methods is understanding the land use, and microbial and chemical processes
that influence fate and transport of fecal indicators from various sources to receiving streams. To better
understand current water quality and predict water quality changes as development occurs, we initiated a
water quality monitoring program. Four creeks within the Watauga River watershed in Northeast
Tennessee are routinely monitored using a targeted sampling approach to determine the extent and
sources of fecal pollution. These creeks are monitored based on the existence of pathogen TMDLs or the
expectation that they will require pathogen TMDLs. Boones, Cash Hollow and Sinking Creek are listed
on the State of Tennessee’s 303D list due to pathogen loading, but only Cash Hollow and Sinking Creeks
have TMDLs. Buffalo and Sinking Creek have similar patterns of fecal coliform loading, suggesting that
TMDL development may require multi-year data using a targeted sampling approach instead of a 30-day
geometric mean. Boones and Cash Hollow Creek have elevated fecal coliform concentrations (1115 and
733 CFU/100ml, respectively) but different land use patterns. Agricultural land use impacts Boones
Creek, while surface runoff and land development impact Cash Hollow Creek. Previous multivariate
statistical analyses have demonstrated that the microbial and chemical parameters influencing water
quality differ between creeks, further suggesting that land use patterns influence microbial contamination.
The objective of this research is to update results of previous reports and apply multivariate statistical
techniques to identify common patterns associating land use to monitored water quality parameters and
various pollution sources. Canonical correlation and canonical discriminatory analyses demonstrate that
sources of impairment are related to land use patterns in the watershed. The application of multivariate
statistical methods to water quality data has helped to identify common patterns associating land use to
monitored water quality parameters and various pollution sources. Combined with a targeted water
quality monitoring program, this data analysis approach is a useful method to identify sources of
impairment and to identify Best Management Practices that will prevent and remediate the effects of rapid
urbanization in the Watauga River watershed.
2008 Appalachian Student Research Forum Page 56 HETEROLOGOUS EXPRESSION AND ELUCIDATION OF BIOCHEMICAL FUNCTION OF TWO
PUTATIVE FLAVONOID GLUCOSYLTRANSFERASE CLONES (PGT2 AND PGT3) FROM
CITRUS PARADISI
M.V.K.P Siddhartha, Daniel K. Owens and Cecilia A. McIntosh
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
Flavonoids are phenolic compounds found naturally in plants and are one of the major classes of plant
secondary metabolites. To date there are 6000 naturally occurring flavonoids in the plant kingdom and
they play a variety of roles in protecting plants against UV light, acting as pigments, for instance
anthocyanins a class of flavonoids, impart red and blue colors to fruits such as strawberries, grapes,
blueberries, and apples and some provide characteristic flavors. Flavonoids are also known for their
antioxidant and antimicrobial properties. Flavonoids have 9 major subclasses synthesized through a core
pathway. 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. Derivatization reactions include hydroxylation, methylation, sulfation, acylation,
methoxylation, glycosylation, and prenylation and Glycosylation is a predominant modification reaction
which includes addition of sugars to flavonoids and these reactions are catalyzed by glucosyltransferases
(GTs). Glycosylation evident in citrus paradisi where 40-70% of the dry weight of young leaf and fruit
tissues is composed of the bitter flavanone diglycoside, naringin.The research focus of our lab is on
regulation of flavonoid metabolism specifically on glycosylation and the GTs catalyzing these reactions.
Citrus paradisi was chosen as a model system as it is known for accumulation of glycosides of several
flavonoid subclasses including flavonone 7-O-glycosides, flavones 7-O-glycosides, flavonol 3-Oglucosides, and chalcone glycosides, flavonol 7-O-glcuosides and the enzymes that are responsible for
these reactions. Current efforts are focused on cloning, expression and characterization of putative
glycosyltransferases from grapefruit. Currently we have 8 putative GTs in hand. The research described
here is designed to test the hypothesis that grapefruit PGT2 and PGT3 clones are flavonoid
glucosyltransferases. PGT2 and PGT3 clones have been cloned into pCD1 vector and transformation has
been achieved using BL21 cell lines. Heterologous expression will facilitate the expression of soluble
proteins. We present here the strategies that are being used for optimizing the expression of soluble
protein such as testing media composition, culture temperatures, and induction duration. Once the
expression of soluble protein is optimized, expressed protein will be screened for flavonoid GT activity
using different substrates. If activity is found, the expressed enzymes will be further characterized.
Characterization will include elucidation of the reaction products, substrate specificity, optimum pH,
activators, inhibitors, isoelectric point, as well as kinetic parameters. Information on efforts to obtain
additional putative GT clones from grapefruit will also be presented.
NORMATIVE SPEECH ACCURACY MEASURES
OF CHILDREN 24-30 MONTHS OF AGE
Marie Fillers, B.S., Amanda Cox, B.A., Amanda Mershon, B.S., and Nancy Scherer, Ph.D.
Dept. of Communicative Disorders, East Tennessee State University, Johnson City, TN
The availability of normative guidelines for early speech development is limited. One measure of speech
accuracy that has gained popularity is Percent Consonants Correct-Revised (PCC-R); however, there is
limited normative data for children under three years of age. Campbell, Dollaghan, Janosky, and Adelson
(2007) used a curve fitting approach to interpret PCC-R data from some existing studies, although the
data at the early ages was limited. The purpose of this descriptive study was to compare PCC-R of typical
24 and 30 month olds to the performance curves developed by Campbell et al. Method: Participants – The
sample included 28 typically developing children, 14 (7 males and 7 females) in the 24-month age group
and 14 (8 males and 6 females) in the 30-month age group. Mean MLU scores for the 24-month group
2008 Appalachian Student Research Forum Page 57 was 1.8 while the 30-month group was 2.5. Procedures – A 100-utterance spontaneous speech sample was
collected using three theme-based play sets containing a controlled number of consonants in initial and
final word positions. Each sample was video recorded, transcribed phonetically by two graduate students
of Speech-Language Pathology at East Tennessee State University, and analyzed for PCC-R and
consonant inventory. Inter-rater transcription reliability on 17% of the sample, was calculated to be
90.5%. Results: Consonant inventory and PCC-R scores were compared between the 24 month and 30
month age groups using t-tests. The consonant inventories and PCC-R scores were statistically larger at
30 months with p<0.05. The expansion in consonant inventories indicated that, by 24 months, nasals (e.g.,
m, n), glides (e.g., w, j), and stop consonants (e.g., p,b,t,d,k,g) have generally been mastered. By 30
months inventory increases consisted of primarily fricatives (e.g., f, s, z, sh) with an increase from 3.1 to
4.5 at 30 months of age. PCC-R scores from this study were then compared to the Campbell et al.
performance curve using t-tests. The data collected in the present study were not statistically different
from the Campbell et al. performance curves (24 months, df= 13, t =-1.5, p=0.15, and 30 months df = 13, t
=-0.56, p=.59). The data in this study support the validity of the performance curves at both 24 and 30
months of age. Discussion: This study provided validation of PCC-R performance curves developed by
Campbell et al. at two age points under three years of age. This finding provides additional support for the
use of the performance curves in interpreting speech accuracy measures during speech assessment of
children under three years of age. While more data should continue to be collected to validate other age
ranges, the performance curves appear to provide accurate normative guidelines for comparison of speech
impairments that will be helpful for speech-language pathologists during their assessments of very young
children. These norms may facilitate early identification of children with speech impairments and
subsequent referrals for early intervention.
2008 Appalachian Student Research Forum Page 58 Division IV – Medical Students
EFFECTS OF CIGARETTE SMOKE EXTRACT ON CYTOKINE PRODUCTION
FROM IL-1BETA-ACTIVATED MAST CELLS
Thomas Soike, Ta-Chang Lin, Kenton Hall, Zong Doa Wu, Yu-Yin Liu, Jonathan Moorman, Guha
Krishnaswamy, and David S. Chi
Dept. of Internal Medicine, East Tennessee State University, Johnson City, TN, 37614
Dept. of Environmental Engineering, National Cheng Kung University, Tainan, Taiwan
Human mast cells are multifunctional cells capable of a wide variety of inflammatory responses and
associated with allergy, asthma, and atherosclerosis. Our previous studies have shown that IL-1β
activates human mast cells to produce selected inflammatory cytokines. We examined effects of cigarette
smoke extract (CSE) on the production of inflammatory cytokines from IL-1β-activated human mast
cells. Mainstream (Ms) and Sidestream (Ss) cigarette smoke were collected onto fiber filters and
extracted in RPMI-1640 medium. Two ml of HMC-1 at 1 x 106 cells / mL were cultured with Ms and Ss
CSE at various concentrations in the presence or absence of IL-1β (10 ng / mL) for 24 hrs. The
supernatants were harvested and assayed for IL-6, IL-8, and MCP-1 by ELISA. The Ms and Ss CSE
(32.5 to 500 g / mL) alone induced either no or trace amounts of cytokines from HMC-1. However,
both Ms and Ss CSE significantly increased IL-6 and IL-8 production (all p < 0.001) from IL-1βactivated HMC-1 in a dose dependent fashion, except Ms CSE at 500 g / mL, which significantly
decreased IL-6 production (p < 0.0001). Contrarily, both Ms and Ss CSE at concentrations from 62.5 to
500 g / mL, but not 32.5 g / mL, significantly inhibited MCP-1 production (all p < 0.05) by IL-1βactivated HMC-1 in a dose dependent fashion. The effects of Ms CSE appeared to be more potent than
that of Ss CSE. The results show that CSE had differential effects on the production of inflammatory
cytokines from IL-1β-activated mast cells, which may partially explain why cigarette smoke contributes
to lung and cardiovascular diseases. (Supported by the Ruth R. Harris Endowment and RDC of ETSU)
BONE MINERAL DENSITY AND CONTENT OF COLLEGIATE THROWERS: INFLUENCE OF
MAXIMUM STRENGTH
J. M. Whittington, E. J. Schoen, L. L. Labounty, J. A. Gentles, J. M. Kraska, A. M. Swisher, J. E. Keller,
M. E. Stone, M. W. Ramsey, R. C. Hamdy, and M. H. Stone
Sport Performance Enhancement Consortium, East Tennessee State University, Johnson City, TN
Bone is a plastic tissue, changing in density and size with different levels of stress. . Furthermore, it
appears that BMD is altered in a site specific manner. However, BMD has not been studied extensively
in all types of athletes, particularly well trained strength-power athletes, such as throwers. The purpose of
this study was to examine the BMD of USA Division I collegiate throwers (shot put, discus, etc.). BMD
was compared to normative data and to different athletes. Measures of whole body maximum strength
and throwing performance were correlated with BMDs. Potential right/left side and sex differences were
examined. Athletes were 4 males, 3 females age 19.9 ± 0.9 years. BMD was measured with a DEXA
Maximum isometric strength was measured using a mid-thigh pull standing on a force plate. Force timecurves were generated during the strength tests. Peak force (PF) and normalized peak force (PFa) were
correlated with BMDs. Comparison indicates throwers have denser bones compared to normative data
and compared to other types of athletes. Male throwers tend to have greater total body BMD than female
throwers (p < 0.05). Dominant arm showed slightly greater BMD compared to non-dominant (p < 0.05).
