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2007 Appalachian Student Research Forum
Abstracts
(Names are listed alphabetically by first author in each Division and Category)
Division 1 – Undergraduates
Biomedical Sciences
RELATIONSHIP BETWEEN COLLEGIATE THROWERS’ ABILITY TO PRODUCE
MAXIMUM FORCE AND THEIR ABILITY TO POTENTIATE
J.G. Calloway, J.M. Kraska , M.A. Kinser, M.E. Stone and M.W. Ramsey. Department of
Kinesiology, Leisure and Sport Sciences, East Tennessee State University, Johnson City,
TN 37614
PURPOSE: This investigation examined the relationship between a collegiate thrower’s ability
to produce maximum force and post-activation (or post-exercise) potentiation.
METHODS: 5 male and 3 female collegiate throwers (mean + SD; age = 19.8 + 0.9 y; weight =
111.7 + 18.9 kg) participated in a specifically designed 4-week strength and conditioning
program. At the end of the 4-week training cycle maximal force and explosive power were
measured with and without a potentiation protocol. The ability to produce maximum force was
determined by measuring the isometric peak force (IPF). On the first day athletes performed
isometric pulls from the power position (mid-thigh) in a custom designed force rack over a force
plate (1000 Hz). Along with the isometric pulls the athletes were instructed to perform an
overhead ball throw. On day two the athletes performed a 1 RM snatch test and a potentiation
protocol consisting of two trials of mid-thigh clean pulls at 60 kg, 100 kg, 120 kg, 140 kg, 160 kg,
and 100 kg. Each trial was separated by approximately one to two minutes of rest in order to
ensure that full recovery occurred between each trial. All trials were performed on a custom
rack placed over a force plate (1000 Hz), two linear position transducers were attached to the
ends of the barbell. Individual force time curve analyses were used to determine the peak force,
peak velocity, peak power, and peak rate of force development for each trial. Peak rates of
force development (RFD) and peak force were determined from the ground reaction forces,
while the velocity data were derived from the vertical barbell displacements collected by the
linear transducers. Power was then calculated from the force values and the velocity data. A
t-Test was used to determine if significant difference existed between pre- and post-potentiation
values that were tested. Correlations were calculated using a pearson-r to determine the
strength of the relationship between peak isometric force (N), RFD (0-250 ms), and the
potentiation of dynamic peak force, peak velocity, and peak power, as well as ball throw (m),
and maximum snatch (kg).
RESULTS: Potentiation values for peak force, peak velocity, peak power, and RFD showed
significant statistical improvement (p < 0.05). When correlating these potentiation values to IPF
the correlations ranged from moderate to strong for peak force (r=0.47), peak velocity (r=0.34),
and peak power (r=0.76). Isometric peak force also correlated strongly with ball throw (r=0.76)
and maximum snatch (r=0.79).
CONCLUSION: These data indicate that the relationship between the ability to produce force
(IPF) and the potentiation ability in athletes are related. Not only will stronger athletes perform
2007 Appalachian Student Research Forum
better, but also their performance can be enhanced to an even greater extent, versus a weaker
athlete, through potentiation.
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THE RELATIONSHIP BETWEEN POSTMENOPAUSAL HORMONE THERAPY AND
THE EXERCISE ELECTROCARDIOGRAM: A RETROSPECTIVE STUDY
Lindsey Perry and Dr. Melessia Webb. Department of Professional Roles and Mental
Health Nursing, East Tennessee State University, CON, Johnson City, TN 37614
Postmenopausal Hormone Therapy (PHT) is a hormone regimen consisting of estrogen or
estrogen-plus-progestin which is administered to females in order to relieve symptoms of
menopause and to prevent osteoporosis. Estrogen has been found to be molecularly similar to
the drug digitalis, a positive inotrope. Consequently, PHT may be associated with significant
ST-segment changes on the electrocardiogram. A retrospective chart review was conducted
with the sample’s inclusion criteria of females, 50 years of age or older who had experienced a
stress test followed by a cardiac catheterization in order to investigate a possible relationship
between PHT and cardiac-stress testing. The patient’s cardiac catheterization results were
used as the reference standard to identify true positive stress test from false positives. The
researchers reviewed 128 medical records and collected the following data: demographics,
laboratory values, medical history, current list of medications, stress test date and results,
cardiac catheterization date and results, and interventions after cardiac catheterization. Results
of this study did not identify a higher rate of false-positive results for stress tests in females
taking PHT than females who were not taking PHT in the study group. However, females with a
lower percentage of coronary artery occlusion of specific arteries, the right coronary artery
(RCA), the left anterior descending artery (LAD), and the left circumflex artery (LCx), were
associated with a false-positive stress test. In addition, the following medical histories were
significant in relation to having a higher rate of false-positive stress tests: (1) no previous
myocardial infarction (MI); (2) no invasive cardiac intervention; (3) peripheral vascular disease
(PVD); (4) no significant vessel disease; (5) not taking a lipid-lowering agent; (6) not taking an
antiplatelet medication; and (7) taking an antiarrythmic medication. It is also noteworthy that
40% (n = 51) of the study population had a false-positive stress test, while only 23% (n = 30)
had a true-positive stress test result. Females, who had a higher rate of true-positive stress
tests, were those with lower HDL cholesterol levels and a higher percentage of occlusion in the
RCA, the LAD, and the LCx. Medical histories associated with a higher percentage of truepositive stress tests were positive history of MI, history of invasive cardiac intervention, having
significant vessel disease, females taking a H2-Blocker, females taking an antiplatelet
medication, and not taking a Proton Pump Inhibitor. Due to the small number of participants,
the effect size was small with a power of 0.169; the researcher recommends that this study be
conducted again with a larger sample size in order to increase the power of the study. The
results may prove useful in future referral for noninvasive diagnostic testing for heart disease in
females.
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NATURAL VITAMIN E ISOFORMS, GAMMA AND DELTA MODULATE TGFβ 2 IN
PC-3 PROSTATE CANCER CELL LINES
Regina Phillips, Chuan Xing Ho, Sarah Whaley, Koyamangalath Krishnan, William Stone
and Sharon Campbell. Department of Internal Medicine, East Tennessee State
University, COM, Johnson City, TN 37614
In early stages of cancer, TGFβ (primarily TGFβ1) inhibits tumor development, but at later
stages of tumor progression, cancer cells evade the growth inhibition by TGFβ. TGFβ is
switched from a tumor suppressor to a tumor promoter. Lu et al. has demonstrated that the
PC-3 cancer cell line exhibits constitutive expression of NF-ĸB, an anti-apoptotic transcription
factor. The constitutive expression of NF-ĸB conveys resistance to Tumor Necrosis Factor
alpha (TNF-α), a natural apoptotic stimuli. Transforming Growth Factor β2 (TGFβ2) has been
identified as a factor mediating the constitutive activation of NF-ĸB. The Alpha-Tocopherol,
Beta Carotene study found that substantially fewer participants taking alpha-tocopherol
developed prostate cancer. Helzlsouer et al. furthered these findings by showing that high
plasma concentrations of gamma-tocopherol enhanced the effects previously seen with alphatocopherol. Further research has shown that vitamin E is able to induce cell death in PC-3 cells
with the effect being greatly increased when cells are treated with both TNFα and vitamin E,
suggesting a possible mechanism of action for TGFβ induced cell death. In this study, the
gamma and delta isoforms of vitamin E were tested for their ability to halt cell proliferation in
PC-3 prostate cancer cell lines in comparison with alpha-tocopherol at 20, 40 and 60 µM. By
western blotting and ELISA assays, it was found that the gamma and delta isoforms are more
potent inhibitors of cell proliferation than alpha-tocopherol. These natural vitamin E isoforms are
able to allow PC-3 cells to overcome the resistance to TNFα-mediated cell death. We further
demonstrate that these isoforms down regulate the expression of TGFβ2, NF-ĸB, and TGFβ
Receptor 1.
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CELL CYCLE REGULATORY PROTEINS IN TASTE CELL TURNOVER IN MICE
Colin Spaulding, Dr. Theresa Harrison and Lorraine Adams. Department of Biology and
Department of Anatomy and Cell Biology, East Tennessee State University, COM,
Johnson City, TN 37614
The cells in mammalian taste buds are known to regenerate throughout life every 10 – 14 days,
but this process of taste receptor cellular turnover, particularly details concerning the regulation
of progenitor cell division and differentiation, is still poorly understood. In all cells, cellular
division and cell cycle exit prior to differentiation is regulated primarily by cell cycle regulatory
proteins (CCRPs) such as cyclin dependent kinases (Cdks), Cdk inhibitors (CKIs), and the
cyclin family of proteins. Studies have shown that normal taste buds express the CKI, p27(Kip1)
(Hirota et al, 2001). Previous experiments in this laboratory (Adams et al, 2006) demonstrated
that animals with null mutations for the p27(Kip1) gene (“knockouts”) have normal taste bud
morphology, but an increased rate of cell turnover, and cyclin D2 is the major D cyclin
expressed in taste tissues. In the present experiments, we wanted to examine in more detail
the expression of p27(Kip1) and cyclin D2 in normal taste buds, and to determine if the absence
of p27(Kip1) in mutants results in changes in the expression of D cyclins and/or other CKIs
which may act to maintain the generally normal phenotype observed in taste buds of p27(Kip1)
knockout mice. We used single and double-label immunohistochemical techniques to evaluate
the expression of D cyclin isoforms and proteins of the Cip/Kip family of CKIs in circumvallate
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papillae of adult wild type and p27(Kip1) knockout mice. Light and confocal microscopy
techniques were used to analyze the investigated proteins in fluorescent-labeled sections of
tissue. Mature taste buds in wild type animals showed between 10 – 15% of their total cells to
be labeled for p27(Kip1). Experiments to quantify the occurrence of p27(Kip1) and cyclin D2
labeled cells in the taste buds of wild type mice revealed averages of 5 – 6 cells per taste bud
labeled for p27(Kip1) and cyclin D2. Extensive overlap was found between labeled cell groups,
such that 55% of p27(Kip1) labeled cells also express cyclin D2, and 78% of cyclin D2 labeled
cells were double-labeled for p27(Kip1). When investigating the possibility of upregulation of
other CCRPs in the p27 (Kip1) null mutants, the results were negative for other members of the
cyclin family (i.e. D1, D3) and, in initial experiments, for other members of the Cip/Kip family of
CKIs (p21, p57). Experiments to date indicate that the hypothesis that normal taste bud
morphology in knockout animals is maintained due to compensatory increases in expression of
other Cip/Kip family CKIs and/or D cyclin proteins is not correct. Other CKIs (i.e. the INK family)
may be more important in regulating taste bud cell numbers. Surprisingly, cyclin D2 continues to
be expressed by some cells in the taste bud despite the fact that they have apparently
withdrawn from the cell cycle. This suggests that, in addition to its role in facilitating transit
through the cell cycle, cyclin D2 may play a different role in maturing and/or mature cells within
the taste bud.
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FANCONI PROTEINS IN DOUBLE STRANDED DNA REPAIR
Robin Woodard, Ph.D. and Henning Kuich. Department of Natural Sciences, University of
Virginia's College at Wise, Wise, VA 24293
Fanconi Anemia is a hereditary form of Aplastic Anemia. Aplastic Anemia is a disorder in which
destruction of bone marrow leads to a deficiency of circulating erythrocytes, leukocytes, and
platelets. As a result, patients exhibit symptoms such as easy bruising, fatigue, and a
decreased ability to fight off infections. In Fanconi Anemia, known mutations lead to a decrease
in chromosomal repair in bone marrow cells and their consequent inefficiency in blood cell
production. The Fanconi complex necessary to repair these chromosomal breaks is composed
of the proteins FancA, FancC, FancG, and FancF. Upon chromosome breakage, FancE
accumulates in the nucleus via an interaction with FancC, which in turn leads to the binding of
these two proteins to the remainder of the Fanconi Complex. Once the components are
assembled, the complex localizes to the chromosome break and initiates a repair pathway.
DNA-PK is a kinase known to be vital to repair of double stranded (ds) DNA breaks as well as
the joining of ds breaks induced during non-homologous recombination. It is composed of 3
subunits. The subunits Ku-70 and Ku-87 are responsible for binding the DNA and form a dimer
upon exposure to ds breaks. This dimer recruits the catalytic subunit (360 kDa), which in turn
activates a ds break repair pathway. This study investigated the relationship between the DNAPK and the Fanconi repair pathway by looking for interactions between proteins of the different
pathways in the presence of double stranded breaks. Upon verification of the binding of Ku-70
and Ku-87 to the simulated double stranded breaks, it was shown that FancC in fact does
interact with the same ds breaks. This interaction could be attributed to interactions with ds
breaks, Ku-70, Ku-87, or any combination of the three. This study was also able to show that
FancA does not seem to directly interact with ds breaks or either of the Ku proteins.
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Natural Sciences & Mathematics
FERMAT’S LAST THEOREM, A COMBINATORIAL APPROACH
Connie Blalock. Depart of Mathematics, East Tennessee State University, Johnson City,
TN 37614. Dr. Debra Knisley, advisor.
Fermat’s Last Theorem is perhaps one of the most famous theorems in all of mathematics. The
theorem states that the equation xn + yn = zn does not have positive integer solutions if n > 2.
Of course, there are many solutions if n = 2, such as 32 + 42 = 52. The case when n = 2 has a
well known geometric interpretation, namely x, y and z are lengths of sides of a right triangle.
Fermat claimed to have proof that no solution exists if n > 2, but he simply did not have room in
the margin to write the proof down. For more than 300 years, mathematicians tried to prove
Fermat’s claim, or find a counter example. Finally, the theorem was proved by Andrew Wiles, a
mathematics professor at Princeton, in 1995. His proof, however, was clearly not the proof that
Fermat claimed to have since it requires mathematics not yet invented at the time of Fermat.
In this work, we approach Fermat’s last theorem using a field of mathematics called
combinatorics and techniques that were known at the time of Fermat. In particular we use
numerical sequences with alphabets of size x, y, z where each sequence is of length n. In so
doing, we demonstrate a combinatorial interpretation rather than a geometric one. This method
provides an alternative interpretation of Fermat’s claim.
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CLONING PGT-4 FOR EXPRESSION AND CHARACTERIZATION AS A PUTATIVE
GLUCOSYLTRANSFERASE IN CITRUS PARADISI (GRAPEFRUIT)
Epling, L. and McIntosh, C. Department of Biological Sciences, East Tennessee State
University, Johnson City, TN 37614
Flavonoids are important molecules in plants, including Citrus paradisi. These compounds are
of significance because of their probable power as antioxidants and the characteristics they give
to plants, including the bitter flavor of grapefruit. Several research projects have identified
naringin as being a possible antioxidant, even at nutritionally absorbable levels. Beyond
antioxidant capabilities, research has shown correlation with anti-proliferation of cells, inhibition
of angiogenesis, inhibition of cellular signaling, and effects on DNA repair enzymes (Gao et. al.
2006). Commercially, it is much harder to sell grapefruit or derived products if there are
especially high levels of the bitter compounds, thus further supporting the need for research in
this area (McIntosh, Personal Communication).
The bitter flavanone naringin accounts for 40-70% of the dry weight of young grapefruit
(Jourdan et. al. 1985). Synthesis of bitter naringin involves a two-step glycosylation of
naringenin (McIntosh and Mansell 1990, McIntosh et. al. 1990, Frydman et. al. 2004). In order
to be able to control this biosynthetic pathway by the modification of enzymes, clones must be
obtained for analysis of structure and function.
This research focuses on characterization of PGT-4, a putative flavonoid glucosyltransferase
obtained viaSMART RACE RT-PCR using degenerate primers designed using the PSPG (plant
secondary product glucosyltransferase) box sequence. (Strong 2005, Roy Sarkar et. al. 2007).
This full length clone contained internal restriction endonuclease recognition sites that
prevented it from being inserted in expression vector in the same way as previous PGT clones
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obtained in the McIntosh Lab. Primers were designed to incorporate a Bsa I restriction site with
Nco I compatible overhang on the five prime end of the sequence and a Xho I site on the three
prime end of the gene sequence for PCR (polymerase chain reaction) modification. These
primers were designed to be used in the same PCR reaction, and this modification was
performed on PGT-4. The PCR product was intermediately cloned into the PCR-4 TOPO
vector. Bsa I and Xho I restriction endonucleases were used to isolate the desired DNA
fragment. Cloning is in progress to insert mPGT-4 (modified) in the PCD1 expression vector for
transformation into BL21 (DE3) RIL e. coli cells and subsequent induction of protein expression.
The expressed proteins will be tested for possible flavonoid glucosyltransferase activity.
---------------------------------------------------------------------------------------------------INTERACTING GALAXIES AND THE EFFECTS ON STAR FORMATION AND
GALACTIC STRUCTURE
Sabrina H. Hurlock, Dr. Beverly J. Smith and Dr. Mark Hancock. Department of Physics,
Astronomy and Geology, East Tennessee State University, Johnson City, TN 37614
By analyzing images of interacting galaxies, we are able to study the effects that galactic
interactions have on the involved galaxies, including new star formation, matter bridges between
the galaxies, and changes in overall galactic structure. We obtain the data by making
observations, via the internet, from the SARA (Southeastern Association for Research in
Astronomy) 0.9 meter telescope at the Kitt Peak National Observatory in Tucson, Arizona. We
observe many different interacting galaxies selected from the Arp Atlas in multiple filters, making
numerous 10-minute observations in each filter. After making the observations, we analyze the
data using the IRAF (Image Reduction and Analysis Facility) program, to remove the bias and
dark current, flat-field the data, and co-add the images. The DS9 software is then used to view
the final compiled images of each galaxy. At this point, we are able to determine the location
and size of new areas of star formation, the size, shape and location of tails and bridges
between galaxies, and the transformation of the structure of the galaxies. This research is
important in helping us to understand the magnitude of the effects of galactic interaction and
what will one day be the fate of our own galaxy.
