ABSTRACTS

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ARTICLES PAGE

Uptake of Nanoparticles of Cerium Oxide and Yttrium Oxide by
Acanthamoeba castellanii (Protozoa) and Daphnia magna
(Crustacea). James R. Palmieri, Geneva Gehring, Catherine
Minichino, and Shaadi F. Elswaifi. 3

ABSTRACTS OF PAPERS, 89th Annual Meeting of the
Virginia Academy of Science, May 25-27, 2011,

University of Richmond, Richmond VA 15

SECTION ABSTRACTS

Aeronautical and Aerospace Sciences 15

Agriculture, Forestry and Aquaculture Science 16

Astronomy, Mathematics and Physics &Materials Science 20

Biology with Microbiology & Molecular Biology 24

Biomedical and General Engineering 39

Botany 41

Chemistry 46

Computer Science 51

Education 53

Environmental Science 56

Medical Science 57

Natural Histoiy & Biodiversity 71

Psychology 81

Statistics 86

Structural Biology, Biochemistry and Biophysics 86

JUNIOR ACADEMY AWARDS 91

Virginia Journal of Science
Volume 62, 1 & 2
Spring 2011

Uptake of Nanoparticles of Cerium Oxide and Yttrium
Oxide by Acanthamoeba castellanii (Protozoa) and

Daphnia magna (Crustacea)

James R. Palmieri 1 , Geneva Gehring, Catherine Minichino, and

Shaadi F. Elswaifi

Department of Microbiology, Infectious, and Emerging Diseases,
Edward Via College of Osteopathic Medicine, 2265 Kraft Drive,

Blacksburg, Virginia, USA 24060

ABSTRACT

Currently, nanoparticles are synthesized and used at an unprecedented rate for
industrial, medical, and research applications. The use of cerium oxide
nanoparticles (CeONP) and yttrium oxide nanoparticles (YtONP) results in
their spread as contaminants into the environment. Once in the environment,
CeONP and YtONP can be taken up by organisms in the food chain where
they may pose a public health risk. In this study we determine whether
Acanthamoeba castellanii and Daphnia magna uptake CeONP or YtONP
from their environment and thereby play a role in the transmission of the
nanoparticles. Using electron microscopy, organisms exposed to the
nanoparticles were examined. Our results indicate that the nanoparticles are
associated with cell and organelle membranes. These findings have
implications for the health risks associated with environmental contamination
by CeONP and YtONP.

INTRODUCTION

In this study we determine whether protists and crustaceans play a role in the
transfer of cerium oxide nanoparticles (CeONP) and yttrium oxide nanoparticles
(YtONP) from the environment to other organisms within the aquatic food chain.
Acanthamoeba castellanii, a common protist, and Daphnia magna , a planktonic
crustacean, are important components in many aquatic ecosystems. Because
acanthamoebae, such as A. casteUanni , are aggressive feeders they consume inorganic
and organic compounds from their environment, thereby serving as a link by
transferring normally unavailable inorganic components to the food chain (Weekers et
al. 1993). D. magna feeds on acanthamoebae and other protists found in lower trophic
levels. Because of the high reproductive potential of D. magna, these planktonic
crustaceans can substantially alter the structure and functioning of microbial food webs
in freshwater ecosystems , such as acidic swamps, freshwater lakes, ponds, rivers and
streams (Guisande 1993). D. magna and other aquatic crustaceans have the ability to
filter particles of a variety of sizes including nanoparticles (Rosenkranz et al. 2009,
Kim et al. 2010, Zhu et al. 2010). The authors hypothesized that CeONP or YtONP in

1 Corresponding author: jpalmieri@vcom.vt.edu

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VIRGINIA JOURNAL OF SCIENCE

the natural environment could be taken up by these organisms. Particles engineered at
dimensions between 1-100 nm, are referred to as nanoparticles. Currently, nanoparticles
are sy nthesized and used for industrial, medical, and research applications. CeONP are
used as diesel fuel additives, in automotive catalytic converters, and are a by-product
of many industrial processes, including the polishing of glass and semi-precious stones.
CeONP also have potential uses for medical applications acting as an antioxidant
(Elswaifi et al. 2009). Yttrium oxide and cerium oxide belong to the rare earth
elements. Yttrium oxide nanoparticles (YtONP) are used in the manufacturing of
cathode ray tubes for computer monitors and televisions and have potential medical
applications due to their ability to act as antioxidants (Schubert et al. 2006, Cotton
2006, Okuyama et al. 2007, and Gilmore et al. 2008). The use of CeONP and YtONP
results in their release into the environment (Biswas and Wu 2005, Chow et al. 2005)
where they may exist in concentrations and forms that are toxic. This release may result
from the process of their synthesis, as a by-product of their use, or from their
indiscriminate disposal after use. Presently, CeONP and YtONP are released into the
environment from diesel engine emissions, from improper disposal of automotive
catalytic converters, and from improper disposal of old TV and computer monitors.
These nanoparticles may then make their way into air, soil, or ground water (Biswas
and Wu 2005, Chow et al. 2005). As with many of the engineered nanoparticles,
CeNOP and YtONP may also have toxic effects on humans and animals (Gatti and
Montanari 2008). The toxicity of CeONP and YtONP has been recently investigated
in vivo and in vitro (Gojova et al. 2007, Gatti and Montanari 2008, Andelman et al.
2009, Hardas et al. 2010). Toxic effects include reduced cell viability, increased
cellular oxidative damage, and apoptosis. Effects also include vascular inflammation
that may lead to pulmonary thromboemblism resulting in stroke or myocardial
infarction. Chronic inflammation may also lead to rare earth pneumoconiosis or lung
cancer (Gojova et al. 2007, Gatti and Montanari 2008, Andelman et al. 2009, Hardas
et al. 2010, Lin et al. 2006). However, the routes of exposure of humans to these
nanoparticles are poorly understood. Examples of routes of exposure of CeONP and
YtONP may be through the contamination of organisms in the food-chain or through
contamination of drinking water (Holbrook et al. 2008). In a typical food chain, there
is usually a maximum of four or five trophic levels, although food chains in aquatic
ecosystems frequently contain more levels than those in terrestrial ecosystems (Pimm
and Lawton 1977). As organisms in lower trophic levels are consumed by those in a
higher level, nanoparticles may become concentrated in top level consumers, level
consumers, namely fish and animals that consume fish, including humans. Many
protists, including acanthamoebae, are voracious feeders of organic and sometimes
inorganic materials as they occupy the bottom of aquatic and some terrestrial
ecosystems (Khan 2009).

Acanthamoeba spp. are free-living amoebae that are ubiquitous in aquatic and
terrestrial ecosystems. Acanthamoebae exist as trophozoites, the feeding stage, and as
cysts. Presently, 23 species of Acanthamoeba are reported and their biology and
pathogenicity reviewed by Marciano-Cabral and Cabral (2003) and Khan (2009). At
least three species of Acanthamoeba have been reported as parasites of animals, and
humans (Marciano-Cabral and Cabral 2003 and Khan 2009). It is unknown whether
CeONP or YtONP particles are taken up and concentrated by protists or by crustaceans
in contaminated aquatic ecosystems. In this report we investigate whether CeONP and

UPTAKE OF NANOPARTICLES

5

FIGURE 1. Scanning electron micrograph showing two trophozoites feeding stages of
Acanthamoeba castellanii.

YtONP maybe incorporated into the protest Acanthamoeba castellanii (Fig. 1) and into
the arthropod Daphnia magna, two integral components representing organisms at two
different trophic levels of the aquatic food chain.

MATERIALS AND METHODS

Exposure of Organisms to Nanoparticles: CeONP and YtONP stock solutions were
prepared by suspension of nanoparticles in distilled water. Samples of the nanoparticles
suspended in distilled water were placed in a vortex apparatus for 5 minutes before and
after preparation of the solution to minimize formation of nanoparticle aggregates.
D. magna cultures were exposed to nanoparticles by the addition of the respective
solution to make a final concentration of 10 pM of CeONP or YtONP. D. magna
viability was determined by observing motility and gill movement. Distilled water
without nanoparticles was used in control samples. CeONP or YtONP were added to
cultures of A. castellanii containing >95% trophozoites, making a final nanoparticle
concentration of 100 nM. Equal amounts of distilled water were used for exposure of
control samples. Cultures were incubated for 24 hours at 25C, washed two times in
Page’s saline solution (Petry et al. 2006), then processed for electron microscopy using
standard techniques. Control groups consisted of organisms treated with only distilled
water. To define the appearance CeONP and YtONP alone, suspensions were used to
prepare samples for observation by transmission and scanning electron microscopy.

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VIRGINIA JOURNAL OF SCIENCE

FIGURE 2. Outer membrane of Acanthamoeba castellanii demonstrating
accumulations of yttrium oxide nanoparticles attached to outer surface. Arrows
indicating yttrium oxide nanoparticles (Bar = 500nm).

The appearance of CeONP and YtONP in cells was compared to those reported in other
studies. Each experiment was performed twice including control groups. There were
sixteen experimental group cultures; eight cultures of A. castellanii and eight cultures
of D. magna. There were four control groups run for each species; two cultures each,
exposed to CeONP for D. magna and A. castellanii and 2 cultures each, exposed to
YtONP for D. magna and A. castellanii.

Electron Microscopy: After exposure of A. castellanii and D. magna to CeONP or
YtONP, specimens were prepared for electron microscopic examination. A. castellanii
was washed two times using Page’s saline solution, centrifuged and fixed in 0.5%
buffered glutaraldehyde at 4C for 72 hours. D. magna was pre-fixed in FAA fixative
solution for 5 minutes at 4C and post-fixed in 0.5% buffered glutaraldehyde at 4C for

UPTAKE OF NANOPARTICLES

7

FIGURE 3. Golgi apparatus of Acanthamoeba castellanii demonstrating cerium oxide
nanoparticles associated with Golgi vesicles. Arrows indicating cerium oxide
nanoparticles (Bar = 500nm).

72 hours. A. castellanii and D. magna were then processed for ultra-thin sectioning and
examination using transmission electron microscopy according to standard methods
used by the Virginia-Maryland Regional College of Veterinary Medicine Morphology
Services Laboratory. Electron micrographs of thin sections of A. castellanii and D.
magna were examined for densely stained nanoparticles of CeONP and YtONP to
determine if nanoparticle uptake had taken place. For scanning electron microscopy,
samples of A. castellanii were isolated and processed according to standard methods
used by the Virginia-Maryland Regional College of Veterinary Medicine Morphology
Services Laboratory.

RESULTS

Our results demonstrate that CeONP and YtONP can be readily taken up by A.
castellanii and D. magna. We have also demonstrated that after uptake, CeONP and
YtONP were associated with cell and organelle membranes in these organisms.
Acanthamoeba castellanii : CeONP and YtONP were observed in association with cell
and organelle membranes of A. castellanii. These nanoparticles were observed on the
outer surface of the cell membrane (Fig. 2) and on the membranes of intracellular
organelles, including Golgi apparatus (Fig. 3) and various vesicles (Fig. 4). Vesicles

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VIRGINIA JOURNAL OF SCIENCE

FIGURE 4. Acanthamoeba castellanii demonstrating yttrium oxide nanoparticles
within organism outer membrane and organelle membranes. Arrows indicating yttrium
oxide nanoparticles (Bar = 500nm).

containing nanoparticles were observed lining the inner surface of the cell membrane
(Fig. 4). In addition, A. castellanii treated with YtONP revealed that the nanoparticles
blanketed the outer cell surface and sometimes appeared as aggregates, some crystalline
and some with ill-defined edges throughout the organism.

Daplmia magna : CeONP and YtONP were observed throughout the tissues of D,
magna. These nanoparticles were observed in multiple organs including reproductive
and digestive, and were also observed in gill filaments. Observation of both cross
sections and longitudinal sections of gill filaments revealed a close association with

UPTAKE OF NANOPARTICLES

9

FIGURE 5. Cross sections of gill filaments (GF) of Daphnia magna showing yttrium
oxide nanoparticles on gill filament borders. Arrows showing yttrium oxide
nanoparticles on gill filament boarders (Bar = lOOnm).

YtONP (Fig. 5 and Fig. 6). YtONP were also observed in the space underlying the gill
filaments (Fig. 6). CeONP were observed at the site of muscular attachment to the
cuticle. At the sub-cellular level, CeONP and YtONP were observed in mitochondria,
nuclei, muscle fibers, vesicles, and within the multilamellar bodies (Fig. 7). CeONP

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VIRGINIA JOURNAL OF SCIENCE

FIGURE 6. Longitudinal section of gill filaments of Daphnia magna demonstrating
yttrium oxide nanoparticles along gill filament borders. Arrows showing yttrium oxide
nanoparticles on gill filament boarders (Bar 500 nm).

and YtONP were observed on the surface of various organelle membranes, on
mitochondrial cisternal membranes, and on the membrane surfaces and internal
contents of vesicles.

DISCUSSION

The observations of nanoparticles in both A castellanii and D. magna indicate that
nanoparticles are associated with cells and their membranes. The association of CeONP
and YtONP with cellular membranes of A castellanii and D. magna , may indicate the
first step in the diffusion of the nanoparticles into the cell. The association of the
nanoparticles with vesicles found along the cell surface (Fig. 4) may indicate that the
nanoparticles are actively transported into the cells by endocytosis or possibly delivered
through membrane channels to vesicles for further breakdown. Alternatively, the
nanoparticles may diffuse passively through the membranes. The presence of proteins

UPTAKE OF NANOPARTICLES

11

' . 4 *,

4 f ■ -; -

J

v

FIGURE 7. Multilamellar body and associated structures of Daphnia magna showing
cerium oxide nanoparticles throughout. Arrows showing cerium oxide nanoparticles
(Bar = lOOnm).

on the cells surface of A. castellanii may play a role in enhancing the uptake of
nanoparticles by coating the nanoparticles and enhancing phagocytosis (Andelman et
al. 2009, Lynch et al. 2009). Both CeNOP and YtNOP were associated with cell and
organelle membranes. However, the amount and distribution of YtONP in cells
appeared to be greater than that for CeONP.

Acanthamoebae and planktonic crustaceans are important components in aquatic
food chains (Jurgens 1994, Zollner et al. 2003). Acanthamoebae interact directly with
bacterial and fungal decomposers, and with autotrophic and heterotrophic
microplankton. Our observation of YtONP in the gill filaments of D. magna (Fig. 5.
and Fig. 6) supports the ability of these crustaceans to filter particles in the nanoscale
range. The presence of D. magna may influence the food chain structure because they

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consume microplankton from lower trophic levels. In turn they are consumed by fish,
making D. magna a key factor in the propagation of trophic cascades from bacteria to
fish (Jurgens 1994). It is possible in the natural environment that nanoparticles could
be transferred to Daphnia following the ingestion of acanthamoebae containing
nanoparticles; fish could then feed on these Daphnia. Fish containing nanoparticles
may transfer them directly to humans who consume the fish, or indirectly through the
feeding of forge-fish to livestock, which are then consumed by humans.

The ability of A castellanii and D. magna to uptake CeONP and YtONP from their
environment has implications on the organisms’ ability to deliver nanoparticles to
higher trophic levels, including human who consume fish species. The potential for
transfer of CeONP and YtONP to humans through the food chain poses a significant
public health risk (Biswas and Wu 2005, Chow et al. 2005, Gojova et al. 2007, Gatti
andMontanari 2008, Gojova et al. 2009) that requires further investigation. The ability
of A castellanii to uptake CeONP and YtONP also has implications for transmission
of microbial diseases. Acanthamoebae are ecologically important as natural grazers that
feed on organic and inorganic matter in the soil and water, thereby contributing to the
natural recycling of nutrients and minerals. Acanthamoebae also consume bacteria,
fungi, and other protists. Therefore, acanthamoebae are important for ecological
balance and help regulate microbial populations in nature (Sinclair et al. 1981, Foster
and Dormaar 1991, Kreuzer et al. 2006, Khan 2009). The ability of CeONP and YtONP
to act as cellular antioxidants, to protect cells, and to elongate their lifespan
(Rzigalinski 2005, Schubert et al. 2006, Rzigalinski et al. 2006, Elswaifi et al. 2009)
means that these effects may apply to bacteria engulfed by the acanthamoebae.
Therefore, environmental contamination by these nanoparticles may lead to the
development of bacteria that resist killing in the phagosome and become “superbugs”
(Elswaifi et al. 2009) thereby posing a further risk to the environment and to human
health. On the other hand, bacteria may be directly exposed to the nanoparticles in the
environment, then, if consumed by humans, those bacteria may become more resistant
to killing by the immune system or by antibiotics (Elswaifi et al. 2009).

CONCLUSIONS

In conclusion, our results demonstrate that CeONP and YtONP can be readily taken
up by a protozoan (A castellanii) and a planktonic crustacean (D. magna), two integral
components in an aquatic food chain. We have also demonstrated that after uptake,
CeONP and YtONP are associated with cell and organelle membranes. These finding
have implications for the health risks associated with environmental contamination by
CeONP and YtONP.

AUTHORS' CONTRIBUTIONS

JRP conceived of the study, helped design the study, participated in growing the
organisms and exposing them to nanoparticles, participated in interpreting the results,
and drafted the manuscript. SFE conceived of the study, helped design the study,
participated in interpreting the results, and assisted in drafting the manuscript. CM
prepared Acanthamoeba for electron microscopy and collected and photographed
specimens. GG prepared Daphnia for electron microscopy and collected and
photographed specimens. All authors read and approved the final manuscript.

UPTAKE OF NANOPARTICLES

13

ACKNOWLEDGMENTS

The authors thank Kathy Lowe, Morphology Lab Supervisor, at the Virginia

Maryland College of Veterinary Medicine for preparation and examination of samples

for electron microscopy. The authors acknowledge the Edward Via College of

Osteopathic Medicine for providing funding and laboratory facilities for this project.

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ABSTRACTS

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ABSTRACTS OF PAPERS, 89th Annual Meeting of the
Virginia Academy of Science, May 25-27, 2011,
University of Richmond, Richmond VA

Aeronautical and Aerospace Sciences

U S. NAVY AVIATION EVENTS RELATED TO THE HAMPTON ROADS,VA
AREA . M. Lerov Spearman, Langley Research Center, Hampton, VA & Robert
W. Heath, Canon of Virginia, Newport News, VA. The Navy acknowledged a
role for aviation in September 1910 when Capt. Washington I. Chambers was
designated as the officer in charge of aviation matters. Soon Glenn Curtiss, a U S.
pioneer in aviation, began meeting with Chambers in an effort to convince the Navy
of the value of aircraft for Naval use. In November 1910 Eugene Ely took off in a
Curtiss airplane from a wooden platform built over the bow of a ship in the harbor at
Hampton Roads, VA. In 1921 the Army Air Service at Langley Field, VA conducted
bombing tests off the coast of Virginia in which Army bombers successfully sank a
captured German battleship. The tests demonstrated the powerful striking force of air
power. The Navy recognized the need for fleet defense and expanded the
development of aircraft carriers In July 1919 the Navy acquired a ship designed to
launch and recover airplanes at sea. The ship was commissioned at Norfolk, VA in
March 1922 as the USS Langley (CV-l) the Navy's first aircraft carrier. In October
1922, the first take off and landing were made from the CV-l near Hampton
Roads, VA. In November 1922, the first catapult launching was made from the CV-l
located at anchor in the York River near Hampton Roads. In 1933 the first ship
designed and constructed as an aircraft carrier, the USS Ranger (CV-4), was launched
at the Newport News ship yard. Since then, 28 aircraft carriers have been constructed
for the U.S. Navy at that shipyard and Navy aviation was well underway.