Furthermore, BMD is related to PF (r = 0.68) and PFa (r = 0.56). Throwers have greater BMD’s than non-
2008 Appalachian Student Research Forum Page 59 athletes or most other types of athletes. However, throwers showed only a small indication of sidedness.
These observations likely stem from their training program (whole body heavy loading).
CHILDHOOD OBESITY AND TELEVISION:
A CORRELATION IN JOHNSON COUNTY, TN
Wanda Couch, Megan Cruze, Jessica Jones, Kyle Little, Kristie Markland, Andrew McCormick, Mary
McCormick, Katherine Rochelle, Ryan Ward, Dr. Jim Shine and Dr. Judith Anderson. Rural Primary
Care Track
Dept. of Family Medicine, Quillen College of Medicine, East Tennessee State University, Johnson City,
TN
Whether you are in rural Appalachia or in a major city, obesity is an escalating problem in our society
today. The growing problem of obesity in Johnson County, Tennessee, led the Health Council to
specifically identify childhood obesity as an area of concern in their community. Because of this
increased emphasis on learning about this trend and a desire to decrease it throughout Johnson County, a
group from ETSU was approached regarding studying and implementing a program to hopefully reverse
this epidemic. The project that was implemented by the group from ETSU was based on the results from
the data collected in the spring semester of 2006. The group hoped that through our project two goals
would be accomplished. The first goal of the project was to inform the community about the appropriate
amount of time that doctors recommend children spend watching television, 2 hours. The second goal of
the project was to suggest and inform the children of the many alternative activities to watching television
that are available in their community. The project enacted by the ETSU group was loosely based on a
project started in California that encouraged children to reduce the amount of time they watched
television. From this, the slogan “Break Free from T.V.” was used throughout our projects duration. In
order to evaluate the effect of the educational program, an initial survey was distributed to children asking
three basic questions: 1) Do you believe watching TV makes you gain weight, 2) How much TV. is ok to
watch each day, 3) Please list 3 things you do instead of watching TV. or playing video games. After
collecting this survey, our group began trying to inform everyone of the community about the doctor
recommended amount of time a child should spend watching TV. This information was distributed
through fliers, newspaper articles, skits, morning announcements, and talking with people of the
community. At the conclusion of the project, the survey was redistributed to the same children, and the
answers were compared to the initial survey. The results from the analysis of the surveys indicated that
the goals of the project had been accomplished. The children surveyed appeared to be more
knowledgeable about the recommended amount of time to watch TV. Through the course of this project,
it was realized that informing children about the relationship between TV and gaining weight was an
obtainable goal. While this conclusion was good for the project, the next step will be accessing the
feasibility of continuing this project in the future with the hope of decreasing the obesity problem in the
Johnson County community.
2008 Appalachian Student Research Forum Page 60 ACCESS TO OBSTETRIC CARE IN RURAL NORTHEAST TENNESSEE: ASSOCIATION WITH
BIRTH OUTCOMES
Heather N. Wright, B.S., Beth A. Bailey, Ph.D., Laura K. Jones Cole, M.S., M. A.
Tennessee Intervention for Pregnant Smokers (TIPS)
Dept. of Family Medicine, East Tennessee State University, Johnson City, TN
Background A nationwide downward trend in the number of rural obstetric providers has created
considerable barriers to prenatal care for pregnant women residing in underserved areas. This limited
access to prenatal and delivery services, compounded by socioeconomic, environmental and cultural
factors, has the potential to limit health information pregnant women receive, complicate pregnancy and
delivery, and adversely affect birth outcomes. This study sought to illuminate differences in birth
outcomes between two Northeast Tennessee counties with disparate access to obstetric care. Methods
Birth records from the two delivery hospitals in Washington County, TN were used to compare birth
outcomes of patients who resided in Johnson County (identified by the state of Tennessee as a medically
underserved area, with no obstetric providers or delivery services) and Washington County (a recognized
urban area with multiple obstetric practices and delivery hospitals). T-tests and chi-square analyses were
used to compare the women from the two counties on the birth outcomes of interest, including newborn
birth weight, birth length, gestational age, and incidence of low birth weight and preterm birth. Results
During the two year study period (2006-2007), complete delivery records were available for 2220 women
from Washington County, and 114 women from Johnson County. Compared to those from Washington
County, babies born to women from Johnson County had significantly lower birth weights, decreased
birth lengths and shorter gestational durations. In addition, Johnson County babies were twice as likely to
be low birth weight and more than four times as likely to be extremely preterm. There was no significant
difference in smoking status between the two counties. Discussion Consistent with current literature,
these results suggest that ready access to prenatal care may play an important role in birth outcomes in
Southern Appalachia. Low birth weight and preterm delivery are high priority public health concerns and
are associated with increased risk of infant mortality and long-term health impairments. Efforts to
ameliorate these problems in rural areas of the South should address issues related to local availability of
obstetric and delivery services.
2008 Appalachian Student Research Forum Page 61 Division V – Residents / Post-Doc Fellows
EOSINOPHILLIC GRANULOMA OF THE LUNG
"A MIMICKER OF MALIGNANCY"
Adel M. EL-Abbassi, Ryland P. Byrd, and Thomas M. Roy
Dept. of Internal Medicine, East Tennessee State University, Johnson City, TN
Eosinophilic granuloma , pulmonary histicytosis X, Lanerhans cell granulomatosis are the synonyms for
an uncommon interstitial lung disease. We are presenting a case of 52 year old man with 40 pack-year
history of cigarette smoking presented to the ER for progressive shortness of breath on exertion,
occasional dry cough, chest tightness, weakness and easy fatigability. CT scan of chest was done and
confirmed multiple small scattered masses in both lungs. PET scan showed increased uptake in the lung
nodules. Bronchoscopy was performed and Cytological analysis of BAL yielded no Diagnosis. FNA was
done as a next step but was not conclusive. Patient underwent wedge resection of the LUL and biopsy
was compatible with Langerhans cell histiocytosis. This case represents and highlights the association of
smoking to eosinophilic granuloma, and the presentation which can mimic malignancy with positive PET
scan.
A NOVEL PELVIC SURGICAL SIMULATION MODEL
Foulk, Brooke E.; Herrell, Howard E.; Hemphill, William K.; Sims, J. Paul; Eason, Martin J.; and, Olsen,
Martin E.
Dept. of Obstetrics and Gynecology and Center for Experiential Learning, Quillen College of Medicine,
East Tennessee State University, Johnson City, TN
Pelvic surgery can be very challenging to accurately and safely teach. Procedures take place in a small
and deep cavity, which can be difficult for learners to accurately view and understand the correct steps in
surgical technique. The anatomy in the female pelvis is also very intricate and complex, making
dissection very complex to novice surgeons with limited experience. Even to the expert gynecologic
surgeon, pelvic surgery can be complicated. For this reason, attempting to teach pelvic surgeries to
students and residents can be dangerous using living patients. Our purpose in creating a novel highfidelity pelvic surgical simulator was to make this process safe, teachable and reproducible, with the
ability for teachers to easily monitor and critique students. Material scientists along with our simulation
experts and obstetrics and gynecology faculty have devised a high-fidelity pelvic model, specifically for
surgical simulation. This novel model fits into our established patient simulator and represents a realistic
pelvis, including the female anatomic organs, layers of the abdominal wall, an anatomically correct
circulation system which can be adjusted to flow at different rates and pressures, and a system to monitor
and record learners performing procedures in real-time. By developing a simulation that has realistic
anatomy, including gynecologic organs, a circulatory system with actual perfusion, the layers of the
abdominal wall including skin, subcutaneous adipose tissue, fascial layers, muscle, peritoneum and other
anatomic landmarks, the instructor is able to teach a step-wise approach to many gynecologic surgeries.
The model has a realistic look and feel, making the learning experience much more believable and
accurate. A video output worn by the learner records what the learner is viewing and allows teachers to
observe each move made by the surgeon and critique both correct and incorrect maneuvers. This process
has enabled students and residents to “get their hands dirty” without having to step foot in a real operating
room, accruing operating time and costs, and risking harm to living patients. With this pelvic surgical
model, we are able to simulate multiple pelvic surgeries for various etiologies, including ectopic
2008 Appalachian Student Research Forum Page 62 pregnancy, different types of ovarian cysts, abdominal and vaginal hysterectomy, tubal ligation,
oophorectomy, and salpingectomy. This allows endless numbers of surgical possibilities, patients, and
time. Learners can avoid the pressures of operating room with living patients and can be closely
monitored and instructed without the constraints of time, money, and their limited experience. Procedures
are easily reproducible, with adjustments made as needed for each learner’s specific needs. This novel,
high-fidelity pelvic simulator model provides endless possibilities for gynecologic surgical training.
A WEB-BASED ELECTRONIC FETAL MONITORING TUTORIAL
Howard Herrell, Brooke Foulk, Martin Eason, Imran Quraishi
Dept. of OB/GYN and the Center for Experiental Learning, Quillen College of Medicine, East Tennessee
State University, Johnson City, TN
Interpretation of electronic fetal monitoring has traditionally been taught to learners through a formal,
lecture-based didactic, with examples of fetal tracings presented in the form of static images showing a
representative portion of a tracing. To date, there have been no non-commercial, web-based attempts at
modernizing this aspect of obstetrical education. We sought to develop a web-based tutorial integrating
video recordings of tracings, interactive teaching modules, case-based learning, and interactive quizzes.
We acquired more than one hundred video recordings of fetal tracings of normal and interesting cases
from the archives of the Johnson City Medical Center related to obstetrical cases from 2003–2006. These
videos served as the basis for the development of core teaching modules, quizzes, and cases. A narrative
was constructed discussing the history of fetal monitoring, its physiologic basis, and methods of
interpretation based on currently accepted guidelines. These were constructed into a website
(http://www.ob-efm.com) and improvements were continuously made based on feedback from peer
reviewers and medical students/residents who were asked to trial the website. Learners have provided
individual feedback that they prefer this web-based, interactive approach over our previous traditional
lecture-style didactic. Research is currently underway to objectively assess short- and long-term retention
of subject matter by novices exposed to either the traditional didactic or to the website. We have observed
more uniformity with the fetal monitoring terminology guidelines suggested by the National Institute of
Child Health and Human Development and better communication between providers. We are currently
working to validate these observations. We propose that this pedagogic method, which relies on the
learner to self-direct through the material at his or her own pace and ties the material with real-to-life
examples of the material being studied, better prepares the learner to interpret fetal tracings, document the
interpretation, and intervene where necessary as compared to a traditional lecture. We have observed
better initial and long-term retention of the material. Additionally, this format can free-up traditional
lecture time and give the learner instant feedback in the form of online quizzes. Research is underway to
validate these conclusions. This resource has also been directly tied to our lab-based simulation program
and representative fetal tracings/cases have been used as a back-drop for obstetric simulations.
HIGH-FIDELITY SIMULATION AS AN ADJUNCT
TO PHYSICIAN IMPAIRMENT TRAINING
Howard E, Herrell, Martin J. Eason, and Martin E. Olsen.