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IN SITU VASCULAR ENDOTHELIAL GROWTH FACTOR LOCALIZATION
Kevin Long. Department of Health Sciences, East Tennessee State University, Johnson
City, TN 37614
Vascular Endothelial Growth Factor is the primary molecule that acts upon endothelial cells,
triggering the growth of new blood vessels. It is the only known growth factor that acts
exclusively on endothelial cells. This factor can be found in the ovaries of all mammals and
plays a pivotal role in the estrous cycle. VEGF is known to be more prevalent at different parts
of the estrous cycle, but it is not known in which part of the ovaries VEGF production originates.
We have attempted to locate the source of Vascular Endothelial Growth Factor in the ovaries of
hamsters and mice. Since we could not locate the growth factor itself, we had to find the mRNA
that encodes for VEGF production. The required process for such situ hybridization localization
is in situ hybridization. We began this procedure by using template DNA to create an RNA
probe through transcription reactions. The probe was made from a vector known as PGEMIII
and was cut using EcoRI and SP6 restriction enzymes. Once the probe was made, it was used
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in the staining process of in situ hybridization in order to stain the microscope slides with the
ovarian tissue. Because our probe was comprised of RNA, many precautions had to be made
in order to keep RNase enzymes out of our solutions and equipment. These enzymes would
degrade the probe, rendering it useless for our study.
We attained limited results, but explored several new techniques in discovering the prevalence
of VEGF throughout the cycle. We improved our methods through a significant amount of
troubleshooting and research. The study will continue and with what we have learned and been
able to improve upon, it should lead to extensive findings about VEGF throughout the cycle.
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ASSESSMENT OF PUTATIVE GLUCOSYLTRANSFERASE PGT-2 EXTRACTED
FROM CITRUS PARADISI THROUGH CLONING, SEQUENCING AND KINETIC
ANALYSIS
Shannon McConnell, Daniel Owens and Cecilia McIntosh. Department of Biological
Sciences, East Tennessee State University, Johnson City, TN 37614
Flavonoids, a class of secondary metabolites, are a kay focus in plant biochemistry. Flavonoids
are polyphenolic compounds consisting of fifteen carbons; two benzene rings attached by a
three carbon chain. There is considerable interest in flavonoids with respect to their antioxidant
effects and their contribution to human health. Flavonoids are characterized by their molecular
structure and research is focused on the common pathway that synthesizes them. With over
5,000 different flavonoid metabolites present in plants, research is focused on the core/common
pathway that synthesizes flavonoids. Not much is understood or known about regulation of
modification reactions where the synthesis of flavonoid derivatives are concerned, but
glyosylation is one of the most common modifications that occur in the synthesis of flavonoids.
Glycosylation is a chemical reaction in which glycosyl groups are added to a substrate to
produce glycosylated derivatives. The focus of the research was to analyze/evaluate putative
glucosyltransferase PGT-2 from grapefruit leaves using techniques such as cloning, sequencing
and experiments to test enzyme activity. Over the course of evaluating PGT-2, it was cloned
into Topo and PCD1 vectors. The success of PGT-2 being inserted into these vectors was
assessed using plasmid isolations along with evidence visualized in DNA gels and verification
through sequencing. After confirming that PGT-2 had been inserted in PCD1 it was transformed
into BL21 DE3 RIL cells and further evaluated by induction experiments at varying temperatures
to evaluate protein production. Current efforts are focused on optimizing induction conditions to
increase levels of soluble protein and testing expressed protein for flavonoid GT activity.
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REGULATION OF SABP2: A CRITICAL COMPONENT OF THE PLANT INNATE
IMMUNITY
Poonam Sheth, Daniel F. Klessig and Dhirendra Kumar. Department of Biological
Sciences, East Tennessee State University, Johnson City, TN 37614 and Boyce
Thompson Institute, Ithaca, NY 14853
SABP2 has been identified as a receptor for SAR (a form of innate immunity) in plants (Kumar &
Klessig, 2003). RNAi mediated silencing of SABP2 makes the plant susceptible to pathogen
infection and the plant produces methyl salicylate (MeSA) in its response (Farhad et. al., 2005).
MeSA is an ester and is hypothesized to move through the membranes and is also a volatile
compound. SABP2 is required in the uninnoculated systemic tissue to convert the inactive
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MeSA into active salicylic acid (SA) by methyl esterase activity. The active SA is critical for
basal, local and systemic acquired resistance and therefore it is important to know the
regulation of SABP2. A cell suspension culture of Nicotiana tabacum were freshly inoculated by
diluting 1:10 in MS media from the cultures that were previously established. The cultures were
grown at 20-25º C on a shaker maintained at approximately 130 rpm. After 3-4 days the cells
were treated with MeSA, MeJA, nitric oxide donor (GSNO). Cells were collected at various time
intervals and used for RNA isolation. Total RNA was isolated using Tri-reagent (Sigma) and
1µg of the RNA was used for cDNA synthesis. Total cDNA was used for amplification of SABP2
and other genes using PCR. PCR products were analyzed by separating on a agarose gel. To
confirm the results northern analysis is being carried out using a non radioactive probe (SABP2
cDNA) using the kit from GE Healthcare Life Sciences. For Western analysis polyclonal
antibodies (produced in rabbit) against the recombinant SABP2 will be used. All the exciting
results of this ongoing experiment will be presented. This study will help us to understand the
regulation of a critical component of plant immunity.
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Social & Behavioral Sciences
THE EFFECTS OF NICOTINE ON CONDITIONED PLACE PREFERENCE IN A
RODENT MODEL OF PSYCHOSIS
Jennifer Abraham, Yoko E. Ogawa, Elizabeth L. Cooper and Russell W. Brown.
Department of Psychology, East Tennessee State University, Johnson City, TN 37614
Increases in sensitivity of the dopamine D2 receptor has been shown to play important roles in
behavioral abnormalities in schizophrenia. Past studies from this laboratory have shown that
neonatal treatment with the drug quinpirole (a dopamine D2/D3 agonist) results in long-term
increases of sensitivity of the D2 receptor that persists throughout the animal’s lifetime. A major
health problem within the schizophrenic population is an increased incidence of smoking
cigarettes, which contain the psychoactive and addictive ingredient nicotine. This study was
designed to analyze the effects of nicotine on conditioned place preference (CPP) in both early
postweanling and adolescent rats that were neonatally treated with quinpirole. All rats were
admininistered a daily intraperitoneal injection of Quinpirole HCl (dose: 1 mg/kg) for the first
three weeks of development (P1-21). Rats were weaned from the female dam at P21. There
were two conditioned place preference shuttle boxes used in this study: A two-chamber and a
three-chamber apparatus. The two chamber was a black/white shuttle box constructed of
plywood that was 30 x 18 x 20 in that either had only a black and white chamber. The threechamber apparatus was constructed in a similar fashion but had a gray chamber separating the
black and white chamber. In experiment 1, beginning on 23 or 31 days of age, animals were
conditioned by administering animals nicotine tartarate (0.8 mg/kg free base) in the nonpreferred environment (white compartment), whereas saline was given in the black
compartment for eight consecutive days, and controls received saline in both compartments.
Controls were administered saline was given in both compartments. Results demonstrated that
regardless of age, rats neonatally treated with quinpirole demonstrated a significant preference
for the black compartment, but nicotine alleviated this preference after eight days of
conditioning. Rats neonatally treated with saline and conditioned with nicotine demonstrated a
stronger preference for the white compartment. In Experiment 2, rats were conditioned in a
similar fashion but in a three chamber apparatus, and a similar result was found: neonatal
quinpirole produced a significant preference for the black compartment in both early
postweanlings and adolescents that was alleviated by nicotine. These results appear to show
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that neonatal quinpirole, that results in priming of the D2 receptor, produces a significant
increase in stress that is alleviated by nicotine.
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OUTCOMES OF THE APPALACHIAN TEACHING PROJECT
Erika Adams. Department of Family and Consumer Sciences, East Tennessee State
University, Johnson City, TN 37614
The Appalachian Teaching Project began in 2000 having in mind bridging the gap between
communities and institutions of higher education. The colleges and universities of Appalachia
set about to engage with their communities through specific projects designed to stabilize the
Appalachian region and contribute to keeping the region’s youth committed to their
communities. Developing community projects that will have a continuing presence and a lasting
influence, the thirteen participating schools in nine states which have joined the project
contribute through community based research projects and service learning which is partially
funded by the Appalachian Regional Commission. This poster project was designed to highlight
the numerous outcomes of Appalachian Teaching Projects of the 2004-2005 year. Research
data was collected through interviews with participating students and faculty, tours of project
sites, and attending presentations and conferences. The outcomes of the various projects are
as diverse as the Appalachian population they represent.
------------------------------------------------------------------------------------------------------------------DANGEROUS DRIVING AND FUNDAMENTALISM, SPIRITUALITY, AND
RELIGIOSITY
Amber Cline, Chris S. Dula, Robert B. Russell, Kevin Gilmer, Jodie A. Lamb and Julia T.
Sutherland. Applied Psychology Laboratory, East Tennessee State University, Johnson
City, TN 37614
Risk-taking is a dimension of dangerous driving (Dula & Geller, 2003). Risky driving is a
problem because in 2002 alone, over 42,000 people were killed in traffic crashes on U.S. roads
(NHTSA, 2003). An interesting set of unpublished pilot data suggested that drivers perceived
levels of spirituality, as assessed by a single question, was related to how often people reported
wearing safety belts in their vehicles. To explore this concept more in-depth, measures of
fundamentalism, spirituality, and religiosity, were given to undergraduate participants along with
the Dula Dangerous Driving Index (DDDI, Dula & Ballard, 2003) and a demographic
questionnaire assessing a wide variety of traffic safety behaviors and history of traffic crashes.
With only a single item of spirituality having previously shown a mild, but significant, positive
correlation with reported safety belt use, no specific hypotheses were made with regard to
fundamentalism or religiosity as they pertain to risky driving behaviors. It was hypothesized that
higher levels of perceived spirituality would be related to lower reported risky driving as
measured by the DDDI, and a lower incidence of reported dangerous driving behaviors. A total
of 117 undergraduate students participated in an online survey of traffic safety behaviors and
personality constructs. Results are discussed and future research directions are proposed in
this poster presentation.
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SOCIAL DESIRABILITY BIASES AND SELF-REPORT MEASURES OF
DANGEROUS DRIVING
Kevin Gilmer, Chris S. Dula, Robert B. Russell, Amber Cline, Jodie A. Lamb and Julia T.
Sutherland. Applied Psychology Laboratory, East Tennessee State University. Johnson
City, TN 37614
Adolescents and young adults are particularly prone to unintentional injury by motor vehicles
crashes. Healthy People 2010 noted that the death rates associated with motor vehicle-traffic
injuries are highest in the age group 15 to 24 years (U.S. Department of Health and Human
Services, 2001). Thus, the college population is a particularly apropos venue to study traffic
safety issues. Measures of driver risk do exist, such as the Dula Dangerous Driving Index
(DDDI, Dula & Ballard, 2003), the Propensity for Angry Driving Scale (PADS, DePasquale,
Geller, Clarke, & Littleton, 2001), and the Driving Behaviour Inventory (DBI, Gulian, Matthews,
Glendon, Davies, & Debney, 1989). A problem all these measures have in common is an
obvious item content whereby test takers can readily discern how to answer in a positive
manner. This study looks at the construct of Social Desirability, a tendency for people to
answer survey items in a manner more positive than is likely the case. Social desirability was
hypothesized to be negatively correlated with the DDDI and the PADS. A total of 117
undergraduate students participated in an online survey of traffic safety behaviors and
personality constructs. Results are discussed and future research directions are proposed in
this poster presentation.
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A CROSS-CULTURAL COMPARISON OF MINIMUM LEGAL DRINKING AGE LAWS
AND ACTUAL DRINKING BEHAVIOR
Kristina Hollowell. Department of Psychology, East Tennessee State University,
Johnson City, TN 37614
It is no secret that underage drinking exists and is prevalent throughout the United States.
Binge drinking is considered a common social problem within our culture, especially among
underage drinkers (Miller et al., 2006). The United States’ minimum legal drinking age (MLDA)
is 21; one of the highest in the world. This leads one to assume that one of the country’s
solutions to prevent drinking is to implement laws forbidding one to consume alcohol until they
reach an age approved by the government. However, the law itself does not necessarily
guarantee obedience, but rather imposes consequences for use and limits availability of alcohol
(Wagenarr, Toomey, & Lenk, 2005). I approached my research with a cross-cultural
perspective exploring the relationship between MLDA and alcohol consumption behavior
comparing East Tennessee and Amsterdam. The United States and the Netherlands have very
different governmental policies concerning alcohol (International Center for Alcohol Policies,
1998). In The US a person is not allowed to purchase or consume alcohol until the age of 21.
In contrast, in the Netherlands a person is allowed to purchase beer and wine at the age of 16
and liquor at 18. The participants for my research were all between the ages of 18 and 20 years
old; those from Amsterdam could drink legally while those from the United States could not. My
measurement consisted of a survey containing 18 questions that asked the participants about
their alcohol consumption, consequences of drinking behavior, family alcohol problems,
perception of colleagues’ consumption, and their perception of their own consumption. If the
United States’ laws are effective in preventing underage drinking, then the U.S. participants
should drink less and consequently have fewer negative consequences than the Amsterdam
participants who had been able to drink legally anywhere from two to four years at the time they
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2007 Appalachian Student Research Forum
filled out the survey. There were 82 participants from Vrije Universiteit and 118 participants
from East Tennessee State University. Over 80% of the participants from both universities were
considered to be drinkers. At ETSU 44% were binge drinkers while 53.6% at Vrije binge drank.
ETSU participants reported a significantly higher rate of negative consequences in their
personal relationships than Vrije participants. While only 13.0% of Vrije participants claimed
other students’ negatively impacted their life on or around campus, 40.2% of ETSU participants
agreed. Students from both universities reported a similar rate of negative effects upon their
jobs and school work. Both set of participants perceived their colleagues to drink more than
what was actually reported. The results agree and contradict the original hypothesis. Although
Vrije participants did report more binge drinking than ETSU participants, the ETSU participants
reported more negative consequences of their drinking behavior. Despite all of the attention
focused on alcohol consumption, colleges and universities have failed to control binge drinking
rates (Glassman, 2002). Weschler and colleagues (2002) showed that approximately 44% of
the participants in their study reported binge drinking, a rate almost identical to previous years.
Regardless of the legality status adolescents often drink. It is my belief that it is important to
teach healthy drinking habits, not abstinence from alcohol.
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METHYLPHENIDATE (RITALIN) EXPOSURE TO ADOLESCENT D2-PRIMED RATS
ELIMINATES THE INITIAL HYPOACTIVE RESPONSE TO NICOTINE IN
ADULTHOOD
Andrew B. Hughes, A. Brianna Sheppard, Ian D. Longacre and Russell W. Brown.
Department of Psychology, East Tennessee State University, Johnson City, TN 37614
Past studies from our laboratory have shown that neonatal quinpirole (dopamine D2/D3 agonist)
treatment produces increases in sensitivity of the dopamine D2 receptor, an effect that has
some similarities with attention deficit/hyperactivity disorder. A common drug typically used to
medicate ADHD is Ritalin (methylphenidate), a drug that has stimulant properties and abuse
potential. Additionally, studies have shown that children that are medicated with Ritalin show a
significant increased propensity to smoke cigarettes. In this study, we analyzed whether
adolescent rats neonatally treated with quinpirole will demonstrate a significant increase in
locomotor sensitization when methylphenidate is administered in adolescent in D2-primed and
non D2-primed rats. Rats were administered quinpirole, a dopamine D2/D3 agonist from
postnatal days (P1-21) and raised to adolescence (P29). Rats were habituated to a locomotor
arena, and subsequently administered methylphenidate (Trade name: Ritalin; 5 mg/kg) or saline
every other day for three weeks. Additionally a nicotine challenge was performed at postnatal
day (P60), in early adulthood, to analyze the effects of nicotine on D2-primed rats exposed to
methylphenidate in adolescence. On overall activity counts, results showed that
methylphenidate produced rapid sensitization, and D2-primed animals that received
methylphenidate began to demonstrate a significant decrease in activity during the third week of
testing, suggesting increases in stereotypic behavior and immobility. In fact, a significant
increase in immobility was observed during the third week of testing in D2-primed animals
administered methylphenidate, supporting this claim. On the nicotine challenge, D2-primed
animals exposed to methylphenidate as adolescents demonstrated no hypoactive response to
initial nicotine exposure, and in fact demonstrated a significant increase in activity compared to
all other groups on the nicotine challenge. These results suggest that D2-priming combined
with methylphenidate exposure may lead to elimination of the hypoactive response to initial
nicotine, and may be suggestive as to the mechanism of the increased propensity of
methylphenidate-treated ADHD patients to smoke cigarettes.
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2007 Appalachian Student Research Forum
THE FUTURE IMPLICATIONS OF INDOOR UV TANNING AT A YOUNG AGE
Leslie King, Preston Visser, Joel Hillhouse and Lana McGrady. Department of
Psychology, East Tennessee State University, Johnson City, TN 37614
Indoor UV tanning is a significant health-risk for the development of skin cancer in young adults.
Research has shown that it is strongly related to the development of all types of skin cancer
including melanoma, the most lethal form of skin cancer. Despite this, the popularity of indoor
tanning is strong and growing in young people. The prevalence of indoor tanning is consistently
found to be highest among young adult females. This project sought to examine the risks
associated with indoor tanning at a young age by comparing groups of individuals who first
indoor tanned before and after chosen ages. We surveyed 174 female college students from a
regional university (East Tennessee State University) as a part of a larger research project.
Using a software program that allows for complete online surveying, subjects were given a
survey assessing both their history of indoor UV tanning as well as their intentions to indoor UV
tan in the future. In the study we divided the subjects into two groups based on the age of their
first indoor UV tanning experience. We then compared the two groups to the amount of times
they reported indoor UV tanning in the past twelve months. We found a significant difference
between individuals who first indoor UV tanned at age 16 or below compared with those who
first indoor UV tanned at ages 17 or above (p=<.01). This suggests that a female is more likely
to indoor UV tan at a greater frequency in the future when she is exposed to it at an earlier age.