SOME NACA/NASA AERODYNAMIC RESEARCH RELATED TO SECURITY
AND DEFENSE. M. Leroy Spearman. NASA-Langley Research Center. Hampton,
VA . The use of aircraft by European nations in World War I resulted in concern that
the U.S. was lagging in aviation developments. This lead to an act of the U.S.
Congress in 1915 that established the National Advisory Committee for Aeronautics
(NACA) with the charge to conduct aerodynamic research. The research began at
Langley Field, VA in the early 1920's and over the years this research transformed
low-speed, wood and fabric, propeller-driven airplanes into high speed, all-metal, jet
and rocket propelled airplanes. This lead to the fields of supersonic and hypersonic
flight and provided for access to space. The need for space research lead to the
establishment of the National Aeronautics and Space Administration (NASA ) in July
1958.The nucleus of the NASA was the existing NACA with the charge expanded to
include space research. The Space Act does specify that the NASA should interact
with other agencies on matters affecting security and defense, During the cold war era
following World War II, the preponderance of Soviet Union military equipment was
perceived as a threat. Accordingly, at NASA-Langley a research activity was
undertaken to provide experimental and analytical information to assess a variety of

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VIRGINIA JOURNAL OF SCIENCE

foreign aircraft and missile concepts of concern to our security and defense. Over the
years research information has been acquired for eleven aircraft, twelve surface-to-air
missiles, five air-to-air missiles, two air-to-surface missiles, two surface-to-surface
missiles, five ballistic missiles, two anti-tank missiles, four reentry vehicles, and one
wing-in-ground effect vehicle.

TRACING THE GROWTH OF U.S.AIRPOWER AND THE AIRCRAFT
INVOLVED. M. Lerov Spearman. Langley Research Center, Hampton, VA The
U.S. military first recognized a role for aviation in 1907 with the establishment of
an Aeronautical Division in the U.S Army Signal Corps. The U.S Navy
acknowledged a role for aviation m 1910 The Army procured an airplane from the
Wright Brothers in Augustl908 and began flight tests at Ft. Meyers, VA. A year later
an improved Wright Flyer was accepted by the Army as ‘Airplane No. 1 ’ Another
American designer, Glenn Curtiss, was becoming well known and in 1911 a Curtiss
airplane was the second airplane to be sold to the U.S. Army. The first airplane to be
built for the U.S. Navy was a Curtiss airplane 1911. The first appropriation for military
aeronautics was in the 1912 War Department for $ 125,000 . From these funds, orders
were placed for three Wright airplanes and two Curtiss airplanes. War broke out in
Europe in 1914 and it was apparent that a disparity existed between European progress
and U.S. foot- dragging in the field of military aviation. In 1915 the U.S. Congress
approved legislation to create a research facility to enhance aeronautical programs.
In 1916 property was purchased just north of Hampton , VA that provided a flying
field for aeronautical research and flight testing. Named Langley Field, a portion of
the field was set aside for construction of the newly created National Advisory
Committee for Aeronautics (NACA). The U.S. entered World War I in 1917 and did
learn from flying and building European airplanes. Significant growth began in the
1920's with trainers, bombers, first Navy fighters and Army pursuits, observation
airplanes, the first monoplanes, attack airplanes, first jet aircraft in early 1940’s, first
twin-engine fighter jets, swept-wing concepts, advanced supersonic designs such as
the Century series of fighters - a total of over 60 military aircraft designs appeared
from 1940 to 1980. Currently some aircraft using stealth technology are being
deployed.

Agriculture, Forestry and Aquaculture Science

YIELD AND RIPEN ING OF GREENHOUSE TOMATOES IN RESPONSE TO
DAYTIME TEMPERATAURE. Mark Kraemer & Frangoise Favi, Agricultural
Research Station, Virginia State University, Petersburg, VA 23806.
Greenhouse-grown tomatoes are a high-value crop for 1 united-resource producers in
the mid-Atlantic states. Increased heating costs in recent years have encouraged
many to reduce growing temperatures. Measurement of photosynthetic rates
indicated lower temperatures were feasible and some professional growers
recommended them. However, little actual data was available in the published
literature. Thus, we evaluated tomato yield differences between a warm and cool
greenhouse sections. The average daytime temperature in the warm section
averaged about 2°C greater than the cool greenhouse. Night temperatures were
similar. Each section had 18 tomato plants (var. Trust) arranged in three rows.

ABSTRACTS

17

The experiment was run for 3 months in the fall of2009 and again in 2010, with warm
and cool sections reversed. The results indicated that total fruit yield was the same
for both sections but that fruit ripened faster in the warm section. It is
recommended that growers allow temperatures to rise during sunny days by reducing
venting and using C0 2 generators. Increased heating costs during cold, sunny
periods could be repaid with earlier harvests.

LOCATION OF THRIPS (THYSANOPTERA: THRIPIDAE) ON SOYBEAN
SEEDLINGS AND IMPLICATIONS FOR SAMPLING. J.A. Samler 1 , D A.
Herbert', S. Malone 1 , D. Owens', T.P. Kuhar, & C. Brewster 2 . 'Department of
Entomology, Virginia Tech Tidewater Agricultural Research and Extension Center,
Suffolk, VA 23437.department of Entomology, Virginia Tech, Blacksburg, VA
24061. Thrips feeding can cause significant injury to soybean seedlings resulting in
stunted growth and delayed maturity. Determining accurate numbers of thrips on
plants is necessary for making control decisions. Currently, sampling is based on
assessing populations on whole plants which veiy labor intensive. This research was
conducted to determine where thrips are located on soybean plants and whether
subsamples of plant material are adequate for monitoring populations. Using a
stratified random sampling method, soybean plants were randomly selected and
systematically sectioned into three parts: the top three unopened leaflets (Terminal);
the first fully expanded trifoliate (Trifoliate); and the remainder (Remainder) of the
plant which was cut at the base just above the soil surface. Thrips larvae and adults
were washed from each plant sample and the leaf area was measured. Results showed
that the soybean Terminal has the highest density of thrips larvae (ja = 1.412) while
the Remainder of the plant has the highest absolute count (p. = 4.515) when density is
not a factor. There was no correlation between the number of larvae and leaf area in
any of the plant sections (Terminal: r = -0.0086, p = 0.9628; Trifoliate: r = -0.2742, p
= 0.1289; Remainder: r = -0.1158, p = 0.5280). Larval counts from the Remainder of
the plant had the strongest correlation with whole plant counts (r = 0.895, p =
<0.0001). Implications are that whole plant samples are the most accurate even
though labor intensive.

INVESTIGATING THE ROLE OF OLFACTION IN THE HOST PLANT
SELECTION OF HARLEQUIN BUG (MURGANTIA HISTRIONIC), PEST OF
COLE CROPS. Wallingford, A.K., T.P. Kuhar and P.B. Schultz. Dept, of
Entomology, Virginia Tech. Harlequin bug (HB) is a piercing-sucking pest of cole
crops. Adult and nymphal feeding causes cosmetic injury to vegetables sold as
greens and, if left untreated, HB infestations can stunt or kill a plant. There is
potential for managing HB by planting a preferred or more attractive host plant, or
trap crop, to divert insect feeding away from the protected cash crop This study
evaluated several species of brassicales and a non-host control for HB host preference
and thereby the suitability for use as a trap crop Caged choice tests show that HB
adults prefer to feed on mustard (Brassica juncea ‘Southern Giant Curled’) over,
arugula (Eruca sativa), bean (Phaseolus vulgaris), collard (B oleracea ‘Champion’),
rapim (B. rapa ‘Roquette’) and rapeseed (B. napus ‘Athena’). Olfactometer choice
tests showed that male HB are attracted to the odors from the leaves of both bean
(non-host) and mustard (preferred host) over clean air, but when given a choice

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VIRGINIA JOURNAL OF SCIENCE

between the two, males orient to mustard over bean. This is not the case for females
as they do not orient to odors from mustard or bean, and show no preference for the
odors of one over the other. Male and female HB both orient to odors produced by
the combination of odors from males feeding on mustard leaves (presumably emitting
an aggregation pheromone) over mustard leaves alone, while neither orient to males
feeding on bean versus bean alone and do not orient to males alone versus clean air.
Implications for trap crop implementation are discussed.

POTENTIAL USE OF CHLOROPHYLL FLUORESCENCE TO IDENTIFY
DIFFERENCES IN TRANSPIRATION EFFICIENCY IN PEANUT. Jacqueline F.
Hawkins & George T. Byrd, School of Natural Sciences & Mathematics, Ferrum
College. Ferrum, VA 24008. Production of peanut (Arachis hypogaea L.) often
occurs in areas prone to drought, therefore genotypic selection should be based on
efficient use of water. This study examined the potential use of chlorophyll
fluorescence for assessing genotypic variation in peanut transpiration efficiency
(TE), defined as the weight of diy matter per unit of water transpired. The
experimental hypothesis was the rate of decline in chlorophyll fluorescence in peanut
leaves would differ under different watering regimes and among genotypes and these
differences would be correlated with TE, Since reports show TE to be correlated with
specific leaf weight (SLW), this trait was also assessed. Seventeen genotypes were
grown in closed containers in a soil-sand mix and subjected to two soil moisture
regimes -well-watered and water-deficit. Peanut genotypes differed in TE, however,
TE did not differ between well-watered and water-deficit conditions. Transpiration
efficiency was positively correlated with SLW across water treatments (> = 0.75).
Correlation between TE and chlorophyll fluorescence, however, was not significant.
Although chlorophyll fluorescence has shown promise to indicate water-deficit stress
responses in species like cotton and sorghum, peanuts showed little difference in the
decline in chlorophyll fluorescence under well-watered and water-deficit conditions.
Further research is needed to determine the relationship between chlorophyll
fluorescence and traits associated with drought tolerance in peanut. (Supported by a
Faculty Development Grant, Ferrum College).

POTENTIAL FISH PATHOGENS FOR CAGE AQUACULTURE IN VIRGINIA.
David Crosby, Virginia Cooperative Extension, Virginia State University, PO Box
9081, Petersburg, VA 23806. Many farm pond owners raise catfish in cages for
personnel consumption or to sell as a farm commodity to local markets in Virginia.
These farmers typically buy catfish fingerlings that are sold into Virginia from out of
state commercial catfish fingerling farms. What fish farmers do not realize that there
is a potential disease risk involved with caged catfish. The typical bacterial pathogens
that cage producers would encounter are Aeromonas and Columnaris. These are quite
common in the aquatic environment and one of tire leading causes of disease
problems in cages. A new strain of Aeromonas hydrophila that has originated in
Alabama is highly pathogenic. This strain has been observed in Virginia. A fish
health study conducted at VSU during 2007 to 2009 found that catfish fingerling from
commercial farms had various external parasites. These catfish typically had
Trichodina, proliferative gill disease and Ligictaluriadus on the gills. In one case,
highly pathogenic bacteria, Enteric Septicemia of Catfish, which is common to

ABSTRACTS

19

catfish farming, was found during the study. This study also found that fish were
exposed to endemic pathogens such as Tch and Trichophyra. By stocking commercial
produced catfish a farmer increases the risk of bringing in pathogens that would cause
a disease outbreak, or result in an epizootic from an endemic pathogen(s), or worst
case scenario introduce a new potential pathogen to the fish farm.

THE WHY, WHERE, AMD WHEN OF CEREAL LEAF BEETLE (OULEMA
MELANOPUS L.). Chris Philips 1 , Ames Herbert 1 , Tom Kuhar 1 , Dominic Reisig 2 ,
and Sean Malone 1 . 1 Department of Entomology, Virginia Tech, Blacksburg, VA.
and ~ Department of Entomology, North Carolina State University, Plymouth, NC.
Because of the difficulty in predicting when and where cereal leaf beetle outbreaks
will occur many growers in the Southeast apply insecticides on a calendar basis rather
than using a threshold-based IPM approach. Our challenge is to develop new
information and procedures that will encourage growers to reevaluate the way they
are approaching spring-time insect control in wheat, and consider adoption of the
IPM approach. Using small plot trials in 2010 and 2011, we found that a number of
t insecticides were effective at controlling CLB when applied at the right time. We
also found that an IPM approach was more effective at controlling CLB than an
automatic application. In addition, large scale surveys indicated that CLB was not
randomly distributed on a field and regional scale. Finally, degree day information
was used to create a predictive model for when peak CLB egg laying will occur.
This model was used to predict the average calendar date of peak eggs for each
population as well as for the average of all populations. In 2010 and 2011, the model
predicted the average egg peak within 2 days and 1 day, respectively. Larval peaks
occurred approximately 16 days after egg peak in 2010 and 17 days after egg peak in
2011. Given the accuracy of 182 DD, historical weather information was used to
create a predictive map of when areas will exceed 182DD. This map predicted the
calendar date for each area in Virginia within 4 days.

OPTIMAL GROWTH CONDITIONS ACROSS ORGANIC HIGHBUSH
BLUEBERRY CULTIVARS AND SOIL TREATMENTS. Braydon P Hoover,
Denay M. Fuglie, & Roman J Miller, Eastern Mennonite Univ., Harrisonburg, VA
22802. Four growth parameters including height, primary stalk diameter, relative
‘bushiness’, and volume of plant cylinder were determined for four soil treatments
and five highbush blueberry cultivars at Knoll Acres Organic Blueberry' Farm in the
autumn of 2010. Planter’s Choice and pine straw composts had significantly higher
values than both horse and sheep manure in height (75.5±2.7) and bushiness
(1171±140 cm 3 ), stalk diameters (8.20± 1.60 cm) and plant cylinder volumes (381±37

'j t

dm ) respectively. Chandler had significantly higher values in both the stalk diameter
(8±0.36 mm) and bushiness (976.38±172.8 cm 3 ) parameters. Duke bushes had
significantly lower plant cylinder volume (1146834=14639 cm 3 ) than all other
cultivars and Bluecrop bushes had significantly greater average heights (77.85±2.6
cm). The photosynthetic rates of Jersey bushes were found to be significantly higher
than Bluecrop, however, no significant difference was found to indicate that soil
treatments affected photo synthetic rate.

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VIRGINIA JOURNAL OF SCIENCE

EFFICACY OF A NATIVE BEE FOR POLLINATION OF EASTERN
ORCHARDS. Melanie Cutter & Mark Kraemer, Agricultural Research Station,
Virginia State Univ., Petersburg, VA 23806. The blue orchard bee, Osmia lignaria
Say, has been shown to be an excellent pollinator of tree fruits in western North
America. A subspecies of this bee is widespread in eastern North America and may
be similarly effective. The objective of our study was to determine pollen
preference of this bee with respect to nest placement. Sets of 4 nest shelters were
placed within the orchard, an adjacent hardwood forest, and along the border between
orchard and forest. Each shelter contained a wood block with 24 nest holes. Prior
to apple bloom, each nest shelter was provided with 3 nests of adult bees that were
ready to emerge. Weekly images of nests were taken to date the completion of new
nests. Pollen samples were collected from individual nest cells in early May, after
the end of apple bloom. Random samples of 300 pollen grains per bee nest cell were
identified to species using a scanning electron microscope. The results indicated
that the bees preferred to collect pollen from nearby Eastern redbud (Cercis
canadensis L.) trees than the orchard fruit trees. Placement of nest shelters did not
make a significant difference in pollen preference. We concluded that eastern
redbud trees near orchards could reduce the pollination efficacy of O. lignaria within
eastern orchards.

Astronomy, Mathematics and Physics
with Materials Science

OPTICAL CHARACTERIZATION OF SELF-ASSEMBLED

POLYMER/MICROPARTICLES THIN FILMS. Daniela M. Topasna & Gregory
A. Topasna . Department of Physics and Astronomy, Virginia Military Institute,
Lexington, VA 24450. Thin films are an important component in many optical and
electrical devices, including optical filters. We fabricated multiple layer thin films
of sodium salt of poly( styrene sulfonate)/titania using a layer-by-layer self-assembly
method. The optical properties of these films were determined based on the
calculations of a theoretical numerical model that we developed. The UV-V1S-NIR
measurements of these films performed in the 300-2400 nm range confirmed the
relationship between the type and number of layers and were consistent with the
modeled transmittance values at specific wavelengths.

DETERMINATION OF PHOTON ARRIVAL RATES AND S/N
RELATIONSHIPS FOR THE VIRGINIA MILITARY INSTITUTE’S 0.5 METER
TELESCOPE. Gregory A Topasna & Daniela M. Topasna, Virginia Military
Institute, Lexington, VA 24450. We determined the photon arrivial rate for unfiltered
CCD images using the Virginia Military Insitute’s 0.5 meter telescope. Observations
of the open clusters NGC 1502 and M 44 were made using an Alta U6 CCD camera
with different integration times. Aperture photometry was performed on stars of
known visual magnitude and, using the standard CCD equation, the signal-to-noise
ratio was determined for the different integration times. The photon arrival rate for
each star was then determined from these data and the CCD equation. As a function
of magnitude m the photon arrival rate is approximatly equal to left 9 ' 26-771 / 2 - 4 ) Th e

ABSTRACTS

21

airmasses of the cluster images differed by ~ 1 and our analysis showed that arrival
rates were not strongly dependent on this airmass difference.

LOVE WAVE PROPAGATION IN A POROUS ELASTIC LAYER UNDER
RIGID BOUNDARY. Shishir Gupta & Dinesh K. Majhi, Dept, of Applied
Mathematics, Indian School of Mines, Dhanbad-826 004, India. In this paper,
propagation of Love waves in a porous elastic layer under a rigid boundary over a
porous elastic half space has been considered . Pores contain nothing of mechanical or
energetic significance. The study shows that such a medium transmits two types of
Love waves. The first front depends upon the change in volume fraction of the pores.
The second front depends upon the modulus of rigidity of the elastic matrix of the
medium and is the same as the Love wave in an elastic layer under a rigid boundary
over an elastic half-space. It is found that the phase velocity of Love waves is
considerably influenced by porosity. It is also observed that the first wave front is
faster than that of shear wave in the porous medium due to change in volume fraction
of the pores

THERMALLY DRIVEN ATMOSPHERIC ESCAPE: MONTE CARLO
SIMULATIONS FOR TITAN’S MULTI-COMPONENT ATMOSPHERE. Orenthal
J. Tucker & Robert E. Johnson, University of Virginia, Charlottesville, VA., 22904.
Kinetic Monte Carlo model simulations of thermal escape from Titan’s upper
atmosphere are used to reproduce the Cassini INMS density measurements of N 2 and
CH 4 without requiring significant loss rates as proposed by recent continuum models.
In the kinetic model intennolecular collisions between N 2 , CH 4 and H 2 are described
using temperature dependent cross sections and allowing the exchange of internal
energy as w r ell as translational between molecules. Fits to INMS density data for N 2
obtained when Titan was in Saturn’s plasma sheet indicate H 2 has a Jeans parameter
~3 at the exobase. The kinetic Monte Carlo simulations obtain an H 2 loss rate ~1
xl0 2S H 2 /s which is essentially the Jeans rate. The results from multi-component
kinetic simulations are used to examine the change in the H 2 escape rate when Titan is
not in the plasma sheet to when it is in the plasma sheet using average magnetospheric
conditions to obtain the energy deposition by the flux to 0+ and H+ into Titan’s
upper atmosphere.