Dept. of Obstetrics and Gynecology and the Center for Experiential Learning, College Of Medicine, East
Tennessee State University, Johnson City, TN
We sought to develop a high-fidelity simulation of an obstetric or gynecologic emergency that combines
both knowledge of the diagnosis and management of the emergency with education regarding physician
2008 Appalachian Student Research Forum Page 63 impairment in order to emphasize the core competencies in our curriculum. Using high-fidelity
mannequins in our simulation laboratory, we developed an obstetric and a gynecologic emergency
simulation that could be combined with our physician impairment curriculum. The obstetric emergency
involves the learners being presented with sustained fetal bradycardia remote from delivery, necessitating
emergency cesarean delivery, while the gynecologic emergency requires the learner to diagnose ruptured
ectopic pregnancy while instituting medical resuscitation and ordering emergent surgical management. In
each scenario, after the learner has decided that the patient needs to proceed to the operating room, a
participant arrives at the scenario who acts as an impaired attending physician. This impaired physician
attempts to disrupt patient care and block the learners’ attempts to take the patient to the operating room.
Realism is added through scripted irrational behavior, dishevelled appearance, and whiskey dabbed on the
shirt collar as an olfactory queue to the learner. The learners are required to (1) diagnosis, manage, and
understand the severity of the emergency; (2) identify physician impairment; and (3) counteract the
impaired physician and call for other supervisory assistance in less than ten minutes. In the subsequent
debriefing, the learners meet with an expert in physician impairment and learn the signs and types of
physician impairment and strategies to prevent it; they are educated about programs to assist and monitor
impaired physicians; and they receive training in due process and the rights and responsibilities afforded
to impaired physicians. As a secondary objective, the management of the obstetric or gynecologic
emergency is reviewed. Variants of this simulation have been used to train and evaluate OB/GYN
residents, clerks, and prospective resident applicants. Learners at all levels have reported their confidence
bolstered in the management of the emergency and their knowledge greatly enhanced with the “surprise”
secondary goal of physician impairment training. High-fidelity simulation of obstetric and gynecologic
emergencies provides an excellent opportunity to train residents and students in the management of
secondary learning objectives involving the core competencies not previously thought adaptable to highfidelity simulation. Physician impairment is only one possibility and a variety of other scenarios involving
professionalism, interpersonal and communication skills, systems-based practice, practice-based learning
and improvement, as well as medical knowledge and patient care have been successfully tied to highfidelity simulation in this manner at our institution.
POST-PRIMING MANIPULATION OF DISCORDANT ANTIVIRAL T CELLS
Subhadra Nandakumar1, Akaike, T.2, Mege, R.M.3 and Uday Kumaraguru1
1
Dept. of Microbiology, College of Medicine, East Tennessee State University, TN
2
Dept. of Biomolecular Engineering, Institute. of Technology, Yokohama, Japan,
3
INSERM, Institut du Fer a Moulin, Paris, France.
Memory T cells that have encountered the antigen earlier are maintained in a proliferative state and are
subjected to aging limitations of proliferating cells and eventually undergo “replicative senescence”.
Conditions for such accelerated senescence of T cells are more conducive in latent infection with HSV
that periodically reactivates stimulating a T cell response every time. The key features of such T cells are
their oligoclonality, functional impairment and acquisition of new phenotype. Their accumulation results
in contraction of repertoire, loss of co-stimulatory molecule, decrease or change in cytokine production
and strongly correlates with severe manifestations of diseases. Hence, we propose that understanding the
mechanisms regulating their ontogeny, change in phenotype, acquisition of novel function and means to
rejuvenate will be pivotal to development of future therapies. Our analyses, using in vitro generated antiHSV senescent T cells and in vivo generated T cells from latently infected and periodically reactivated
mice have shown phenotypic changes different from normal memory T cells. This includes among others
increased expression of NK inhibitory receptors (KLRG1), MHC unrestricted killing, aberrant cytokine
production, and reduced proliferative ability. Cadherins described recently as the ligand for KLRG1 are
also down regulated in chronic viral infections. Our investigations besides characterizing the functional
defect in the responding T cells also looks at modulation of KLRG1hi T cells with their cognate ligands
2008 Appalachian Student Research Forum Page 64 the N and E cadherins. The results from our study seem to suggest that the responding T cells have
changed their phenotype, are less poly-functional, and lose their MHC restricted behavior and becoming
potentially hazardous leading to self destruction. Interventionist manipulations and signaling through the
KLRG1 or other inhibitory receptors with their ligand can help in modulating the functional behavior and
result in the rescue of dysfunctional T cells.
IDENTIFICATION, RECOMBINANT EXPRESSION, AND BIOCHEMICAL CHARACTERIZATION
OF A FLAVONOL 3-O-GLUCOSYLTRANSFERASE
FROM CITRUS PARADISI
Daniel K. Owens1 and Cecilia A. McIntosh1,2
1
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
2
School of Graduate Studies, East Tennessee State University, Johnson City, TN
Flavonoids are a diverse group of natural products that are ubiquitous in higher plants with over 6000
naturally occurring compounds having been identified. These compounds are associated with a number of
essential physiological roles in planta and have agriculturally and pharmacologically significant activities.
Glycosylation is a predominant modification reaction in flavonoid biosynthesis affecting the solubility,
stability, and subsequent availability of compounds. Glycosylation of flavonoids also has an impact upon
the quality of foods and food products. This is of particular interest in citrus species where flavonoid
glycosides convey taste characteristics making flavonoid glucosyltransferases interesting targets for
biotechnology applications. In this work, a C. paradisi (grapefruit) glucosyltransferase gene was
identified, cloned, and introduced into the pET recombinant protein expression system utilizing primers
designed against a predicted flavonoid glucosyltransferase gene (AY519364) from C. sinensis (orange).
The encoded C. paradisi protein is 51.2 KD with a predicted Pi of 6.27 and is 94.5% identical to the C.
sinensis homologue. The enzyme glycosylated only the flavonol aglycones Quercetin (Km=67 µM;
Vmax=20.45 pKat/µg) , Kaempferol (Km= 12 µM; Vmax= 11.63 pKat/µg), and Myricetin (Km= 33 µM;
Vmax= 12.21 pKat/µg) among a number of tested compounds representing various flavonoid subclasses.
Glycosylation was shown to occur at the 3 position within the flavonoid backbone based on HPLC and
TLC analysis with certified reference compounds. The optimum pH was determined to be 7.5 with a
pronounced buffer effect noted for reactions performed in Tris-HCl buffer. The enzyme is inhibited by
the metal ions Cu2+, Fe2+, and Zn2+ as well as UDP (Ki = 69.5 µM) which is a product of the reaction.
Treatment of the enzyme with a variety of amino acid modifying compounds suggests that cysteine,
histidine, arginine, tryptophan, and tyrosine residues are important for activity. The thoroughly
established biochemical functions and characteristics for this C. paradisi flavonol 3-O-glucosyltransferase
add to the growing base of flavonoid glucosyltranferase knowledge, and will be utilized to further
investigate structure and function relationships for these types of enzymes.
WHAT IS THE BEST TREATMENT OF CHRONIC FECAL RETENTION?
Zia Rahman, M.D., and Fereshteh Gerayli, M.D.,
Johnson City Family Medicine Residency Program, Johnson City, TN
Chronic Functional Constipation (CFC) with or without encopresis is a common pediatric problem, which
is distressing to the child and family. High quality randomized controlled trials ( RCTs) on the
management of CFC are lacking. The objective of this study was to identify the best available evidence
on the management of this chronic condition. A structured search of Medline, BMJ Clinical Evidence,
Cochrane Library and National Guideline Clearinghouse between 1966 and Feb 2007 was performed by
2008 Appalachian Student Research Forum Page 65 the authors and the medical school librarian. Our search located seven relevant RCTs and two relevant
systematic reviews (SRs.). The Best available evidence for treatment of chronic functional constipation in
children is on use of osmotic laxatives). Increased fiber is the most commonly recommended first step in
the management of CFC, however, there is only limited evidence to support this approach (SOR: B, based
on one RCT). There is insufficient evidence on the use of stimulant laxatives (SOR: A, based on a
systematic review). There is no evidence on use of bulk forming laxative. Minimal evidence supports use
of mineral oil as a stool softener. There is limited evidence to support the use of behavioral intervention
and no evidence that biofeedback or anorectal manometry adds any benefit to conventional treatment in
the management of CFC in children with fecal incontinence. Increased fluid intake is usually suggested,
but there is limited evidence to support this dietary modification. In conclusion, the best evidence for
treatment of chronic functional constipation (CFC) in children is with osmotic laxatives (Strength of
Recommendation [SOR]: A, based on six fair to good quality randomized controlled trials [RCTs]).
EVALUATION OF INFLUENCE OF FLUORIDE IN DRINKING WATER OR AS A DIETARY
SUPPLEMENT OF PREGNANT WOMEN RECEIVING PRENATAL CARE AT ETSU OB/GYN ON
DENTAL HEALTH OF THEIR OFFSPRING AND IT’S IMPACT ON INCIDENCE OF PRETERM
LABOR.
Olga Sarkodie , Martin Olsen, Kevin Breuel,
Dept. of Obstetrics and Gynecology, College of Medicine, East Tennessee State University, Johnson City,
TN
The United States spends more than twice as much on healthcare as the average of any other of developed
nations. Yet more than 41 million Americans have no health insurance, and even more have no dental
insurance. In theory, primary teeth could benefit from prenatal exposure to fluoride. There are some
encouraging trends in existing data, but administration of dietary fluoride supplements to pregnant women
is not recommended at this time. Topical fluoride in tooth-paste is one of methods that are responsible
for a significant drop in the level of cavities since 1960. According to ADA (American Dentistry
Academy), systematic fluorides such as community water fluoridation and dietary fluoride supplements
are effective in reducing tooth decay. These fluorides provide topical as well as systemic protection
because of presence of fluoride in saliva. Fluoride can prevent tooth decay by strengthening teeth to
block acid from penetrating the enamel. The most commonly recommended level of fluoride in drinking
water is 0.7-1.2 parts fluoride per million parts water. Water fluoridation has been proven to reduce
inexpensive means of obtaining the fluoride necessary for optimal tooth decay prevention, but not
everyone lives in a community with a centralized, public, or private water source that is fluoridated. Of
the roughly 513 registered water districts in Tennessee about 155 choose not to fluoridate. Although in
modern research there are controversies regarding necessity of fluoridation CDC (United States Center of
Disease of Control and Prevention) recognizes the fact that public water fluoridation is one of 10 great
health achievements of the 20th century. We feel that current standards of providing pregnant women with
dental care are sometimes not sufficient. We think there is a connection in between dental health of
pregnant women and their future children as well as it (dental health) may influence on outcome of
pregnancy such as preterm labor.We started this project anticipating to clarify the role of prenatal
exposure to fluoride in drinking water or as a supplement in incidence of preterm labor and dental health
of offspring. Our study failed to show statistically significant difference between two groups of patients
(group A—Patients who delivered prematurely, and group B--those who delivered full term.), with
regards of use of Fluoride. Further research is needed to clarify the importance of dental health on the
outcome of pregnancy and dental health of offspring.