This may have significant implications in potential interventions by encouraging the investigation
of the efficacy of targeting younger female populations for pre-behavioral treatment.
Importantly, we also found a significant difference between average number of times
participants reported indoor UV tanning based on whether or not they first indoor UV tanned
before the age of 18 (p=<.01; mean difference of 18.8). Data such as this may be crucial in the
debate concerning illegalizing indoor UV tanning for persons under the age of 18 as it
demonstrates that females indoor UV tan significantly more in the future if they first indoor UV
tan before the age of 18.
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EARLY WOODLAND CERAMIC TYPOLOGY AND CULTURE CHRONOLOGY IN
UPPER EAST TENNESSEE
Lucinda M. Langston and Jay D. Franklin. Department of Sociology and Anthropology,
East Tennessee State University, Johnson City, TN 37614
Based on research and excavations at Phipps Bend on the Holston River, Robert Lafferty
(1978, 1981) developed a model for Early Woodland ceramic typology and chronology for upper
East Tennessee. The utility of the model has never been evaluated beyond Phipps Bend,
however. In this paper, we examine the Early Woodland ceramic assemblages from several
sites in upper East Tennessee in an attempt to place them within the chronological framework
developed by Lafferty. Where possible, we use radiocarbon assays as an independent means
of assessing the efficacy of the model.
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2007 Appalachian Student Research Forum
AGE AND GENDER DIFFERENCES IN THE VIRTUAL MORRIS WATER MAZE
FROM 12-25: A FRESH LOOK AT STRATEGIES
Matthew F. Lazenka and Dr. Russell W. Brown. Department of Psychology, East
Tennessee State University, Johnson City, TN 37614
The Morris water maze (MWM) has been utilized as one of the primary behavioral neuroscience
tasks in rodents and is an excellent test of spatial memory function (Gerlai, 2001). Two primary
versions of the MWM have been utilized: A place version, in which the platform is stationary,
and a match-to-place version in which the platform moves to a new location on each day of
training. Recently, a virtual version of the MWM has been developed to test human participants
on this task (Astur, et al., 1998). In this version of the MWM, an environment is presented on a
computer screen that includes a depiction of a three-dimensional extramaze cue environment.
Participants must navigate through the environment using arrows on a keyboard to a hidden
platform. Past studies have reported significant gender differences on the place version of the
vMWM (Driscoll, et al., 2005), but there has not been any reports on the match-to-place version
of the vMWM. Similarly, research has also failed to address the differences between
adolescents and adults utilizing the match-to-place version, as well as, the effects of gender
stereotype threats.
Participants for experiment one were students (12-13 years old) from a public school located on
the campus of East Tennessee State University (ETSU). Participants for experiment two were
college students (18-25 years) recruited from Psychology and Sociology classes at ETSU. The
computer simulation consists of a pool located in the center of a square room with various
landmarks surrounding the pool. For experiment one, adolescents were read a script
discussing how to perform the task, mentioning that they would have to swim in search of a
hidden platform located under an opaque water surface and that the experiment would consist
of 20 training trials. In experiment two, the same script was read and another was read
revealing that either men or women were superior in the task alluding that this superiority was
due to evolutionary significance.
Results from the first experiment showed that adolescent males had reduced latency, heading
error and path length. In reference to paths, adolescent males and females did not differ. Both
preferred to alternate between a circle and zig-zag strategy; however, their paths are more
closely related to those of >60 year olds than college-aged students. This may be a factor
paralleling the developing hippocampus with the aging hippocampus.
In experiment two, mentioning male superiority seems to eliminate the gender difference while
mentioning female superiority does not entirely. Surprisingly, no significant difference was
found within genders. Compared with the strategies previously found with college-aged
participants, males differed in their choice of strategy. Males in experiment two circled more
than males in that study (Click & Brown, unpublished). This may be influenced by mentioning of
a strategy.
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2007 Appalachian Student Research Forum
NICOTINE SENSITIZATION IN ADOLESCENT AND ADULT RATS D2 RECEPTOR
PRIMED AS NEONATES: GENDER DIFFERENCES AND NEUROTROPHIC
FACTORS
Ian D. Longacre, Marla K. Perna and Russell W. Brown. Department of Psychology, East
Tennessee State University, Johnson City, TN 37614
We have demonstrated in past work that neonatal quinpirole (D2/D3 receptor agonist) treatment
to rats produces long-term priming of the dopamine (DA) D2 receptor that persists throughout
the animal's lifetime. In Experiment 1, the offspring of six male-female Sprague-dawley (SD) rat
breeder pairs were administered one daily i.p. injection of quinpirole (1mg/kg) or saline from
postnatal days 1-21 (P1-21) and raised to adolescence. After three consecutive days of
habituation to the locomotor arena, beginning on P31 all animals were administered one i.p.
injection of 0.5 mg/kg free base nicotine tartarate every other day for three weeks. Results
showed that neither D2-primed or non D2-primed adolescent female rats demonstrated nicotineinduced hypoactivity early in training, and D2-primed adolescent females showed a significant
decrease in activity as compared to females administered nicotine in week 2 and 3 due to
increased stereotypic behavior. D2-primed male adolescents demonstrated more rapid
sensitization to nicotine than non D2-primed males. In Experiment 2, offspring of seven breeder
pairs were given the identical neonatal drug treatment as in Experiment 1 but raised to
adulthood (P60). Although initial hypoactivity did not differ across groups, D2-primed adults rats
administered nicotine demonstrated significantly more robust sensitization than controls given
nicotine during the third week of testing. In both Experiments 1 and 2, one day after testing was
complete brain tissue was taken and the nucleus accumbens and frontal cortex were analyzed
for brain-derived neurotrophic factor (BDNF). In adolescents, nicotine produced a significant
increase in BDNF in the nucleus accumbens that was unaffected by neonatal quinpirole
treatment. In adults, Neonatal quinpirole treatment produced a significant decrease in nucleus
accumbens BDNF in adult rats that was alleviated by nicotine. These results suggest that
sensitization to nicotine is more robust in D2-primed animals, but sensitization is dependent
upon both age and sex. Additionally, it does not appear that BDNF plays a significant role in
nicotine sensitization in either adolescents or adults, which is contrary to findings with other
psychostimulants such as cocaine and amphpetamine.
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GENDER DIFFERENCES IN STIGMA AGAINST TATTOOS
Benjamin A. Martin, Sonia L. Coney and Dr. Chris S. Dula. Department of Psychology,
East Tennessee State University, Johnson City, TN 37614
Tattoos have grown in popularity in recent years, mainly brought to the forefront by celebrities
and “reality-television” shows involving various tattoo shops. One theory for explaining the
proliferation of fleshworks is that with the demise of the mythology of Christianity in the popular
imagination (Mercury, 2000). The purpose of the present study was to evaluate gender
differences in stigma against tattooing, and two measures of adventurousness; men were
expected to have less stigma associated with tattooing and were also expected to be more
adventurous. It is notable that previous studies have basically adhered to the quasi-interview
method, which may be a limitation to the studies in that it often leads to desirable responding.
For this pilot study, 58 undergraduates (9 men and 49 women) participated for modest extra
credit. Participants completed a set of surveys that included attitudes regarding tattooing, the
Adventurousness Scale (van der Zee & van Oudenhoven, 2000), a version of the EPQV
modified to measure adventurousness (Eysenck et al., 1985), and a short demographic
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2007 Appalachian Student Research Forum
questionnaire. Men scored higher on the stigma measure than women but this difference was
not significant. Significant gender differences were found for scores on both the
Adventurousness and EPQV Scales. The impact of a non-significant gender difference in
responses to the stigma measure, eludes to an overall social taboo against tattooing. Further
research will be aimed at how familiarity with the tattoo process affects overall stigma scores.
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NICOTINE SENSITIZATION IN ADOLESCENT BETA ARRESTIN-2 KNOCKOUT
MICE: IMPLICATIONS FOR MECHANISMS OF NICOTINE ADDICTION
Daniel M. Noel, Jennifer A. Correll, A. Brianna Sheppard and Russell W. Brown.
Department of Psychology, East Tennessee State University, Johnson City, TN 37614
This study was designed to sensitize adolescent Beta Arrestin-2 knockout (BA-2 KO) mice to
the psychostimulant nicotine. Activation of the dopamine D2 receptor has been shown to
cause G protein-coupled receptor kinase-dependent receptor phosphorylation, and a robust
translocation of Beta-arrestin to the cell membrane and profound receptor internalization by the
cell membrane. Thus, it appears that Beta Arrestin-2 plays an important role in cell signaling
from the dopamine D2 receptor in response to stimulants. Sensitization to nicotine as
measured through locomotor activity in rats has been shown to be an excellent behavioral test
for dopaminergic activity and is a clinically relevant test for addiction to drugs. Induction of
sensitization has been referred to as the locomotor activity produced by administration of the
drug, and expression of sensitization has been referred to as the locomotor activity response
after a period of abstinence. Both induction and expression of sensitization was measured in
the present study. Finally, nicotine addiction typically begins in adolescence, and research has
shown adolescent and adults demonstrated significantly different behavioral responses to
nicotine. In this study, 3-4 week old adolescent BA-2 KO and wild type C57/Bl6 mice were
habituated to a square locomotor arena for one 30 min session. The following day, animals
were administered either nicotine tartarate (s.c, 0.5 mg/kg free base) or saline 10 min before
being placed into the locomotor arena for seven (Experiment 1) or 14 consecutive days
(Experiment 2). A drug-free abstention period of seven days followed in both experiments, at
the end of which animals received a nicotine challenge (0.5 mg/kg free base). In Experiment 1,
results showed that BA-2KO mice demonstrated significantly decreased levels of activity as
compared to wild type animals during habituation, and also demonstrated an initial hypoactivity
to nicotine as compared to wild type controls that received nicotine. In fact, BA-2KO mice
treated with nicotine remained in a hypoactive state throughout the first 6 days of sensitization
training compared to saline-treated BA2 KO mice as well as wild type controls. However, by
seven days, wild type controls receiving nicotine demonstrated equivalent levels of activity to
saline controls, and thus did not demonstrate sensitization to nicotine. On the nicotine
challenge, however, BA 2 KO mice demonstrated significantly decreased activity levels
compared to all other groups, and thus failed to express sensitization to nicotine. In Experiment
2, both BA-2 KO mice and wild type controls demonstrated sensitization to nicotine, although
the knockout mice demonstrated significantly lower levels of activity. On the nicotine challenge,
BA-2 KO mice did not demonstrate sensitization to nicotine, and in fact demonstrated
significantly lower levels of activity. Therefore, it appears that Beta arrestin-2 molecule is
necessary for induction of sensitization to nicotine, but does not play a role in expression of
nicotine sensitization. These results appear to indicate the importance of the BA-2 protein in
locomotor sensitization to nicotine in adolescence.
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2007 Appalachian Student Research Forum
AN INVESTIGATION OF PERFECTIONISM, OPTIMISM, AND SELFREINFORCEMENT AMONG COLLEGE ATHLETES
Sabrina D. Rohr and Kevin L. Burke. Department of Kinesiology, Leisure, and Sport
Sciences, East Tennessee State University, Johnson City, TN 37614
Perfectionism and optimism are common characteristics found in college athletes (Gould &
Diefenbach, 2002). However, self-reinforcement is not as commonly studied in sport. Athletes
at a southeastern university (N=91) completed the Frost Multidimensional Perfectionism Scale
(Frost, Martin, Lahart, & Rosenblate, 1990), Life-Oriented Test Revised (Scheier, Carver, &
Bridges, 1994), and Frequency of Self-Reinforcement Questionnaire (Heiby, 1983) during either
practice or study hall sessions. Athletes from the following seven sport teams participated:
men’s tennis, women’s golf, women’s softball, women’s basketball, men’s baseball, women’s
soccer, women’s volleyball, and men’s golf. Results indicated a slightly negative relationship
between perfectionism and optimism (r= -.060). With an alpha level of .01 used for all statistical
test, the correlation between perfectionism and optimism was found not to be significant,
p=.569. Also, the results indicated perfectionism and self-reinforcement had a negative
relationship and was found to be significant (r= -.321, p=.002). Conversely, a positive
relationship was found between optimism and self-reinforcement with a correlation of r=.306
and found to be significant, p=.003. Although the correlation between perfectionism optimism
was negative, the athletes were found to have high levels of perfectionism (M= 106.35) and
optimism (M=20.91). Likewise, the athletes were found to have high levels of self-reinforcement
(M=20.78). Team sports (M=108.8) indicated higher perfectionism scores than individual sports
(M=98.19). However, levels of optimism (team M=20.34, individual M=22.81) and selfreinforcement (team M=20.7, individual M=21.05) were higher in individual sports than team
sports.
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ACTIVELY CARING AND PRO-SOCIAL DRIVING
Robert B. Russell, Chris S. Dula, Kevin Gilmer, Amber Cline, Jodie A. Lamb, and Julia T.
Sutherland. Applied Psychology Laboratory, East Tennessee State University, Johnson
City, TN 37614
According to the U.S. Department of Health and Human Services (USDHHS, 2001), motor
vehicle crashes are the leading cause of serious injury in our society. Motor vehicle injuries
remain the most costly and fatal of unintentional injuries. The National Safety Council (NSC,
2001) estimated that motor vehicle crashes were the cause of $169 billion in lost wages,
medical expenses, and administrative costs. Motor vehicle crashes are the leading cause of
death for people ages 1 to 33 (NSC, 2001). While there is a good deal of literature exploring the
effect of personality on dangerous driving, it is likely that personality traits also affect positive
driving behaviors, or at least attenuate dangerous driving behaviors. Driving behaviors that
endanger or have the potential to endanger have been considered as lying on a behavioral
spectrum of dangerous driving, including three dimensions: 1) intentional acts of aggression
toward others, 2) negative emotions experienced while driving, and 3) risk-taking (Dula &
Ballard, 2003; Dula & Geller, 2004). This study looks at the construct of Actively Caring, a
tendency for some people to go out of their way, or beyond the call of duty, to be helpful to
others. Actively caring was hypothesized to be negatively correlated with all dangerous driving
variables. A total of 117 undergraduate students participated in an online survey of traffic safety
behaviors and personality constructs. Results are discussed and future research directions are
proposed in this poster presentation.
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2007 Appalachian Student Research Forum
Division II—
Graduate Students 1 - 2 years
Arts & Humanities
SNAPSHOTS OF THE HISPANIC IMMIGRANT COMMUNITY IN APPALACHIA
Vivian Gonzales Gladson. Department of Sociology & Anthropology, East Tennessee
State University, Johnson City, TN 37614
The proposed poster presentation focuses on my ethnographic fieldwork with Hispanic
immigrant community in Appalachia. Images include brief ethnographic descriptions that
illustrate aspects of everyday life and work among Hispanics in Johnson City and nearby
communities. My work centers primarily on secondary migration, or the process by which
Hispanics have come to Appalachia from other cities in the U.S. The poster presentation
includes issues, such as migrant seasonal labor; concerns about documentation and
immigration laws; gender and children; social problems within the Hispanic community; and
religious belief folklore. Broader themes connecting these issues are processes of
acculturation, concepts of "home" and identity, and the continuity of tradition.
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Natural Sciences & Mathematics
ISOLATION AND IDENTIFICATION OF THE PUTATIVE GLUCOSYLTRANSFERASE
PGT5 FROM CITRUS PARADISI
J.K. Cooke, D. Owens, and C. McIntosh. Department of Biological Sciences and
Department of Health Sciences, East Tennessee State University, Johnson City, TN 37614
The isolation of the putative glucosyltransferase clone PGT5 and its resolution from other
glucosyltransferases from Citrus paradisi (grapefruit) is described. This clone was obtained by
designing two primers based on the sequence of the liminoid glucosyltransferase found in Citrus
unshiu. These primers were designed so that PCR products would encompass the signature
Plant Secondary Product Glucosyltransferase (PSPG) domain. Forward primer JC2 was taken
from an area of the sequence approximately 50 base pairs ahead of the PSPG box. Reverse
primer JC4 was taken from an area of the sequence approximately 100 base pairs after the
PSPG box toward the 3’ end. A PCR reaction with cDNA from C. paradisi yielded a fragment
approximately 500 base pairs in length. This fragment was cloned into TOPO vector and
plasmids were isolated from the cells, digested using EcoR1, and sent for sequencing. Analysis
of sequences confirmed the presence of the PSPG box and showed >90% homolysis with the
liminoid GT in C. unshiu. New primers CU5R and CU6F were designed approximately 75 base
pairs from the 5’ and 3’ ends respectively. These primers were used in separate PCR reactions
combined with SMARTRACE universal primers in order to "walk out" and obtain additional
sequence for this putative GT. These PCR products have been cloned into TOPO vector and
isolated, and sequencing results are pending.
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2007 Appalachian Student Research Forum
A COMPUTATIONAL CHEMISTRY STUDY OF SPIN TRAPS
Tande Jacob Fosso. Department of Chemistry, East Tennessee State University,
Johnson City, TN 37614
Many defects in physiological processes are due to free radical damage: reactive oxygen
species, nitric oxide and hydroxyl radicals have been implicated in the parthenogenesis of
cancer, diabetes mellitus, and rheumatoid arthritis. Also there is growing evidence that aging is
due to free radical mediated regulatory processes that result in altered gene expression. The
effects and defects of these radicals are studied using biological probes commonly known as
spin traps. In this study, theoretical calculations are carried out on the two main types of spin
traps (DMPO and PBN) at the density functional theory level (DFT). The energies of the
optimized structures of the spin traps and the hydroxyl radical adduct are calculated at the
DFT(B3LYP)/6-31G(d), hyperfine calculations in gaseous and aqueous phases are carried out
at the DFT (B3LYP)/6-31G(d)/6-311G(2df,p). The dielectric effect on the performance of the
spin trap is determined using the polarized continuum model. Calculations show a localization
of spin densities in both cases. However, DMPO spin traps are shown to be more stable and
more interactive in aqueous environment.