THE MEANING OF EINSTEIN’S EQUATION. Emory F. Bunn, Physics
Department, University of Richmond, Richmond, VA 23173. The Einstein field
equation, relating spacetime curvature to the matter stress-energy' tensor, is one of the
fundamental laws of physics, but, due to the level of mathematical machinery
required to express it, it is generally omitted from the undergraduate curriculum.
However, it is possible to express the entire content of this equation in relatively
simple geometric terms, without the apparatus of tensor calculus. To be specific, the
Einstein equation relates the rate of change of the volume of a small ball of
initially-comoving test particles to the density and pressure of matter in the ball’s
interior.

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VIRGINIA JOURNAL OF SCIENCE

THE COSMIC MICROWAVE BACKGROUND RADIATION. Emory F. Bunn,
Physics Department, University of Richmond, Richmond, VA 23173.
Observations of tire cosmic microwave background radiation are among the chief
causes of the rapid advance in our understanding of cosmology over tire past 20 years.
Microwave background temperature anisotropy measurements have confirmed the
presence of acoustic waves in the early Universe, providing support for the standard
gravitational bistability paradigm and allowing us to deduce the properties of acoustic
waves in the early Universe and hence to measure cosmological parameters.
Attention is now shifting to measurements of microwave background polarization.
Polarization measurements can shed light on several aspects of cosmology, but
arguably the most exciting is the prospect of detecting “B modes,” which could
provide direct evidence of an inflationary epoch. Although existing data are in most
ways remarkably consistent with the standard cosmological model, some apparent
anomalies have been noted on large scales. The significance of these anomalies is
hotly debated. To settle the question, we need a new data set that probes perturbations
on similar scales but is statistically independent of the existing measurements. Future
microwave background polarization maps may provide such a data set.

ORBITS AND SCALING FOR AN ISOTROPIC METRIC. J. D. Rudmin, Dept, of
Integrated Sci. and Tech., James Madison University, Harrisonburg VA 22807.
Scaling of physical quantities shows the symmetries of an isotropic metric, including
extension to the equivalence principle. For example, invariance of Planck's constant
under gravitational scaling provides consistency of general relativity with quantum
mechanics. Invariance of charge provides consistency with electromagnetism.
Transitivity of scaling eliminates the traditional need for a globally preferred
reference frame Rather, diagonalization of the metric yields local rest frames.
Scattering, orbital period, and precession offer ways to distinguish an isotropic from a
Schwarzschild metric.

AN ANNUAL PATTERN IN WATER TEMPERATURE DIFFERENTIALS
BETWEEN ADJACENT REGIONS OF THE CHESAPEAKE BAY, VIRGINIA.
T.C. Mosca III, Dept, of Mathematics, Rappahannock Community. College, and W.
C. Coles, Div. of Fish and Wildlife - Dept, of Planning and Nat. Res. Each winter
the Atlantic Ocean, lower Chesapeake Bay, and the major tributaries cool to about
the same temperature. In the spring, water farther from the ocean warms faster,
and the difference in temperature between the bodies grows. The maximum
difference occurs in spring near the ocean, in summer farther upstream. Warming
occurs by the mfluence of solar radiation at the surface, the rate of which is the
same in all locations. The ocean snnply has more water per unit of surface area,
and warms more slowly. The lower regions of Chesapeake Bay and the rivers will
warm more slowly than the upper regions because ocean water circulates in these
parcels. The lag in warming is attributable to the exchange of energy with tire
Atlantic Ocean, even at the farthest distances examined in this study. The different
timing of the lags in the maxnmim temperature differences that occur in the upper
reaches (June vs. April) is an artifact of distance from the ocean. The only area
where this pattern changes is in the lower portion of the Rappahannock River,
probably due to a ledge across the mouth of the river, where depths quickly become

ABSTRACTS

23

half that of the nearby deep zone. We examined the lags as differences in
long-term average monthly temperatures of regions of the Bay and tributaries. This
feature of energy transfer is probably present in other estuaries, because a defining
feature of an estuary is density driven circulation.

EFFECTS OF THE CORIOLIS FORCE ON THE LARGE-SCALE
ATMOSPHERIC TURBULENCE. lordanka N. Panayotova, Department of
Mathematics and Statistics, 4700 Elkhom Ave., Old Dominion University, Norfolk,
VA 23529. Earth’s rotation causes an inertial force, known as the Coriolis force.
The Coriolis force is small but its effects become noticeable for motions occurring
over large distances and long periods of time (large-scales motions). Well known
examples of large-scale formations in the atmosphere are cyclonic/anticyclonic
systems, or jet-streams and atmospheric waves. On the other hand the large-scale
geostrophic turbulence is a chaotic three-dimensional nonlinear motion of the fluids
that are near to the state of geostrophic and hydrostatic balance. A simple
three-dimensional model of the tropopause dynamics is a model with introduced a
quasi-horizontal interface separating regions of homogeneous potential vorticity of
different values. The quasi-geostrophic (QG) approximation to this model reduces
dynamics of the flow to quasi-two-dimensional turbulence. In this way, the
three-dimensional flow is entirely modeled by the horizontal advection of potential
temperature on the interface (surface quasi-geostrophic (sQG) model). Including
weakly non-linear dynamics and the meridional variation in the Coriolis parameter
((3-effect) will result in the so called (3-sQG+l numerical model. Here this model is
applied to study the effects of tire Earth’s rotation on the large-scale atmospheric
turbulence. The numerical simulations capture some important dynamical
characteristics of the large-scale atmospheric dynamics known from observations, in
particular the meridional asymmetries in eddies spatial and time scales, and their
orientation.

UNCERTAIN GROUND: MAPPING ERRORS THROUGH THE POM-SAT
MODEL OF PALEOCLIMATE RECONSTRUCTION. Marshall G. Bartlett, Dept,
of Physics, Hollins University, Roanoke, VA 24020. Borehole temperature-depth
profiles contain information about ground surface temperatures (GST) a region has
experienced in the past and provide complementary information to the surface air
temperature (SAT) record of climate change. The borehole method of climate
reconstruction assumes the dominant heat transport mechanism in the upper few
hundred meters of the earth's crust is conduction; mathematically, conduction is a
compressive (information losing) mapping. Consequently, the solution obtained in
non-unique. One robust means of dealing with the non-uniqueness problem is to
limit the number of parameters sought in the solution space. However, even when
only a single parameter (the pre-observation mean GST, or POM) is sought in the
inversion, a certain amount of a priori information must be assumed. 1 am interested
in how uncertainties in this a priori information are mapped into uncertainties in the
solution space. 1 perfonn a Monte Carlo analysis to investigate how uncertainties in
our model of the ground thermal regime, SAT observations, and the background
geothermal gradient are mapped into the solution space of the POM-SAT method of

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VIRGINIA JOURNAL OF SCIENCE

climate reconstruction from borehole data. Results indicate that uncertainties in the
SAT time series and thermal parameterization of the ground are reduced by an order
of magnitude, while uncertainties in the geotherm are magnified by an order of
magnitude in the solution-space.

Biology

with Microbiology and Molecular Biology

THE EFFECTS OF BACTERIOPHAGE INFECTION ON BACILLUS ANTHRACIS
DETA STERNE AND BACILLUS THURLNGIENSIS. Catherine A Johnson &
Lynn O Lewis, Department of Biological Sciences, University of Mary Washington,
Fredericksburg, VA 22401. Bacillus anthracis and Bacillus thuringiensis are soil
dwelling bacteria that are capable of forming endospores. These bacteria go through
the processes of sporulation and germination to form the endospores and vegetative
cells. Both species of bacillus can be found m various soil samples, along with
species of bacteriophages that target each bacterium. The goal of this research was to
isolate one temperate bacteriophage for Bacillus anthracis Delta Sterne (Delta
Sterne) and one temperate bacteriophage for Bacillus thuringiensis A1 Hakam (A1
Hakam) and then determine die effects the bacteriophages have on the sporulation
and germination processes for Delta Sterne. A temperate bacteriophage was isolated
for each bacillus species, with the one diat infects Delta Sterne being called Texas 1
4CI. It was able to cross infect A1 Hakam, but the A1 Hakam temperate phage was
unable to cross infect Delta Sterne. There were no apparent morphological
differences between colonies of infected Delta Sterne and uninfected Delta Sterne
through sporulation and germination. However, it appears that with successive
sporulation and germination, the infectivity of the bacteriophage for Delta Sterne
increases from 1 plaque at 10' 3 dilution, to 32 plaques at 10' 3 after a second
sporulation and germination. (Supported by UMW Undergraduate Student
Research Grant. )

CHARACTERIZATION OF MY COBACTERIOPHAGE VENKMAN. Thien T.
Phan & Lynn O. Lewis, Department of Biological Sciences, University of Mary
Washington, Fredericksburg, VA 22401. Bacteriophage are abundant in the
environment and have co-evolved with the bacteria they target. Through soil
collected, we were able to have a better understanding of how mycobacteriophage
interact with their host, Mycobacterium smegmatis. One bacteriophage (Venkman)
was chosen for genomic sequencing. Restriction enzyme digests were performed to
compare the phage’s DNA to other known bacteriophage DNA. Electron
microscopy was performed to examine the morphology of the Mycobacteriohpage.
By using different bioinformatics programs, we determined that the
Mycobactenophage Venkman is in the F cluster and within the F1 subcluster. The
genome size is approximately 50 kbp - to 60 kbp, but is unfinished at this time.
Further investigations are still being performed on the genome of Mycobactenophage
Venkman. (Supported by UMW Undergraduate Student Research Grant.)

ABSTRACTS

25

POST TRANSLATIONAL MODIFICATION OF A BACTERIAL PROTEIN IN
HUMAN CELLS. Moushimi Amaya , Ancha Baranova & Monique van Hoek,
Department of Systems Biology, George Mason University, 10900 University Blvd.,
Manassas, VA 20110. This study is an on-going investigation into the post
translational modification, the prenylation, of a Francisella tularensis protein in
human cells. Prenylation is a lipid modification, whereby either a farnesyl or
geranylgeranyl group is added to a nascent protein harboring a C-termmal CAAX
motif, which results in subsequent targeting of the mature protein to the cell
membrane. F.tularensis is a gram negative bacterium that causes the disease
tularemia and can potentially be used a bioweapon due to its low infectivity dose
(10-15 organisms) upon inhalation. Bioinformatical analysis using the Prenylation
Prediction Suite program revealed a single F.tularensis protein that could potentially
undergo prenylation. The proposed prenylated protein is 13kDa in size and is of
unknown function. Human alveolar lung epithelial cells (A549) are the model system
chosen to test for prenylation of the Francisella protein. Preliminary Western blot
experiments have confirmed that the protein of interest is expressed in A549 cells.
We aim to detennine if prenylation does indeed occur in A549 cells. We further aim
to investigate, by use of inhibitors, which type of prenylation is undertaken:
famesylation or geranylgeranylation. We also aim to determine the localization of the
potentially prenylated Francisella protein in A549 cells by employing confocal
microscopy analysis.

FUNCTIONAL GEMOMIC ANALYSES REVEAL COMPLEX
TRANSCRIPTIONAL REGULATORY NETWORKS MEDIATING DENDRITIC
ARCHITECTURE. Eswar Prasad R. Iver 12 . Srividya Chandramouli Iyer 12 ,
Ramaknshna Meduri 12 , Dennis Wang' 2 and Daniel N Cox 12 ’ 'School of Systems
Biology, George Mason University, Manassas, VA 20120, USA/Krasnow Institute
for Advanced Study, George Mason University, Fairfax, VA 22030, USA, ’Present
Address: Yale University, New Haven, CT 06520, USA Elucidating the molecular
mechanisms controlling dendrite development is key to understanding the pivotal
role these structures play in influencing synaptic integration and neural function.
Despite significant advances in this field, genetic pleiotropy remains a significant
impediment to investigating such complex developmental processes. To circumvent
this problem, we have applied class specific neuron transcriptional expression
profiling coupled to an in vivo RNAi functional validation screen in order to dissect
the molecular bases of Drosophila class IV dendritic arborization (da) neuron
dendritogenesis. Microarray analyses reveal transcriptional regulation as one highly
enriched biological and functional category with 420 transcription factors
significantly expressed in class IV neurons. Among these, we identify roles for 268
genes in mediatmg a broad spectrum of functions mcluding dendritic field coverage,
branching, routing, and tiling. Collectively, our analyses provide a more
comprehensive framework of the role complex transcriptional networks play in
directing distinct aspects of class specific dendrite morphogenesis.

CUT MEDIATED TRANSCRIPTIONAL REGULATION OF THE COPII
SECRETORY PATHWAY DIRECTS CLASS SPECIFIC DENDRITE
MORPHOGENESIS IN DROSOPHILA. Snvidva C. Iver . Eswar P R. Iyer,

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VIRGINIA JOURNAL OF SCIENCE

Ramakrishna Meduri, Madhu Karamsetty, & Daniel N. Cox, School of System
Biology, Krasnow Institute for Advanced Study, George Mason University, Fairfax
VA 22030. Elucidating the molecular mechanisms controllmg dendrite
development is key understanding how neuronal morphologies arise and how they
function in achieving synaptic integration and neuronal function. Recent studies
demonstrate select secretory pathway genes act in preferentially affecting dendritic
growth. Phenotypic analyses of sec31 mutants reveal a reduction in dendritic
branching implicating the COPII secretory pathway in regulating dendritic
complexity. Furthermore, gain-of-function (GOF) analyses indicate sec 31
differentially effects dendritic complexity in distinct da neuron subclasses.
Microarray analyses, quantitative RT-PCR and immunohistochemistry experiments
reveal that overexpression of the homeodomain transcription factor Cut upregulated
expression levels of the COPII-mediated secretoiy pathway genes as well as another
key transcription factor, CrebA. Moreover, simultaneous expression of Cut coupled
with RNAi knockdown of CrebA suppressed the Cut GOF phenotype indicating that
CrebA functions as a downstream effector of Cut mediated transcriptional regulation
in da neurons. Consistent with this regulatory relationship, overexpression of CrebA
in da neurons likewise leads to higher expression levels of components of
ER-to-Golgi transport. Collectively, these findings provide novel insight into the
role of transcriptional regulation of the COPII-mediated secretory pathway in
mediating class specific dendrite morphogenesis.

NEUROPEPTIDE AF-INDUCED ANOREXIA IS ASSOCIATED WITH
CHANGES IN HYPOTHALAMIC CHEMISTRY IN SPRAGUE-DAWLEY
RATS. Brandon A. Newmyer and Mark A. Cline. Dept of Biol, Radford University,
Radford VA 24142. We recently demonstrated that NPAF’s anorectic effect is
associated with changes in hypothalamic chemistry in nuclei associated with satiety
perception in Cobb-500 chicks. In order to elucidate whether this effect was
conserved through divergent evolution, NPAF was centrally administered to
Sprague-Dawley rats and food intake as well as NPAF-associated changes in brain
chemistry were observed NPAF reduced food intake in rats at similar doses and
magnitudes as it did in chicks and also affected hypothalamic chemistry. Similar to
chicks, central NPAF was associated with increased neuronal activation in the
magnocellular region of the paraventricular nucleus These data support that NPAF's
anorectic effect is conserved in a mammalian model and thus may be a logical target
utilize in the treatment of human eating disorders.

EFFECTS OF COMBINED VITAMIN C & E TREATMENT ON PLAQUE
FORMATION IN ALZHEIMER'S DISEASE. Anum K. Shaikh & Deborah A.
O’Dell, Dept, of Biol., University of Mary Washington, Fredericksburg VA 22401.
Alzheimer’s disease (AD) is characterized by the inflammation and (3-amyloid
plaques in the brain. The improper cleavage of amyloid precursor protein (APP)
which leads to (3 amyloid plaques may result from inflammation. We studied the
effects of these Vitamins E and C on inflammation and plaque formation in a
transgenic mouse model of AD. Mice were divided into three experimental groups
that received Vitamin C, Vitamin E or both Vitamin C and E for a period of 34 weeks
beginning at 14 weeks of age. Levels of tumor necrosis factor alpha (TNF-a) and

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27

(3-amyloid in homogenized brain preparations were measured using a direct ELISA
method. The number and size of the plaques in sections of formalin fixed and
paraffin embedded cerebral cortex was compared between the groups. The levels of
(3-amyloid were marginally significantly lower in the combined Vitamin C and E
treatment group compared to the control (p=0.056). Vitamin E treatment alone did
not lower (3-amyloid relative to the control. Vitamin treatments also show reduced
levels of inflammation as measured by levels of TNF-a compared to the control with
Vitamin C and Vitamin C&E showing die lowest levels. There was no significant
difference noted in comparison of die number and size of plaques between all the
groups. These results show that Vitamin C and E may afford the greatest protective
effects against AD. (Supported by: Chi Beta Phi Honorary Society, The Virginia
Academy of Science, Univ. of Mary Washington Undergrad Research Funds).

A SYSTEM GENOMICS APPROACH FINDS CANDIDATE GENES FOR
NON-INSULIN DEPENDENT DIABETES MELLITUS Lataisia Jones & Glenn
C. Harris, Dept of Biology, Virginia State University, VA 23806. Non-insulin
dependent diabetes mellitus (NIDDM) is one of the most significant chronic human
diseases, affecting over 20 million people hi the United States (7% of the population).
NIDDM is associated with obesity and characterized primardy by insulin resistance
and unpaired insulm production. Evidence has been budding that NIDDM is a
multifactorial complex disease, with many different genes and gene-gene and
gene-environment interactions potentially contributing to the physiological
symptoms. Here we demonstrate a systems genomics approach to finding likely
candidates for NIDDM. A review of linkage analysis studies found a region on
human chromosome 01 significantly associated with NIDDM symptoms. Syntenic
regions in rats and mice were identified and subjected to a single nucleotide
polymorphism (SNIP) analysis. Only one gene was found in the mouse and rat
syntenic regions that exhibited functional SNiPs in symptomatic models. A
functional SNiP was defined as producing an amino acid change in the resulting
protein. The gene, Tchhll, has not been previously implicated as a candidate gene
for NIDDM or obesity. Future projects are described that will attempt to
characterize the function of Tchhll in NIDDM phenotypes.