2008 Appalachian Student Research Forum Page 66 AN UNUSUAL CAUSE OF HYPOTENSION IN A PATIENT WITH ANTICOAGULATION
Gerson David Valdez Figueroa, M.D., Gerson Teran, MD, Roger Dean Smalligan
Dept. of Internal Medicine, East Tennessee State University, Johnson City, TN
Objectives:
1- Identify spontaneous rectus sheath hematoma as an unusual complication of anticoagulation in
elderly patients.
2- Recognize Carnett’s sing as a simple clinical finding helpful to distinguish intrabdominal causes
of abdominal pain than abdominal wall pathology.
CASE : A 77 year old female with past medical history of HTN , DM presented to the ER with an
hypertensive crises, vomiting and confusion on initial workup sodium : 124, troponin I : 0.21 which
prompted the administration of aspirin , metoprolol and anticoagulation with enoxaparin after blood
pressure stabilization, on second day of hospitalization cardiology was consulted and adenosine stress
test was scheduled , during the stress test patient BP : dropped from 161/51 to 70/50 mm Hg ,patient
became confused ,and complained of nausea and diffuse abdominal pain ,PE: BP:70/50 , HR: 68 x’ ,
RR:22 x’, T: 97.2 F, General alert oriented , in acute distress , HEENT unremarkable, neck supple no
JVD no bruits, lungs clear ,CVRR no murmurs rubs nor gallops, abdomen supple tender over the RLQ, a
12 x5 cms mass over the same area , Carnett’s sing was present ,BS present no organomegaly, EXT:
pulses symetrical, neurological exam no focal patient was placed of IV fluids , BP medications holded as
enoxaparin , ECG showed no abnormalities ,and cardiac troponin I :0.02 ng/ml , CBC showed HB : 7.1 ,
HT:31%( 10 the day before) , WBC: 19800,N:70 % ,PLT : 306000 , patient was started on IV fluids and a
CT scan of the abdomen showed a right rectus sheath hematoma and pelvic hematoma , patient received 3
Units PRBC , surgery was consulted and they opted for conservative management , patient was
monitored closely , her hemoglobin stabilized after transfusion and serum sodium returned to normal
limits after NS infusion patient had and uneventful clinical course.DISCUSSION:This case depicts an
unusual but life threatening condition ; rectus sheath hematoma it is a rare condition, linked to age
,systemic anticoagulation, intrabdominal injections, abdominal wall strain, minor trauma , pregnancy as
risk factors, our patient had received anticoagulation for 24 hrs ,and developed an intrabdominal mass
within hours , clinical manifestations include: abdominal wall pain( 84%) and abdominal mass(
Fothergill’s sing) , decrease in hematocrit, ecchymosis 69% , Carnett’s sing ( increase in the abdominal
pain when patient lift the head and shoulders off the examination table) can help in the diagnosis, the
diagnosis is confirmed by abdominal USG or CT scan of the abdomen, Treatment is mostly expectant,
including blood transfusion and pain management ,some cases require intravascular embolization or
surgery ,Although outcome is usually good ,there are reports of fatal cases .Physicians must be alert of
this rare but life threatening condition specially in elderly patients with anticoagulation.
METASTATIC ADRENOCORTICAL CARCINOMA PRESENTING WITH DIPLOPIA.
Gerson David Valdez, M.D. and Gerson Ariel Teran, M.D., Roger D Smalligan M.D., MPH
Dept. of Internal Medicine, East Tennessee State University, Johnson City, TN
Objectives :
1-To remind internist useful clinical findings that can point an unusual cause of Cushing’s syndrome
such as adrenocortical carcinoma .
2- Remind an unusual presentation of an unusual neoplasm .
2008 Appalachian Student Research Forum Page 67 CASE: A 43-year-old woman with a history of hypertension and diabetes mellitus presented with 5months of amenorrhea, acne , and hirsutism, 3 months of abdominal pain with a 9 pound weight loss, and
2 weeks of diplopia. PMH: HTN 2 years, DM 14 months. Family/social history: unremarkable and
negative for smoking, alchohol or illicit drugs. Medications: metformin, lisinopril, glipizide, furosemide.
PE: BP: 161/89, HR 96, RR 18, alert/oriented, cushinoid facies, neck with buffalo hump, abdomen obese
with striae, no hepatosplenomegaly, skin with abundant acne and hair over thorax, neurological exam
showed left sided sixth cranial nerve palsy. Laboratory Hgb 14.5, WBC 11,000, creatinine 0.9, sodium
136, potassium 4.4, calcium 9.1, random cortisol 39 (5-22), AM cortisol 37 (5-22), urine cortisol 193
(<45), ACTH 7 (10-46), TSH, DHEA 15.3 (1.9-7.6), DHEA SO4 872 (35-430), testosterone 237 (10-70),
normetanephrine 0.31 (<0.5), metanephrine 0.2 (<0.9), serum aldosterone 28 (<31), prolactin 6.5 (2.8-29),
FSH 0.3 (3-33.4), LH 0.1 (1.5-9.3). Abdominal CT revealed a left adrenal mass (11.4x7.4cm) and
hypodense nodules on the liver and extensive retroperitoneal and mesenteric lymphadenopathy. Head
MRI showed a 1.2x0.8x1.2cm mass in the cavernous sinus and adjacent to the pituitary gland compatible
with metastases. Liver biopsy showed adrenocortical cells by with specific stains compatible with
adrenocortical carcinoma. The patient received radiation therapy for the brain mass and palliative
chemotherapy with Mitotane. Ten days later the patient requested comfort care after developing
intractable nausea/vomiting and prerenal acute kidney injury and she died shortly afterwards.
DISCUSSION: Adrenocortical carcinoma is a rare neoplasm with an incidence of 0.02-2 cases per
million per year. In 53 % cases the tumor is functional with the most common presentation being
Cushing’s syndrome, occasionally along with virilization (10%)as in this case. Our patient presented
with a cranial nerve palsy secondary to a cavernous sinus metastasis . The fact that she paradoxically lost
weight and had associated virilization raised suspicions of adrenocortical carcinoma as the underlying
cause of her hypercortisolism. Thirty-four percent patients with adrenocortical carcinoma have
metastasic disease at the time of the diagnosis with the most frequent sites being liver, lymph nodes, lungs
and rarely the bones. To our knowledge this is the first case reported of intracranial metastasis producing
an isolated sixth cranial nerve palsy. Our patient received radiotherapy as a palliative measure. Further
palliative measures have not been standardized in this setting . Our patient developed a gastric
hematoma, nausea, vomiting and prerenal acute kidney injury. Physicians should remember to consider
hypercortisolism in cases of intractable hypertension and diabetes and also to consider adrenocortical
carcinoma if there is an unexplained weight loss in this setting.
NON-PHARMACOLOGICAL THERAPEUTIC OPTIONS IN TREATMENT OF INSOMNIA
Umesh Vyas, M.D., Hetal Brahmbhatt, M.D.
Dept. of Psychiatry and Behavioral Sciences, James H Quillen College of Medicine, East Tennessee State
University, Johnson City, TN
Educational Objectives: At the conclusion of this presentation, the participant will be able to know
different non-pharmacological therapeutic options available for use in their practice especially for chronic
insomnia. Introduction and Hypothesis: Insomnia is the most frequent sleep disorder. It is a symptom, and
is defined as “chronic inability to obtain the amount of sleep needed for optimal functioning and wellbeing”. Prevalence estimates indicate one third of adult population reports insomnia. Prevalence is higher
among women, elders and patients with medical or psychiatric disorders. Insomnia is said persistent if it
lasts from one to six months, and chronic if it lasts more than six months. Untreated insomnia is
associated with significant morbidity and mortality. Thorough assessment of insomnia is essential in
choosing the most appropriate strategy for management. If a cause of insomnia is identified, initial
treatment should be directed at specific factor. Since insomnia is a chronic condition, long-term and safe
treatments are warranted. Non-pharmacologic options have been available for decades, but lack of
physician awareness and training, difficulty in obtaining reimbursements and questions about efficacy
have limited their use. These therapies offer the greatest potential for long term gains in preventing
2008 Appalachian Student Research Forum Page 68 recurrence in insomnia. Pharmacological options are most useful for acute insomnia. They are commonly
used but long term use of hypnotics can become complicated by drug tolerance, dependence or rebound
insomnia. Method: Pubmed.gov was searched by using pre-determined key word: Insomnia. Results:
Non-pharmacological interventions produce reliable and durable clinical benefits in the treatment of
primary insomnia, insomnia associated with medical or psychiatric conditions and insomnia in elders.
Additional research is still needed to develop and validate treatment algorithms that would optimize
outcomes and reduce morbidity. Conclusion: Non-pharmacological therapies should always be a
component in treatment of Insomnia.
2008 Appalachian Student Research Forum Page 69 Oral Presentations
Graduate Students
A TEMPORAL AND SPATIAL ANALYSIS OF NORTH AMERICAN AVIAN REDISTRIBUTIONS
IN RESPONSE TO CHANGES IN CLIMATE
Kenneth W. McDonald1,3, M. Phillip Nott2,3, Justin T Peyton1,3, Thomas C. Jones1,3, Tashauna A.
Gilliam1, Mckayla B. Johnson1, & Laura L. Catron1
1
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
2
The Institute for Bird Populations, Point Reyes Station, CA
Understanding the response of organisms to climate variables is important to assessing the impact of
projected changes in climate. Our analysis of North American bird banding records of select avian species
over a known period of climatic change demonstrates that the distribution and change of avian
morphological characters and population parameters correspond over space and time to perturbations in
climate variables indicative of overall changes in climate. Furthermore we suggest hypotheses regarding
the redistribution and evolution of avian species relative to current models of forecasted global climate
change.
LONG-TERM GEOCHEMICAL IMPACTS
FROM PERMANGANATE OXIDATION: FIELD-SCALE ASSESSMENT
Kelly Moore and Michelle Crimi, Ph.D.
Department of Environmental Health, East Tennessee State University, Johnson City, TN
In-situ chemical oxidation (ISCO) using permanganate is a remediation technology that has been used
under a wide range of geological conditions for the destruction of hazardous contaminants in soil and
ground water. The most common contaminants treated by permanganate ISCO are chlorinated solvents
(i.e. tetrachloroethylene (PCE), trichloroethylene (TCE), dichloroethylene (DCE), and vinyl chloride
(VC)). Soluble manganese, a byproduct of the oxidation reaction, is a concern post-treatment because it
can exceed the US EPA secondary drinking water standard of 0.05 mg/L. Another concern is chromium,
found as a co-contaminant or naturally-occurring at a site, which can be oxidized by permanganate to a
more toxic form (hexavalent chromium). Permanganate oxidation can also cause significant increases or
decreases in groundwater pH and also leads to significant increases or in oxidation-reduction potential.