Spin density map of PBN hydroxyl radical adduct
-------------------------------------------------------------------------------------------------------------------CHARACTERIZATION OF G.S.INT1, A GROUP IIC INTRON FROM THE
THERMOPHILLE GEOBACILLUS STEAROTHERMOPHILUS
H. Sun, S.E. Moretz and B.C. Lampson. Department of Health Sciences, East Tennessee
State University, Johnson City, TN 37615 and NIH, Rockville, MD
Group II introns are small segments of DNA that reside in the chromosome of bacteria or the
genome of organelles of primitive eukaryotes. These elements have two very interesting
properties. First, they are retrotransposons that can move to a new location in DNA via a
reverse transcription mechanism. Second, they form a large ribozyme that mediates selfsplicing of the intron from pre-m RNA. Recently, a group II intron type protein with similarity to
reverse transcriptase was discovered in the thermophile Geobacillus stearothermophilus strain
10 (Vellore et al., 2004. AEM 70: 7140). The purpose of this work is to clone and characterize
the intron that codes for this protein. Because it is found in a thermophile, this intron (and
ribozyme) may have novel properties and structural changes that allow it to function at hot
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2007 Appalachian Student Research Forum
temperatures. Numerous copies of the intron protein gene are present in the chromosome of
strain 10, but absent from a related ATCC 12980 strain, based on a Southern hybridization
experiment. Three separate copies of the entire intron plus flanking exon DNA were cloned
(and their DNA sequenced) from a HindIII restriction fragment, plasmid library of the strain 10
genome. Because the intron has inserted into different positions in the chromosome the exact
junctions of the intron were identified. The intron, designated G.s. Int1, is about (the size varies)
1890 base pairs and codes for a bacterial type C intron ribozyme. Analysis of the nearly
completed genome sequence of strain 10 identified at least seven additional copies of the
intron, one being a truncated copy. DNA primers, designed to anneal to exon DNA that flanks
the intron, were used to detect in vivo splicing of the intron in G. stearothermophilus cells. The
intron was cloned into a T7 expression plasmid to allow inducible expression in Escherichia coli.
Splicing experiments using these constructions are in progress.
Keyword: Geobacillus stearothermophilus; Reverse transcriptase; Group II intron
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Social & Behavioral Sciences
TYPICALLY DEVELOPING CHILDREN DISPLAY TWO PRELINGUISTIC ACTS PER
MINUTE BEFORE FIRST WORDS
Jessica Brown, Megan Ringley, Tiffany Barber, Chantal Newell and Dr. Kerry ProctorWilliams, PhD, CCC-SLP. Department of Communicative Disorders – Speech-Language
Pathology, East Tennessee State University, Johnson City, TN 37614
The goals of this longitudinal study were to define the transitional rates per minute for
protoimperatives, protodeclaratives, behavioral regulation, joint attention, social interaction,
canonical vocalizations, and total communicative acts at both the prelinguistic and one-word
stages of language development. Specifically, we asked: Are the rates of these behaviors the
same in both unstructured and uninstructed parent-child free-play and book-reading activities as
in those the more structured samples collected with clinicians in Wetherby, Cain, Yonclas, and
Walker (1988)? This study examined 29 typically developing children at 7, 10, 13, and 20
months during structured and uninstructed parent-child interactions. Unstructured and
uninstructed conditions create a more natural setting for the child and mother, allows for more
variability in parent’s responsiveness, and is more consistent with diagnostic procedure in the
field of speech language pathology. The following categories statistically significant increased
from the prelinguistic to one-word stages and yielded medium effect sizes: total communicative
acts, protodeclaratives, and canonical vocalizations. The average rate of total communicative
acts used by participants in the prelinguistic stage was 1.11 acts per minute. The average rate
of total communicative acts used by participants in their first one word stage was 2.12 acts per
minute. This shows that children must use somewhere between 1.11 and 2.12 acts per minute
before transitioning into the one-word stage. As well, correlations were conducted to see if
rates of total communicative acts and use of canonical vocalizations at early ages predicted
later production of words. The sample included a slightly smaller sub-sample of 27 children
because two subjects never reached the one word stage. Our results indicated significant
relationships between rates of prelinguistic communicative acts and later word production. Our
findings generally coincide with those of Wetherby et al. (1988) in that typically developing
children display around one communicative act per minute for the prelinguistic stage and that
this rate is necessary for transition to the one word stage. Our data predicts that children need
to display an overall increase in communicative act functions, with the most critical being
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2007 Appalachian Student Research Forum
protodeclaratives, canonical vocalizations, and total communicative acts. Future research
should replicate this study using a larger sample size with a more economically and ethnically
diverse population to represent more adequately the population as a whole. Future studies
need to incorporate longer unstructured activities for mother and child participation in order to
elicit a wider range of communicative acts. The results from this study will provide speechlanguage pathologists with normative guidelines about transitions from the prelinguistic stage to
the one-word stage. This information can assist in assessment and therapy of languageimpaired children.
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AMPHETAMINE SENSITIZATION IN A RODENT MODEL OF PSYCHOSIS
Zackary A. Cope, A. Brianna Sheppard, Ian D. Longacre, Marla K. Perna, Kimberly N.
Thompson and Russell W. Brown. Department of Psychology, East Tennessee State
University, Johnson City, TN 37614
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 D2 receptor priming. An
increase in D2 receptor sensitivity is consistent with several behavioral disorders, including
schizophrenia. Additionally, a past study from a collaborating laboratory has shown that acute
amphetamine (street name: Speed) administration produces a 5-fold increase in dopamine
release of D2-primed rats, suggesting that the dopamine system in the brain, which mediates
positive reinforcement, is hypersensitive in psychosis. Congruent with this findings, individuals
that are diagnosed with schizophrenia demonstrate a 4-5 fold increase in abuse of drugs in the
psychostimulant class, of which amphetamine belongs, which suggests that increases in
dopamine activity in response to amphetamine may be more reinforcing to schizophrenics than
to the normal population. 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 using a software program
purchased in our laboratory (AnyMaze, Stoelting, Wood Dale, IL). Results showed that D2primed rats receiving amphetamine demonstrated a significant increase in locomotor activity
across all seven days of testing relative to all other groups. This is consistent with past findings
demonstrating the D2-primed rats administered amphetamine demonstrated significant
increases in dopamine levels, as increases in dopamine levels in the brain typically positive
correlate with significant increases in locomotor activity. Controls receiving amphetamine also
demonstrated a significant increase in activity over days. Interestingly, D2-primed rats given
saline also sensitized to the environment, 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 will be assayed in the coming months.
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2007 Appalachian Student Research Forum
EVALUATION OF THE EFFICACY OF AFTER SCHOOL PROGRAMS WITH AT RISK
CHILDREN
Heather R. Hyder and Chris S. Dula, PhD. Department of Psychology, East Tennessee
State University, Johnson City, TN 37614
The purpose of this study was to evaluate the efficacy of an after school program with children
at-risk for poor behavioral outcomes. Participants in the program were referred by school
guidance counselors, classroom teachers, the Department of Children’s Services and Juvenile
Court Systems. Referrals were largely for lower socioeconomic status children and based on
poor academic performance, socially deviant or defiant behavior, and potentially unsafe home
environments. Research has suggested that socially deviant behavior, family history of problem
behavior, divorce, child abuse and neglect, average or below average academic proficiency,
truancy, absenteeism, economic deprivation and higher community crime rates are positively
correlated to problem behaviors in children (OJJDP Model Programs Guide). The after school
program implemented a standardized program known as Life Skills Training, which focused on
improving decision making skills related to socially acceptable behaviors at home and in school,
resisting negative peer influences, refusing drugs, and avoiding delinquent behavior. Data were
collected by the host community organization and were analyzed by the present authors as
independent consultants. With appropriate qualification due to issues with a large number of
missing data points and a lack of demographic data, involvement in Life Skills Training
produced an increase in knowledge about risky situations and behaviors and an increase in
positive attitudes and intentions to resist negative influences and to choose positive behaviors.
The evaluation of whether such programs are successful is essential for the effective design
and implementation of future programs. Future directions will examine the conditions in which
after school programs are most successful and what amount of time in a program produces the
most desirable outcome.
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VOICE ONSET TIME AS A CLINICAL INDICATOR OF HYPERFUNCTIONAL VOICE
DISORDERS
Brandon Phillips. Department of Communicative Disorders, East Tennessee State
University, Johnson City, TN 37614
Voice onset time (VOT) is a measurable time interval that can be used to measure glottal
pulsing after the oral release of a stop consonant. VOT is an important measure for
distinguishing stop consonant voicing and place of articulation during speech across a variety of
languages. This measurement is relatively easy to make and displays the complexity of the
coordination between the supralaryngeal and laryngeal mechanisms. Given that VOT reflects
the timing differences between supralaryngeal articulation and phonatory gestures, previous
studies have examined if VOT can reflect laryngeal dysfnuction. Tyler and Watterson (1991)
compared the VOTs of individuals with hyperfunctional voice disorders and individuals with
normal laryngeal function, and reported that the individuals with laryngeal hyperfunction
displayed longer VOTs for voiced stops than the normal laryngeal function group. However,
several extraneous variables were not controlled, including rate of speech, gender of the
participants, and vowel context, each of which have been shown to influence VOT values
(Ryalls et al., 1997; Whiteside & Irving, 1997; 1998; Robb et al,. 2005). Thus, the purpose of
this study was to examine the VOTs of individuals with hyperfunctional voice disorders when
compared to the VOTs of individuals with normal voices within English stop consonants while
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2007 Appalachian Student Research Forum
controlling for rate, gender, and vowel context. Based on the results of previous research, it
was hypothesized that individuals with hyperfunctional disorders would display longer VOTs for
voiced stops than individuals without hyperfunctional voice disorders.
Recordings were made of 10 adults between the ages of 18-55 with normal laryngeal function
and 10 adults between the ages of 18-55 with hyperfunctional voice disorders. Participants
were classified as hyperfunctional or normal by undergoing a diagnostic voice assessment,
including auditory-perceptual measures and acoustic measures. Once the participants were
divided into two separate groups, they were each instructed to read six short phrases containing
English stop consonants in the word-initial position (ex., “Peek at a peacock.”). The stop
consonants contained within the phrases were the analyzed for VOT. A mixed analysis of
variance was used to determine any between group and/or within group differences.
Results indicated that both the hyperfunctional group and the normal group produced similar
VOT’s across all English stop phonemes; thus, no significant differences in VOTs were reported
between the groups. The current results differ with previous research; however, the current
methodology used was much more strident than previous studies. It is important to mention that
laryngeal gestures involved in the contrast between voiced and voiceless stop consonants is
only one of many coordinated muscular movements reflected in VOT. Thus, it seems that VOT
cannot be used as a reliable measure of laryngeal function by itself.
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NICOTINE-CONDITIONED HYPERACTIVITY IN A RODENT MODEL OF PSYCHOSIS
Brianna Sheppard and Russell W. Brown, Department of Psychology, East Tennessee
State University, Johnson City, TN 37614
Nicotine, the addictive psychostimulant drug in cigarettes, has positive reinforcing properties
that often become associated with environmental and contextual cues. The schizophrenic
population in particular exhibits especially high rates of smoking, which is nearly three times
that of the general population. The high incidence of smoking among these patients raises the
possibility of an underlying neurochemical mechanism involving schizophrenia and nicotine use
in which the midbrain dopaminergic system, also referred to as the mesocorticolimbic pathway,
has been implicated. This high incidence of smoking may be further exacerbated by the
presence of nicotine-associated cues. Past studies have shown that a nicotine-conditioned
context can elicit locomotor hyperactivity in a rodent model of nicotine addiction during a
nicotine-free trial. However, the role of context in nicotine addiction has not been examined in
an animal model of schizophrenia. The aim of this investigation was to determine the ability of
a nicotine-conditioned context to elicit locomotor hyperactivity in a neonatal quinpirole animal
model of schizophrenia. Sprague-Dawley rats were treated with either saline or the dopamine
D2 receptor agonist quinpirole from postnatal days (P) 1-21 to create long-term D2 receptor
supersensitivity that persists throughout the animal's lifetime, a phenomenon known as D2
priming. On P29, the beginning of early adolescence in the rat, animals were habituated to a
locomotor arena for three consecutive days. On each day of habituation, all animals were
given an injection of saline and placed into a locomotor arena ten minutes later, and each
habituation session lasted for ten minutes. This was done to allow animals to adjust to
surroundings of the arena. On P32, animals were injected i.p. with either nicotine (0.5 mg/kg
free base) or saline and placed in the locomotor arena 10 minutes later. Results showed that
D2 primed rats that received nicotine demonstrated a significant increase in activity compared
to all other groups, suggesting that the dopamine system may have a hyperactive response to
nicotine in D2-primed animals. Interestingly, on the drug-free test, these animals were also
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2007 Appalachian Student Research Forum
more active compared to all other groups, suggesting that nicotine may have stronger
associative properties in D2-primed as compared to non D2-primed animals.
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RELATIONSHIP BETWEEN PARENTAL ATTITUDES TOWARD WEIGHT/WEIGHT
CONTORL AND CHILD FEEDING PRACTICE
Liang Wang, Amit Bodhani, Tiejian Wu and Karen Schetzina. Department of Public
Health, East Tennessee State University, Johnson City, TN 37614
Childhood obesity is a significant and growing health problem in the US. Secular trend in the
prevalence of child obesity in the US suggest that children have become substantially heavier
over the last three decades, and as a result their risk for a number of health problems including
hypertension and type II diabetes is increasing. Parents are key agents to develop a home
environment that fosters healthful eating and physical activity among children and adolescents.
Parents can shape their children’s dietary practices, physical activity, sedentary behaviors, and
ultimately their weight status in many ways. The purpose of this study was to investigate
parental attitudes toward weight/weight control and their associations with child feeding
practice/behaviors. Study subjects included 60 mothers or other primary child care givers of
children 5-11 years old who were attending a pediatric clinic for health care. Parental attitude
toward weight/weight control was assessed using the Dieting Belief Scale (DBS) and child
feeding practice was assessed using the Child Feeding Questionnaire (CFQ). Preliminary data
analysis indicated a significant association between parental attitudes toward weight/weight
control and certain child feeding practice/behaviors.
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2007 Appalachian Student Research Forum
Division III—
Graduate Students 2+ years
Biomedical Sciences
IDENTIFICATION OF A POSSIBLE FUNCTIONAL CRE-ELEMENT FOUND IN THE
PROMOTER OF THE α7 NICOTINIC RECEPTOR SUBUNIT AND ITS ASSOCIATION
WITH SCHIZOPHRENIA
Michelle Chandley, Chris Newell, Tracy Wilson, John Beddies and Dr. Barney Miller.
Departments of Psychiatry and Anatomy/Cell Biology, East Tennessee State University,
COM, Johnson City, TN 37614
Objective: The identification and characterization of a possible Cyclic-AMP response element
(CRE) within the promoter region of the α7 subunit gene.
Abstract: Insight regarding the complex etiology of schizophrenia may lie in one of its most wellknown symptoms, auditory hallucinations (AH). One abnormality that has been described in
schizophrenia, that may contribute to AH, is the lack of auditory gating, i.e. the inability to filter
background stimuli and focus on a single auditory. This gating deficit can be demonstrated by
attenuation of P50, which is a measurable brainwave evoked by external auditory stimuli. The
neural pathway associated with P50 involves the α7 subunit of the acetylcholine nicotinic
receptor (α7nAChR), which has high affinity binding to nicotine, and has been implicated in
normal auditory gating, suggesting a link between schizophrenia and this receptor. A7nAChR’s
involvement is further supported by the high incidence of tobacco use in the diagnosed
schizophrenic population, between 85% and 90%, more than triple the normal use. Nicotine upregulates the expression of α7nAChR, perhaps explaining the seemingly self-medicating
behavior. In addition, diminished post-mortem brain and blood levels of the α7 nicotinic
receptor have been reported in schizophrenic patients. Transcriptional regulation of the
α7nAChR subunit of the nicotinic receptor is poorly understood. Identifying the transcriptional
events that lead to expression of α7 in the central nervous system could uncover alterations in
these control mechanisms which might establish a bridge between the symptoms (such as AH)
and a genetic etiology. A cyclic-AMP response-like element is located in the proximal promoter
region of the α7nAChR gene. The purpose of this study was to assess whether this promoter
region acts as a functional CRE (cyclic-AMP response element) and to determine if
polymorphisms flanking this region, which have been found in schizophrenic patients, have any
effect on transcription of the gene. Electrophoretic Mobility Shift Assay (EMSA) was performed
using radioactively-labeled oligonucleotides representing the putative CRE region of the α7
promoter and Cyclic-AMP response element binding protein (CREB). Controls included a
purchased, concensus CRE-element oligonucleotide, a mutated CRE-element oligonucleotide,
and a concensus Activating Protein 2 (AP2) DNA transcription site. Reporter gene constructs
were created using the α7nAChR promoter to drive expression of luciferase protein. Chromatin
crosslinking/immunoprecipitation assays (CHIP), using antibodies for CREB, were used to
isolate DNA bound in vivo to CREB, which were then subjected to PCR using primers specific to
the α7 promoter region. Phosphoimaging of the EMSA gels demonstrated a shift in the mobility
of the labeled oligonucleotides in the presence of CREB protein. Luciferase reporter gene
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2007 Appalachian Student Research Forum
assays using mutated regions of the promoter demonstrated a decrease in gene expression
compared to consensus promoter sequences. CHIP assay/PCR analysis of chromatin isolated
using CREB antibodies displayed a 400bp amplified band which migrates with the same MW as
the band amplified from genomic DNA. The region of DNA in the α7 promoter analyzed in this
study could be a previously uncharacterized regulatory promoter element (CRE) in the gene for
the α7 subunit of the nicotinic receptor. By characterizing the transcriptional process of α7, new
advances in molecular medicine within this pathway may lead to a more effective treatment for
schizophrenia.