CHROMATIN STATE MAPPING IN NEUROBLASTOMA CELLS IDENTIFIES
GENES POTENTIALLY IMPORTANT FOR MAINTAINING PLUR1POTENCY
Shaili Shah & Melissa A. Henriksen, Dept of Biol., University of Virginia,
Charlottesville VA 22903. Neuroblastoma is a common childhood cancer that arises
from the neural crest The clinical diversity seen in neuroblastoma is due to its
cellular heterogeneity. The Intermediate (T-type), highly malignant, cell differentiates
into both a non-malignant (S-type) cell and a less malignant Neuroblastic (N-type)
cell. The multipotent I-type cell has been identified as a cancer stem cell due to its
ability to self-renew and differentiate. Since epigenetic mechanisms underlie the
phenotypic differences among these three cell types, chromatin state mapping was
used to identify potential bivalent genes involved in cancer stem cell multipotency
and differentiation. Of the 6 genes identified, one gene, MSX2, was examined further
due to its known role in triggering apoptosis during neural crest development. Since
this gene was not expressed in I-type cells, the potentially inappropriate silencing of

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this gene was reversed by over-expression in I-type cells. Ongoing experiments
focus on using inducible lentiviral based methods to infect the cancer stem cell and
quantify cell death in the cancer stem cell in comparison to the other, differentiated,
neuroblastoma cells. These preliminary over-expression experiments suggest that
MSX2 may also trigger apoptosis of the cancer stem cell. The aberrant chromatin
pattern at MSX2's promoter may cause inappropriate gene silencing, and future
experiments will determine how crucial this gene is for maintenance of the
neuroblastoma cancer stem cell. Understanding the epigenetic mechanisms that
govern genes implicated in cancer stem cell maintenance may offer novel therapeutic
targets for cancer treatment.

MELANOMA-ASSOCIATED SUPPRESSION OF DENDRITIC CELL
MATURATION/ACTIVATION DEPENDS ON TUMORIGENICITY. Kristian M.
Hargadon . Osric A. Forrest, & Pranay R. Reddy, Dept, of Biol., Hampden-Sydney
College, Hampden-Sydney VA 23943. The accumulation of data demonstrating
immunity to cancer over the last two decades has sparked significant interest in the
field of tumor immunology. However, while the existence of anti-tumor immunity
is promising, tumor cells have frequently been reported to induce anti-tumor
immune dysfunction. Despite the overwhelming significance of this problem, the
basis for this immune dysfunction is often poorly understood. Dendritic cells play
critical roles in both innate and adaptive immunity, and their numerous functions are
tightly linked to their maturation and activation status. Here, we characterize the
murine dendritic cell line DC2.4 as a model for studying dendritic cell maturation
and activation, and we evaluate the influence of melanoma tumor cells on these
processes. Exposure of DC2.4 cells to the Toll-like receptor ligand
lipopolysaccharide induces both maturation and activation of these cells,
characterized by upregulation of costimulatory molecule expression and
proinflammatory cytokine/chemokine production. This maturation and activation is
suppressed by soluble factors derived from both the highly tumorigenic B16-F1 and
the poorly tumorigenic D5.1G4 murine melanoma cell lines. Interestingly, the
extent of DC2.4 immunosuppression by these melanomas correlates with their
tumorigemcity, suggesting a vital role for dendritic cell/tumor cell interactions in the
regulation of anti-tumor immunity and tumor outgrowth.

PATTERNS OF TRANSIENCY, SEX BIAS, AND BODY WEIGHT IN
OPEN-HABITAT RODENT POPULATIONS. Stephen E, Rice & Robert K. Rose.
Department of Biological Sciences, Old Dommion University, Norfolk, Virginia
23529-0266. As the most numerous and diverse order of the Class Mammalia,
rodents often have served as a common basis for study and modeling biological
concepts. Dating from the writings of Robert Collett in the late 19th Century,
rodents are believed to live their lives within a small area, with juvenile males the
most likely to disperse. To test these assumptions we examined long-term
capture-mark-release data from grid populations with attention to the weights at first
capture and the transiency/residency patterns of herbivorous rodents studied in
old-fields in Virginia, Illinois, and Kansas. Although few studies report the
proportions or sexes of juveniles among first-captured animals, many studies have
indicated that nearly half of tagged rodents are never seen again. In order to

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29

evaluate the evidence that different species of rodents are philopatric, we examined
the sex ratios and body weights of newly tagged old-field rodents and their patterns of
transience or residency. Transience was associated with season for Virginia and
Kansas cotton rats. Both populations had significant proportions of adults and
displayed an association of population groups. Annuals in breeding condition were
male-biased for Virginia and Kansas. The breeding individuals in Virginia were
independent of residency status, whereas Kansas breeders were associated with a
high level of transience, 59.33%. Transience was found to be significant for the
Virginia population, 54.41%, but not for the Kansas population, 51.00%. No
difference in body weight was found between residents/transients for either
population. Seasonal sex biases were found in both populations.

THE LIFE HISTORY FEATURES AND ECOLOGY OF TENODERA SINENSIS
AND T. ANGUSTIPENNIS MANTISES IN EASTERN VIRGINIA. Cory A. Gall
& Robert K. Rose, Department of Biology, Old Dominion University, Norfolk, VA
23529. Praying Mantises, Tenodem sinensis and T. angustipennis, are generalist
predators located at the top of the arthropod food chain. These species inhabit eastern
North America, and are common among old-field vegetation. Virginia is considered
the southern limit of their natural range, with few field studies of these two mantises
coexisting. My study was conducted on the Wildlife Refuge located on Virginia’s
Eastern Shore and is focused on their life history, movement, and species interaction
in this region. Previous studies have shown that T. sinensis hatches earlier in the
spring, giving it the ability to become larger in size than T. angustipennis . My initial
results support these findings. In addition, I investigated the species ratio and sex
ratio for both species throughout their lifecycle. I found that T. sinensis dominated for
the majority of the year, but at the end of fall the species ratio was nearly 50:50 Past
research has indicated T. sinensis and T. angustipennis hatch with 50:50 sex ratios;
however, male presence are found to be greater until sexual maturity is reached, then
females become more numerous. However, my results illustrate that T. sinensis and
T. angustipennis sex ratios fluctuated throughout the life cycle, with T. sinensis and T.
angustipennis females are dominating at different times in their growth season. My
study also supports past research that females are more stationary to measured plots,
while males often moved off measured plots.

A SMALL MAMMAL COMMUNITY IN A CHANGING LANDSCAPE IN
SOUTHEASTERN VIRGINIA, 2005-2011. Jana Eggleston & Robert K. Rose.
Department of Biological Sciences, Old Dominion University, Norfolk, VA 23508.
In 2005 we began a monitoring program of the small mammal community on an old
field site in southeastern Virginia. This site is a part of the Nature Conservancy
Stewardship of lands adjacent to the Great Dismal Swamp National Wildlife Refuge,
and a part of a greater wetland habitat restoration plan. An assortment of trees was
planted on the site, drainage was altered, and vegetation succession was allowed to
progress. We hypothesized this small mammal community would change from
herbivorous old field species to those of forested wetlands, and that the numerically
dominant species would change as succession progressed. Our study site was an 8 x
8 grid, withl2.5m intervals, and with two modified Fitch traps per station. We
trapped for three days each month, averaging 4600 trap nights per year. Results to

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date show the general decline of all old field species, the virtual disappearance of Mus
musculus and Oryzomys palustris, stable densities of Reithrodontomys humulis, a
shift in dominance from Microtus permsylvanicus to Sigmodon hispidus , and most
recently, the arrival of Peromyscus leucopus, a forest species.

THE EFFECTS OF VISUAL AND CHEMICAL CUES ON MALE MATE CHOICE
IN EASTERN MOSQUITOF1SH ( GAMBUSIA HOLBROOKI). Nouman J. Rana &
Lisa Horth, Dept, of Biol. Old Dominion Univ., Norfolk, VA 23529. Previous studies
have illustrated that pheromones releases by gravid females western mosquitofish
(Gambusia affinis) increased male sexual activity during mating selection. However,
little is known about the activities during mating selection in eastern mosquitofish
{Gambusia holbrooki). In this study, we conducted experiments to determine (1)
whether pheromones from gravid female mosquitofish induce different male
activities as compared to non-gravid females and (2) if male mosquitofish prefer both
a chemical and visual cues during mate selection. Our preliminary results indicate
that pheromones released by gravid females are different from non-gravid females. In
addition, gravid females induce a more active response from males during mate
selection than non-gravid females. Also, males prefer to have both visual and
chemical cues during the mate selection process. These results suggest that males are
more likely to pick a female that they can visually see and one that has recently given
birth as compared to one that cannot be seen and has not recently given birth.

EXPLORING THE ROLE OF THE C-TERMINAL REGION OF APQ12 IN THE
REGULATION OF CELL CYCLE PROGRESSION AND CELL SEPARATION
James R. Oliver & Michael J Wolyniak, Department of Biology, Hampden-Sydney
College, Hampden-Sydney, VA 23943 The Saccharomyces cerevisiae peripheral
nuclear membrane protein Apql2 is believed to play a critical role in the regulation of
membrane fluidity and, ultimately, the way in which cells divide and separate. To
further investigate the proposed role of Apql2 in these processes, we performed a
series of site-directed mutagenesis experiments that attempted to delete the
transmembrane domains of the protein as well as generate C-terminal truncations via
nonsense point mutations. This allowed us to investigate if there was a specific
domain of the Apql2 sequence that is necessary for its putative role in the regulation
the cell cycle processes and cell separation. Our findings indicated that while Apql2
is, as a whole, a dispensable protein with respect to cell viability, the C-terminal
region of the protein may provide a fine-tuned regulation of cell morphology, with
different domains of this region acting in an antagonistic relationship to determine
cell shape. Since we also found that cells deleted for Apql2 were slower in overall
growth rates, our findings suggest that the C-terminal region of Apql2 may
contribute to the efficiency of yeast cell cycle progression through the regulation of
cell shape during the cell division process.

ISOLATION AND CHARACTERIZATION OF A NEW CAENORHABDITIS
SPECIES TO BE USED IN EVOLUTIONARY AND DEVELOPMENTAL
STUDIES. Borwyn A. Wang, Mejgan Mukhtarzada . Erin Haynes & Theresa M.
Grana, Department of Biological Sciences, LTniversity of Mary Washington,

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31

Fredericksburg VA 22401. The worm community of researchers is actively seeking
a sister species for comparative genomics analysis for the identification of a C.
elegans sister species The purpose of this project was to isolate, identify, and
characterize several new Cctenorhabditis species that are phylogenic relatives of the
model organism Caenorhabditis elegans. Soil samples were collected in
Fredericksburg, VA as well as in King George County in which nematodes were
isolated and stocks were established on agar plates. Isolated nematodes were tested
for model organism characteristics such as the ability to survive freezing, starvation,
and male/female/hermaphrodite classifications. Nomarski microscopy was used to
film mouth parts, pharyngeal bulbs, vulvas, and tails to further compare each
species. Additional studies include filming embryo development and sequencing a
part of the rDNA encoding the 18s ribosomal subunit to construct phylogenic trees.
This study was supported by UMW Undergraduate Research Grants and a VAS
undergraduate grant.

EFFECT OF CpG OLIGONUCLEOTIDE AND LL-37 ON PROSTATE CANCER
CELL GROWTH AND INVASION. April Lao 1 , Angela Gupta 1 . Maria Craig 2 , &
Paul Deeble 1 , Departments of 'Biology and 2 Chemistry and Physics, Mary Baldwin
College, Staunton, VA 24401. CpG oligonucleotides, short pieces of DNA with
characteristics of bacterial DNA, have recently been considered as an alternative to
chemotherapy. Some studies found CpGs to stimulate anti-tumor immune responses
with none of the side effects associated with traditional chemotherapy. However,
CpGs effects are weak, and previous results are contradictory, with anti-tumor effects
seen in ovarian cancer in mice but increased invasion and decreased growth in PC-3,
LNCaP, and DU-145 prostate cancer cell lines. LL-37 is an antimicrobial
DNA-binding peptide that increases the body’s immune response to CpG. LL-37 is
found throughout the body and in higher amounts in breast, lung, and prostate cancer
tumors. The combination of CpG and LL-37 has recently been shown to enhance the
anti-tumor effects seen with CpG in ovarian cancer in mice. In this project, the effect
of CpG in combination with LL-37 on prostate cancer cell growth and invasion was
investigated. The addition of LL-37 was hypothesized to enhance the effects seen
with CpG (i.e. decreased growth and increased invasion), Experiments utilized
androgen-independent PC-3 cells and androgen-responsive LNCaP cells. Results
indicate the combination of CpG and LL-37 increases growth in PC-3 cells and
increases invasion in LNCaP cells. These results could indicate that CpG
oligonucleotides in combination with LL-37 should not be used as a therapy against
prostate cancer.

NEUROENDOCRINE DIFFERENTIATION AND ITS EFFECTS ON
CHEMORESISTANCE IN PROSTATE CANCER CELLS. Alex Kelly , Sophia
Stone, Caitlin Combs, & Paul Deeble, Department of Biology, Mary Baldwin
College, Staunton, VA, 24401 Neuroendocrine (NL) differentiation, induced by
forskolin (Fsk) and 3-isobutyl-1 -methylxanthine (TBMX), was analyzed in a prostate
cancer progression model using androgen-responsive LNCaP and
androgen-independent PC-3 prostate cancer cells. NE differentiation was quantified
by measuring neuritic branch points and mean process length, and LNCaP cells were

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VIRGINIA JOURNAL OF SCIENCE

found to exhibit a higher level of NE characteristics when compared to PC-3 cells.
Cell viability in response to various chemotherapeutics, including Docetaxel and
Rapamycin, was measured in undifferentiatied and NE differentiated cancer cell lines
using an MTT assay. NE differentiation was found to protect LNCaP cells from cell
death induced by chemotherapeutic agents.

EFFECTS OF PHENOLIC ACIDS ON OVIPOSITIONAL SELECTION IN THE
CABBAGE WHITE BUTTERFLY, PIER1S RAPAE. Jessica L. Brav . Mary E.
Lehman & Amanda J Lentz-Ronning, Department of Biological and Environmental
Sciences, Longwood University, Farmville VA 23909. Pieris rapae oviposit on
plants that are most suitable for larval growth and survival. The presence of a variety
of plant secondary metabolites may be one factor in determining ovipositional
choices. Phenolic acids are secondary metabolites that are widespread among plants,
but their effects on Pieris rapae have not been well characterized. Wisconsin Fast
Plants were treated with four different phenolic acids (1.0 mM concentrations) to see
if they would defer or .stimulate oviposition. Female butterflies were provided a
choice of whether to lay eggs on the control plants sprayed with deionized water or
the plants sprayed with a phenolic acid. P-coumaric acid significantly stimulated
oviposition, whereas salicyclic acid and protocatechuic acid had no significant effect.
With the removal of one extreme outlier from the data set, the stimulatory effect of
femlic acid was also significant. Ferulic acid and p-coumaric acid are very similar in
chemical structure and preliminary experiments suggest that p-coumaric acid
increases larval growth.

EFFECTS OF HOST PLANT ALLELOCHEMICAL AND NUTRIENT STATUS
ON THE CABBAGE WHITE BUTTERFLY, PIERIS RAPAE. Kristen. S Walker .
Mary E. Lehman & Amanda J. Lentz-Romiing, Department of Biological and
Environmental Sciences, Longwood University, Farmville VA 23909. Pieris rapae
is known to detect the chemical content of plants that would be best suited for
offspring development. Plant allelochemical and nutritional status w^ere manipulated
to determine the main and interactive effects on ovipositional choices of Pieris rapae.
Wisconsin Fast Plants were supplied with Hoagland’s nutrient solution at
concentrations of I/2X (control) or 1/8X (reduced nutrient treatment). Plants were
sprayed with deionized w'ater (control) or 1.0 mM p-coumaric acid (PCO;
allelochemical treatment). The effects of PCO and nutrient concentration were first
assessed individually and then in an interaction experiment. Females and plants were
placed in light boxes and eggs were counted after a 24- hour period. A significantly
higher number of eggs w r ere deposited on the 1/2X control plants (mean ± SE: 21.7 ±
5.5) compared to the 1/8X reduced nutrient treatment (9.5 ± 2.8). Females oviposited
significantly more eggs on the PCO-treated plants (35.3 ± 3.7) than on the DI water
controls (25.2 ± 4.4). In the interaction experiment, only the main effect of nutrient
concentration was significant. PCCTs lack of significance and the lack of a significant
interaction may be due to the low number of replicates (N=6) Overall, results suggest
that female Pieris rapae can distinguish the nutrient status of plants and that this is a
stronger determinate of ovipositional choice than host plant allelochenucals.

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THE ROLE OF INTERFERON GAMMA IN EXPRESSION OF KLF4 IN LUNG
FIBROBLASTS. H. Rushdi 1 . E. Ouraishi 1 . S Qureshi 1 , J. Forrest 1 , S.D. Nathan & G
Grant 1 , 'SSB GMU, Manassas VA, 2 IHV1 Inova Fairfax, Falls Church, VA.
Kriippel like 4 Factor (KLF4) is a member of the zinc finger transcription factor
family which is involved in cells’ transcriptional response to multiple critical stimuli,
such as DNA damage, growth stnnulation and differentiation. KLF4 activity is
cell-type dependent acting as both an inducer and repressor of gene expression. It is
predominantly associated with post-mitotic terminally differentiated/ing cells,
playing a major role in cell homeostasis by induction of p21 and inhibition of p53.
However KLF4 can also act as a tumor promoter in cells that have bypassed the p21
checkpoint. Idiopathic pulmonary fibrosis (IPF) is a terminal lung disease propagated
by the fibroblasts. IPF fibroblasts over-express KLF4 in vivo. Therefore we
investigated the potential role of KLF4 overexpression in IPF by inducing its
expression with Interferon gamma (INFy).IPF and normal fibroblasts were seeded at
2.5x1 Cf cells per 60 mm dish in 10% DMEM at 37°C in a 5% C0 2 incubator. Cells
were serum starved for 16 hours to synchronize prior to addition of INFy at 0,200 and
400 Units per ml for 24 hours. RNA was extracted (RNeasy Kit Qiagen) and cDNA
generated (iscript BioRad) from ljjg of total RNA. Real Time Quantitative PCR
was carried out for KLF4 induction using ribosomal 18S as a normalization control.
INFy exposure of IPF and normal fibroblasts resulted in increased expression of
KLF4 protein which may influence the proliferative and survival behavior of these
fibroblasts in the IPF lung.

INVESTIGATIONS OF THE EFFECT OF THYMOSIN-BETA4 ON LUNG
FIBROBLAST SURVIVAL. S Qureshi'. J Forrest 1 H, Rushdi 1 , E. Quraishi',S.D.
Nathan 2 & G Grant 1 . ^SB, GMU, Manassas VA 2 IHVI Inova Fairfax, Falls Church
VA. Idiopathic Pulmonary Fibrosis (IPF) is a fatal lung disease, with no therapy and
no cure. IPF is propagated by over abundant pulmonary fibroblasts which deposit
excessive extracellular matrix (ECM) causing damage to the alveoli. The excessive
ECM interferes with gaseous exchange and ultimately results in organ failure
Thymosin Beta 4 (T(34) is a tiny cellular protein with hormone like properties.
Initially identified as a cytoskeletal protem, it has been recently identified with
wound healmg, cell migration, angiogenesis, and anti-apoptosis. T(34 is upregulated
in IPF cells compared to normal lung fibroblasts. In this experiment we investigated
the effect of T(34 to prevent hydrogen peroxide (H 2 O 2 ) induced apoptosis in prnnary
IPF and normal fibroblasts. IPF and normal fibroblasts were exposed to lethal doses
of H 2 0 2 in the presence and absence of T[34 for 24 hours in Dulbecco's Modified
Eagle Medium (DMEM) containing 10% fetal calf serum m a 96 well format. After
24 hours of exposure cell survival was determined by the Acid Phosphatase and MTT
cell proliferation assay. Tp4 exposure increased cell survival in both normal and IPF
cells. Results indicate that T|34 has the potential to play a significant role in
preventing apoptosis in IPF lung fibroblasts. This cytoprotective property of T|34
may contribute to the overall abundance of fibroblasts in the IPF lung, a hallmark and
contributing factor of this disease. Further investigations into the action of this
protein may lead to therapeutic and biomarker discoveries which may aid in
preventing and treating IPF.