Often, large quantities of permanganate are injected into the subsurface without considering the possible
long-term effects on ground water quality. While theoretical information exists (guidance documents), no
comprehensive and integrated review of field case study data has been conducted to assess the role
residual manganese has on groundwater quality. A comprehensive review was conducted of data from 30
sites where permanganate has been injected. Data sets include pre- and post-oxidation measurements of
metal concentrations (manganese and chromium), pH, and ORP, along with the remediation design
approach (i.e. oxidant concentration, mass delivered, and injection method). Data were normalized based
on monitoring date and distance from the target treatment zone (TTZ). A two-way ANOVA was used to
compare pre- and post-ISCO values over time and distance. Several correlation analyses were run to
identify relationships between manganese concentrations and other parameters and with the design
approach. Results show that in most cases manganese concentrations decrease with distance from TTZ
and with time. Concentrations return to pre-ISCO levels by 48 weeks post-ISCO. Results also show that
hexavalent chromium increases significantly after injection of permanganate. However, after
2008 Appalachian Student Research Forum Page 70 approximately 6 to 20 weeks, concentrations return to pre-ISCO levels. Other results indicate that
permanganate ISCO can increase or decrease pH levels depending on site characteristics (i.e. media type).
These levels usually return to normal by 32 to 52 weeks post-ISCO. ORP levels increase after
permanganate injection. After 20 to 32 weeks, the levels begin to decrease and by 48 to 52 weeks the
levels are very negative. The levels usually remain more negative than the pre-ISCO levels. Manganese
and chromium concentrations and ORP levels appear to be correlated with the mass and concentration of
permanganate delivered. Whereas, pH levels seem to be more positively correlated with the site
characteristics. The results of this research have several important benefits. One benefit is the providing
of field-scale evidence to support guidance based theoretical recommendations. Another benefit is the
determination of impacts that manganese can have on ground water quality. This knowledge will help
ISCO practitioners make more informed decisions related to system design and implementation. This will
in turn reduce the likelihood of adverse effects to public and environmental health.
DEVELOPMENT OF A GIS PROTOCOL TO PRIORITIZE WATER QUALITY IMPROVEMENTS IN
RESERVOIR EMBAYMENTS IN TENNESSEE
T. Shannon O’Quinn, Yongli Gao, and Jessica Buckles
Dept. of Physics, Astronomy, and Geology, East Tennessee State University, Johnson City, TN
To simplify water quality improvement in reservoirs, it has been suggested that efforts should be focused
on smaller and more manageable units such as reservoir embayment areas. Embayments are prime
locations to locate marinas, parks, beaches, and residential homes. Current data and information on
reservoir embayments in Tennessee will be assembled into a GIS-based database. GIS-based data models
have been developed to identify specific characteristics of embayments that influence water quality.
Embayments of 11 main reservoirs have been mapped and digitized in ArcGIS. Initial characterization
criteria include water quality, embayment area/watershed area ratio, embayment area/reservoir area ratio,
flux between embayment and main reservoir, and stream influence on embayments. A characterization
process has been developed for Chickamauga and Boone reservoirs to identify and prioritize embayments
that are most likely to be affected by watershed restoration efforts. This protocol is being expanded to
prioritize water quality improvements in reservoir embayments in Tennessee.
ETSU HEALTH CORPS
Mihir Patel,
Department of Public Health, East Tennessee State University, Johnson City, TN
The aim is to deliver preventive health care to undeserved rural Appalachian population by ETSU
medical students, student nurses, medical residents and medical fellows under the supervision of ETSU
medical faculty. The project is designed to respond to the original mission of ETSU Quillen College of
Medicine (Teague/ ranston Act, i.e., federally supported medical schools for medically undeserved
areas.) The project intends to evaluate a model of rural preventive health care. Special emphasis is placed
on a trans–generational social system (Health Corps Concept) in which young professionals come to the
rescue of elderly medically undeserved individuals living in the Appalachian countryside. The viability
of the project depends on voluntarism and philanthropy. Important support will be sought from the state
(ETSU), local business (ambulances supported by automobile dealerships, school districts, churches,
drugstore chains (Walgreens, CVS), department stores (Wal-mart pharmacies). medical device industry
and Pharma industry. Because preventive medicine is the crucial component of a viable national health
care system, it is hoped that grant support at eh national level will be achievable. Although we will
2008 Appalachian Student Research Forum Page 71 initially obtain philanthropic support , this project will be critically evaluated for its medical economic
viability independent of any grant support.
CONTROL OF MnO2 PARTICLES DURING PERMANGANATE-ISCO THROUGH USE OF
CHEMICAL STABILIZATION AIDS
Mark Quickel1, Michelle Crimi2, Saebom Ko3
Dept. of Environmental Health, East Tennessee State University, Johnson City, TN
In situ chemical oxidation (ISCO) is a remediation technique used in the treatment of soils that contain
organic contaminants. Permanganate (MnO4-) is a common oxidant used in the ISCO process. During
remediation, manganese dioxide (MnO2) particles form as a by-product of the reduction of permanganate.
These particles have been shown to deposit in and around permanganate injection wells. The deposition
of MnO2 over time may alter the flow of permanganate, preventing effective oxidant distribution and
ultimately reducing contact with contaminants. Based on laboratory experimentation, groundwater
chemistry is known to impact the behavior of MnO2, thus their transport and deposition in the subsurface.
Soil composition is also expected to affect their transport and deposition via surface chemistry and
filtering effects. The goals of this experimentation are to (1) better understand the influence of various
media types on transport and retention of MnO2 and (2) to improve transport and inhibit deposition of the
particles through the use of stabilization aids. To test the effects of different porous media compositions,
the following media types were employed in these studies: sand, sand with clay (montmorillonite), sand
with organic matter, and sand with iron oxides (goethite).
The stabilization aid, sodium
hexametaphosphate (HMP), was identified in batch experimentation as an effective additive to prevent
settling, stabilize particles in solution, and reduce overall particle size. It is presumed that HMP will
facilitate the transport of MnO2 particles in porous media. 1-Dimensional (1-D) transport experiments
were carried out in two stages to evaluate the control of MnO2 particles with permanganate ISCO. The
first stage involved the characterization of the movement of MnO2 through the various media types. The
1-D columns were wet packed with the selected media. Trichloroethylene (TCE), a common organic
contaminant, was injected at the influent end of the columns, and simulated groundwater was passed
through the columns. Permanganate was then introduced and passed through the columns to generate and
transport MnO2 in situ. Samples of the effluent were collected and analyzed for total solids, total
dissolved solids, and total suspended solids. An extraction was carried out on both the effluent and
sections of the column. The extraction allowed for the separation of water extractable and barium
extractable Mn, and dissolved any retained MnO2 for quantification. All conditions were then repeated
with the addition of sodium hexametaphosphate. With the addition of HMP, it was found that particle
retention was decreased for each media type. Particle retention, directly in the NAPL source zone,
decreased by 85% in sand + goethite media, 53% in sand + clay media, 47% in sand + organic carbon
media, and 25% in sand media. The decrease in particle retention can lead to better oxidant distribution
and contact time with the contaminant, which translates to more efficient ISCO treatment.
SKELETAL MORPHOLOGY AND SYSTEMATICS OF LATE PLIOCENE ALLIGATOR SP
Jeremy B. Stout
Dept. of Biological Sciences, East Tennessee State University, Johnson City, TN
The American alligator (Alligator mississippiensis) is one of only two species of alligator in the modern
world. It is only distantly related to the other extant species (A. sinensis), separated geographically by
approximately 11 000 km and up to 25 million years of evolution. Much closer relatives to A.
2008 Appalachian Student Research Forum Page 72 mississippiensis are known from the geologic past of North America. A disparity exists, though, in the
fossil record between A. mississippiensis and its closest relative, the late Miocene – early Pliocene A.
mefferdi. While A. mississippiensis is well known from the early Pleistocene and later, no Alligator was
known from the middle and late Pliocene until the discovery of Haile 7G, a new late Pliocene (~ 2 Ma)
site from Alachua County, Fl. North American fossil Alligator from this gap in the geologic record will
undoubtedly provide invaluable clues to the evolution and historical biogeography of the modern
American alligator lineage. This research seeks to examine the available Alligator material from this site
and apply it to the diagnostic skeletal (primarily cranial) character matrix established by Brochu, 1999
and 2004*. This will afford the specimens species-level identification and confirm one of the three
possible hypotheses: a.) that the Haile 7G alligators represent the modern A. mississippiensis, b.) that they
belong to a known fossil taxon (eg. A. mefferdi), or c.) that the specimens cannot be easily placed within
known taxa, and therefore will either become the holotype of a new species or exhibit a suite of characters
from more than one species. The character data will then be applied to a cladistic program (PAUP 4.0) to
place the fossil alligators into a phylogenetic perspective. At present, one large skull is on loan to the
ETSU Museum of Natural History and is being prepared by the author. Based on the preliminary
findings of this specimen, the available character data suggests A. mississippiensis, though the alternate
hypotheses cannot yet be ruled out.
* Poster and/or oral presentation(s) provide complete citation.
ETHNIC DIFFERENCES IN RISK AND PROTECTIVE FACTORS
FOR DEPRESSION
Preston Visser1, Jameson Hirsch1, Angela Jones1, and Elizabeth Jeglic2
Dept. of Psychology, East Tennessee State University, Johnson City, TN.
2
Dept. of Psychology, John Jay College of Criminal Justice, New York, NY.
1
Risk and protective factors for depression are well-documented; however, less is known about risk factors
and protective characteristics among ethnically and racially-diverse groups. There is a particular lack of
research on the function of protective factors across ethnic groups. We examined the association between
positive psychological characteristics, including trait hope and religiosity, and depressive symptoms in an
ethnically diverse sample of college students (N= 407; Black = 24.5%, Hispanic = 39.2%, White =
17.4%, Asian= 5.4%). We also investigated anger and hopelessness, which are more traditional risk
factors for depression. Participants completed the Beck Depression Inventory II (BDI-II), the Beck
Hopelessness Scale (BHS); Trait Hope Scale, the Multidimensional Anger Inventory (MAI) and the Brief
Multidimensional Measure of Religious/Spirituality (BMMRS). We hypothesized that higher scores on
measures of hopelessness and anger and lower scores on measures of religiosity and trait hope would be
related to increased levels of depressive symptoms. Data were analyzed using correlation techniques and
a series of multivariate linear regressions, statistically controlling for age and gender. In correlation
analyses, greater levels of hopelessness and anger were significantly associated with increased depressive
symptoms, and higher levels of trait hope were significantly negatively associated with depressive
symptoms (for all models, p<.001). In a multivariate model examining all cases, regardless of ethnicity,
hopelessness, anger and trait hope were significant predictors of depressive symptoms; anger and
hopelessness predicted increased depressive symptoms, whereas trait hope was associated with decreased
symptoms of depression. We also stratified our sample by ethnicity. Anger was significantly related to
depressive symptoms in Blacks (β= .237), Hispanics (β= .312), Whites (β= .497), and Asians (β= .803; all
ps<.001). With the exception of Asian participants, trait hope was negatively related to depressive
symptoms. In Blacks and Hispanics hopelessness was a significant risk factor for increased depressive
symptoms (β= .460 & .382, respectively). In Black individuals, a marginally significant trend toward
significance existed for the association between increased religiosity and decreased depressive symptoms
2008 Appalachian Student Research Forum Page 73 (p= .059); this relationship warrants further examination. Our findings have implications for future
research and therapeutic interventions. Researchers should more closely examine how potential
etiological influences of depression may vary across ethnicities. For example, it may be insightful to
determine why hopelessness in Blacks and Hispanics is strongly predictive of depression while trait hope
strongly predicts depression in Whites. Investigation of cultural characteristics and how they relate to the
development of depressive symptoms may help researchers to better understand the complex interactions
between risk and protective factors of depression. In the treatment of depression, clinicians should
promote protective cognitive and emotional characteristics while reducing the impact of traditional risk
factors. Although risk and protective factors for depression are largely similar across racial and ethnic
groups, differences that exist may be of therapeutic usefulness.