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A CANNABINOID (WIN 55,212-2) INDUCES APOPTOSIS IN PANCREATIC TUMOR
CELLS VIA CB1-DEPENDENT AND CB1-INDEPENDENT MECHANISMS
Theresa Pickle and Douglas Thewke, Department of Biochemistry and Molecular
Biology, East Tennessee State University, COM, Johnson City, TN 37614
Pancreatic cancer is the fourth leading cause of cancer related deaths. In 2007, an estimated
37,170 Americans will be diagnosed with cancer of the pancreas and an estimated 33,370
Americans will die 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 9-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 and immunoblotting 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 receptor-specific antagonist, AM251, but not by a CB2 receptorspecific 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 or CaPan1 cells. In conclusion, the results presented here demonstrate that several
pancreatic tumor cell lines express cannabinoid receptors and that a cannabinoid, Win 55,2122, potently inhibits the viability of these pancreatic tumor cells, at least in part, by inducing
apoptosis via CB1-dependent and CB1–independent mechanisms.
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2007 Appalachian Student Research Forum
HSPA12B IS INDUCED BY HYPOXIA/REOXYGENATION IN VITRO AND IS
PROTECTIVE AGAINST MYOCARDIAL ISCHEMIA/REPERFUSION INJURY IN VIVO
Rebecca Steagall, Fang Hua, Chuanfu Li, Nilanjana Maulik and Zhihua Han, Department
of Biochemistry and Molecular Biology, East Tennessee State University, COM, Johnson
City, TN 37614 and Molecular Angiogenesis and Cardiology Laboratory Department of
Surgery, University of Connecticut Medical Center, Farmington, CT 06030
Hypoxia and oxidative stress associated with Ischemia and reperfusion (I/R) represent major
mechanisms of tissue injury and organ failure. Angiogenesis is a major protective mechanism
against ischemic injury. Recently, we cloned Heat shock protein A12B (HspA12B), the newest
member of a newly defined sub-family of proteins related to the Hsp70 family that is
predominantly expressed in endothelial cells (ECs) and is required for angiogenesis. Because
of its homology to Hsp70 we proposed that it may represent a specialized molecular chaperone
that is involved in angiogenesis. In the present study, we found that HspA12B expression was
induced by hypoxia and reoxygenation in endothelial cells. We used three models to investigate
the hypoxia-induced HspA12B gene expression and its protective effect against I/R-induced
myocardial infarction conceivably due to HspA12B stimulated angiogenesis. First, in human
coronary artery endothelial cells (HCAECs), knockdown of HspA12B by siRNA resulted in a
significant decrease in hypoxia-induced tubular morphogenesis compared to control. Second,
in the in vivo C57BL/6 mouse model, we found that angiogenesis was induced by overexpression of HspA12B by adenovirus in a Directed in vivo Angiogenesis Assays (DIVAA).
Third, in the in vivo myocardial I/R injury model, the HspA12B adenovirus decreased I/Rinduced infarct size in the myocardium. In conclusion, these results suggest that pro-angiogenic
HspA12B is induced by hypoxia and may contribute to a cardioprotective effect in myocardial
ischemic models. Many studies have shown that pro-angiogenesis treatments protect against
I/R. Alternatively, cytoprotective effect could arise from either protection against apoptosis or
chaperone activity, both of which have been demonstrated for Hsp70s. We show here that
hypoxia induced HspA12B regulated angiogenesis provided protection against I/R injury to the
myocardium. We are in the process of elucidating the mechanisms. Ultimately, overexpression of a therapeutic HspA12B transgene using a vector that will provide expression in
response to an endogenous pathophysiological stimulus such as hypoxia may protect tissues at
risk of ischemia/reperfusion injury.
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HSV-2 CO-INFECTION STIMULATES CHLAMYDIA TRACHOMATIS PERSISTENCE
VIA A NOVEL MECHANISM
J. Vanover, S. Deka, J. Sun, J. Kintner, J. Whittimore and R.V. Schoborg, Department of
Microbiology, East Tennessee State University, COM, Johnson City, TN 37614 and
ViroMed Laboratories/LabCorp, Minnetonka, MN 55343
Epidemiological studies have demonstrated that co-infections of herpes simplex virus type 2
(HSV-2) and Chlamydia trachomatis occur in vivo. Data from a tissue culture model of C.
trachomatis/HSV-2 co-infection indicate that viral co-infection stimulates the formation of
persistent chlamydiae. Several models of chlamydial persistence have been previously
described, including IFN-, IFN-α, IFN-β, and TNF- α exposure and iron, amino acid or glucose
deprivation. We hypothesize that HSV-2 co-infection induced chlamydial persistence is not
mediated by one of these known inducers of persistence. ELISA analyses demonstrate that
accumulation of intracellular ferritin remains similar in co-infected and singly-infected cells,
indicating that viral co-infection does not reduce host intracellular iron stores. Supplementation
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2007 Appalachian Student Research Forum
with excess amino acids, iron-saturated holotransferrin, or glucose during co-infection also does
not restore chlamydial infectivity in co-infected cells, demonstrating that viral-induced
persistence is not mediated by depletion of these nutrients. Previously published studies from
our laboratory demonstrate that HSV co-infection induces persistence in co-infected HEC-1B
cells. Because HEC-1B cells do not produce indoleamine-2,3-deoxygnease (IDO) in response
to IFN-γ, these data suggest that viral-induced persistence is not mediated by IFN- . Several
studies indicate that the chlamydial trp operon is strongly up-regulated when the organisms are
confronted by tryptophan limiting conditions, as occurs during IFN- induced persistence.
However, RT-PCR studies indicate that chlamydial trpA mRNA is not up-regulated during coinfection. Additionally, Luminex assays indicate that IFN-, IFN-α, and TNF- α are not released
from co-infected HeLa cells. Finally, RT-PCR studies demonstrate that IFN-, IDO, and IFN-β
mRNA transcripts are not present in co-infected HeLa cells. Collectively, these data suggest
that the synthesis/release of IFN-, IFN-α, IFN-β and TNF- α is not responsible for viral-induced
persistence. Co-infection with UV-inactivated, replication incompetent HSV-2 reduces
chlamydial infectivity without altering chlamydial genomic DNA accumulation, suggesting that
productive HSV replication is not required. Furthermore, co-incubation of fixed, HSV-2 infected
inducer cells with viable C. trachomatis infected responder cells suppresses production of
infectious chlamydial elementary bodies and stimulates the formation of swollen, aberrantly
shaped reticulate bodies. These data: 1) demonstrate that co-infection induced persistence is
not mediated by any currently characterized persistence inducer and 2) suggest that HSV
attachment to host cell surface receptors can provide the necessary stimulus to alter C.
trachomatis development. Thus, we hypothesize that, during HSV-2 attachment, the actions of
one or more virion proteins trigger a novel host anti-chlamydial pathway that restricts chlamydial
development.
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TRANSCRIPTIONAL REGULATION OF C-REACTIVE PROTEIN EXPRESSION IN
HUMAN HEPATOCYTES
Bhavya Voleti, Prem Prakash Singh and Alok Agrawal. Department of Pharmacology,
East Tennessee State University, COM, Johnson City, TN 37614
The hepatic synthesis of C-reactive protein (CRP) increases in inflammatory conditions resulting
in raised serum CRP levels. The regulation of CRP gene expression in hepatocytes occurs at
the transcriptional level. Previously, we reported that a C/EBP-binding site overlapping a NF-ĸB
p50-binding site (C/EBP-p50-site) on the CRP promoter was critical for IL-6-induced CRP
expression in human hepatoma Hep3B cells. Transcription factors C/EBPβ and p50 occupy the
C/EBP-p50-site in the nuclei containing constitutively active or IL-6-activated C/EBPβ. The aim
of this study was to identify the transcription factors that occupy the C/EBP-p50-site in the
absence of C/EBPβ. Accordingly, we treated Hep3B nuclear extract with a C/EBP-binding
consensus oligonucleotide to generate an extract lacking active C/EBPβ. Such treated nuclei
contain only C/EBPς because the C/EBP-binding consensus oligonucleotide binds to all C/EBP
family proteins except C/EBPς. EMSA using this extract revealed formation of a C/EBPςcontaining complex at the C/EBP-p50-site. This complex also contained RBP-Jĸ, a transcription
factor known to interact with the ĸB sites. RBP-Jĸ
formation of C/EBPςcontaining complexes. The RBP-Jĸς-containing complexes were also
formed at the C/EBP-p50-site on the CRP promoter in the nuclei of primary human hepatocytes.
In transactivation assays, overexpressed C/EBPς abolished both C/EBPβ-induced and (IL-6+IL1β)-induced CRP promoter-driven luciferase expression. These results indicate that under
basal conditions, C/EBPς occupies the C/EBP-site, an action that requires RBP-Jĸ. Under
induced conditions, C/EBPς is replaced by C/EBPβ and p50. We conclude that the switch
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2007 Appalachian Student Research Forum
between C/EBPβ and C/EBPς participates in regulating CRP expression. This process utilizes
a novel phenomenon, that is, the incorporation of RBP-Jĸ into C/EBPς-complexes solely to
support the binding of C/EBPς to the C/EBP-site.
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PROTECTIVE MECHANISMS OF NECROSTATIN-1 ON GLUTAMATE-INDUCED
TOXICITY IN HT-22 CELLS
Xingshun Xu, Chu C. Chua, Jiming Kong, Richard M. Kostrzewa, Udayasankar
Kumaraguru, Ronald C. Hamdy and Balvin H.L. Chua. Department of Pharmacology,
East Tennessee State University, COM, Johnson City, TN 37614
Glutamate, a major excitatory neurotransmitter in the central nervous system, plays a critical
role in nervous system disorders such as stroke and Parkinson’s disease. Recent studies have
suggested that glutamate excess can result in a form of cell death called glutamate-induced
oxytosis. In this study, we explore the protective effects of necrostatin-1, an inhibitor of
necroptosis, on glutamate-induced oxytosis. We show that necrostatin-1 inhibits glutamateinduced oxytosis in HT-22 cells through a mechanism that involves an increase in cellular
glutathione (GSH) levels as well as a reduction in reactive oxygen species production.
However, necrostatin-1 had no protective effect on free radical-induced cell death caused by
hydrogen peroxide or menadione, which suggests that necrostatin-1 has no antioxidant effects.
Interestingly, the protective effect of necrostatin-1 was still observed when cellular GSH was
depleted by buthionine sulfoximine, a specific and irreversible inhibitor of glutamylcysteine
synthetase. Our study further demonstrates that necrostatin-1 significantly blocks the nuclear
translocation of AIF (a marker of caspase-independent programmed cell death) and inhibits the
integration of BNIP3 (a pro-death member of the Bcl-2 family) into the mitochondrial membrane.
Taken together, these results demonstrate for the first time that necrostatin-1 prevents
glutamate-induced oxytosis in HT-22 cells through GSH-related as well as AIF and BNIP3related pathways.
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Social & Behavioral Sciences
AN ANALYSIS OF LOUDNESS PERCEPTION IN PERSONS WHO EXPERIENCE
TINNITUS
Jeffrey A. Py and Marc Fagelson, PhD. Department of Communication Disorders, East
Tennessee State University, Johnson City, TN 37614
The purpose of this study was to examine whether persons with normal hearing thresholds and
tinnitus performed differently on a test of intensity discrimination when compared to results from
persons with normal hearing and without tinnitus. There were two groups of five subjects;
Group 1 consisted of normal hearing patients without tinnitus and Group 2 consisted of subjects
with normal hearing and subjective complaints of tinnitus. The test stimulus consisted of three
trains of five short tone pips. The five tones were presented in one of three patterns; intensity
increasing in 0.5 dB steps (producing a cumulative of 2.5 dB increase), intensity decreasing in
0.5 dB steps (producing a cumulative 2.5 dB decrease), or intensity unchanged. Initial tone
level was varied randomly around a 50 or 70 dB HL standard in order to minimize onset and
offset cues. Verbal judgments were made by test subjects with regard to their perception of a
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stimulus train increase, decrease or no change in stimulus level. Differences between the two
groups were compared by 1) proportion of correct responses and 2) proportion of response
types (either ascending, descending, or unchanged). There was no significant difference
between groups regarding number of correct responses, but there was a significant difference
between groups regarding response type. The results indicated a possible processing
difference with regard to intensity discrimination in that the patients with tinnitus were either
unable to discern, or unwilling to report, a change in signal loudness.
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TRADITIONAL VS COMPUTER-BASED INTERVENTION FOR TREATMENT OF
SPEECH SOUND DISORDERS
Ashley Rice, Julie Eanes, Marsha Mallory and A. Lynn Williams. Department of
Communicative Disorders, East Tennessee State University, Johnson City, TN 37614
Children with speech sound disorders (SSD) comprise the most common communication
disability in preschool and school-aged children (American Speech-Language Hearing
Association, 2000). Traditional treatments of SSDs utilize pictured stimuli while working with
children at a table. Recent technological advances have resulted in the availability of software
programs for treating SSDs using computer-based intervention (CBI). Despite the emergence
of software programs, there is little research that has compared the relative effectiveness and
benefits of these two treatment formats. Such research would be of value in assisting speechlanguage pathologists (SLPs) to make decisions regarding the most efficacious and effective
treatment strategies for children with SSDs. The purpose of this study was to determine the
differences, if any, in treatment outcomes for children receiving a traditional paper format of
therapy versus a computer format. The computer software program Sound Contrasts in
Phonology (SCIP) was utilized to create materials for both table-top and computer conditions.
The SCIP illustrations were printed out for the traditional condition, whereas the computer
conditions operated the SCIP illustrations on the computer screen. Participants for this study
were two males (3 years, 8 months and 4 years, 7 months) with moderate to profound SSD. A
single-subject AB design was used to evaluate the treatment outcomes of each child.
Participants were randomly assigned to treatment conditions. Participant 1 received the
traditional treatment condition and Participant 2 received the CBI condition. One phonological
goal was targeted for each child in the assigned treatment condition. Treatment consisted of
five minimal pair contrasts for each targeted phoneme in both treatment conditions. Treatment
sessions were held twice weekly for 30 minutes in length per session. A generalization probe of
the target sounds in an untrained context was constructed for each participant in order to
measure baseline performance as well as learning throughout the course of treatment. The
results indicate that the computer condition was an effective service delivery format in achieving
improvement for treatment and generalization. In addition, these gains were achieved in less
time (5 sessions) as compared to the traditional condition (12 treatment sessions).
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Division IV—
Medical Students
RELATIONSHIP OF MATRIX METALLOPROTEINASES WITH MARKERS OF FETAL
LUNG MATURITY
Scott Cook, Tiffany Ford, Gary Randall, John Kalbfleisch and Kevin Breuel. Departments
of Obstetrics and Gynecology and Biometry and Medical Computing, East Tennessee
State University, COM, Johnson City, TN 37614
Neonatal respiratory distress syndrome (NRDS) remains a major cause of infant morbidity and
mortality. The primary cause of neonatal NRDS is a deficiency in the production of surfactant
which results in a reduction in surface tension in the alveolar walls and reduced gas exchange
by the lungs. Diagnosis of neonates with fetal lung immaturity allows effective intervention
which reduces the risk of NRDS. Specialized laboratory tests (L/S Ratio, Optical Density (OD)
and PG) used to diagnose lung maturity in amniotic fluid are quite sensitive (95-96%) in
identifying infants with immature lungs whom as a result may develop NRDS. However, these
tests lack specificity (60-68%) which result in falsely diagnosing infants with immature lungs
when they are actually mature. Matrix metalloproteinases (MMPs) and tissue inhibitors of
metalloproteinases (TIMPs) are known to regulate extra cellular matrix remodeling and thus are
important in the morphogenesis of all the fetal organs. The objective of this study was to
determine whether amniotic fluid levels of various MMPs are correlated with L/S ratio, OD or
levels of PG and thus might be used to assess fetal lung maturity (FLM). Seventy-eight,
amniotic fluid samples, previously collected for assessment of FLM, were removed from
storage (-80°C) and processed for analysis of MMP (1, 2, 3, 7, 8 and 12) levels. Levels of
various MMPs were quantified with a Fluorokine MAP multiplex assay kit run on the Luminex
100 instrument. L/S and OD measures were related to MMPs with single and multiple
regression analysis. Additionally, mean MMPs were compared for presence (Mature) and
absence (Immature) of PG banding subgroups. P = 0.05 or smaller was used for statistical
significance. The L/S and OD were always positively correlated with MMP3 levels (P<0.01)
irrespective of whether the other MMPs are included in the regression analysis. Mean levels of
MMP3 (270±40 pg/ml) and MMP7 (19,447±3565 pg/ml) in amniotic fluid samples with PG
present were higher (P<0.05) when compared to samples without PG (142±32 and 6,031±1659,
respectively). In conclusion, MMP3 and MMP7 may be candidates for further study in an
attempt to identify a predictor of FLM that is both sensitive and specific.
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BINDING OF C-REACTIVE PROTEIN TO IMMOBILE IMMUNE COMPLEXES
Christopher L. Cropsey, Madathilparambil V. Surech, Sanjay K. Singh and Alok Agrawal.
Department of Pharmacology, East Tennessee State University, COM, Johnson City, TN
37614
C-reactive protein (CRP) participates in the first line of defense against pathogens. The exact
mechanism through which CRP functions in vivo has not been defined yet. In vitro, CRP has
been shown to bind to Fcγ receptor IIa (also called as CD32). CD32 is present on the cells of
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the immune system and is the receptor for IgG-containing immune complexes. In this study, we
explored CRP-CD32 interaction in an ELISA-based solid-phase binding assay using
recombinant CD32-coated plates. CRP bound to CD32 in a dose-dependent manner.
Surprisingly, when CD32 was blocked with anti-CD32 IgG antibodies, the binding of CRP to
CD32 was not inhibited, indicating a possible interaction between CRP and Cd32-anti-CD32
complexes. Accordingly, when normal huma IgG-coated plates were used in the assay, we
observed a dose-dependent binding of CRP to igG. The inability of the binding of a mutant form
of CRP to IgG further showed the specificity of CRP-IgG interaction. CRP also bound to
immune complexes prepared from BSA and Anti-BSA antibodies, indicating that the presence of
the antigen did not affect the binding of CRP to IgG. CRP did not bind to immune complexes in
the fluid-phase. The capability of CRP to interact with immobile immune complexes suggests
that CRP may also bind to immune complex-occupied CD32-bearing cells and modulate
intracellular signaling an subsequent inflammatory responses triggered by the binding of
immune complexes to CD32. Our findings have implications for the functions of CRP in the
antibody-mediated clearance of pathogens and in immune complex-mediated pathologic
conditions.