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VIRGINIA JOURNAL OF SCIENCE

THE DROSOPHILA SPECTRAPLAKIN SHORT STOP DIFFERENTIALLY
REGULATES CLASS SPECIFIC DENDRITE MORPHOGENESIS. James
Boddu . Sarah A. Trunnell, & Daniel N. Cox, School of System Biology, Krasnow
Institute for Advanced Study, George Mason University, Fairfax VA 22030.
Disorders of the nervous system can often be attributed to developmental
abnormalities occurring during neurogenesis, winch affect the morphology, and
ultimately functionality of neurons. The Drosophila peripheral nervous system
(PNS) provides an excellent model system in which to elucidate the molecular
mechanisms governing dendrite morphogenesis. Spectraplakins are an
evolutionarily conserved family of cytoskeletal cross-1 inking proteins that link the
actin and microtubule cytoskeletons. Cytoskeletal structure and organization are key
mediators of neuronal shape, and by extension neuronal function. To address the role
of Spectraplakins in dendrite morphogenesis, we focused on the sole known
Spectraplakin gene in Drosophila referred to as short stop (shot). Here we
demonstrate that shot is present in all classes of dendritic arborization (da) neurons
where it exerts differential effects on class specific dendrite morphogenesis.
Comparative morphological analyses reveal shot is required to restrict dendritic
complexity among the simpler class I and II da neurons, whereas shot is required to
promote dendritic complexity among the more complex class III and IV da neurons.
These results suggest dendrite morphogenesis is subject to context-dependent
regulation mediated via shot.

INVESTIGATING THE ROLE OF RNAi REGULATION IN CLASS-SPECIFIC
DENDRITE MORHPOGENES1S. Mvuraian Rubaharan , Eswar P.R. Iyer, & Daniel
N. Cox, School of System Biology, Krasnow Institute for Advanced Study, George
Mason University, Fairfax VA 22030. Dendrites function as the primary sites of
synaptic and/or sensory input and integration in the developing nervous system The
initiation and maintenance of dendritic arbors determine both the number and ty pe of
inputs they receive and thus is a critical determinant in establishing functional neural
networks. Despite their functional importance, the molecular mechanisms governing
cell-type specific dendrite morphogenesis remain largely unknown. One emerging
mechanism involves the regulation of differential gene expression in neuronal
subclasses via the evolutionarily highly conserved RNA interference (RNAi)
pathway. The Drosophila melanogaster, peripheral nervous system, an excellent
genetic and morphological model system for studymg class specific dendrite
morphogenesis, was used to perform loss-of-function (LOF) and gam-of-function
(GOF) phenotypic studies to understand the role(s) of ten key genes essential for
mediating RNAi in the fruitfully. These studies provide novel mechanistic msight
into the role RNAi pathway plays in controlling class-specific da neuron dendrite
morphogenesis. The broader implications of these studies are related to the relative
contribution of this conserved pathway in controlling gene expression at the
class-specific neuronal level in mammalian systems and how this regulation
ultimately contributes to acquisition of distinct neuronal morphologies that underlie
the establishment of complex neural networks.

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35

FORAGING AT THE SNAIL DINER: THE EFFECTS OF PARASITE
INFECTION ON SNAIL FORAGING BEHAVIOR. Sabrina Brooks 1 . Jeremy M.
Wojdak 2 & Lisa K. Belden 1 , ’Virginia Tech, Department of Biological Sciences,
Blacksburg, VA 24060, 2 Radford University, Department of Biology, Radford, VA
24142. Snails are common hosts of parasitic flatworms (trematodes). Snails
typically feed at dawn and dusk, and during this time they are especially vulnerable to
predators. Parasite infections can alter this feeding pattern, which can increase the
probability of infection or transfer of die parasite to tire next host. We examined how
infection by Echinostoma trivo/vis can affect the foraging behavior of Helisoma
trivolvis snails, hi separate trials, we examined foraging behavior in (1) snails
infected as first intermediate hosts and (2) snails either exposed or not exposed to
cercariae (second intermediate host infection). We used two assays to assess
foraging behavior. First we examined how the frequency of snails on the tiles
changed over twelve hours and the rate at which the snails in the different infection
groups reached the tiles. The second assay compared the amount of lettuce
consumed within 25 hours among snails in the different infection groups. We did
not see a difference in proportion of snails on the resource tiles between cercariae
exposed and non-exposed snails. However, for the first intermediate host snails, the
proportion on the tiles was higher after 120 minutes and remained higher until the end
of the trial. For the lettuce trials, there was no difference in the amount of lettuce
consumed among the three categories of snails. While there were some differences
in foraging behavior between first and second intermediate host snails in our first
assay, we did not see dramatic difference in foraging based on infection status in
Helisoma snails.

SUSCEPTIBILITY OF PSEUDOMONAS AERUGINOSA BIOFILM TO
ALPHA-HELICAL PEPTIDES: D-ENATIOMER OF LL-37. Scott N. Dean .
Barney M Bishop & Monique L. van Hoek, Department of Systems Biology, George
Mason University, 10900 University' Blvd., Manassas, VA 20110. Pseudomonas
aeruginosa is a highly versatile opportunistic pathogen and its ability to produce
biofilms is a direct impediment to the healing of wounds and recovery' from infection.
Interest in anti-microbial peptides has grown due to their potential therapeutic
applications and their possible use against antibiotic resistant bacteria. We tested the
human AMP, EL-37, and tire effect of a protease-resistant LL-37 peptide mimetic, the
peptide enantiomer D-LL-37, for anti-microbial and anti-biofilm activity against P.
aeruginosa. The CD spectra of D- and L-LL-37, and the trypsm resistance of
D-LL-37 was confirmed. The helical cathelicidin from the cobra Naja atra
(NA-CATH), and synthetic peptide variations (ATRA-1, ATRA-2,
NA-CATH:ATRA1-ATRA1) were also tested. Although the cobra cathelicidin and
related peptides had strong anti-microbial activity, they did not mhibit Pseudomonas
biofihir formation. D-LL-37 inhibited attachment, promoted Pseudomonas motility,
and decreased biofilm formation by altering the rate of twitching and downregulating
the expression of the biofilm related genes, rhl/i and rhIB. D-LL-37 protected
Galleria mellonella in vivo against Pseudomonas infection, while
NA-CATH:ATRA 1-ATRA 1 peptide did not. This study shows D-LL-37 is able to
promote bacterial twitching motility and inhibit biofilm formation, and protect

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against infection, and suggest that this peptide may be a critical component for the
development of new treatments for P. aeruginosa infection,

CHARACTERIZATION OF DOWNSTREAM EFFECTORS MEDIATING CUT
TRANSCRIPTIONAL REGULATION OF CLASS-SPECIFIC DENDRITE
MORPHOGENESIS. Luis Sullivan , Eswar P R. Iyer, Madhu Karamsetty, & Daniel
N. Cox, School of System Biology, Krasnow Institute for Advanced Study, George
Mason University, Fairfax VA 22030. Neuronal form dictates function and in a
circuitry as complex as the human brain the post-synaptic properties of the neuron are
established in large part by dendritic morphology. Transcriptional regulation has
emerged as a pivotal mediator of class specific dendrite morphogenesis; however, the
downstream effectors of these transcription factors remain largely unknown as are the
cellular events that direct morphological change. Recent studies have implicated the
Drosophila homeodomain transcription factor Cut and it’s vertebrate homolog in
mediating dendrite morphogenesis in the peripheral and central nervous systems. To
characterize putative transcriptional targets of Cut regulation, a genetic suppressor
screen has been performed in which Cut overexpression has been coupled with target
gene-specific in vivo RNAi knockdown. Preliminary analyses have identified >400
genes that represent potential direct targets of Cut regulation in Drosophila dendritic
arborization (da) neurons. Here we report the discovery of target genes that either
suppress or enhance Cut-mediated effects on da neuron dendritic morphology. The
molecules uncovered in our screen cover a broad range of biological
functions. Collectively, these analyses reveal novel transcriptionally regulated
pathways and cell biological processes essential to the specification of class specific
dendritic morphologies.

THE DETECTION OF MYCOPLASMA SPECIES CONTAMINATION IN
CULTURED HUMAN CELLS. Amy Yu . Pranvera Ikonomi, Nadine Kabbani,
Ancha Baranova & Aybike Birerdinc, College of Science, George Mason University,
Fairfax, VA. Mycoplasma can contaminate eukaryotic cell cultures. It can alter
reproducibility of experiments or completely destroy the cell line. In most of the cell
culture labs, only one or a few species of Mycoplasma are systematically monitored.
The novel Universal Mycoplasma Detection Kit from American Type Culture
Collection (ATCC) provides a way to detect over 60 species of Mycoplasma. AIM:
To perform field test of die Universal Mycoplasma Detection kit usmg continuous
cell cultures. Methods: The kit contains reagent for a PCR reaction using universal
primers tiiat are specific to the 16S rRNA coding region in the mycoplasma genome,
allowing to specifically amplify Mycoplasma DNA. If a cell culture is contaminated,
after gel electrophoresis, the presence of a 464-bp amplicon is detected. In our
experiment, 8 PCR reactions were performed for each cell culture, including sample
A, B and C provided by die kit as reference examples of different levels of
mycoplasma contamination; two test samples; positive control; purified Mycoplasma
DNA, positive control with lysate to confirm that inhibition of PCR by the cell lysate
did not occur, and negative control. Results: Cell culture samples from Dr. Kabbani’s
and Dr. Van Hoek’s lab were negative for Mycoplasma. Sample B, Sample C,
positive control and positive control with lysate all showed a band at 464-bp. Sample
A, all test samples and negative control showed no bands. This confirms the absence

ABSTRACTS

37

of Mycoplasma in the cell cultures and proves that ATCC kit is reliable, easy to use
and can be recommended for introduction to the market.

ADAR FACILITATED RNA EDITING IN HUMAN PLASMACYTOID
DENDRITIC CELLS (PDC). A. Sharma 1 , K. Doyle 1 , M. Connors 4 , A.
Patamawenu 4 , P. Gillevef 1 , A. Birerdinc 12 , & A. Baranova 1,2 , 1 School of Systems
Biology, George Mason University, Fairfax VA 22030, 2 Betty and Guy Beatty Center
for Integrated Research, Inova Health System, Falls Church, VA 22042, J
Microbiome Analysis Center, George Mason University, Manassas, VA 20100,
4 National Institute of Health, Bethesda, MD 20892. Adenosine (A) to Inosine (I)
RNA editing is facilitated by enzy mes known as ADAR (Adenosine Deaminase that
Act on RNA), ADARs specifically recognize double stranded RNA structures or
RNA duplex structures as their substrates. Inosine is translated as Guanosine, since
most enzymes recognize Inosine as Guanosine. Examples of physiological ADAR
editing are edits to neuronal Glutamate and Serotonin receptor transcripts Here we
set to find out whether ADAR-editing in human PDCs (Plasmacytoid Dendritic Cell)
is limited to TER7, or whether it covers other known ADAR targets, including other
TLR receptors, FLNA, IGFBP7, KCNA1, GABRA3, and CYFIP2 Site specific
primers around previously known edited sites were designed using NCBI primer blast
and then tested on cDNA derived from universal RNA and adipose tissue cDNAs
from purified PDC cells will be used as templates for PCR amplification, tagged,
purified and subjected to Multitagged (MTPS) pyrosequencmg on Roche GS-FLX
instrument. If successful, these experiments will be the very first demonstration that
RNA editing activity is present in PDCs and acts on physiologically important gene
targets.

USP22 ACTIVITY REQUIRED FOR EFFICIENT nriRNA 3’ PROCESSING OF
STAT1 REGULATED GENES. Edmond Chipumuro and Melissa A Henriksen
Department of Biology, University of Virginia, Charlottesville VA 22904. Histone
modifications represent a major mechanism of eukaryotic transcriptional regulation
at multiple levels. There is increasing experimental evidence that histone H2B
monobiquitination (ubH2B) and its deubiquitination are both involved in gene
activation Recent findings have suggested that, the ubiquitin specific peptidase 22
(USP22) regulates c-MYC and p21 expression and is required for appropriate cell
cycle progression, However, a mechanistic understanding of USP22 function in gene
expression remains elusive. Here, we utilized the rapid and transient STAT1
activation of the JRF1 gene to characterize roles of USP22 RNAi mediated depletion
of USP22 significantly increased /FF/-associated levels of ubH2B and down
regulates 1RF1 expression. Importantly, USP22 knockdown consistently resulted in
inefficient mRNA 3’ processing of the 1RF1 gene and led to increased read through
RNA polymerase II. Furthermore, depletion of USP22 resulted in lower occupancy of
RNA Pol II phosphorylated at serine 2 (CTDS2) and cleavage and polyadenylation
specificity factor 73 (CPSF 73) on tire JRF1 gene body and it’s read through. Taken
together, our results suggest that, USP22 activity is required during IRF1
transcription to regulate co-transcriptional processing of the mRNA.

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DEVELOPMENT OF A SIMPLIFIED METHOD FOR DETECTING NUCLEAR
IMPORT. Adil Quraish & Stephen Gallik, Ph . D., University of Mary Washington.
Nuclear import is an important cellular process through which proteins enter the
nucleus through the nuclear pore complex. It is commonly detected and studied using
sophisticated protocols too time-consuming, complicated and expensive to be
routinely used in the undergraduate teaching or undergraduate research laboratory.
The specific objective of the study reported here is to develop a simplified method for
the detection of nuclear import that can be more easily used in an undergraduate
setting. The simplified approach taken here involves homogenization of rat liver,
incubation of the homogenates with commercially-available fluorescently-labeled
proteins, isolation of the rat liver nuclei from the homogenates using a standard
Optiprep density gradient centrifugation techniques, followed by the
fluorescent-microscopic inspection of the isolated nuclei. Results showed nuclei
isolated from homogenates incubated with fluorescently-labeled proteins conjugated
to a 13-amino acid nuclear localization signal (NTS) fluoresced, showing nuclear
import of the fluorescent protein, while nuclei isolated from homogenates incubated
with fluorescently-labeled proteins lacking anNLS did not fluoresce.

THE EFFECT OF VEGF ON ADIPOSE STEM CELL MIRGRATION. Laila
Almahdali & Kathryn E. Loesser-Casey, Dept, of Biol., University of Mary
Washington, Fredericksburg VA 22407. Stem cell research is one of the cutting
edge ways scientists and clinicians hope to treat human diseases in the future, but
stem cells are difficult to obtain in sufficient quantities. In order for stem cells to be of
any use following a myocardial infarct or similar injury, they must be able to migrate
to the place of damage. It has been found that growth factors such as Vascular
Endothelial Growth Factor (VEGF) can assist in the migration pathway by
stimulating endothelial growth, migration and blood vessel formation It is also found
the VEGF is secreted by adipose stem cells, leading to the possibility that VEGF may
be necessary for stem cells release or migration. The goal of this experiment was to
determine if treatment w ith 50ng/ml VEGF increased migration and whether the age
of the stem cells affected migration. Cells were collected from both young (10 to 12
week) and old (over 30 week) CF-1 mice and used for migration assays using Boyden
blind-well chambers The number of cells that had migrated towards media with or
without VEGF was counted and data evaluated using a one-sample T-test. Cells
isolated from the younger mice appeared to show an increase in cell migration
towards the VEGF-containing media; however statistics mdicates no significant
change in cell number over the control. When cells from adult mice were tested we
saw a decrease in cell migration, but no statistics can be performed on this data
because only one assay was performed due to very slow cell growth. We plan to
repeat these experiments, changing the cell number used for each migration and
mcreasmg the concentration of VEGF to get more consistent results.

ABSTRACTS

39

Biomedical and General Engineering

DIFFERENTIATING MANIFESTATIONS OF SOPITE SYNDROME AS
PRODUCED BY INERTIAL MOTION AND VECTION. Brittany N. Neilson & J.
Christopher Brill, Department of Psychology, Old Dominion University', Norfolk VA
23529. Sopite syndrome is a polysymptomatic manifestation of motion sickness
that presents itself as marked drowsiness despite receiving an adequate night’s sleep,
relaxation, and lack of motivation Previous studies evaluate sopite syndrome using
either a motion device or a vection device. The present research aims to differentiate
the symptoms of sopite syndrome produced by actual motion versus perceived
self-motion (i.e., vection) in order to better identify the psychophysiological markers
of sopite syndrome. Twenty-five undergraduate psychology 7 students participated in
the pilot study. An analysis of covariance was used to evaluate participant’s
self-reports on motion questionnaires, physiological measurements, and brain
activity across three motion conditions (inertial motion, vection, and control) holding
traits of sleepiness and motion sickness constant. Participants in the motion
condition reported mcreased symptoms (M = 20.75, SD = 23.95) compared to
participants in the vection (M= 8.53, SD = 25.45) and control conditions {M= 5.53,
SE = 24.25), but this effect was not statistically significant at p< .05, F( 2, 19) = 2.87,
p = .081, rf = .20, due to insufficient sample size. Implications for future research
will be discussed.

SEGMENTATION OF INTERFACE PRESSURE MAPS: THE USE OF A
CONNECTED ELLIPSE MODEL TO IDENTIFY AREAS AT RISK FOR
PRESSURE ULCER DEVELOPMENT. Anathea A. Pepperl and Paul A Wetzel,
Department of Biomedical Engineering, Virginia Commonwealth University,
Richmond, VA 23284. Mary Jo Grap, Department of Adult Health and Nursing
Systems, Virginia Commonwealth University', Richmond, VA 23298. In order to
study pressure ulcer development, it is useful to analyze pressures at specific
high-risk areas, or regions of interest (ROIs). A contour-based connected ellipse
model was used to segment ROIs. A user selected the approximate centers of ROIs.
An Expectation-Maximization algorithm refined the center locations and defined an
ellipse at each ROT. The connected ellipse model was then iteratively refined so as
to minimize (1) the distance between connected ellipses and (2) the distance between
the image contour and the edge of the closest ellipse. Ten pressure images were
collected from four patients in intensive care units at the Virginia Commonwealth
University Health System (VCUHS). The distance between ROT peak pressures
generated by automatic and manual segmentations was defined as the error, The
average errors (in inches) for each ROI were: 6.64, left scapula; 7.76, right scapula;
4.54, left trochanter; 5 31, right trochanter; 0, left heel; 0.85, right heel. The average
error was 4.18 inches. Visual inspection showed that this error was caused by the
peak pressure lying just beyond the contour of the ellipse. The connected ellipse
model is an effective method for segmenting pressure images. The model may be
further refined through the inclusion of intensity data.