UP-REGULATING DEATH RECEPTOR 5 VIA TUNICAMYCIN ADMINISTRATION INCREASES
ANTI-HSV IMMUNE RESPONSE
Stacie N. Woolard, Subhadra Nandakumar, and Uday Kumaraguru
Dept. of Microbiology, College of Medicine, East Tennessee State University, Johnson City, TN
Tunicamycin is a naturally occurring antibiotic that induces endoplasmic reticulum stress; thus enhancing
susceptibility to TNF-Related Apoptosis Inducing Ligand (TRAIL) mediated apoptosis. In previous
tumor research, Tunicamycin has been implicated to improve cancer therapies by sensitizing tumor cells
to apoptosis. Tunicamycin in a dose-dependent manner up-regulates DR5, one of the two death receptors
for TRAIL; enabling the control of malignant cell growth. However, the cooperative relationship between
Tunicamycin and TRAIL has yet to be examined in the course of Herpes Simplex Virus (HSV) immune
induction. Preliminary data from our laboratory points to the involvement of TRAIL in antigen
presentation. Dendritic Cells (DCs), the professional antigen presenting cell, will present antigen to T
cells to induce an immune response. However, upon infection, dermal DCs migrate to peripheral lymph
nodes to transfer the viral antigen to the professional APC, which is lymph node resident. Within the
lymph nodes, Natural Killer cells expressing TRAIL induces apoptosis in the antigen bearing DCs;
allowing the apoptotic transfer of viral antigen for CD8 T cells cross-priming. We propose that the
expression of DR5 can be manipulated by administering a controlled concentration of Tunicamycin in a
viral infection as well. Cloned Dendritic Cells (DC2.4) and splenocytes were co-cultured with various
concentrations of Tunicamycin and analyzed every eight hours to determine the optimal concentration
and time point of DR5 up-regulation. Using this kinetics information, we determined the optimal timing
and dosage of Tunicamycin requirement for the up-regulation of DR5 on DCs in vivo. With this
information, injections of Tunicamycin can be administered in vivo to modulate the quality of CD8+ T
cell function. Concluding, Tunicamycin is able to up-regulate the death receptor for TRAIL-mediated
antigen transfer elevating the quality of an anti-viral response.
2008 Appalachian Student Research Forum Page 74 Medical Residents and Post-Doctoral Fellows
TRACHEAL METASTASIS OF LUNG CANCER: A RARE COMPLICATION
Venkataramanan Gangadharan MBBS, Muhanad Al Hasan MD, Ryland P. Byrd, Jr. MD
Pulmonary/Critical Care Division, Dept. of Internal Medicine, East Tennessee State University, Johnson
City, TN
A 77 year-old man complained of shortness of breath on moderate exertion and hemoptysis about 2-3
times per week measuring to less than a teaspoon at each instance. He was diagnosed with squamous cell
cancer of the lung one year prior to these complaints and had undergone successful resection of the tumor.
His stage at surgical resection was Stage 1B (T2N0M0). The surgical margins and lymph nodes were free
of malignancy. He had no adjuvant chemotherapy or radiation treatments. The patient was afebrile and his
vital signs were normal. His physical examination was significant only for a post-thoracotomy scar. There
was no wheezing, stridor, or other airway noises present on auscultation of the chest. His chest
radiographs demonstrated post-surgical changes with no new findings. Computerized tomography of his
chest documented a small pleural effusion and tracheal mucosal irregularities. Fiberoptic bronchoscopy
demonstrated variable sized masses in the trachea but no bronchial abnormalities. On further examination
it was noted these masses were friable in nature and easily bled on manipulation. Histopathology
examinations of biopsies obtained at fiberoptic bronchoscopy were returned as squamous cell carcinoma.
The patient was subsequently referred to undergo brachytherapy. Lung cancer is one of the most
commonly diagnosed malignancies and is the most common cause of cancer related death in the United
States. Its presentation varies from small asymptomatic lung nodules discovered serendipitously, to
symptomatic masses, to symptoms related to metastatic disease or paraneoplastic syndromes. As with
many malignancies, metastasis from lung cancer is possible. Lung cancer typically metastasizes to the
brain, bone, and adrenal gland. Endobronchial metastasis from a primary lung cancer is unusual with a
reported incidence of 2-18%. Metastasis to the trachea from a primary lung cancer is even rarer. The
reported incidence is approximately 0.6%. Symptoms from tracheal metastasis include: cough, dyspnea,
hemoptysis, and respiratory distress from tracheal obstruction which may mimic asthma. A careful history
and thoracic examination is essential in order to provide appropriate and timely investigations and
interventions to reduce morbidity. This report should, therefore, alert health care providers to the
possibility of tracheal metastasis. It is hoped that early recognition of tracheal metastasis may afford the
patient better and more effective treatment.
A CASE OF VENTRICULAR TACHYCARDIA TERMINATED
BY CAROTID MASSAGE
Sumit Kalra, MD, Romsford Brenya, MD, Rajendra Kadari, MD, Terry Forrest, MD
Department of Internal Medicine, East Tennessee State University, Johnson City, Tennessee
Initial evaluation of a broad complex tachycardia includes a differentiation between ventricular
tachycardia and supraventricular tachycardia with aberrant conduction. Vagal maneuvers are a time
honored method for this differentiation based on the fact that supraventricular tachycardias are slowed or
terminated by enhanced vagal tone whereas ventricular tachycardias are not affected. We present an
unusual case of ventricular tachycardia which was terminated by carotid massage. An 88-year-old
Caucasian male with a past medical history of chronic bronchitis, diabetes, hypertension and
hypothyroidism was admitted with respiratory tract infection. On telemetry, he was noticed to have a
broad complex tachycardia at a rate of 140 beats/minute. The patient denied any chest pain or shortness of
2008 Appalachian Student Research Forum Page 75 breath associated with this heart rhythm and was hemodynamically stable. His physical examination was
normal and his laboratory profile did not reveal any electrolyte abnormality. He denied any past history of
coronary artery disease, angina, syncope, aborted sudden death or any cardiac arrhythmia. The differential
at this point included ventricular tachycardia Vs supraventricular tachycardia with aberrant conduction.
On carotid massage, his rhythm converted to sinus rhythm but changed back to broad complex
tachycardia as soon as the carotid massage was stopped. A 12 lead EKG revealed broad complex
tachycardia with AV dissociation, fusion beat, and capture beat all pointing towards a diagnosis of
ventricular tachycardia. A carotid massage was attempted again and the rhythm cardioverted back to sinus
rhythm again only to revert back to ventricular tachycardia on stopping the carotid massage. He was
admitted to the Cardiac Care Unit and successfully cardioverted with intravenous amiodarone. Later, an
adenosine myoview stress test and echocardiogram were performed and were reported as normal. A
diagnosis of idiopathic ventricular tachycardia was made in the absence of ischemic heart disease and a
structurally normal heart.
About 10% of ventricular tachycardias originate in structurally normal hearts and are idiopathic. A
majority of these are believed to originate in the outflow tracts of the heart (left or right ventricular
outflow tract ventricular tachycardias, OTVTs) and can be recognized based on the 12 lead EKG. It has
been proposed that the mechanism of these OTVTs may be cyclic AMP (cAMP) mediated. The beta
adrenergic receptor stimulation activates the myocardial cell adenylate cyclase thus increasing the
intracellular cAMP concentration. cAMP then activates protein kinase A leading to an increased
intracellular calcium concentration which subsequently causes an increase in the inward sodium current
(delayed afterdepolarizations). When a delayed afterdepolarization reaches a threshold, it generates an
action potential and leads to a triggered ventricular tachycardia. Acetylcholine (through vagal maneuver)
is an inhibitor of adenylate cyclase and acts on M2 muscarinic receptors on the myocardial cell to
decrease the concentration of cAMP. This in turn attenuates the afterdepolarization through the
mechanism described above and leads to the termination of triggered activity. Our patient had an EKG
most consistent with Outflow tract ventricular tachycardia (Subtype: Aortic Cusp Ventricular outflow
tract tachycardia) based on previously published EKG criteria. We believe that the OTVT in our patient
was probably cAMP mediated and therefore terminated by vagal stimulation as described above. Our case
reminds internists that although vagal maneuvers are a useful tool in the differentiation between
supraventricular and ventricular origin of broad complex tachycardias, they are by no means diagnostic.
AN EXCEPTIONAL VARIANT OF SINGLE CORONARY ARTERY ANOMALY, PRESENTING
WITH ACUTE ST SEGMENT ELEVATION MYOCARDIAL INFARCTION
Georges El Khoury and Lucien Abboud
Depts. of Internal Medicine and Cardiology, College of Medicine, East Tennessee State University,
Johnson City, TN
Congenital coronary artery anomalies are infrequently seen during coronary angiography occurring in
0.06 % to 1.3 %. Some variants are totally benign and are discovered incidentally in the catheterization
laboratory, however others may present with sudden cardiac death. Single coronary artery is one subtype
which is often associated with other congenital cardiac malformations most of which are benign. We
report a 75 y o male patient with an exceptional variant of single coronary artery anomaly with right
coronary artery arising from the distal left circumflex system who presented with ST Elevation
Myocardial Infarction of the anterolateral wall secondary to Left Anterior Descendant artery occlusion.
The patient has no previous cardiac history. Would the patient has survived if he had the lesion involving
the Left circumflex artery or would it be a case of undiagnosed coronary artery anomaly presenting with
sudden cardiac death?
2008 Appalachian Student Research Forum Page 76 ELEVATED IGE, EOSINOPHILIA AND PERSISTENT AIRWAY OBSTRUCTION
Christopher W. T. Miller and Guha Krishnaswamy
Dept. of Allergy and Immunology, College of Medicine, East Tennessee State University, Johnson City,
TN
The prevalence of Strongyloides parasitosis in the East Tennessee belt is strikingly high, and may be
masked by atypical clinical or laboratory features. A 67-year-old female patient presented with a recent
worsening of her baseline intermittent cough and dyspnea, associated with a number of nonspecific
complaints (rhinorrhea, nasal congestion, and sneezing). Pertinent past medical history included severe
chronic obstructive pulmonary disease, diabetes mellitus type 2, and a history of non-small cell lung
cancer treated with radiation therapy 13 months before the current presentation. Physical exam was
essentially unremarkable apart from some mild wheezing bilaterally. Work-up yielded a normal white
blood cell count with an elevated eosinophil percentage varying from 16% to 21%, along with an elevated
C-reactive protein level of 70.7. Allergic assays were essentially negative for major offenders and stool
studies were repeatedly negative for ova and parasites. Serum immunoglobulin assays revealed an
extremely elevated IgE level of 4998.5. The patient was assessed by several specialists without reaching
a definitive etiology despite extensive work-up (including bone marrow biopsy, chest computed
tomography, HIV, hepatitis screen, autoimmune markers, heavy metal screen, TSH and cortisol levels).