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INHIBITORY EFFECTS OF BAICALEIN ON PRODUCTION OF SELECTED
INFLAMMATORY CYTOKINES BY IL-1Β-ACTIVATED HUMAN MAST CELLS
Justin R. Sigmon, Chia-Jung Hsieh, Kenton Hall, Guha Krishnaswamy, Tuanzhu Ha,
Chuanfu Li and David S. Chi. Departments of Internal Medicine and Surgery, East
Tennessee State University, COM, Johnson City, TN 37604
Human mast cells play an integral role in the inflammatory response by mediating the
production of inflammatory cytokines. Baicalein (BAI), a compound isolated from the traditional
Chinese medicinal herb Scutellaria baicalensis Georgi, has been shown to have antiinflammatory effects. We examined the effects of BAI on the production of inflammatory
cytokines, IL-6, IL-8, and MCP-1 from IL-1β-activated human mast cells (HMC-1). Two mL of
HMC-1 at 1 x 106 cells/mL were cultured with BAI (at predetermined non-toxic concentrations) in
the presence or absence of IL-1β (10 ng/mL) for 24 hrs. The supernatants were harvested and
assayed for cytokines by ELISA. BAI alone did not induce cytokine production from HMC-1.
However, BAI (15 and 30 µM) significantly decreased production of IL-6 and MCP-1 (p<0.0004)
from IL-1β-activated HMC-1. BAI at all tested concentrations (1.8 to 30 µM) significantly
decreased the production of IL-8 (all p<0.009) from IL-1β-activated HMC-1 in a dose dependent
manner. The results show that BAI had inhibitory effects on the production of selected
inflammatory cytokines, most notably IL-8, and may be useful in the development of novel antiinflammatory therapies. (Supported by the Chair of Excellence in Medicine, the Ruth R. Harris
Endowment, and RDC of ETSU.)
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Division V—
Residents/Post-Doctoral Fellows
REVIEW OF LITERATURE ON PROTEINURIA IN PREECLAMPSIA
Rachna Bharti, MD. Department of Family Medicine, East Tennessee State University,
COM, Johnson City, TN 37614
BACKGROUND INFORMATION: The research comparing 24 h urine protein to urine
protein/creatinine ratio in women with suspected preeclampsia has been ongoing since 1990
(most recent study published in December 2006), however many physicians are not comfortable
implementing the use of spot urine protein/creatinine ratio for diagnosing preeclampsia.
OBJECTIVES OF THE STUDY: To review the current literature comparing 24 h urine protein
and urine protein/creatinine ratio methods and draw conclusions about the significance and
validity of the findings of the research. To identify the most appropriate method for diagnosing
preeclampsia.
METHODOLOGY: Over 20 articles were retrieved via Medline regarding the methods for
diagnosing proteinuria, more than half of which focused specifically on pregnant women. The
outcomes of the studies in the articles were then reviewed for statistical significance. Studies
were identified as either supportive or non-supportive of the protein/creatinine ratio for diagnosis
of preeclampsia as compared to 24 h urine protein collection.
RESULTS: Nearly all of the research articles reviewed support the use of urine
protein/creatinine ratio as an alternative to 24 h urine protein for diagnosis of preeclampsia.
Only one study done in 2003 found that the protein/creatinine ratio should not be used an
alternative to the 24 h urine protein. Although this study found a significant relationship between
the two methods (p<.0001), the correlation coefficient was cited as being too low (r2=0.41).
However, another study in the same year showed a very strong correlation between the two
methods (r2=.929). A majority of the studies agreed that the protein/creatinine ratio method was
simpler, faster, and could be a more practical alternative for assessment of proteinuria.
CONCLUSION: The research supports that the urine protein/creatinine ratio is nearly
equivalent to the 24 h urine protein analysis. The urine protein/creatinine ratio may be used as
reliable method for diagnosing preeclampsia in pregnant women.
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SURGICAL SIMULATION OF COLD KNIFE CONIZATION AND SUCTION DILATION
AND CURETTAGE PROCEDURES USING A PATIENT SIMULATOR AND PAPAYA
MODEL
Foulk, Brooke E., Eason, Martin J. and Olsen, Martin E. Department of Obstetrics and
Gynecology, East Tennessee State University, COM, Johnson City, TN 37614
Objective: Our purpose was to equip residents in performing gynecologic surgical procedures
using a patient simulator and papaya model. Some training institutions have used papayas,
cantaloupes or other fruits as uterine models for practice of various surgical procedures. We
proposed to further the realism of this design and take advantage of this learning tool in
combination with our institution’s innovative patient simulator lab to better represent actual
gynecologic patients and surgical procedures.
Background: Similar to Paul and Nobel’s 2005 description,1 we planned to teach and practice
suction dilation and curettage with the papaya model. However, we incorporated our
NOELLE™ Birthing Simulator for a more realistic experience. Also, we added to this the
simulation of a cold knife conization on the model prior to the suction dilation and curettage.
Methods: Using the papaya as a uterine model, the resident performed two surgeries. The
papaya is placed into the NOELLE™ Birthing Simulator, with its stalk representing the cervix
inside the vaginal vault.
1. A cold knife conization was performed by the learner using standard surgical
instrumentation, with removal of a small, cone-shaped section from the simulated cervix.
2. Next, a suction dilation and curettage was undertaken inside the papaya, which represents
the intrauterine cavity. The fruit’s internal contents and seeds, which have a consistency similar
to products of conception, were removed. Injection of red dye prior to the procedure allows this
material to demonstrate an appearance similar to products of conception.
Results: Resident participants of all training levels have undertaken this simulation. Residents
found the educational experience to be valuable and an effective teaching tool which prepared
them for and realistically simulated actual surgeries on human patients.
Conclusions: This papaya model enables the learning curve to rapidly increase in a controlled
setting, with unlimited “patients” and time. This allows interns and even students to act as
primary surgeon in a practical experience not typically provided at their levels of training. It is
well-suited for increasing the learner’s familiarization with the techniques and instrumentation of
the simulated procedures.
Discussion: We agree with Paul and Nobel’s suggestion “that simulation is an effective first step
in teaching uterine aspiration procedures.” The simulation provides an accurate surgical
experience with the potential for competency evaluation and pre- and post-procedure testing
and safety analysis. The risks involved with live patients and the costs associated with a realtime operating room, staff, anesthesia, and instruments are avoided.
1. Paul, M. and Nobel, K. Papaya: A Simulation Model for Training in Uterine Aspiration, Fam Med 2005;
37 (4):242-4.
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2007 Appalachian Student Research Forum
ALVEOLAR HEMORRHAGE: A RARE, BUT UNDERDIAGNOSED COMPLICATION
OF TREATMENT WITH GLYCOPROTEIN IIB/IIIA INHIBITORS
Said B. Iskandar, MD, Ehab S. Kasasbeh, MD and Philip D Henry, MD, FACC. Department
of Internal Medicine, Division of Cardiovascular Disease, East Tennessee State
University, COM and Veterans Affairs Medical Center, Mountain Home, TN, USA
Objective: Alveolar hemorrhage (AH) is a very rare complication of treatment with GP IIb/IIIa
inhibitors. Hemoptysis, a constant sign, lacks in specificity, and may occur in confounding
syndromes such as pulmonary edema, pulmonary infarction and pneumonia. Here, we
performed a large scale retrospective analysis to define the incidence and risk factors of AH in
the setting of GP IIb/IIIa inhibitors therapy.
Methods: We reviewed for the period extending from August 1998 to January 2005
consecutive histories of acute myocardial infarction patients receiving coronary arteriography
and treatment with either eptifibatide or abciximab. Concomitantly admitted acute myocardial
infarction patients not treated with GP IIb/IIIa inhibitors were reviewed and served as a control
group. Potential co-variates including pulmonary disease, pulmonary hypertension, smoking,
and use of other anticoagulant or anti-platelet agents were evaluated.
Results: Six of 1,810 patients (0.33%) receiving eptifibatide and five of 3,648 patients (0.14%)
receiving abciximab exhibited typical symptoms and signs of AH. Contrarily, only one of 4,136
patients (0.025%) receiving no GP IIb/IIIa inhibitors presented with similar symptoms and signs.
Statistically significant differences were found between control patients and patients receiving
eptifibatide alone (P=0.004). There was also a significant difference between untreated patients
and those receiving eptifibatide and abciximab (P=0.017). No differences were found between
eptifibatide and abciximab-treated patients (P=0.19) or between abciximab and untreated
controls patients (P=0.105).
Conclusions: AH is a very rare complication of treatment with GP IIb/IIIa inhibitors. Its
incidence ranged from 0.14% in patients treated with abciximab to 0.33 % in those receiving
eptifibatide. Compared to control group, patient treated with GP IIb/IIIa inhibitors had a
statistically increased risk for AH.
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FATAL IATROGENIC HYPERPHOSPHATEMIA: A GROWING DISASTER
Dima Nassour, Kosseifi Semaan, Byrd Ryland and Roy Thomas, IV. Department of
Internal Medicine, East Tennessee State University, COM, Johnson City, TN 37614
Fleet Phospho Soda is a widely used colorectal laxative, for which the safety profile has been
clearly questioned. Its inadvertent use, however, may be associated with a plethora of adverse
effects, some of which may be life-threatening. We are reporting an elderly patient who was
given sodium phosphate enemas, which resulted in marked hyperphosphatemia and
hypocalcemia followed by his demise.
Case presentation: A 77 year old man who was admitted to the hospital for an acute diffuse
abdominal distention, nausea, non-bloody vomitus. His past medical history include coronary
artery disease, remote history of cerebrovascular accident, congestive heart failure,
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2007 Appalachian Student Research Forum
hypertension, vascular dementia, chronic renal disease with last previous creatinine levels being
within normal limits, non-insulin dependent diabetes mellitus. His past surgical history was
significant for remote open cholecystectomy, transurethral retrograde prostatectomy.
Medications included Aspirin/Dipyridamole, Furosemide, Glyburide, Metoprolol succinate,
Omeprazole, Tamulosin and Nitroglycerine tansdermal patch. On initial presentation, patient
blood pressure was 139/87 mm of Hg, heart rate of 78 per min, respiratory rate 22/min, and
temperature 97.5 °F. On physical exam, patient was awake, demented. Head and neck
examination revealed dry oral mucosa otherwise within normal. Cardiovascular and pulmonary
exam was also within normal. His abdomen was distended, with sluggish bowel sounds and
diffuse tenderness on palpation. Rectal exam was normal with the absence of stools in rectal
vault. Admission laboratory data were consistent with acute pancreatitis, creatinine 1.8 (Normal
0.8-1.5 mg/dL), cholestatic liver disease with total bilirubin level 2.7 (Normal 0.2-1 mg/dL).
Patient was kept NPO and gastric decompression was performed. Flat and upright abdominal
radiograph revealed ileus with possibility of partial small bowel obstruction. Subsequently, he
was given a fleet phosphate enema every 4 hours for a total of 6 units. CT- scan of the
abdomen and pelvis without intravenous contrast showed evidence of acute interstitial
pancreatitis. Over 24 hour period, patient's condition deteriorated, to become hypotensive and
in respiratory distress. Patient was transferred to the medical intensive care unit, and initial
resuscitative measures were initiated. Shortly after that, he coded and died. Pre-mortem blood
work up results came back post-mortem, revealing a calcium level of 4.8 (Normal 8.4-10.2
mg/dL) that was previously normal on admission and a phosphorus level of 16.0 (2.7-4.5 mg/dL)
with normal amylase and lipase levels. We assume that our patient died from iatrogenic
complications of hyperphosphatemia, hypocalcemia leading to cardiopulmonary arrest and
demise.
Conclusion: Since fleet phosphate enemas are widely used for various indications, physicians
from all specialties need to be alerted regarding their injudicious use that can lead to an
increase in morbidity and mortality. A discussion about the etiology, pathogenesis, clinical
presentation, diagnosis and treatment of hyperphosphatemia will also be provided.
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SPONTANEOUS SPLENIC RUPTURE ASSOCIATED WITH STREPTOCOCCAL
VIRIDANS ENDOCARDITIS
Dima Nassour, Imran Samnani and Dafer Haddadin. Department of Internal Medicine,
East Tennessee State University, COM, Johnson City, TN 37614 and Medical Service,
James H. Quillen VAMC, Johnson City, TN
Splenic infarction in infective endocarditis (IE) is not uncommon, though abscess formation is
less common. Mycotic aneurysm (MA) can be the end result of arterial wall weakening caused
by septic emboli. MA leading to spontaneous splenic rupture is a rare complication of splenic
infarction due to embolism. Streptococcus viridians IE complicated by splenic rupture and
cerebral embolization in the same patient is extremely rare. We report a case of spontaneous
splenic rupture and ischemic stroke associated with Streptococcus viridans endocarditis along
with a literature review.
Case presentation: A 46-year-old Caucasian male patient presented with unsteady gait,
headaches, multiple falls, fever, non-specific abdominal pain, weight loss and loss of appetite of
three week duration. Three months prior to this presentation, patient had dental cleaning. His
past medical history was significant for sexually transmitted diseases in the past, negative HIV
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2007 Appalachian Student Research Forum
test done 6 months before presentation, smoking disorder and remote history of intravenous
drug use along with marijuana and cocaine abuse. Admission vital signs were stable besides
temperature of 101 F. Significant findings on physical examination revealed a soft
systolicejection murmur best heard at the left sternal border, grade III/VI. Significant
neurological examination revealed gait disturbance, decreased motor power in right arm and
numbness in right leg. Rest of physical examination was within normal limits. Initial laboratory
data revealed white cell count of 5200, 92% segments, platelet counts of 197,000, hemoglobin
10.1 g/dl, creatinine 1.0 mg/dl, elevated sedimentation rate (64 millimeters in the first hour) and
C-Reactive protein (29 mg/L), albumin 2.3g/dl. Serology tests for syphilis, hepatitis A, B, C
viruses and HIV were negative. His blood cultures were positive for Streptococcus viridans.
Brain magnetic resonance imaging including angiogram showed occlusion of the right insular
branch of the MCA along with severe atherosclerotic narrowing of the distal first portion of the
right middle cerebral artery with right insular gyrus and corona radiata white matter ischemia.
Additional ischemia involved the medial superior left cerebellum. CT abdomen showed small
bilateral pleural effusions and a 3.0 cm lesion in the superior margin of the spleen consistent
with infarct. Trans-esophageal echocardiogram showed a large 1.7 cm aortic valve (AV)
vegetation with significant AV prolapse and severe aortic regurgitation. A diagnosis of AV
endocarditis with embolisation phenomenon to brain, spleen was made. Therapy with
ceftriaxone and gentamicin was started. While patient was in the process of work-up prior to AV
replacement surgery, he started complaining of left pleuritic chest pain associated with left sided
abdominal pain and fever of 100 F. Repeated laboratory data showed worsening leukocytosis
up to 30,000, creatinine up to 3.3 mg/dl and drop in Hgb of 5 g/dL. Repeat CT abdomen
showed new splenic hemorrhage with hematoma formation and hemoperitoneum. He received
supportive care with blood products. Angiogram of abdominal vessels showed bleeding
aneurysm in the splenic artery. Patient underwent splenectomy with operative findings of
severely lacerated ruptured spleen with splenic artery aneurysm. Histopathology confirmed the
mycotic aneurysm in the splenic artery. He was discharged after a prolonged hospital course.
IE, not only, affects the heart but also other organs through other mechanisms, including
embolisation and immunological phenomena. Recognizing IE and its complications, with its
salient features, is very important when treating such patients for better outcome.
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IDENTIFICATION AND CHARACTERIZATION OF PUTATIVE FLAVONOID
GLUCOSYLTRANSFERASES FROM CITRUS PARADISI
Daniel K. Owens and Cecilia A. McIntosh. Department of Biological Sciences and School
of Graduate Studies, East Tennessee State University, Johnson City, TN, 37614
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 actions that are agriculturally
and pharmacologically important. The addition of sugars is a predominant modification reaction
in flavonoid biosynthesis, particularly in grapefruit where up to 70% of the dry weight of very
young fruit consists of flavanone glycosides. Glycosylation has a number of effects upon the
solubility, stability, and subsequent availability of metabolic products. The glycosylation of
flavonoids also has an influence upon the quality of foods and food products. In particular,
flavonoid glycosides convey taste characteristics in citrus making flavonoid glucosyltransferases
interesting targets for biotechnology applications in these species. To investigate glycosylation
of flavonoids in Citrus paradisi, putative glucosyltransferase clones are being isolated from
young leaf cDNA employing BD SMART™ RACE reactions with degenerate primers designed
against the plant secondary product glucosyltransferase (PSPG) box signature motif. Complete
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2007 Appalachian Student Research Forum
gene sequences are identified by primer walking, cloned into the Novagen pET® system for
recombinant protein expression, and purified by metal affinity chromatography. Ultimately, the
recombinant enzymes will be tested for substrate usage and thoroughly characterized by
biochemical assay. While new putative glucosyltransferases continue to be sought, PGT3 has
been established in the pET® system and the production of soluble recombinant protein
verified. Efforts to further develop metal affinity chromatography and begin biochemical assays
are currently underway.
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PREVALENCE OF PREMENSTRUAL DYSPHORIC DISORDER (PMDD):
UNITED STATE NATIONAL COMORBIDITY SURVEY REPLICATION (NCS-R)
Avani Prabhakar MBBS MPH, Merry N. Miller, MD and Deepak Prabhakar MD MPH.