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VIRGINIA JOURNAL OF SCIENCE

DEVELOPMENT OF A NOVEL BLOOD ANALOG WITH PHYSIOLOGICAL
SHEAR-THINNING AND VISCOSITY CHARACTERISTICS. S. J. Warren, C. E.
Taylor, and G. E. Miller, Dept. Biomedical Engineering, VCU. Mock loops are a
integral part of the testing and evaluation of cardiac assist devic. In order to obtain
near physiological data in a mock circulatory loop, a compressive fluid similar to
human blood is needed. This project will test the properties of an oil base fluid, in
order to match the viscoelastic properties of Human blood. The work presented was
based on the following research questions: Does tire oil based fluid show shear
thinning properties closest to those of human whole blood? Does the oil based fluid
correlate with tire viscoelastic curve of human whole blood better than other blood
analogs currently in use? All viscosity data was collected on an Anton Paar
Rheometer with a parallel plate and 500 micron gap. The test was a shear sweep from
.001 to 1000 l/s. The work presented two candidates; one that is optical clear and
the other is rheological more accurate. The components are pure and commercially
validated for mechanical properties, which releases the end user from having to
revalidate the constructed solution. Choice of fluid construction was designed to
prevent large dilution factors, to provide a robust method for fabricated a
shear-thinning blood analog for cardiac assist device evaluation.

CONTROL MODEL AND PERFORMANCE OF A COMPLIANCE CHAMBER
FOR SIMULATING ARTERIAL SYSTEMIC COMPLIANCE. C. E. Taylor and G.
E. Miller, Dept, of Biomedical Engmeermg, VCU. Utilizing mock circulatory loops
as a bench test method for cardiac assist technologies provides that the system must
be capable of reproducmg the circulatory conditions that would exist physiologically.
Of particular interest is the ability to determine compliance delivered by the system,
and the ability to change the compliance in real-time. The latter capability allows for
continuous battery testing of conditions without stopping the loop operation when
changmg the compliance chamber settings. The compliance chamber presented
utilizes a natural latex rubber membrane separating the fluid and air portions of the
device. Change in system compliance is affected by the airspace pressure, which
creates more reaction force at the membrane to the fluid pressure. A pressure sensor
in the fluid portion of the chamber and a displacement sensor monitoring membrane
position allow for real-time inputs to the control algorithm. A predefined numerical
model correlates the displacement sensor data to the volume distention of the
membrane. The PI controller tuning was achieved by creating a computational
model of the compliance chamber using Simulink Simscape toolboxes. It was found
that the resulting control architecture was capable of maintaining a compliance set
point, and allowed for changes to the compliance without stopping the pulsatile flow
of the mock circulatory loop.

THE EVOLUTION OF AN EVENT DETECTION ALGORITHM. Pallavi
Ramnarain and Dr. Paul A. Wetzel, Dept, of Biomedical Engineering, Virginia
Commonwealth University, Richmond, VA The purpose of this presentation was to
explore the methodological processes behind the development of event detection
algorithms. Signal acquisition imposes the first set of limitations on event detection.
After that an iterative process is initiated. The first step is to identify the event of
interest. Then it must be defined using finite, quantitative characteristics that

ABSTRACTS

41

distinguish it from the remainder of the signal. Then an approach to detection can be
developed. The approach must be validated and then separately evaluated for its
accuracy. Limitations are identified and quantitatively defined, and so the process
continues. To illustrate this example data from a study examining preterm infant
bottle feeding competency was used. The focus was placed on the detection of bottle
sucking activity. Multiple approaches to event detection were explored.

A Patient Specific Computational Model of Adult Acquired Flatfoot
Deformity. E.M. Snratlev & J.S. Wayne, Dept, of Biomedical Engineering &
Orthopaedic Surgery, VCU, Richmond, VA 23284. Computational modeling
offers advantages over MRI and plane-film x-ray alone in the quantification of adult
acquired flat foot deformity, (AAFD). As such, a population of solid body models
was created for the purpose of replicating common radiographic tests used in the
quantification of degenerating flat foot morphology. This was accomplished by
acquiring a sample of sub-millimeter resolution MRI scans from a group of
clinically diagnosed Stage II AAFD sufferers prior to surgery. The scans were
imported into the medical imaging package, Mimics (Materialise, Ann Arbor MI),
wherein the bony surfaces were isolated from the surrounding tissues and
triangulated to form a solid bodies for each bone in the foot. Subsequently, the
bodies were imported into SolidWorks (SolidWorks Corp., Concord MA) and
templates recreating an oblique anteroposterior film x-ray view were constructed.
This template allowed the talo-1 sl metatarsal angle (T1MT) and talo-navicular
coverage angle (TN) to be calculated for each of the AAFD models as well as a
non-flat foot cadaver-based model that served as the control. The normal model
had a T1MT=5.35° and a TN=0.76°. The AAFD models had a mean T1MT=8.28
±1.66° and a TN=5 15±4.65°. While these values are lower than those in the
literature for T1MT and TN, the values for T1MT were within ranges reported
clinically. Additionally, the sample set was able to capture the characteristic
increase in the two angles for AAFD feet over those of non-afflicted feet. Further
refinement of these models could lead to better classification of AAFD patients with
the ultimate goal of improving the treatments prescribed.

Botany

GROWTH AND NUTRIENT ACCUMULATION RESPONSES TO
PHOSPHORUS DEFICIENCY IN CELLULOSE SYNTHASE MUTANTS OF
Arabidopsis thaliana. Rob Harbert, Roanoke College. Cellulose synthase
mutants of Arabidopsis thaliana are affected not only in the overall amount of
cellulose they are able to produce, but also in the carbohydrate composition of their
cell walls. These alterations in the composition of the cell wall may also impact
the functions of other components of the cell wall and plasma membrane. The
cellulose synthesis mutants JE100, rswl-1 , and prcl, as well as the Columbia and
Wassilewskija (WS) wild types, have been used to see if changes in cell wall
composition have any effect on how these mutants perform in phosphorus deficient
conditions. The results indicate that JE100 plants respond differently to

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phosphorus than the wild types or possibly the other mutants. The results suggests
further, that cell wall alterations may have an effect on how nutrients enter roots, or
how growth responses are processed. Ward et. Al. (2008) suggested that the
phosphorus deficiency dependent growth responses occurred as a result of iron
over-accumulation through an interaction that is not well understood at this tune.
This observation was used to test whether or not the cell wall alterations in the
JE100 plants affect how nutrients get across the cell wall and into the root. To do
this, phosphorus and iron content was measured in dried plant tissues for Columbia
and JE100 plants grown on tire four phosphorus conditions on ATS media. It was
observed that while the JE100 plants follow the same iron accumulation trend as
Columbia, the JE100 plants accumulated more of both nutrients in all scenarios than
the Columbia plants do. While this observation does not explain the JE100 plant’s
relative insensitivity to phosphorus deficiency, it does imply that the JE100
mutation alters the cell wall in such a way that nutrient uptake and accumulation
appear to be affected. It is, however, unclear what the nature of this interaction is,
or if this affects how nutrient uptake across the cell wall (the apoplast) is affected.

STATUS AND TRENDS OF PHYTOPLANKTON POPULATIONS IN VIRGINIA
WATERS AND CHESAPEAKE BAY. H.G. Marshall, T.A. Egerton, M. Semcheski,
& M. Lane. Dept. Biological Sciences, Old Dominion University, Norfolk, VA
23529-0266. Results of monitoring phytoplankton populations the past 25 years was
presented with emphasis placed on bloom producing species in Virginia tidal
tributaries. Many of these blooms have been associated with reduced oxygen levels in
these waters and fish kill events. Potentially harmful taxa, including known toxin
producers, of major concern in these waters include Cochlodinium polykridoides,
Alexandrium monffatum, Karlodinium veneficum, Microcystis aeruginosa,
Chattonella subsalsa , and Prorocentrum minimum. C. polykrikoides and K.
veneficum blooms were becoming more common, with A. monilatum and C. subsalsa
blooms of more recent development regarding their regional impact. P. minimum
has shown a significant long-term trend of increased abundance over the last 2
decades within the lower Chesapeake Bay. Algal blooms by a variety of other taxa
also occurred seasonally. A survey of 19 Virginia lakes in 2010 also indicated
presence of several potential toxin and bloom producing species at 8 of these lakes.
These were Anabaena spiroides, Aphanizomenon flos-aquae, Cylindrospermopsis
raciborskii, Limnothrix redekei, Microcystis aeruginosa , and M. wesenbergii. In
summary, algal blooms are common in Virginia waters and Chesapeake Bay. They
are produced by a diverse group of algae, some of which are toxin producers. There
is evidence that several of these algae have expanded their range and duration of
bloom formation; including the presence of bloom producing invasive species
entering Virginia waters and the Chesapeake Bay. Funding support for this study
came from VDEQ and VDH.

COMMUNITY-MEDIATED CONSERVATION IN THE SOUTH PACIFIC: HOW
HUMAN INFLUENCE OVER THE PAST MILLENNIA HAS SAVED A RARE
TREE SPECIES FROM POSSIBLE EXTINCTION. Helen K, McKmnev 1 . Ryan
D. Huish & Joseya Mateboto% Department of Biology, Hollins University,
Roanoke, Va, 2 Fiji Forestry Department, Kadavu Island, Fiji. Community-mediated

ABSTRACTS

43

management strategies for indigenous resources, employed by various communities,
have shown success on local levels despite overall trends of resource decline. While
the history of sandalwood trade in the South Pacific is rife with over exploitation,
ancient cultural practices and modem management techniques for native Santahmi
yasi by Fijian and Tongan communities may have actually enabled S', yasi s survival
despite the continuing threat of over harvesting, and habitat destruction. Remaining
wild S', yasi stands lack genetic insularity typical of island populations, suggesting
significant migratory events, such as the pre-historic movement of seeds and
seedlings across and between Tonga and Fiji via trade, and marriage practices as
documented by early explorers to these regions. Furthermore, current strategies
employed by a Fijian village in the management of S. yasi reveal foundational
principles that can guide the development of effective management of other
endangered economic resources and curtail common problems of implementation.
The inclusion of all community members, in the benefits and responsibilities of
resource tenure and stewardship, enables better prevention of poaching, and fiscal
community interests. Further research and application of these, and similar practices
in other areas, may help resolve current management challenges to the preservation of
culturally and economically valuable species around the world.

ANTIMICROBIAL PROPERTIES OF THE TONGAN MEDICINAL PLANT
SYZYG1UMCOR YNOCARPUM: BIOASSAYS IN SUPPORT OF

ETHNOBOTANICALLY-GUIDED RESEARCH. Erika Schoolev & R. D. Huish,
Dept of Biology, HU, Much of the traditional medicine of Tonga specifically treats
infections. In a previous study, Syzygium corynocarpum not only showed significant
inhibition rates of Staphylococcus aureus in laboratory bioassays, but was also the
plant most frequently referenced by Tongan healers for use in treating infections.
This correlation alludes to the benefits of ethnobotanically-guided drug research.
Furthermore, in medicinal preparations, Tongan healers specifically use the young
leaves of S. corynocarpum , which are a characteristic red-purple color changing to
dark green at maturity. Extracts of mature leaves and bark, as well as the young
leaves, are being tested for inhibition rates of S. aureus to compare efficacy and
strengthen the case for ethnobotanically-guided research in the search for novel
drugs.

CURRENT POPULATION DYNAMICS OF A RARE SANDALWOOD TREE
(SANTALUM YASI) IN FIJI AND TONGA. Ryan D. Huish '. L. Thomson 2 , T.
Faka’osi 3 , H Likiafu 3 , M. Tuiwawa 4 , J. Mate bo to \ & D. Little 6 , 'HU, 2 Secretariat of
the Pacific Community, 3 Tongan Ministry of Forests, 4 USP, 5 Fiji Dept, of Forestry,
6 NYBG. Valued internationally for the aromatic oil found within its heartwood,
Sandalwood {Santa/um, Santalaceae) is one of the most heavily exploited groups of
plants across its range—Oceania, Southeast Asia, and India. One of the most valued
sandalwood species is Santalum yasi, harvested extensively in Fiji and Tonga. To
help aid in conservation efforts for this now rare tree, population dynamics, current
species distribution, and ecological threats were mvestigated. It was concluded that
the few remaining wild stands display discontinuous size class structures, are under
regenerative stress, and that the natural distribution has diminished significantly,
even to local extinction in some areas. Several threats appear to be affecting the

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population dynamics of S. yasi , including destructive harvesting and invasive
species. It is suggested that Forestry and governmental efforts focus on the
promotion of local involvement in assisted natural regeneration of wild stands
through community-mediated conservation.

A METHOD FOR LONGLEAF PINE TIMBER IDENTIFICATION AND ITS
APPLICATION TO RELICTS IN SOUTHEASTERN VIRGINIA. Thomas
Eberhardt 1 , Philip Sheridan 2 . Arvind Bhuta\ 'USDA Forest Service, Pineville, LA,
"Meadowview Biological Research Station, Woodford, VA, College of Natural
Resources & Environment, Virginia Tech. Longleaf pine ( Pinus pahistris P. Mill.)
cannot be distinguished from the other southern pines based on wood anatomy alone.
A method involving measurements of pith and second annual ring diameters,
reported by Arthur Koehler in 1932, was revisited as an option for identifying
longleaf pine timbers and stumps. Cross section disks of longleaf, loblolly (P. taeda
L.), and shortleaf (P. echmatci P. Mill.) pines were measured and the diameters of
their piths and second annual rings plotted against each other. Longleaf pine could
be differentiated from the other two pine species demonstrating that a method
established with trees harvested more than 70 years ago is still applicable to standing
timber today. No evidence was found to suggest that growth rate differences impact
method applicability . In those situations where the second annual ring is intact, but
not the pith, very large second annual ring diameters (> 40 mm) may provide an
adaptation of the method to identify timbers with a lower probability of being
longleaf pine. In addition to the identification of very old lightwood stumps as part
of a longleaf pine restoration effort, both methods may be applied to timber
identification in historic structures and the niche forest products industry involving
the recovery and processing of highly prized longleaf pine logs from river bottoms.
Measurements from relicts sampled in this study yielded results consistent with the
purported range for longleaf pine in Virginia.

A CENSUS OF PURPLE PITCHER PLANT, SARRACENIA PURPUREA L., IN
MARYLAND AND VIRGINIA. Philip M. Sheridan, Meadowview Biological
Research Station, Woodford, VA Sarraceniapurpurea L. is a rare wetland plant in
Virginia and a threatened species in Maryland, with two potential subspecies in the
region. A census was performed of existing populations, all known historical data
on the species compiled, reasons for the species demise investigated and dates of
extinction predicted. Bloom phenology was examined to see if climate change may
have influenced bloom period. Only 31% of purple pitcher plant (4 of 13) sites are
extant on the western shore of Maryland and District of Columbia while 33% (14 of
42) remain m Virginia, with respective total populations of 46 and 513 clumps.
Causes of regional extirpation include beaver flooding, succession, and
development. Predicted pitcher plant population extmction dates, based on trend
line froml30 years of data, are 2015 (Maryland) and 2055 (Virginia). Disturbance,
especially natural fire, played an essential role m maintaining purple pitcher plant
historically in Maryland and Virginia. Sarracenia purpurea blooms May 8 - June
12 in Maryland and Virginia with a peak May 18-20. Peak bloom period of S.
purpurea may have shifted as much as a week from historical dates, perhaps due to
climate change.

ABSTRACTS

45

STUDIES OF THE HEPATICAE AND ANTHOCEROTAE ALONG HAZEL RUN
IN ALUM SPRING PARK, FREDERICKSBURG, VA - IMPLICATIONS OF
CLIMATE CHANGE . Stephen W. Fuller, Biology Department, University of Mary
Washington, Fredericksburg, VA 22401. Climate plays a large role in determining
the distribution of species, especially through physiological thresholds of tolerance to
temperature and precipitation. With general vvarmmg trends, the Tolerance
envelopes’ are expected to migrate pole-ward, and upward in elevation. Studies of the
biological impact of climate change have focused on the abundance and distribution
of species; changes implying impact of these abiotic factors. More reliable
geographical shift information can be ascertained by studying more sedentary
species, considering local extinctions and new records. Fredericksburg experienced a
prolonged period with little rainfall during the last half of August and September
2010. The drying effects of this period were compounded by relatively wanner
temperatures. During the fall of 2008 and 2010 exhaustive sampling was used, from
the top of the stream bank to the center of the stream, within which all plant materials
were surveyed for the presence of liverworts and homworts. Despite the paucity of
rain during some of the warmest months of the growing season (warmer than during
the previous study in 2008), the liverworts did not seem to have suffered. There was a
single place where Conocephalum conicum plants had dried to the point of having
dead edges along the thallus. Evidently the higher temperatures and paucity of
precipitation were not inhibitory to these species’ continued presence and success in
this habitat.

EXAMINING POLAR AUXIN TRANSPORT EVOLUTION IN BRYOPHYTE
GAMETOPHYTES. B T. Piatkowski . G.A, Bader & D B. Poli Department of
Biology, Roanoke College, Salem, VA 24153. Specialized cells in vascular and
non-vascular plants can move auxin unidirectionally in a process termed polar auxin
transport. Data collected from the liverwort, Riccia flu items, suggests that simple
diffusion is utilized for the transport of auxin into the cell. The hornworts,
Phaeoceros Iaevis and Anthoceros agrestis , and the moss, Rhizommum sp. , displayed
facilitated diffusion influx. All four species displayed facilitated diffusion as the
method of auxin efflux, Rhizommum showed sensitivity to the influx inhibitor
1-naphthoxyacetic acid and the efflux inhibitor 1-N-naphthylphthalamic acid
suggesting that bryophyte gametophytes contain protein carriers similar to those used
in PAT in vascular plants but that the process may not be unidirectional and
specialized yet.

EXAMINING POLAR AUXIN TRANSPORT IN THE MOSS GAMETOPHYTE
OF DICRANIUMSCOPARIUM. Jessica N, Branning & DorothyBelle Poli. Dept, of
Biol., Roanoke College, Salem VA 24153. Polar auxin transport is a process
understood most in higher plants. Understanding where a physiological mechanism
evolved requires examining the same process across the entire plant kingdom. Poli et
al. (2003) examined polar auxin transport across the bryophyte sporophytes and
learned that hornworts exhibit simple diffusion, liverworts exhibit facilitated
diffusion, and mosses show active transport. However, examining polar auxin
transport using traditional agar block methods has been limited in gametophytes. This

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study examines the moss Dicranium scoparium, From Bader et al (unpublished) it is
expected that this terrestrial moss gametophyte should show facilitated diffusion.
(Supported by: Jeffress Memorial Grant).

Chemistry

DOES C0 2 WARM THE ENVIRONMENT? TC DeVore & D. Havey,
Deptartment of Chemisdtry and Biochemistry, MSC 4501, James Madison
University, Harrisonburg VA 22807 A series of laboratory exercises have been
developed that let students investigate three postulates related to global wanning: 1 .
burning fossil fuels creates C0 2 , 2. C0 2 absorbs TR radiation, and 3 increasing the
amount of C0 2 in the atmosphere increases the temperature of the environment. To
test the first postulate, students use GC-MS and FTIR to compare the amount of CO?
in a sample of automobile exhaust to the amount of C0 2 in ambient air. A
vibrational and a rotational analysis of the spectrum for C0 2 and preparing a Beer’s
Law plot for C0 2 are used to test postulate 2. The Boltzmann distribution to
determine the rotational temperature of the CO? in the cell is used to test postulate 3.
By measuring the temperature of several different concentrations of C0 2 , the effect of
the concentration of C0 2 on the temperature inside the cell is established. If all parts
of this project were done, students would leam about the combustion of
hydrocarbons, the 1R spectra and rotational analysis of linear tri-atoimc molecules,
Beer’s Law, the measurement of rotational temperatures based on the Boltzmann
distribution and the Greenhouse Effect.