A presumptive diagnosis of hyper-IgE syndrome was established. However, a serum immunoglobulin G
assay for Strongyloides returned positive at 14.7, leading to a diagnosis of strongyloidiasis. The patient
was treated with three courses of ivermectin (15 mg single-dose regimens over 3 months). The patient’s
respiratory symptoms greatly improved, with a corresponding decrease in laboratory findings to nearnormal values. This case reiterates the need for a high level of suspicion in patients with suggestive
symptoms of strongyloidiasis even in the face of negative stool assays.
MOLECULAR LINK BETWEEN TOLL LIKE RECEPTORS,
OBESITY AND RISK OF CANCER
Subhadra Nandakumar, Stacie Woolard and Uday Kumaraguru
Dept. of Microbiology, College of Medicine, East Tennessee State University, Johnson City, TN
According to the National Cancer Institute statistics, the incidence of new cases of cancer in 2007 is about
1,444,920 and it was estimated that there were about 559,650 cancer related deaths in the US. This
number is much above the average cases in the developing and under-developed countries. Recent studies
propose a link between obesity and susceptibility to cancer. Oddly obesity is a major problem affecting
66% of the American population as reported by CDC. Hence, it is tempting to hypothesize that obesity
and cancer development may correlate with exposure pattern to infection that engages Toll like receptors
(TLRs). We tested our hypothesis in C57Bl6 mice. These mice were housed in two different
environments, one an ultra-clean environment, where cages, bedding, food and water needed for the mice
were sterilized. The other was the dirty environment, which was simulated by administering the mice with
cocktail of TLR ligands (containing antigenic components of virus, bacteria and fungi) periodically.
These mice were later subjected to tumor challenge and were analyzed for various immunological
parameters. Our findings show that the mice growing in “dirty” environment had significantly lower
weights and were able to resist tumor challenge much better than those growing in “clean” environment.
Based on the outcome of this study we can conclude that TLR activation by its ligands results in an
increased lipid metabolism leading to the prevention of obesity and heightened immune preparedness.
This increases the ability of the host to resist the tumor challenge. We demonstrate that the activated
phagocytic ability, mediated by key cytokines as the mechanism of this resistance.
2008 Appalachian Student Research Forum Page 77 PHILADELPHIA-CHROMOSOME POSITIVE ACUTE LYMPHOBLASTIC LEUKEMIA:
LONG-TERM SURVIVAL IN AN ADULT JEHOVAH’S WITNESS
Vivekananda Pattabiraman, MD, Harsha G. Vardhana, MD, Stephen J. Smith MD, and Koyamangalath
Krishnan, MD
Dept of Internal Medicine, Quillen College of Medicine, East Tennessee State University, Johnson City,
TN
Key words: Philadelphia-positive ALL (Ph+ ALL), Stem Cell Transplantation (SCT), Tyrosine kinase
Inhibitor (Imatinib) . Background: Adult Ph+ ALL has a poor prognosis. We present a case of Ph+ ALL
who belongs to the Jehovah’s faith and exhibited prolonged survival with modified conventional
chemotherapy along with long-term treatment with a tyrosine-kinase inhibitor. Case Report: A 31 yr old
man who belonged to the Jehovah’s faith was diagnosed with Ph+ ALL. His religious beliefs precluded
blood transfusion and hence he was not considered suitable for SCT. He had anemia and
thrombocytopenia at diagnosis. and was therefore treated with attenuated doses of induction
chemotherapy (along with supportive treatment with GM-CSF, Erythropoietin, amino-caproic acid)
followed by usual doses of consolidation and maintenance chemotherapy for 2 years.Imatinib was started
along with maintenance chemotherapy and continued indefinitely. We have been successful in achieving
80month survival (until Feb 2008) along with clinical remission of 60 months. Conclusion: Adult Ph+
ALL is known to be a malignancy with poor prognosis. SCT is regarded as the treatment of choice but
survival rates are far from satisfactory. Conventional chemotherapy along with Imatinib is capable of
inducing long-term clinical, hematological and molecular remission and also better survival when
compared to SCT. Tyrosine Kinase inhibitors are a promising therapeutic option for patients unable to
undergo SCT. References:
Garcia et al: The hyper CVAD regimen in adult ALL: Hematology
Oncology clinics Of North America 14,1381-1386 Thomas DA et al: Treatment of Ph+ ALL with
Hyper-CVAD and imatinib mesylate: Blood 2004 Jun 15;103(12):4396-407
EFFECTS OF VITAMIN E ISOTYPES ON KG-1 LEUKEMIC CELLS GENE EXPRESSION, CELL
GROWTH AND PROLIFERATION
Nakhle Saba, Sarah G. Whaley, Koyamangalath Krishnan and Sharon Campbell
Depts of Internal Medicine and Medical Oncology, College of Medicine, East Tennessee State University,
Johnson City, TN
Background: Vitamin E is a fat-soluble vitamin that exists in eight different isoforms: four tocopherols
(alpha-, beta-, gamma- and delta-), and four tocotrienols (also alpha-, beta-, gamma- and delta-). Studies
are underway to determine whether vitamin E, through its ability to limit production of free radicals,
might help prevent or delay the development of cancer. Other studies suggest that vitamin E may
modulate signal transduction pathways to prevent or reverse the effects of cancer. In our study, we
examined the effect of vitamin E isotypes on some gene products involved in acute leukemia such as
HOXA9 (Homeobox A9), PBX1 (Pre-B cell leukemia transcription factor 1), E2A-PBX1, HOXB6,
CDX2, NUP98, CYCLIN D1 and CYCLIND3, as well as on in-vitro leukemic cell growth and
differentiation. HOXA9 is a transcription factor with a central role in both hemopoiesis and leukemia. A
high level of HOXA9 is a characteristic feature of acute myeloid leukemia (AML) and may be sufficient
to cause this disease. PBX1 is involved in a form of pre-B-cell acute lymphoblastic leukemia (B-ALL).
E2A-PBX1 is a protein resulting from a chromosomal translocation t(1;19)(q23;p13.3) which involves
PBX1 and E2A genes. E2A-PBX1 transforms cells by constitutively activating transcription of genes
2008 Appalachian Student Research Forum Page 78 regulated by PBX1 or by other members of the PBX protein family. NUP98 encodes for a 98 kilodalton
(kDa) O-linked glycoprotein, a component of the nuclear pore complex; Translocation t(7;11) (p15;p15)
in AML fuses the genes for NUP98 and class I homeoprotein HOXA9. NUP98-HOXA9 fusion protein
may promote leukemogenesis. CDX2, a member of the caudal-type homeobox family is aberrantly
expressed in most cases of acute myeloid leukemia and promotes leukemogenesis. Monoallelic
expression of CDX2 is observed in 90% of patients with AML and may perturb hematopoiesis by
affecting HOX gene expression. HOXB6 Collaborates with other genes in generation of AML. Methods:
KG-1 cells were treated with 10 and 20 micromolar (μM) gamma tocotrienol (γT3) and delta (δT3)
tocotrienol, and 25 and 50 μM gamma tocopherol (γT) and delta tocopherol (δT) for 72 hours. Western
Blot analysis was performed on the cell lysates using HOXA9, PBX1, E2A-PBX-1 HOXB6, CDX2,
NUP98, CYCLIN D1 and CYCLIND3 antibodies. MTT assays were done separately using
concentrations starting from 5 μM up to 80 μM of all four isoforms (γT, δT, γT3 and δT3). Results: δT3
and γT3 have downregulatory effect on HOXA9, PBX1 and CDX2 genes when used at 10 and 20 μM
concentrations for 48 hours treatment; γT and δT have downregulatory effect on HOXA9, PBX1, E2APBX-1, HOXB6, CDX2, CYCLIN D1 and CYCLIND3, and an upregylatory effect on NUP98 when used
at 50 μM concentration for 72 hours treatment. MTT assay showed that all four isoforms (δT3, γT3, γT
and δT) have a potent inhibitory effect on KG-1 leukemic cell growth and proliferation.
ELICITING PATIENTS’ CANCER STORIES AND HEALTH DISPARITIES WITH AN
INTEGRATED PATIENT-CENTERED STORYTELLING INTERVIEW, AND THE USE OF THESE
VIDEO RECORDED STORIES TO EDUCATE AND TRAIN HEALTH CARE PERSONNEL IN
EFFECTIVE PATIENT PHYSICIAN COMMUNICATION
Harsha G. Vardhana, MD, Forrest Lang, MD, Joseph Sobol, PhD, Fred Tudiver, MD, and Koyamangalath
Krishnan, MD
Depts. of Internal Medicine and Family Medicine, East Tennessee State University, Johnson City, TN
In Southern Appalachia, a number of factors result in high rates of cancer incidence and mortality. We
decided to explore and document the factors responsible for this disparity through a qualitative research
method. The project involves a set of patient interviews conducted over time with cancer patients that
focus on key oncology-care transition points. The interviews integrate patient centered communication
(an emphasis on the patient’s ideas, concerns and expectations) with elements of the story-telling
tradition. The stories they communicate will be video recorded and distributed to medical audiences
through a module. In the initial phase of this American Society of Clinical Oncology (Young
Investigators Award) and Appalachian Center for Translational Research Disparities (ACTRID) funded
study the investigators interviewed and analyzed a pilot cohort of cancer patients. The initial beta testing
began by interviewing six patients, each selected from the University cancer center. The recorded,
interviews lasted between 1.5 to 2.5 hours. At this point in our beta trial, we will present and discuss our
interview template that incorporates important elements of oncology care, patient-centered
communication skills and the story telling tradition, initial “stories and perspectives” (in the form of
videos) from the beta test that identify potential sources of factors influencing health care disparities and
common themes which have evolved in our interviews highlighting the similarities and differences in the
experiences and preferences of cancer patients. From our Patient-Centered/Storytelling Beta trial we are
beginning to identify certain emerging themes. The themes provide moving video-audio records of
patients’ experience and reactions to a set of common communication issues in cancer care. Examples
include: 1) Jargon and Medical Obfuscation. Patients preferred that their physicians not talk to them in
medical terms; 2) Understanding of prognostic communications. Appalachian patients prefer not to make
decisions in regards to their cancer care based on numbers given in terms of response rates and overall
survival from their oncologist; 3) Nursing support roles. Nurses played a crucial role in terms of offering
emotional support to cancer patients; 4) Examples of insensitive communication about bad news. Patients
2008 Appalachian Student Research Forum Page 79 expressed concerns that health care providers appeared emotionally distant during the communication of
bad news; 5) Emotion handling-the good the bad and the ugly. Patients have provided vivid stories in
which physicians and nurses have ignored patients’ feelings, and also stories of great human compassion.