Department of Internal Medicine and Department of Psychiatry and Behavioral Sciences,
East Tennessee State University, COM, Johnson City, TN 37614 and Department of
Epidemiology, University of North Texas Health Science Center,
Fort Worth, TX 76107
Although premenstrual worsening of mood disorder has been reported in medical literature
since the days of Hippocrates, it was not until 1931 when Robert Frank coined the term
premenstrual tension syndrome. In October 1998 medical community accepted a term called
Premenstrual Dysphoric Disorder (PMDD) as a distinct clinical entity. Women who have major
depressive disorder (MDD) through out the month are by definition excluded from the diagnosis
of PMDD or PMS. The concept of Premenstrual Exacerbation of MDD (PMED) has not been
recognized in DSM-IV, but has been reported in literature informally. DSM-IV Task Force on
Nomenclature and Statistics decided to include PMDD as an example of a mood disorder not
otherwise specified and put the criteria in the appendix. Objective of this study is to investigate
the spectrum of premenstrual depressive syndrome that occur across the women’s reproductive
life span, estimate the prevalence of PMDD, Dysphoric PMS, and PMED in the United States
Population using National Comorbidity Survey: Replication 2001-2003 (NCS-R) Data. We used
the NCS-R data to assess the prevalence rates, NCS-R was a fairly comprehensive survey and
its data is representative of the US trends. Survey investigators used the World Health
Organization (WHO) Composite International Diagnostic Interview (CIDI) questionnaire, a fully
structured instrument designed for use by trained interviewers, where diagnosis is based on
DSM-IV criteria. The prevalence of self-reported PMDD among US household-residing adult
females in their reproductive age group was estimated to be 1.9%, when a previous episode of
MDD was included in the analysis the prevalence rate rose to 2.8%, 0.9% of women report
PMED. The prevalence of dysphoric PMS is estimated to be 10.8%. Amongst Caucasians
prevalence of PMDD, dysphoric PMS, PMED, was, 2.2%, 11.5%, 0.9% respectively, for African
Americans prevalence of PMDD, dysphoric PMS, PMED, was,0.6%, 7.9%, 0.8% respectively
and for other race categories the prevalence of PMDD, dysphoric PMS, PMED, was, 1%, 8.5%,
1.1% respectively. Approximately 25% of adult females suffering from PMDD and 25% of those
suffering from dysphoric PMS received professional treatment at least once in the previous 12
months at the time of the interview, while only 11% of adult females suffering from PMED
received professional treatment at least once in the last 12 months at the time of the interview.
If we extrapolate this data on the US population according to the 2000 census, there are roughly
150 million adult females in their reproductive age in the US suffering from PMDD and around
750 million adult females in their reproductive age suffer from dysphoric PMS. Strict definition
of PMDD underestimates the prevalence. Our criterions were in sync with the DSM-IV PMDD
definition, but have their own limitation due to the methodology of data collection. The data has
been collected retrospectively, cross-sectionally, and thus prospective measurement of
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2007 Appalachian Student Research Forum
premenstrual symptoms in two consecutive months is not achieved in this data. Our analysis
demonstrates that spectrum of premenstrual depressive syndrome is widely prevalent in women
of reproductive age.
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RISK OF BREAST AND OTHER CANCERS DUE TO HYPERPROLACTINEMIA
CAUSED BY ANTIPSYCHOTICS (NEUROLEPTICS) OR OTHER MEDICATIONS: A
LITERATURE REVIEW
Umesh Vyas, MD. Department of Psychiatry, East Tennessee State University, COM,
Johnson City, TN 37614
BACKGROUND: Breast cancer is the most common cancer in females, and is the second most
common cause of death. There are several factors which increase a woman's risk for the
development of breast cancer. Some reports suggest that neuroleptics and other dopamine
antagonists increase the risk of breast cancer due to hyperprolactinemia. There are other
reports which suggest that they may decrease the risk of cancer especially in the rectum, colon
and prostate. Additionally, there is evidence that patients with Parkinson's disease have lower
rates of breast cancer and other types of malignancies.
OBJECTIVE: A literature review was performed to extract evidence and to evaluate risk or
benefit from hyperprolactinemia caused by these medications.
METHOD: Pubmed.gov and other online resources were searched by using pre-determined
key words.
RESULTS: Most studies report no increased risk of breast cancer associated with use of these
medications. Only one study reported a positive correlation between neuroleptic induced
hyperprolactinemia and an increased risk of breast cancer. Other studies report inconclusive
data.
CONCLUSION: At this time we do not have definitive data suggesting increased risk of breast
cancer secondary to hyperprolactinemia caused by antipsychotics. Thus, further studies are
desirable. Author will discuss this literature review in detail in poster presentation.
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2007 Appalachian Student Research Forum
Division VI—
Case History
SPONTANEOUS RUPTURE OF SPLEEN: DIAGNOSTIC DILEMMA, A CASE
REPORT
Rajesh Shivaji Kadam, MD, Hetal K. Brahmbhatt, MD and Avani Prabhakar, MBBS MPH.
Department of Internal Medicine and Department of Psychiatry and Behavioral Sciences,
East Tennessee State University, COM, Johnson City, TN 37614
Spontaneous rupture of the spleen has been defined as rupture of the spleen in the absence of
either trauma or any splenic pathology that has been reported after apparently minor insults
such as coughing and vomiting. To the date there are only two cases of spontaneous rupture of
spleen that have been reported in the literature and cough was supposed to be the underlying
culprit. The authors report a case of suspected spontaneous rupture of spleen secondary to
cough in a 54 year old Caucasian female, who recovered well after appropriate diagnosis, and
timely splenectomy. A fifty four year old Caucasian female was brought in by emergency
medical services. Initial encounter revealed an intubated and unresponsive female who was in
shock. Abdomen was mildly distended with hypoactive bowel sounds, and no mass or
organomegaly was appreciated. Lab data was remarkable for hemoglobin of 8.9 gram per
deciliter, hematocrit of 27.6. CT scan of head done at the time of admission showed no acute
abnormality, and chest x ray was unremarkable. CT scan of Abdomen showed large
hemoperitoneum secondary to a ruptured spleen. Patient was emergently taken to Operation
Theater just as CT scan was completed, there she underwent emergent abdominal exploratory
surgery where a ruptured spleen with a large subcapsular hematoma was found, and prompt
splenectomy was performed. Patient survived the surgery and had a slow post operative
recovery. Culture results from blood, sputum, and urine were all negative, and further serum
tests provided no evidence of acute viral infection but still Epstein Barr virus (EBV) serology
report was pending, which later on turned in positive, patient had no history of any viral infection
or malignancy. Splenic histopathology was suggestive of some degree of reactive hyperplasia
although the significance of this finding was uncertain. There are 5 criteria identified for the
diagnosis of spontaneous splenic rupture that, there should be no history of trauma to the
spleen, no evidence of disease in organs other than the spleen that are known to affect the
spleen, no perisplenic adhesions or scarring of the spleen, spleen should be normal on both
gross inspection and histologic examinations and convalescent, acute phase sera should not
show any significant rise in viral antibody titers suggestive of recent infection with viral types
associated with splenic involvement. This case meets the first 4 criterion and the EBV serology
is at best 91% specific therefore the authors believe that a spontaneous rupture of spleen is still
a possibility until we completely rule out EBV infection. If truly spontaneous this will be the third
case of spontaneous splenic rupture secondary to coughing. The clinician must be alert to the
possibility of occult splenic rupture because the consequences of delayed diagnosis and
treatment of this condition are grave and potentially fatal. Fortunately for our patient who
recovered very well, splenic rupture was diagnosed promptly and she was taken to surgery for a
successful splenectomy.
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A CASE OF CEFTRIAXONE INDUCED NEUTROPENIA
Avani Prabhakar, MBBS MPH and Hetal K. Brahmbhatt, MD. Department of Internal
Medicine and Department of Psychiatry and Behavioral Sciences, East Tennessee State
University, COM, Johnson City, TN 37614
Neutropenia has been reported in the literature as one of the rare but serious complication of
ceftriaxone therapy. Nine cases to the knowledge of these authors have been reported to date,
two of these cases have been reported secondary to prolonged ceftriaxone therapy, both of
which resulted in secondary infection due to neutropenia. We report reversible neutropenia and
transient elevation of liver transaminases; rare complications from ceftriaxone therapy that
developed in a patient who presented to emergency room with altered mental status and
received 2 grams of intravenous ceftriaxone. With in twelve hours of ceftriaxone administration,
patient developed severe neutropenia with absolute neutrophil count of 100cells/deciliter. The
patient subsequently required hospitalization for observation under neutropenic precautions, the
neutropenia and elevated transaminases resolved after conservative treatment and cessation of
therapy. A case report is prepared of this unusual clinical scenario and the authors recommend
that the clinicians must be vigilant to the possibility of this rare but serious side effect of this
commonly used antibiotic because the consequences of delayed recognition and management
are grave and potentially fatal. Fortunately the patient in this case scenario recovered very well,
ceftriaxone induced neutropenia was identified promptly and she was monitored under
neutropenic precautions, moreover neutropenia was transient and leukocyte count returned to
baseline with in 24 hours. In general neutropenia may develop in 15% of patients receiving high
dose of beta-lactam antimicrobials a group of antibiotics to which ceftriaxone belongs. In
particular neutropenia develops in patients treated for ten days or more, but only 4.2% develop
infection secondary to it and neutropenia completely resolves after few days of drug withdrawal
in most of the cases. There are a couple of mechanism suggested in the literature for betalactam induced neutropenia, the first one is direct toxic effect of beta-lactam antibiotics on
granulocyte progenitor cells and the second one is presence of leucoagglutinins complement
fixing antibodies and antigranulocyte antibodies. The consequences of severe neutropenia
include life threatening bacterial infections, thus it is mandatory to monitor blood counts serially
in patients who receive ceftriaxone in a high daily dose, a total high dose or are treated for a
long duration.
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ENTEROSCOPIC RETRIEVAL: A POSSIBLE TREATMENT MODALITY FOR
DISPLACED ESOPHAGEAL STENT
Raja S. Vadlamudi, MD, MPH and Mark F. Young, MD. Department of Internal Medicine,
East Tennessee State University, COM, Johnson City, TN 37614
Esophageal Stent migration is an important complication (6% to 18%). Stent migrations are
usually treated with conservative measures. We present a case of stent migration where the
stent is removed transorally using enteroscope.
A 48 year old male smoker with intermittent dysphagia to solids and GERD underwent EGD with
biopsy that showed squamous cell carcinoma of the distal esophagus. Exploratory laparotomy
showed metastatic involvement of the celiac lymph nodes. He underwent chemo and radiation
therapies without any complications. Later esophagectomy with splenectomy was done and he
developed an esophageal anastomotic leak. An esophageal deficit of 1.5 cm was identified on
EGD and was reduced using surgical clips. A silicone coated Boston Scientific Polyflex®
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esophageal stent was also placed at the anastomotic site. Few days before the proposed stent
removal, he developed abdominal discomfort. The abdominal x-rays showed possible distal
migration of the stent in to the left upper quadrant. He underwent enteroscopy and the stent,
initially found in the jejunum, was pulled into third part of the duodenum. For lack of facility to
use forceps with enteroscope, a pediatric colonoscope was used to remove the stent transorally
using rat toothed forceps without any complications (Figure 1 & 2). Neither small bowel defects
nor distal esophageal deficits were noted on repeat enteroscopy.
This case suggests the use of enteroscopy with rat tooth forceps for retrieval of migrated
esophageal stents depending on the site of displacement and thus contradicting the use of only
conservative measures for stent migration.
Figures:
Figure 1: Third Part of the Duodenum with
the Esophageal Stent
Figure 2: Stent Attached to Rat Toothed
Forceps (Solid Black Arrow)
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2007 Appalachian Student Research Forum
Post-Baccalaureate Oral Presentations
B. CARROLL REECE OF BUTLER, TENNESSEE: HIS EDUCATION AND
FORMATIVE YEARS
Margaret Carr, MALS. East Tennessee State University, Johnson City, TN 37614
One aspect of this project is to consider the educational history of the Butler community in
Johnson County, Tennessee, and the effects of that experience on one individual – B. Carroll
Reece. Reece became the Congressional Representative from Tennessee’s first district in
1921, a post he held for nearly 40 years. Reece, born in 1889 in Butler, graduated from
Watauga Academy in 1911 and also attended Carson-Newman College and New York
University. His career included service in the U.S. Army during World War I, and he served as a
teacher, principal, and professor in schools such as elementary schools in Carter County and at
New York University. The educational history of the Butler Community in Johnson County,
Tennessee – particularly the Watauga Academy – and the academic career of Carroll Reece
serve as an illustration of the development of education in the region. In much of Northeast
Tennessee, public education was poorly funded and standards of instruction were not well
established. Private academies filled some of this void, as did the Baptist-run Watauga
Academy, which was the last incarnation of two earlier private academies. Information was
collected from the Reece papers housed in the Archives of Appalachia at ETSU, from the Butler
Museum in Butler, and from media from the time period covered (Elizabethton Star and other
periodicals); this information was integrated into data compiled from published materials
regarding the history of education in the region and the Watauga Academy, in particular. The
research sheds new light on the education and career of Carroll Reece and examines the
influences of his community and educational institutions.
--------------------------------------------------------------------------------------------------------------------CHARACTERIZATION OF THE PATHWAY LEADING TO THE SYNTHESIS OF
SALICYLIC ACID IN PLANTS UNDER ATTACK BY PATHOGEN
Alexander Eddo and Dhirendra Kumar. Department of Biological Science, East
Tennessee State University, Johnson City, TN 37614
Salicylic acid (SA) a plant hormone mediates plant defense against pathogens, accumulating in
infected and uninfected distal leaves in response to the attack. Two pathways have been
implicated in SA production and one of them the Phenylalanine ammonia-lyase (PAL) pathway
has now been suggested to play only a minor role in plant defense. The other pathway leading
to SA synthesis is through isochorismate synthase (ICS) (an enzyme localized in plant plastids).
Both PAL and ICS pathway synthesize SA from chorismate - the end product of the Shikimate
pathway. In plants and bacteria this pathway is responsible for biosynthesis of aromatic amino
acids. Plants which either fail to accumulate SA or do not produce SA are unable to fight the
pathogen and are susceptible to pathogen infection (nahG transgenic plant and the SID2,
Arabidopsis mutant plant respectively). However mutant plants which accumulate high levels of
SA are shown to be more resistant and exogenous application of SA induces the production of
pathogenesis related (PR) defense proteins and enhanced resistance. Some of the plants have
Resistance genes encoding for R-proteins. These R proteins can in some cases recognize the
avirulence (avr) proteins introduced by pathogen into the plant host cell leading to resistance
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2007 Appalachian Student Research Forum
response. Absence of corresponding ‘R’ protein against the avr protein allows the pathogen to
manipulate the host defenses as it wants. In case of tobacco resistance protein (N) which
recognizes the avr from tobacco mosaic virus (TMV), the interaction results in ~50 fold increase
in SA levels. Signaling pathway following this interaction leading to SA biosynthesis is not fully
understood. In order to characterize this biochemical pathway we have done RT-PCR based
expression profiling of the known genes of this pathway. This was done to determine the
component which receives the initial signal of the positive interaction of the R and avr protein.
Resistant and susceptible tobacco (model plant) plants were infected with TMV and were used
in this study. Results of this work will indicate which enzymes are involved in the signaling to
make SA when resistance is mounted against infection. Better understanding of the plant’s
natural defense mechanism will help us to sensitize it under normal conditions so that a faster
response can be mounted under actual pathogen attack leading to better resistance.
--------------------------------------------------------------------------------------------------------------------YEAST TWO-HYBRID ANALYSIS OF PROTEIN-PROTEIN INTERACTIONS
INVOLVING A MICROSPORIDIA ADAM (A DISINTEGRIN AND
METALLOPROTEASE) PROTEIN
Carrie E. Jolly and J. Russell Hayman. Department of Microbiology, East Tennessee
State University, COM, Johnson City TN, 37614
Microsporidia are spore-forming, obligate intracellular fungal-like protists typically associated
with opportunistic infections in immunocompromised individuals. Our previous research has
demonstrated a direct relationship between adherence of microsporidia spores to the surface of
host cells and infectivity in vitro. As for many other pathogenic organisms, adherence of
microsporidia to the host cell surface may be a prerequisite to infection. In an effort to better
understand adherence, we have turned our attention to determining what microsporidia proteins
may be involved in this process. Through examination of the Encephalitozoon cuniculi genome
database, we identified a gene encoding a protein with sequence homology to members of the
ADAM (a disintegrin and metalloprotease) family of type I transmembrane glycoproteins. ADAM
proteins are involved in a variety of biological processes including cell adhesion, proteolysis, cell
fusion, and signaling. Through the use of immunoelectron transmission microscopy, we have
determined that the microsporidial ADAM protein or MADAM is localized to at least three distinct
regions of Encephalitozoon intestinalis spores. These areas include the surface exposed
exospore, the plasma membrane, and the polar sac-anchoring disc complex (a bell-shaped
structure at the spore apex involved in the infection process). Immunofluorescence assays
show that MADAM is also located on the extruded polar tube, which is the organelle used to
infect the host cell. Since ADAM proteins are known to be involved in cell adhesion and
proteolysis, we employed a yeast two-hybrid system to identify potential proteins capable of
interacting with MADAM. Of the cDNA clones analyzed thus far, several contain an insert that
encodes for polar tube protein 3 (PTP3). Additional yeast two-hybrid analyses demonstrate that
the interaction between MADAM and PTP3 passes the most stringent selection process,
whereas, other polar tube proteins (PTP1 and PTP2) do not. Current studies are underway to
confirm that PTP3 interacts with MADAM by co-immunoprecipitation assays. Further
characterization of the interactions that occur between MADAM and PTP3 may lead to a better
understanding of the events that occur during microsporidia host cell infection.