DESIGN, SYNTHESIS AND INTERACTION STUDIES OF SULFATED
ALLOSTERIC MODULATORS OF THROMBIN. Preetpal Singh Sidhu . Aiye
Liang, Akul Y. Mehta, May Abdel Aziz, Qibmg Zhou, & Umesh R. Desai, Dept, of
Medicinal Chemistry, Virginia Commonwealth University, Richmond, VA-23219.
We have synthesized a library of potent, small, aromatic allosteric inhibitors of
thrombin. The sulfated benzofuran scaffold was designed from the structure of
sulfated lignin and contained charged, polar and hydrophobic substitutions at the
2,3,5,6 positions, which provide diversity to the library of 15 sulfated monomers and
13 sulfated dimers. Due to presence of negatively charged sulfate and carboxylate
groups, these molecules are completely water soluble. Synthesis of the library of
benzofurans derivatives was achieved through a multiple step, highly branched
strategy, which culminated with the introduction of sulfate group(s) using
microwave-assisted chemical sulfation reaction. From the library of 28 potential
inhibitors, 11 inhibited thrombin with reasonable potency. Structure activity
relationship studies showed sulfation at the 5-position of the benzofuran scaffold was
essential for targeting thrombin. The /-butyl derivative was found to be the most
potent with an /C 50 of 7.3 pM under physiologically relevant conditions.
Michaelis-Menten kinetic studies showed that these inhibitors do not affect the
affinity of chromogenic substrate for thrombin ( Km ) but greatly reduce the maximal
velocity (V max ) of reaction, indicating the allosteric mechanism of inhibition. The
results of plasma clotting assays suggest that these inhibitors prolong both the

ABSTRACTS

47

activated partial thromboplastin time and prothrombin time, of which the former is
more affected in a manner similar to the heparins and hence affect the intrinsic
pathway more effectively.

DEVELOPMENT OF A METHOD FOR WATER PURIFICATION UTILIZING
THE ELECTROCHEMICAL PROPERTIES OF POROUS CARBON
MATERIALS. Mikhail M, Goldin 1 . B.M. Grafov 2 , A.D. Davydov 2 , Mark M.
Goldnr’, & V.A. Kolesnikov 4 , 'Dept, of Bio. and Chem., Liberty University;
2 Frumkin Inst, of Phys. Chem. and Electrochem; 3 Sklifosovky Inst, for Emergency
Medicine; 4 Mendeleev Univ. of Chem. Tech, of Russia. The traditional methods for
decontamination of water have limitations and are relatively expensive to use. The
goals of the present work were to develop a prototype electrofilter. AG-3 brand
activated carbon samples were electrochemically pre-modified to initial open-circuit
potentials (OCP) between -775 and +475 mV and then placed in aqueous 2.20 M
r-butanol and 0.100 M CuSCC solutions. Concentrations were measured on an SRI
Instruments 3IOC gas chromatograph for Ebutanol and on a Genesys lOuv
spectrometer at 808 nm for Cir . Effective charge transfer for each experiment was
calculated using the equation n = (C-AE)/(F-AT), where n - effective number of
electrons, C(F)- differential capacitance of the sorbent electrode, A E (V) - OCP
change during the experiment, F (=96500 C mol' 1 ) - Faraday’s constant, AT (mol) -
net adsorption. The effective charge transfer for f-butanol (n <0.1) did not vary
appreciably with potential, which pointed to a mechanism not involving
electrooxidation or electroreduction, n for Cu 2+ was near 2 for potentials more
negative than -470 mV and gradually decreased until +475 mV, where it reached n =
0 indicating a major contribution from Faradaic electroreduction Preliminary
microbiological results also indicated a dependence of bacteria counts on carbon
potentials This study is supported by funds from the Center for Research and
Scholarship Fund of Liberty University.

MEASUREMENT OF DOPAMINE TRANSPORTER ACTIVITY IN THE
LARVAL Drosophila CNS USING FAST-SCAN CYCLIC VOLTANMETRY AT
CARBON-FIBER MICROELECTRODES. T, Vickery & B.J Venton, Dept, of
Chem., University of Virginia, Charlottesville, VA Drosophila melanogaster is
homologous to mammals in primitive neurobiology making it an advantageous model
system in which to study the dynamics of dopamine regulation. Here we report a new
method to measure dopamine transporter activity in isolated, Drosophila ventral
nerve cords A carbon fiber microelectrode is implanted into a larval ventral nerve
cord, while a micropipette is simultaneously implanted approximately 10 pm away.
Dopamine is pressure ejected into the neuropil region, rich with dopaminergic
neurons, and dopamine diffusion and clearance observed. Various concentrations of
dopamine were applied and maximal uptake (V max ) was calculated. Clearance of
exongenously-applied dopamine significantly decreased in the neuropil region
following treatment with 50 uM cocaine, a dopamine transporter inhibitor. Dopamine
clearance also significantly decreased in fumin mutants lacking a functional
dopamine transporter. The effect of acute cocaine application on dopamine clearance
is in good agreement with previous real-time, endogenous dopamine clearance
measurements in Drosophila larvae as well as measurements made in adult

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Drosophila. This method further validates the use of Drosophila as a model system to
study dopamine regulation and the role of the dopamine transporter in the underlying
mechanisms of drug addiction. This work was funded by NIH R01MH085159.

SUBSTITUTION VS. ELIMINATION - THERMODYNAMICS VS. KINETICS.
Charles M. Bump, Department of Chemistry, Hampton University. The competition
between substitution and elimination reactions is central to the study of organic
chemistr>'. It is a challenge to both the classroom teacher and to the student. When
reaction mechanisms are described in textbooks, generalizations are made such as
“Primary alkyl halides react with strong base / nucleophiles principally by an S N 2
mechanism and secondary alkyl halides react with strong base / nucleophiles
primarily by an E2 mechanism. Such generalizations correctly predict the reaction
product and mechanism, but leave the student without a clearly defined chemical
principle for the reaction path. We describe a systematic analysis of some
thermodynamic and kinetic parameters of substitution and elimination reactions
using the semi-empirical program MOPAC 2009. Free enthalpies of formation of
starting materials and products as well as free enthalpy of formation of substitution
and elimination transition states form the basis of the analysis.

CALCULATIONAL ORGANIC CHEMISTRY - A SUPPLEMENT TO
TRADITIONAL WET CHEMISTRY EXPERIMENTS. Charles M. Bump .
Edmund M. Ndip, Godson C. Nwokogu & Michelle K. Waddell, Department of
Chemistry, Hampton University. The ready availability of calculational chemistry
software and its widespread use in graduate education and industry presents a clarion
call for calculational chemistry to be taught with other fundamental lab techniques
such as distillation and recrystalhzation. The web site “Molecules in 4-D” (
http://wetche.cmbi.ru.n1//organic// ) provides an easy-to-use graphical interface to the
MOPAC semi-empirical calculation engine with pre-defined starting materials to be
edited for investigations by “users”, not just theoreticians. We will describe our
record-keeping requirements, procedures, and results in using this free resource for
teaching calculational chemistry with representative examples from the first and
second semester of the organic chemistry sequence. We will demonstrate that
calculations are fast enough that they can be conducted as real-time class
demonstrations by the teacher or assigned by the teacher to small groups or
individuals who report back to the class in five to ten minutes.

0 2 DEPENDENCE of SINGLET OXYGEN YIELDS FROM NATURAL
DISSOLVED ORGANIC MATTER (DOM). Charles M, Sharpless . Department
of Chemistry, University of Mary' Washington, Fredericksburg, VA. Natural
dissolved organic matter (DOM) is a heterogeneous mixture produced by decay of
plant and plankton biomass. Irradiating DOM in aqueous solution produces a suite
of photoreactants including singlet oxygen (’0 2 ), superoxide (0 2 ‘ and its dismutation
product, H 2 0 2 ), hydroxyl radical (OH), solvated electrons, and DOM triplet states
( J DOM*). These photoreactants can degrade aquatic pollutants and likely play a
part in the conversion of DOM to inorganic carbon (i.e., C0 2 ). Much of the
chemistry involves 0 2 , and it is important to establish how the rates of the various
reactions depend on dissolved 0 2 concentrations. Such studies can also provide

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49

insight into the DOM photochemical mechanism, allowing determination of such
properties as the excited state lifetimes of DOM precursors to various photoreactants.
This talk covers relevant background and presents the results of experiments showing
that DOM ! 0 2 quantum yields vary in approximate proportion to dissolved 0 2 over
environmentally relevant concentrations (DO ~ 50 to 250 DM). Modeling '0 2
production as arising from diffusion based reaction between DOM* and 0 2 indicates
that ’DOM* has a lifetime of approximately 5 to 10 s, regardless of the DOM
source.

ALUMINUM’S CONTROL IN THE BLUING OF HYDRANGEA SEPALS. Henry
D. Schreiber . Andrew H. Jones, Corinne M. Lariviere, Kelly M. Mayhew, & Judith
B. Cain, Department of Chemistry, Virginia Military Institute, Lexington, VA 24450.
The pigment in Hydrangea macrophyUa sepals is the anthocyanin
delphinidin-3-glucoside. The color of this pigment provides a red color to the sepals,
as the detphinidin-3-glucoside is in its flavylium cation structure. When grown in
acidic soil, hydrangeas readily assimilate aluminum which results in blue sepals. A
chemical model has been developed that attributes the blue color to AL + complexing
with delphinidin-3-glucoside in its quinoidal base anion structure along with a
stacked flavylium cation structure The role of A1 ’ + is to aid in H + transfer and act as
an anchor for this stacked complex The chemical model also predicts that the bluing
plateaus once a specific excess of Al’ + over the delphinidin-3-glucoside is achieved.
In the natural system, the color change from red-to-blue with aluminum can be
duplicated spectrophotometrically by this A1 3 ^-del phiinidin-3-g 1 ucoside complex
without recourse to co-pigments. The threshold aluminum content in hydrangea

-j i

sepals is 40 pg A1 /g fresh sepal for bluing to occur; this threshold is relatively
cultivar independent, despite the cultivar sepals having different
delphinidin-3-glucoside contents. More aluminum than the threshold did not enhance
the hue or intensity of the blue color of the sepal, in agreement with the chemical
model.

COMBINING COMPUTATIONAL AND EXPERIMENTAL APPROACHES TO
INVESTIGATE THE CATALYTIC MECHANISM OF GLYOXYSOMAL
MALATE DEHYDROGENASE- PART I. H, Guterres . S. Hedrick & E. Bell.
Deptartment of Chemistry, University of Richmond, Richmond, VA 23173.
Geometty optimization calculations have been used to find the minimum total energy
and structure of the transition state during the conversion of malate to oxaloacetate
catalysed by the enzyme glyoxysomal Malate Dehydrogenase, The proton abstraction
is suggested to precede hydride transfer due to the activity of the arginine residues
around the active site. To test this, two truncated models were built; model A
included Histidine 220, malate and NAD+, and model B included these three
moieties from model A with the addition of three arginine residues around the active
site, R124, R130, and R196. Gaussian 03W was used to build the structure of the
compounds, arrange their atoms, and run the geometry optimization calculations. For
model A, the result shows a single imaginary frequency that vibrates to the hydride
transfer. In contrast, for model B there is a single imaginary frequency that vibrates to
the proton transfer. The result suggests that the arginine residues around the active

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site play important roles in facilitating the proton transfer. In silico mutation of these
arginine residues is being correlated with the effects of in vitro site directed mutations
to ascertain the precise roles of individual arginines in binding and catalysis.

INVESTIGATING DIRECT PHOTOLYSIS MECHANISMS OF POLYCYCLIC
AROMATIC HYDROCARBONS IN NON-POLAR SOLVENTS. Jonathan S.
Williams & Charles M. Sharpless, Department of Chemistry, University of Mary
Washington, Fredericksburg, VA 22401 Polycyclic aromatic hydrocarbons (PAHs)
are toxic components of oil spills whose fate is often controlled through
photochemical reactions. PAH photolysis mechanisms in hydrophobic media are not
known, hampering efforts to predict their removal rates from oil in the event of a spill.
One hypothesis is that singlet oxygen ('0 2 ) generated by excited state PAHs is an
important intermediate that contributes to PAH photolysis. In order to test this
hypothesis, we are studying whether PAHs undergo a self-sensitized photolysis
mechanism involving '0 2 in solvents of similar polarity to oil. We employed a '0 2
probe, a-terpinene, to determine whether '0 2 is produced during irradiation of four
PAHs: benzo[a]pyrene, benzo[e]pyrene, benz[a]anthracene, and chrysene. Loss of
a-terpinene in irradiated PAH mixtures revealed that '0 2 is sensitized in hexane.
Additionally, [ 0 2 quantum yields were measured in hexane using a Stem-Volmer
analysis, yielding previously unknown values for benzo [a]pyrene and
benzo[e]pyrene at 0.84 and 0.70 mol Es' 1 , respectively. To determine if this
self-sensitized mechanism was occurring, we investigated whether the direct
photolysis quantum yield (4> D ) was linearly dependent on PAH concentration as
predicted by our kinetic model. Experiments showed non-linear plots of (|) D with
increasing PAH concentration, suggesting that another mechanism is contributing to
PAH photolysis in non-polar media.

Determination of an HPLC Method for the Quantitation of
Estradiol and its Seven Degradation Products, j. Chris Maxwell &
Geoffrey C. Klein, Department of Molecular Biology and Chemistry, Christopher
Newport University, Newport News, VA. Estradiol is an endocrine disrupting
compound that mimics endogenous estrogens found in humans and can directly
interact or interfere with and alter the normal growth, reproduction, or other
physiological function of an organism. The need for a highly sensitive and accurate
technique to measure the presence and concentrations of endocrine disrupting
compounds has generated several analytical approaches for the analysis of these
compounds. This work aims to determine an analytical method for the separation and
quantitation of estradiol and its 7 main degradation products. The HPLC method
determined to facilitate the most ideal separation used a 15-cm Cl 6 Polar Advantage
column (Dionex Corporation) with an isocratic mobile phase composed of 10%
tetrahydrofuran, 63% Phosphoric Acid Dilution (0.085%), and 27% Acetonitrile at a
constant flow rate of 1.5 mL/min for a total run time of 60 minutes. The detection
method used was UV/Vis spectroscopy which measured absorbance at a wavelength
of 225 nm. Each compound was identified individually using the developed method
but peak overlap of two compounds does not support the development of a clear
separation of estradiol and its degradation products. Future research will aim to

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51

create a more complete separation where each component is indicated by a distinct
non-overlapping peak and will explore the kinetics of estradiol photodegradation
under various environmental conditions.

Computer Science

A MINIMAL WORKING CONFIGURATION FOR THE ASTERISK VOIP
SERVER Luke D Acree & Robert M Marmorstein, Dept, of Computer Science,
Longwood University, Farmville, VA 23909. We present a fixed-point algorithm
for reducing a large set of configuration 1 ines to a minimal working configuration, A
minimal working configuration is die smallest set of configuration lines drat allow an
application to provide a pre-deflned level of service. We apply our algorithm to the
Asterisk voice-over-IP server using a baseline of one SIP and one analog phone
connection. Using the algorithm, we are able to reduce a configuration with more
than eighty files and 12, 000 lines of configuration to a simpler system containing
only four files and fifty-six lines. The algorithm finds a local minimum
configuration which, in practice, is usually die smallest working configuration set.
Reducing the number of lines of configuration makes teaching and training easier and
also makes it easier to customize a server to provide exactly the desired functionality
without introducing security flaws.

BIT PARALLEL SEARCH ALGORITHMS OF MOTIFS IN DNA USING TVCA.
S.V. Providence, Deptartment of Computer Science, Hampton University, Hampton,
VA 23668. Time-varying cellular automata (TVCA) are a form of cellular automata
CA where the transition functions vary over time or are time dependent. The DNA
motif search problem is a well-known problem in bio-informatics. Essentially it is
the problem of searching for patterns in biological sequences. Two classical
approaches utilize computation theory to exploit the power of deterministic finite
automata (DFA). A thud approach uses a filtering technique. CA is novel in motive
searching. We view a CA as an aggregate of NFAs or cells where the NFA transitions
are modified to account for neighboring cell states. There is a large body of literature
on CA and large body of literature on NFA in motif searching. In this paper we
extend work done in a previous paper by the author and the classical approach and
explain the usage of a TVCA in motif searching further.

CAN ILLEGAL AND UNETHICAL USE OF TECHNOLOGY BE EFFECTIVELY
DETERRED 1 ? Angela B. Hayden, Department of Computer Science, Hampton
University, Hampton VA 23668. Modern society is advancing at such a high rate of
speed that it is virtually impossible for an individual to keep up with the latest
technological advancement. The worst part is the rampant disregard for ethics and
ethical behavior when using technology. In an attempt to stay current with latest
technology, many individuals engage in illegal and unethical practices such as illegal
downloading of music or software piracy These activities can be seen as socially
acceptable behaviors. However, as computer scientist, we are well aware of the laws
and policies warning against such practices. Therefore, we ask ourselves, what is the

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best method for preventing illegal or unethical activity? Would the knowledge of the
laws, the possible execution of a penalty, a high probability of being caught, or an
enforcement of the penalty decrease the likelihood of a person engaging in illegal and
unethical activities?

ON E-MAIL REPUTATION. Bruce C. Chittenden, Computer Science, Governor’s
School for Science and Technology, New Horizons Regional Education Centers,
Hampton, VA 23666. Spam has been a serious problem for several years.
However, over the last couple of years and more recently in the last six months the
Internet Service Providers have been taking aggressive action to counter spam.
These actions include the creation of Blacklists to block Spammers’ IP Addresses, a
Spam Button on their e-mail User-Interfaces for the user to report spam. Feedback
Loops for e-mail senders so they may unsubscribe unhappy users, requiring e-mail
Senders to sign their e-mails, and other aggressive measures. An entire industry' has
emerged around e-mail reputation with such services as creating and managing
Sender Reputation Scores, maintaining Blacklists and Whitelists, reporting tools to
examine Mail Exchange Servers records, tools to create the infrastructure for signed
e-mails, and e-mail Service Providers that claim high deliverability rates. This paper
examines the actions that should be taken to improve and maintain an e-mail
reputation.

POPULATING THE MACONACHY CUBE Robert A. Willis Jr. Department of
Computer Science, Hampton University Hampton, VA 23668. The purpose of this
presentation is to explam the essential elements of tire Maconachy Cube as seen
above. The Maconachy Cube is a graplncal representation of possible security
scenarios one must account for in Information Assurance. Each element of the cube
can be seen as representing a triplet of indices in a three dimensional array. For
instance the triplet 1, 5, 2 can be seen are representing the transmission, non
repudiation, technolog)' tr iplet. The Maconachy Cube has been widely adopted as a
paradigm for Information Assurance excellence, education, curricula development
and for providing a “multidimensional view required to implement robust LA
programs”. This information assurance model covers every aspect of information
security. The presentation will also outline the development of the Cube using both
independent student research and a Software Development class. Keywords: Triplet,
Information Assurance, Maconachy, Information States, Security Countermeasures,
Security Services.