6) Role of spirituality and religion in dealing with cancer. In addition to publishing our data, we will
develop and distribute a module to help practicing oncologists to communicate more effectively with
patients. This will enable, physicians to reach common ground with their patients regarding cancer
treatment plans. By achieving better patient-physician relationship and integration of the patient’s
personal and cultural beliefs with evidence based cancer treatment we hope to reduce cancer related
health care disparity.
REM SLEEP BEHAVIOR DISORDER (RBD) AS AN EARLY MARKER FOR DEVELOPMENT OF
NEURODEGENERATIVE DISEASES: A LITERATURE REVIEW
Umesh Vyas MD, PGY III,
Dept. of Psychiatry and Behavioral Sciences, James H Quillen College of Medicine, East Tennessee State
University, Johnson City, TN
Introduction and Hypothesis: RBD is a parasomnia characterized by intermittent loss of
Electromyographic atonia normally present during REM sleep and emergence of purposeful complex
motor activity associated with vivid dreams, and there is enactment of dreams. Normally, a diffuse
hypotonia of muscles occur during REM sleep which inhibits motor enactment of dreams. This condition
was first described by Schenck CH et al in 1986. It usually affects older males, and can be either
idiopathic or symptomatic of various underlying disorders, in particular neurodegenerative disease; in
later case RBD may be prodromal symptom of neurological disease. RBD can be acute which is almost
always induced by medications (Multiple medications can induce or aggravate RBD symptoms and
should be avoided in these patients), or chronic which can be idiopathic but most patients will eventually
exhibit signs and symptoms of variety of degenerative neurologic disorder. Sleep disorders have a high
prevalence especially in patients with Parkinson’s disease (PD) of particular interest is RBD. An
idiopathic form may predict later development of Parkinson’s disease or other neurodegenerative
disorders. Prevalence of RBD in general population is 0.5%. Its prevalence is 38% for Parkinson’s
disease, 90% for Multiple System Atrophy, and 95% for Dementia with Lewy bodies. Early diagnosis of
RBD is of paramount importance because of existence of effective treatment and for preventing injuries
to patient and bed partner. A Polysomnogram (PSG) is necessary to diagnose RBD, showing absence of
REM sleep atonia and related abnormal behavior. Method: Author has performed review of articles in
pubmed.gov by using key words: REM sleep Behavior Disorder. Conclusion: RBD often antedates
development of a neurodegenerative disorder. Evidence of RBD in PSG can be used to prescreen high
risk patient for development of neurodegenerative disorders, especially in high risk patient population.
Close follow up of patients with idiopathic RBD could enable early detection of neurodegenerative
disease. This finding may be of great interest when early effective treatment strategies and
neuroprotective agents become available.
2008 Appalachian Student Research Forum Page 80 2007 Award Winners
Division I – Undergraduates
Biomedical Sciences
First Place: Regina Phillips, ETSU. Faculty Sponsor – Dr. Sharon Campbell, Dept. of Internal
Medicine. Natural Vitamin E Isoforms, Gamma and Delta Modulate TGFβ in PC-3 Prostate
Cancer Cell Lines
Second Place: Jonathan Calloway, ETSU. Faculty Sponsor – Dr. Michael Ramsey, Dept. of
Kinesiology, Leisure & Sport Sciences. Relationship Between Collegiate Throwers' Ability to
Produce Maximum Force and Their Ability to Potentiate
Natural Sciences & Mathematics
First Place: Leslie Epling, ETSU. Faculty Sponsor – Dr. Cecilia McIntosh, Dept. of Biological
Sciences. Cloning PGT-4 for Expression and Characterization as a Putative Glucosyltransferase
in Citrus Paradisis (Grapefruit)
Second Place: Connie Blalock, ETSU. Faculty Sponsor – Dr. Debra Knisley, Dept. of
Mathematics. Fermat's Last Theorem, A Combinational Approach
Social & Behavioral Sciences
First Place: Ian D. Longacre, ETSU. Faculty Sponsor – Dr. Russell Brown, Dept. of
Psychology. Nicotine Sensitization in Adolescent and Adult Rats D2 Receptor Primed as
Neonates: Gender Differences and Neurotrophic Factors
Second Place: Andrew B. Hughes, ETSU. Faculty Sponsor – Dr. Russell Brown, Dept. of
Psychology. Methylphenidate (Ritalin) Exposure to Adolescent D2-Primed Rats Eliminates the
Initial Hypoactive Response to Nicotine in Adulthood
Third Place: Leslie King, ETSU. Faculty Sponsor – Dr. Joel Hillhouse, Dept. of Psychology.
The Future Implications of Indoor UV Tanning at a Young Age
Fourth Place: Matthew F. Lazenka, ETSU. Faculty Sponsor – Dr. Russell Brown, Dept. of
Psychology. Age and Gender Differences in the Virtual Morris Water Maze from 12 - 25: A
Fresh Look at Strategies
2008 Appalachian Student Research Forum Page 81 Division II – Graduate Students, 1-2 Years
Natural Sciences & Mathematics
First Place: David Hammond, ETSU. Faculty Sponsor – Dr. Ranjan Chakraborty, Dept. of
Health Sciences. Characterization of Binding & Transport & Initial Identification of the Genes
Involved in the Biosynthesis & Transport of Schizokinen, A Dihydroxamate Siderophore
Produced by Rhizobium Leguminosarum IARI 917
Second Place: Tande Jacob Fosso, ETSU. Faculty Sponsor – Dr. Scott Kirkby, Dept. of
Chemistry. A Computational Chemistry Study of Spin Traps
Arts & Humanities/Social & Behavioral Sciences (combined)
First Place: Jessica Brown, ETSU. Faculty Sponsor – Dr. Kerry Proctor-Williams, Dept. of
Communicative Disorders. Typically Developing Children Display Two Prelinguistic Acts Pre
Minute Before First Words
Second Place: Zachary A. Cope, ETSU. Faculty Sponsor – Dr. Russell Brown, Dept. of
Psychology. Amphetamine Sensitization in a Rodent Model of Psychosis
Third Place: Brandon Phillips, ETSU. Faculty Sponsor – Dr. Christopher McCrea, Dept. of
Communicative Disorders Voice Onset Time as a Clinical Indicator of Hyperfunctional Voice
Disorders
Division III – Graduate Students, 2+ Years
First Place: Michelle Chandley, ETSU. Faculty Sponsor – Dr. Barney Miller, Dept. of
Psychiatry. Identification of a Possible Functional Cre-Element Found in the Promoter of the α7
Nicotinic Receptor Subunit and its Association with Schizophrenia
Second Place: Jennifer Vanover, ETSU. Faculty Sponsor – Dr. Robert Schoborg, Dept. of
Microbiology. HSV-2 CO-Infection Stimulates Chlamydia Trachomatis Persistence via a Novel
Mechanism
Division IV – Medical Students
First Place: Christopher L. Cropsey, ETSU. Faculty Sponsor – Dr. Alok Agrawal, Dept. of
Pharmacology. Binding of C-Reactive Protein to Immobile Immune Complexes
Second Place: Scott Cook, ETSU. Faculty Sponsor – Dr. Kevin Breuel, Dept. of OB/GYN.
Relationship of Matrix Metalloproteinases with Marker of Fetal Lung Maturity
Division V – Residents/Post-Doctoral Fellows
First Place: Rachna Bharti, MD, ETSU. Faculty Sponsor – Dr. Burt Banks, Dept. of Family
Medicine. Review of Literature on Proteinuria in Pre-Eclampsia
Second Place: Umesh Vyas, MD, ETSU. Faculty Sponsor – Dr. Hetal K. Brahmbhatt, Dept. of
Psychiatry. Risk of Breast and Other Cancers due to Hyperprolactinemia Caused by
Antipsychotics (Neuroleptics) or Other Medications: A Literature Review
2008 Appalachian Student Research Forum Page 82 Division VI – Case History
First Place: Avani Prabhakar, ETSU. Faculty Sponsor – Dr. Hetal K. Brambhatt, Dept. of
Psychiatry. A Case of Ceftriaxone Induced Neutropenia
Second Place: Raja S. Vadlamudi, MD, ETSU. Faculty Sponsor – Dr. Mark F. Young, Dept.
of Internal Medicine. Enteroscopic Retrieval: A Possible Treatment Modality for Displaced
Esophageal Stent
Post-Baccalaureate Oral Presentations
First Place: Stacie N. Woolard, ETSU. Faculty Sponsor – Dr. Uday Kumaraguru, Dept. of
Microbiology. Involvement of TNF Receptor Apoptosis Inducing Ligand (TRAIL) in NK Cell
Mediated Anti-HSV Adaptive Immune Response
Second Place: Carrie Jolly, ETSU. Faculty Sponsor – Dr. J. Russell Hayman, Dept. of
Microbiology. Yeast Two-Hybrid Analysis of Protein-Protein Interactions Involving a
Microsporidia Adam (A Disintegrin and Metalloprotease) Protein
Second Place: Alexander Eddo, ETSU. Faculty Sponsor – Dr. Dhirendra Kumar, Dept. of
Biological Sciences. Characterization of the Pathway Leading to the Synthesis of Salicylic Acid
in Plants Under Attach by Pathogen
Resident/Post-Doc Oral Presentations
First Place: Ahmad Halawa, MD, ETSU. Faculty Sponsor – Dr. Ryland Byrd, Dept. of
Internal Medicine. Wegener's Granulomatosis and Purulent Pericarditis: The First Case Report
2008 Appalachian Student Research Forum Page 83 2007 Student Choice Awards
Division I – Undergraduates
Andrew B. Hughes, ETSU. Faculty Sponsor – Dr. Russell Brown, Dept. of
Psychology. Methylphenidate (Ritalin) Exposure to Adolescent D2-Primed Rats
Eliminates the Initial Hypoactive Response to Nicotine in Adulthood
Division II – Graduate Students, 1-2 Years
Zachary A. Cope, ETSU. Faculty Sponsor – Dr. Russell Brown, Dept. of
Psychology. Amphetamine Sensitization in a Rodent Model of Psychosis
Division III – Graduate Students, 2+ Years
Jennifer Vanover, ETSU. Faculty Sponsor – Dr. Robert Schoborg, Dept. of
Microbiology. HSV-2 CO-Infection Stimulates Chlamydia Trachomatis Persistence
via a Novel Mechanism
Division IV – Medical Students
Scott Cook, ETSU. Faculty Sponsor – Dr. Kevin Breuel, Dept. of OB/GYN.
Relationship of Matrix Metalloproteinases with Marker of Fetal Lung Maturity
Division V – Residents/Post-Doctoral Fellows
Umesh Vyas, MD, ETSU. Faculty Sponsor – Dr. Hetal K. Brahmbhatt, Dept. of
Psychiatry. Risk of Breast and Other Cancers due to Hyperprolactinemia Caused
by Antipsychotics (Neuroleptics) or Other Medications: A Literature Review
Division VI –Case History
Avani Prabhakar, ETSU. Faculty Sponsor – Dr. Hetal K. Brahmbhatt, Dept. of
Psychiatry. A Case of Ceftriaxone Induced Neutropenia
2008 Appalachian Student Research Forum Page 84