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THE HOST ADHERENS JUNCTION MOLECULE NECTIN-1 IS DOWN-REGULATED
IN CHLAMYDIA TRACHOMATIS-INFECTED GENITAL EPITHELIAL CELLS
Jingru Sun, Jennifer Kintner and Robert V. Schoborg. Department of Microbiology, East
Tennessee State University, COM, Johnson City, TN 37614
Although Chlamydia trachomatis (serovars D-K) is the most commonly reported bacterial
sexually transmitted disease agent in the US, it is still not clear how chlamydial infection causes
disease. Nectin-1, a member of the immunoglobulin superfamily, is a Ca 2+-independent
homophilic and heterophilic cell adhesion protein that has been implicated in the organization of
E-cadherin-based adherens junctions (AJs) and claudin-based tight junctions (TJs) in epithelial
cells. Nectin-1 also plays key roles in regulating intracellular signaling molecules. For example,
activation of Cdc42 and Rac through c-Src by nectin-1 regulates cell-cell adhesion, gene
expression, and cell polarization. Using Western blot analyses, our laboratory has
demonstrated that accumulation of host cellular nectin-1 is significantly decreased in C.
trachomatis serovar E infected non-polarized HeLa cells. Nectin-1 was decreased by as much
as 85% by 48 hours post infection (hpi); this decrease was sustained until 72 hpi. Similar
results were also obtained using polarized HeLa cell cultures. Indirect immunofluorescence
assay further supported the fact that accumulation of nectin-1 is down-regulated by C.
trachomatis infection. Down-regulation of nectin-1 in C. trachomatis-infected cells is prevented
by chloramphenicol exposure, demonstrating that C. trachomatis protein synthesis and/or
replication is required for this effect. To determine whether persistent chlamydial infection also
reduces nectin-1 accumulation, C. trachomatis infected HeLa cells were exposed to penicillin G
and harvested at 48 hpi. Nectin-1 levels decreased in the presence of penicillin G, indicating
that nectin-1 accumulation can indeed altered by persistent chlamydiae. Interestingly, Ncadherin-dependent cell-cell junctions can be disrupted by C. trachomatis infection, as reported
by Ramsey KH et al. 2002. Taken together, these data suggest that C. trachomatis may disrupt
adherens junctions, at least in part, by diminishing nectin-1 expression. Because other
investigators have observed release of chlamydia-infected epithelial cells in vitro (from
monolayers) and in vivo (from tissues), we speculate that chlamydia modulated down-regulation
of adhesion molecule and subsequent disruption of host cell adherence may be involved in
chlamydial dissemination or pathogenesis.
--------------------------------------------------------------------------------------------------------------------TRANSCRIPTIONAL REGULATION OF C-REACTIVE PROTEIN EXPRESSION IN
HUMAN HEPATOCYTES
Bhavya Voleti, Prem Prakash Singh and Alok Agrawal. Department of Pharmacology,
East Tennessee State University, COM, Johnson City, TN 37614
The hepatic synthesis of C-reactive protein (CRP) increases in inflammatory conditions resulting
in raised serum CRP levels. The regulation of CRP gene expression in hepatocytes occurs at
the transcriptional level. Previously, we reported that a C/EBP-binding site overlapping a NF-ĸB
p50-binding site (C/EBP-p50-site) on the CRP promoter was critical for IL-6-induced CRP
expression in human hepatoma Hep3B cells. Transcription factors C/EBPβ and p50 occupy the
C/EBP-p50-site in the nuclei containing constitutively active or IL-6-activated C/EBPβ. The aim
of this study was to identify the transcription factors that occupy the C/EBP-p50-site in the
absence of C/EBPβ. Accordingly, we treated Hep3B nuclear extract with a C/EBP-binding
consensus oligonucleotide to generate an extract lacking active C/EBPβ. Such treated nuclei
contain only C/EBPς because the C/EBP-binding consensus oligonucleotide binds to all C/EBP
family proteins except C/EBPς. EMSA using this extract revealed formation of a C/EBPς-
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2007 Appalachian Student Research Forum
containing complex at the C/EBP-p50-site. This complex also contained RBP-Jĸ, a transcription
factor known to interact with the ĸB sites. RBP-Jĸ
formation of C/EBPςcontaining complexes. The RBP-Jĸς-containing complexes were also
formed at the C/EBP-p50-site on the CRP promoter in the nuclei of primary human hepatocytes.
In transactivation assays, overexpressed C/EBPς abolished both C/EBPς-induced and (IL-6+IL1β)-induced CRP promoter-driven luciferase expression. These results indicate that under
basal conditions, C/EBPς occupies the C/EBP-site, an action that requires RBP-Jĸ. Under
induced conditions, C/EBPς
β and p50. We conclude that the switch
between C/EBPβ and C/EBPς participates in regulating CRP expression. This process utilizes
a novel phenomenon, that is, the incorporation of RBP-Jĸ into C/EBPς-complexes solely to
support the binding of C/EBPς to the C/EBP-site.
--------------------------------------------------------------------------------------------------------------------INVOLVEMENT OF TNF RECEPTOR APOPTOSIS INDUCING LIGAND (TRAIL) IN
NK CELL MEDIATED ANTI-HSV ADAPTIVE IMMUNE RESPONSE
Stacie N. Woolard, Subhadra Nandakumar, Yagita H and Uday Kumaraguru. Department
of Microbiology, East Tennessee State University, COM, Johnson City, TN-37614 and
Department of Immunology, Juntendo University School of Medicine, Tokyo, Japan
Linking the innate and acquired immune system has been a major topic of research leading to
the examination of certain cells that could facilitate this interaction. Two such cells are the
Dendritic cell (DC), the professional APC, and the Natural Killer (NK) cell, which are recruited
and activated in a viral response. The cross-talk of DCs and NKs has been thoroughly
examined yet the precise role of NK cells in this process has yet to be demonstrated. The DCs
from infected site migrate to the draining lymphoid organ, but transfer the cargo to the lymphoid
resident CD8α+ DC through an undefined process. The CD8α+ DC then process and crosspresent the viral epitope to the T cells. We hypothesized that the NK cells may play a
predominant role in this process. Using PK136 Tg mice (NK cell less mouse) and wild type
C57BL/6 mice we show that the adaptive immune response to HSV was impaired. To further
test if NK cell contributed to this impairment, we looked at NK cell interaction with DC. Our
results suggest the interaction to be reciprocal in that the antigen bearing migratory immature
DC stimulated the NK cells (reduced MHC-I and IL-12). The activated NK cells killed the
immature DC (iDC) resulting in an inflammatory milieu. This in turn stimulates the lymphoid
resident CD8α+ DC to acquire the apoptotic DCs, process and cross-present the viral epitope to
T cells. In addition, the NK cell produced IL-2 and IL-15 act as T cell growth factor. Our studies
on the mechanism suggested that the NK cell used the TNF receptor apoptosis inducing ligand
(TRAIL) to deliver the death signal to the iDC. This was further confirmed in vivo by blocking
the pathway using anti-TRAIL Mab. Hence, we may conclude that HSV infection prevents
maturation of DCs, but upon arrival at the lymphoid organ stimulate the NK cells, they
reciprocate by killing them through TRAIL. The resulting inflammatory milieu then aids the
lymphoid resident DC to take up the antigen bearing dead DC, process and cross-presents the
viral epitope resulting in optimized immune activation.
Acknowledgements: Department of Microbiology, ETSU - Start up Funds and Dr. Wyrick.
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2007 Appalachian Student Research Forum
Resident/Post-Doc
Oral Presentations
WEGENER’S GRANULOMATOSIS AND PURULENT PERICARDITIS: THE FIRST
CASE REPORT
Admad Halawa, MD, Bobby Keith, MD and Ryland P. Byrd, Jr. MD. Department of Internal
Medicine, East Tennessee State University, COM, Johnson City, TN 37614
Wegener's granulomatosis is a systemic vasculitis presents with variable multi-organ
involvement. Different cardiac manifestations, like myocarditis or valvulitis, have been
described before as part of wegener's pathogenesis, but the most common cardiac features
were pericarditis and coronary arteritis. Fibrinous and constrictive pericarditis are well identified
in Wegener's patients who suffered cardiac impairment. To the best of our knowledge, we
present the first case report of purulent pericarditis during an acute Wegener's exacerbation.
Our patient is a young female presented with a typical vasculitis picture including severe
pulmonary nodular infiltrates, arthritis, and neuritis. She also developed a massive pericardial
effusion. Pus was evacuated from the pericardium during pericardectomy and it grew no
organisms. Immunosuppressive therapy dramatically improved the systemic inflammation and
resolved the pulmonary and cardiac defect.
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OLDER GALAXY PAIR HAS SURPRISINGLY YOUTHFUL GLOW
Mark Hancock et al. Department of Physics, Astronomy, and Geology, East Tennessee
State University, Johnson City, TN 37614
A pair of interacting galaxies might be experiencing the galactic equivalent of a mid-life crisis.
For some reason, the pair, called Arp 82, didn't make their stars early on as is typical of most
galaxies. Instead, they got a second wind later in life (about 2 billion years ago) and started
pumping out waves of new stars as if they were young again. Our new observations are from
NASA's Galaxy Evolution Explorer, NASA's Spitzer Space Telescope and the Southeastern
Association for Research in Astronomy Observatory at Kitt Peak, Arizona. Arp 82 is an
interacting pair of galaxies with a strong bridge and a long tail. NGC 2535 is the big galaxy and
NGC 2536 is its smaller companion. The disk of the main galaxy looks like an eye, with a bright
‘pupil’ in the center and oval-shaped ‘eyelids.’ Dramatic ‘beads on a strin’ features are visible as
chains of evenly spaced star-formation complexes along the eyelids. These are presumably the
result of large-scale gaseous shocks from a grazing encounter. The colors of this galaxy
indicate that the observed stars are young to intermediate in age, around 2 million to 2 billion
years old, much less than the age of the universe (13.7 billion years). The pair first burst with
new star formation about 2 billion years ago after swinging by each other. A second close
passage more recently resulted in yet another batch of star formation. The puzzle is: why didn't
Arp 82 form many stars earlier, like most galaxies of that mass range? Scientifically, it is an
oddball and provides a relatively nearby lab for studying the age of intermediate-mass galaxies.
In more popular terms, think of this as an example of arrested development. For some reason,
it took a kick-in-the-pants to get the stars forming recently, whereas most other galaxies of that
mass range formed their stars much earlier (between 4 and 8 billion years ago).
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EFFECTS OF PATHOGEN ASSOCIATED MOLECULAR PATTERNS (PAMPS) ON
ANTI-VIRAL IMMUNE MEMORY
Subhadra Nandakumar and Uday Kumaraguru. Department of Microbiology, East
Tennessee State University, COM, Johnson City, TN 37614
Our immune memory needs to be refreshed from time to time. Immune system of individuals
living in a sanitized society will eventually atrophy and go into immune-amnesia. We know that
some vaccines wear off after a few decades, but upon re-exposure the response is quicker and
better and there is an upward shift in memory frequency. This new set-point is likely higher than
the primary set-point and varies with individuals based on the exposure pattern to other
microbes or its components. Our investigations were designed to test such differences of nonspecific stimulation on anti-viral adaptive immune response by bacteria or its components in
lowering the threshold of activation. We exposed young mice to various TLR ligands in the first
month after birth and analyzed for their ability to resist and survive challenge with virus at the
adult phase. Secondly, pre-immune animals were exposed to TLR ligands in the absence of
cognate antigen to determine the impact on immune memory response. The mice that were in
the simulated dirty environment were better able to resist lethal challenge and needed 10 times
more virus to show symptoms of CNS infection with HSV. This was attributed to the upregulation of certain key co-stimulatory molecules (such as 41BB, OX40, CD40) on antigen
presenting cells (APC), modulation of cytokines, cytokine receptors and anti-apoptotic
molecules on APCs and T cells that are known to be critically involved in memory expansion
and maintenance. In addition, pre-immune mice given TLR ligands in the absence of cognate
antigen showed increase in frequency and quality. The frequency was measured in vitro by
HSV specific tetramer and in vivo by BrdU incorporation and CFSE dilution. The quality was
assessed by measuring intracellular expression of cytokines (IFNγ and TNFα) and release of
perforin and granzymes by assessing lysosome-associated membrane proteins (CD107a and
b). The timing, dose and combination of TLR ligands exposure seem to have an impact on the
positive outcome. These studies provide insights into the role of hyper-sanitized environment
versus PAMP mediated signaling in adaptive immune response and long term memory.
Acknowledgements: ETSU-RDC major grant support, Start up Funds, Department of
Microbiology and Dr. Wyrick.
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Poster – Non-Competitive
Biomedical Sciences
CHARACTERIZATION OF BINDING AND TRANSPORT AND INITIAL
IDENTIFICATION OF THE GENES INVOLVED IN THE BIOSYNTHESIS AND
TRANSPORT OF SCHIZOKINEN, A DIHYDROXAMATE SIDEROPHORE
PRODUCED BY RHIZOBIUM LEGUMINOSARUM IARI 917
D. Hammond, E. Storey, B.C. Lampson and R.N. Chakraborty. Department of Health
Sciences, East Tennessee State University, Johnson City, TN 37614
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 a multicomponent transport system. With a very few exceptions iron is a vital
element for the survival of most living cells. This is because iron serves as a cofactor for the
components of 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 microorganism as it forms an insoluble ferric oxyhydroxide polymer.
Siderophore mediated iron transport is one of the ways in which microorganisms acquire iron to
overcome the shortage of iron. 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. We have already characterized the siderophore produced by
Rhizobium leguminosarum IARI 917 to be ‘schizokinen’ and have also tried to identify the genes
involved in its biosynthesis. Random mutations of strain IARI 917 were done using mini Tn5
mutagenesis. Siderophore overproducers and non-producers were detected on CAS (chrom
azurol S) plates. DNA sequence analysis of several non-producer mutants carrying the
transposon revealed a gene similar to 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). 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.
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Natural Sciences & Mathematics
CUTICLE MICROMORPHOLOGY OF TSUGA
Graham Cooke and Yusheng Liu. Department of Biology, East Tennessee State
University, Johnson City, TN 37614
Cuticle micromorphology from leaves of all 8 species of the genus Tsuga, and one cultivated
variant are being studied by both light microscopy and scanning electron microscopy for the first
time. Specimens collected from the ETSU Arboretum exhibit characteristic differences between
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2007 Appalachian Student Research Forum
species at a micromorphological level. Features of the axial and abaxial cuticle are in the
process of being described for all 8 species and one species cultivar. When completed we
expect to use the description of 28 separate characters, 7 from the outer cuticle and 21 from the
inner cuticle. These characters should provide additional characterization of the species within
the genus, and will provide additional detail to aid in the reconstruction of the phylogeny of
Tsuga. This will also provide a basis for the identification of fossilized cuticle. In addition, the
stomatal density will also be quantified in an attempt to reconstruct characteristics
paleoclimates, and will be applied in modeling of future earth climates.
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ROLE OF SALICYLIC ACID METHYL TRANSFERASE IN PLANT DISEASE
RESISTANCE SIGNALING
Tazley Hotz, Poonam Sheth and Dhirendra Kumar. Department of Biological Sciences,
East Tennessee State University, Johnson City, TN 37614
Salicylic acid (SA) has been shown to play an important role in plant defense response against
microbial pathogens. SA is synthesized in the plant chloroplasts from chorismate, the end
product of the shikimate pathway, through two biochemical pathways: Isochorismate synthase
(ICS) and phenylpropanoid (PAL). 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
bacterial and fungal 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 ever-evolving pathogens.
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Social and Behavioral Sciences
VOCAL FUNCTION EXERCISES (VFE) VERSUS RESONANT VOICE THERAPY
(RVT) FOR THE TREATMENT FOR HYPERFUNCTIONAL VOICE DISORDERS
Shelley Bell, Brittany Kidd, Cora Leemkuil, Annie Smith and Christopher McCrea.
Department of Communicative Disorders, East Tennessee State University, Johnson
City, TN 37614
This case study used patient-based treatment outcome measures combined with acoustic
analysis to compare vocal function exercises (VFE) and resonant voice therapy (RVT) for the
treatment of an individual with mild-moderate hyperfunctional voice disorder. The participant
was a 44 year old, first language general American English speaker, with no history of previous
speech-language impairment. The participant underwent six weeks of RVT, followed by a sixweek withdrawal, and six weeks of VFE. Auditory-perceptual and laryngeal imaging data were
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2007 Appalachian Student Research Forum
collected before and after each six-week treatment phase. In addition, acoustic data were
collected during each treatment session. Laryngeal results indicated that the participant
displayed some noticeable changes in vocal fold function across both treatment protocols.
Acoustic results indicated that the participant’s loudness was consistent during pre-treatment,
mid-treatment, and post-treatment measures and that both treatments were effective in
improving the cycle-to-cycle vibration patterns of the participant’s vocal folds. No noticeable
differences between the two treatments were noted; however, according to patient ratings, it
seemed that the RVT was preferred over VHI. Although several questions remain unanswered,
the results of this case study represent evidence to support the clinical utility of RVT and VFE
within hyperfunctional voice disordered patients.
--------------------------------------------------------------------------------------------------------------------WEBSITE USABILITY
Kevin Bowes. Department of Computer and Information Science, East Tennessee State
University, Johnson City, TN 37614
The usability of a website has many benefits, greater respect for the company and designer, a
wider user base, a greater range of supported user devices (Browsers, PDAs, Cell phones), and
easy access to information. The information available on East Tennessee State University’s
College of Business and Technology’s Career Services’ Job listings website was difficult to
access and maintaining the site was just as difficult. Building a new website, focusing on the
users, to encompass the job listings and other facets of the Career Services department (such
as statistics gathering) was the best method of solving this problem. So, a new website was
built to facilitate these needs. This website is a good starting point toward great user
experience for the Career Services Department and these methods can be used for any data
driven website.
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TEMPORALIZATION OF AN EXISTING ACCREDITATION DATABASE
Matthew Campbell. Department of Computer and Information Sciences, East Tennessee
State University, Johnson City, TN 37614
Storage of accreditation data in a database is an easy and quick solution provided the criteria
for accreditation are unchanging. An accreditation database should be able to store data about
current accreditation criteria and data, as well as previous versions of criteria and data. In order
to do this, the database must be temporalized. Research on Temporal Relational Databases
has progressed greatly since Gadia and Yeung’s generalized model was introduced in the
1980’s. Temporal databases allow for previous versions of data to be placed in the same table
as the current data and to be accessed very easily. For the accreditation database, a valid time
approach was used for tables requiring temporalization. Valid time is the time at which the data
was the current version. All tables in the database were analyzed to see if temporalization was
necessary, and the tables that store the goals for the university, college, and department, as
well as the table that stores the requirements for accreditation by the accreditation organization,
ABET, were the only ones that required temporalization.
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