TRUSTWORTHY COMPUTING MODEL. Yen-Hung Hu, Department of
Computer Science, Hampton University, Hampton, VA 23668. In order for a system
to perform reliably, the mam components of such a system must be installed and
configured properly to provide appropriate operations to satisfy daily needs. These
components include hardware, operating system, network protocol application, and
data. However, even though a system seems to work properly, it doesn’t mean that
this system is trustworthy. A trustworthy system must be able to enforce protection
upon security, privacy, and usability. This research surveys and investigates several
existing mechanisms to enhance trustworthiness of each component, as well as

ABSTRACTS

53

studies their performance. A novel trustworthy computing model has been developed
for enhancing trustworthiness, as well as served as the criteria to assess trustworthy
computing systems.

POWER AWARE PARALLEL AND DISTRIBUTED COMPUTING USING
HETEROGENEOUS SYSTEMS. S. V. Providence and A. Alexander, Department of
Computer Science, Hampton University, Hampton, VA 23668. Energy efficiency is
among the more important issues in supercomputing and high performance
computing. We have a novel means of investigating power consumption on a cluster
computer system. We examine the iozone package which is traditionally used to
measure file system performance in Linux clusters and use it to give indications of
power consumption proportional to file system activity. Typical method use
dynamic voltage and frequency scaling which are describe in this paper. We then
explain our approach and postulate its efficacy.

DISTANCE ANALYSIS OF GENE EXPRESSION PROFILE. USING GLOBAL
SIGNATURES ASDIAGNOSTIC TOOLS. Lei Wang, Gamraju Manyam, Boris
Veytsman, Ancha Baranova. School of System Biology, George Mason University,
Fairfax, VA. Center of Liver Diseases, Inova Fairfax Hospital, Falls Church, VA.
Betty and Guy Beatty Center for Integrated Research, Inova Health System, Falls
Church, VA Department of Bio informatics & Computational Biology, The UT MD
Anderson Cancer Center, Houston, TX. The traditional protein or RNA biomarkers
usually suffer from either low specificity or unsatisfactory reproducibility. Here we
propose a novel approach that involves measuring global distance between entire
gene express profiles of normal and diseased tissues. Psoriasis, a complex
autoimmune disease which pathogenesis is still not fully understood, provides an
excellent model disease in which only a portion of a patient’s skin will be affected,
making it possible to study the difference between lesions, uninvolved skin from
patients and healthy skin from non-diseased controls. To address the performance of
global distances as biomarkers, we analyzed the data generated by microarray
experiments and used Mahalanobis distance as an estimation of the degree of
differentiation. The effectiveness of global signatures was compared with that of
traditional specific signatures, which are defined as highest ranked differentially
expressed genes. We demonstrated that global distances of gene expression profiles
can serve as reliable good classifiers, and therefore it is worth considering to use them
as diagnostic tools.

Education

SERVICE-LEARNING IN ENVIRONMENTAL STUDIES: THREE
SUCCESSFUL APPROACHES FOR UNDERGRADUATE EDUCATION.
Woodward S Bousquet . Environmental Studies and Biology' Departments,
Shenandoah University, Winchester, VA 22601 Over the past three decades,
service-learning in higher education has expanded considerably. More high school
students arrive with service experience, service is built into undergraduate courses,

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and it is added to university mission statements and graduation requirements.
However, to be effective, service-learning must not only meet a community’s needs.
It must also enhance student learning, advance the university’s mission and match
students’ needs and skills. Three successful service-learning approaches used in
Shenandoah University’s environmental studies courses since 1994 involve
preparing booklets and brochures, teaching through teaching stations, and forming
research partnerships. Booklets and brochures have a lasting impact and give
university students a tangible product to show others. Dividing a field trip for school
children into separate teaching stations cuts down on planning time and helps
university students hone their teaching techniques. Forming research partnerships
with local nongovernmental organizations and government agencies can connect
students’ service with actual community needs and provide ongoing opportunities for
future classes. All three of these approaches match student and institutional goals, fit
the structure of their respective courses, and sustain themselves in the curriculum by
building upon past projects’ accomplishments.

SCIENCE ON A SPHERE® AT THE SCIENCE MUSEUM OF VIRGINIA. E, G.
Maurakis , R. Conti, E. Alford, D. Olli. Science Museum of Virginia, 2500 W. Broad
Street, Richmond, VA 23220. Objectives of this program are to use the interactive
six ft diameter sphere and associated visualizations to educate the public about
climate change using inquiry based methods, and evaluate program outcomes. The
program focuses on the National Oceanic and Atmospheric Administration’s
(NOAA) learning goals in climate literacy: the sun is the primary source for earth’s
climate system, climate is regulated by complex interactions among components of
earth systems; life on earth depends on, affects, and is shaped by climate, climate
varies over space and time through natural and human-made processes, and human
activities are impacting the climate system Key messages are climate change is
occurring, detrimental aspects are greater than beneficial aspects, humans are causing
climate change, and humans can solve the problem Funded by NOAA award
NA10SEC0080023 and Science Museum of Virginia,

ON THE USE OF TECHNOLOGIES SUCH AS ALICE, GREENFOOT, AND
SCRATCH TO TEACH COMPUTER SCIENCE TO HIGH SCHOOL STUDENTS.
Bruce C. Chittenden, Computer Science, Governor’s School for Science and
Technology, New Horizons Regional Education Centers, 520 Butler Farm Road,
Hampton, VA 23666. While all of these technologies are very effective at
introducing the student to Computer Science each has its own strengths and
weaknesses and the technology needs to be chosen very carefully based upon several
factors. This paper examines the original design objectives of each of the
technologies, the Computer Science concepts that these technologies present, and
how these technologies are actually being used in the classrooms.

APPLICATION OF 21 st CENTURY LEARNING AND INNOVATION SKILLS
AT THE SCIENCE MUSEUM OF VIRGINIA. E. G. Maurakis. E. Alford, R.
Conti, E. Voelkel, R. Shires, and D. Smith. Science Museum of Virginia, 2500 W.
Broad Street, Richmond, VA 23220. Objectives of this program are to examine the
delivery of science, technology, engineering, and mathematics (STEM) information

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55

using 21 st century learning and innovation skills proposed by the Institute of
Museums and Library Services (TMLS) in programs and exhibitions at the science
museum. IMLS’s skills being implemented and evaluated are critical thinking and
problem solving, creativity and innovation, communication and collaboration, visual
literacy, scientific and numerical literacy, cross disciplinary thinking, and basic
literacy. Laboratoiy experiences (e.g. EcoLab), exhibitions (i.e.. Science
Unplugged), and programs (e.g. Summer STEM workshops) were evaluated using
IMLS 21 st centuiy learning and innovation skills, meeting most of the IMLS skill
categories.

AN INFORMAL SCIENCE APPROACH FOR RENEWABLE AND
NONRENEWABLE ENERGY CONCEPTS. David B.Hagan, Science Museum of
Virginia, 2500 W. Broad Street, Richmond, VA 23220. The Science Museum of
Virginia is a center of informal science education, generally defined as “science
teaching and learning that occurs outside of the formal school curriculum in places
such as museums, the media, and community-based programs.” (NSTA Position
Statement: Informal Science Education). The museum’s project presents
components in several forms of media focused on alternative and conventional
energy sources. These include Question Power , an original six-minute HD video
presenting the challenge of discovering clean, abundant energy sources, Watt Wall , a
large computer-driven museum wall display showing energy demand, and a set of
K-12 curriculum materials. The curriculum materials include a 2’ x 3’ original
detailed map of Virginia showing sites of sources of energy and delivery
mechanisms, including alternative and conventional sources. The curriculum
materials challenge common misconceptions about energy sources. Examples
include: “Wind and solar energy will provide all energy' needs in the near future”, and
“Conventional sources of energy will last indefinitely,” The underlying premise of
the K-12 materials is that every energy source has advantages and disadvantages.
With increasing global demand, currently known clean sources of energy cannot be
expected to serve the world’s needs using present technology in coming decades.
The search for solutions for the future lies with future generations.

BIODIVERSITY AND WATER QUALITY INVESTIGATIONS IN
SCHOOLYARD MESOCOSMS. H. S. Houtz 1 . A. B. Wright 2 & S. E. Marwitz 3 ,
VCU Life Sciences', VCU Dept, of Biology 2 , and Maggie L. Walker Governor's
School’. Mesocosms are experimental water enclosures that support naturally
occurring aquatic communities. Because they provide an easy means to
manipulate variables, mesocosm arrays have been used since the early 1960’s to
address a variety of scientific questions. Although mesocosm research has had a
long history, mesocosms are an underutilized teaching tool that can provide a
practical way for school children to design and implement simple scientific
investigations within school grounds. VCU’s Environmental Outreach Education
team has placed mesocosms in diverse habitats (wetland, forest, open field, etc.) at
VCU’s Rice Center and at 7 Richmond area public schools. Over the past 3 years,
high school and middle school students have measured water quality and monitored
colonization and development of macro invertebrate and algal communities.

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Ongoing student research projects aim to investigate the effects of canopy cover,
habitat isolation, and nutrient additions on community structure. Data have been
shared and analyzed collaboratively among the school systems. Biodiversity and
water quality have varied greatly seasonally and spatially among the sampling
locations, which range from inner-city Richmond to Charles City County, Virginia.
This project was made possible by the Howard Hughes Medical institute (HHMI).

Environmental Science

MODELING FISH SPECIES RICHNESS AND DIVERSITY IN LOWER
PIEDMONT STREAMS. E. G. Maurakis 1 ’ 2 , D. V. Grimes 3 , A. Schutt 4 , and S.
Short. 1 1 Science Museum of Virginia, 2 University of Richmond, 3 Virginia

Department of Environmental Quality, 4 VCU. Objectives were to model fish
species richness and diversity using biotic and abiotic environmental factors in
Quantico Creek (a forested watershed) and Cameron Run (an urban watershed),
Virginia. A total of 17 physical, chemical, and biological factors were used to
construct mathematical models to predict fish species richness and diversity in each
watershed. Fish species richness in Quantico Creek can be predicted using season,
stream order, elevation, river km, stream width and depth, watershed size and percent
undeveloped land cover, Fish species richness in Cameron Run can be predicted
with stream gradient, stream flow, water temperature, and percent undeveloped land
cover. Funded by U.S. Department of Energy Grant DE-FG02-08ER64625,

PRELIMINARY ANALYSIS OF BAY FILTER UNIT SUCCESS IN
FREDERICKSBURG, VA. MICHAEL L. BASS, UNIVERSITY OF MARY
WASHINGTON, FREDERICKSBURG, VA. Virginia Department of Conservation
and Recreation (DCR) evaluates and approves manufactured treatment devices
(MTD’s) deemed reasonable methods of prevention, control and/or treatment of
storm water runoff. Virginia’s stormwater management programs are implemented
under: Virginia Stormwater Management Law and Virginia Stormwater Management
Regulations. DCR maintains tire authority to regulate BMP methods used in Virginia
to control stormwater runoff under the Virginia Technology Assessment Protocol
(VTAP). The assessment protocol deals with the MTD’s that are designed for,
reducing stormwater runoff volume, reducing peak runoff rate and/or and reducing
total phosphorous (TP). The goal of the VTAP regarding runoff quality control is to
determine how much a specific MTD can remove total phosphorous (TP). MTD’s
seeking certification for runoff quality control in Virginia will only be approved for
TP removal at this time, requiring 50% TP removal for influent with TP
concentrations ranging from 0.15 mg/L to 0.5 mg/L. Additional requirements are
80% removal of TSS for influent with TSS concentrations ranging from 100 mg/L to
200 mg/L and > 80% removal of TSS for influent with concentrations greater than
200 mg/L. Baysaver Technologies, Inc has applied for interim approvals to use the
Bay Filter System to meet Virginia requirements for treating stormwater runoff. Flow
through the filter system in gravity-driven and self-regulating. The monitoring
program is intended to demonstrate through field testing that Bay Filter is capable of

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57

removing contaminants from stormwater runoff. The field test will demonstrate the
removal efficiencies attained by the system for TSS, TP, Cu, Zn and other pollutants.
This will then be used to confirm that the system meets stormwater regulations which
require the removal of a minimum 80% of the total suspended sediment load and
treatment of nutrients to the maximum extent feasible. The field testing program will
collect discrete samples from die influent and effluent of the BayFilter. These
samples will be analyzed usmg standard EPA protocols for total suspended solids
(TSS), particle size distribution (PSD), nutrients as well as metal concentrations.
Removal efficiencies will be calculated based on this data using standard scientific
methods. Precipitation and flow records will be aquired during these events as well.
The testing program is anticipated to take 12-18 months to complete and will include
at least 15 qualifying storm events. BayFilter systems to be monitored will treat the
stormwater runoff from Trinity Episcopal Church property in Fredericksburg, VA.
Stormwater runoff from the paved area transports dissolved, colloidal, suspended and
settleable solids in a heterogeneous mixture, which includes metals, organic
compounds and nutrients. These constituents result from atmospheric deposition,
traffic activities, vehicular wear, pavement degradation and deicing, landscape
maintenance and littering. The nutrient load from the site is expected to vary
seasonally.

ASSESSING WATER QUALITY OF CEDAR CREEK USING BENTHIC
MACROINVERTERBRATES. John V, Stevens . Briana L. Barron, James E.
Bisset, Linden E. Lewis, Daniel A. Million, Cory M. Miller, Benjamin S. Sawyer,
Amy L. Smith & Woodward S. Bousquet. Environmental Studies Department,
Shenandoah University, Winchester Virginia 22601. In the spring and fall of 2010,
Shenandoah University researchers assessed the water quality of Cedar Creek in
Frederick and Shenandoah Counties, Virginia. They used the Environmental
Protection Agency’s Rapid Bioassessment Protocols (RBPs) to select sampling sites
and collect specimens In addition to choosing three representative sites on the
creek’s main stem, the researchers selected two locations on ecologically distinct
tributaries: Paddy Run (a cold-water montane stream), and Meadow Brook (a
warm-water valley stream). Analysis was performed with the Virginia Stream
Condition Index (VSCI), a multimetric measure based on the diversity, pollution
tolerance and feeding categories of the invertebrates collected. A majority of the sites
had a VSCI water quality score of >73, placing them in the excellent category. Only
Meadow Brook was rated as severely unpaired (avg. VSCI score = 32.2), most likely
due to surrounding agricultural and residential development. These findings affirm
previous studies that rated Cedar Creek’s overall water quality as among the best in
the Shenandoah Valley region, using chemical, physical and fish community data.
Best management practices should be implemented to protect this beautiful,
high-quality watershed

Medical Science

THE ROLE OF a5 NICOTINIC ACETYLCHOLINE RECEPTORS IN ACUTE
AND CHRONIC ALCOHOL BEHAVIORS IN MICE. Anton J. Dawson & M. Imad

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Damaj. Dept, of Pharmacology & Toxicology, Virginia Commonwealth University,
Richmond, VA 23298. The high incidence of nicotine dependence in alcoholics
suggests that nicotinic acetylcholine receptors (nAChRs) may be involved in both
nicotine and alcohol dependence. Genetic studies suggest that the a5 nAChR-coding
gene is significantly associated in both nicotine and alcohol dependence phenotypes,
but virtually no studies have investigated this receptor subtype with regard to
alcohol's effects in vivo. We hypothesized that the o.5 nAChRs mediate some
acute and chronic behavioral effects of alcohol in alcohol preferring C57BL/6J mice
lacking a5 gene expression (a5 KO mice). We tested their response to alcohol’s acute
effects including initial sensitivity and functional tolerance through the loss of
righting reflex assay, anxiolysis using elevated plus maze, hypothermia, and
locomotor depression. For chronic effects, we chose to study alcohol intake and
preference using two-bottle choice and aspects of alcohol reward using conditioned
place preference. Our results showed that a5 KO mice, indeed respond differentially
to acute and chronic ethanol exposure. These data suggest that a5 nAChRs may
exert differential effects on the acute and chronic effects of alcohol and may,
therefore, represent a potential therapeutic target for the treatment of alcohol and
nicotine co-abuse in the future.

FACTORS RELATED WITH ACTIVE LIVING FOR INDIVIDUALS IN THE
WESTERN REGION OF USA. S. Tiraphat . Q Zhang, J.G. Behr, & L. Shepherd.
Old Dominion University, Norfolk, VA 23529. Adequate physical activity is a
major contributor to a healthy living and prolonging life. Among extreme geographic
diversity' of the US, the disparity in physical activity (PA) has remained substantially
across the region, with the Western part having the highest activity prevalence. It is a
challenge to study and identify what factors influence on active lifestyle of people in
the Western area. Objective: this study is to investigate the influences of individual
and environmental contexts on the odd of being physically active of people in the
Western region. Method data from 2007 BRFSS is obtained and linked with external
environmental data sources from US Census Bureau, USD A, and NORSIS databases.
A two-level logistic regression is analyzed. Outcome variable is meeting PA
recommendation according to CDC guideline (yes / no) Individual-level variables
include demographic factors, socio-economic conditions, lifestyle, and health status.
Environmental -level variables as a county scale include physical activity built
environments (local facilities, green area, and outdoor activity resources),
educational context, crime rate, and natural amenity. Results: At the individual level.
White, male, consuming more fruits and vegetables, more satisfied with life, higher
income, higher education, better health, and younger are significantly positive with
the meeting PA recommendation. At the county level, outdoor activity resources
and educational context are the significant predictors of the meeting PA. This study
emphasizes the advantage of built environments especially outdoor activity resources
on active living of people in the Western part.

REGIONAL GENE EXPRESSION CHANGES ASSOCIATED WITH
INCREASED VOLUNTARY ETHANOL CONSUMPTION DUE TO
DEPRIVATION IN C57BL/6J AND C57BL/6NCRL MICE. J.A, Warner , R.T.
Khisti & M.F. Miles, Virginia Commonwealth University. The alcohol deprivation

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59

effect (ADE) is an increase in ethanol consumption after abstinence and is considered
a rodent model of relapse in human alcoholics. This study used microarray analysis to
identify changes in gene expression in nucleus accumbens, prefrontal cortex, and
ventral midbrain of C57BL/6NCrl mice after ethanol deprivation. Nearly 800 mRNA
transcripts were significantly regulated, including several primary targets of ethanol,
with most regulation in NAC. Analysis revealed over-represented biological
processes, cell signaling pathways, and transcription factor binding sites, as well as
novel coordinately regulated gene networks, with the common theme of
neuroplasticity. Areas highly represented included calcium, sodium, potassium, and
chloride currents, transcription and translation, neuronal remodeling, and control of
cell fate. Components of GABA, glutamate, opioid and serotonin systems were
regulated. Binding sites for diverse transcription factors, including CREB, were
over-represented. Novel gene networks were identified related to gene expression,
protein trafficking, and cellular function and maintenance. Regulated genes likely
alter neuronal function by modulating excitability, protein turnover, and connectivity,
and represent novel targets for further development of pharmacological and genetic
therapies for relapse in alcoholism.

THE CENTRAL MECHANISMS OF GASTRIN-RELEASING
PEPTIDE-INDUCED ANOREXIA. C. R. Dougherty & M. A. Clme Dept, of
Biology, Radford University, Radford, VA 24142. Recently our laboratory
demonstrated that intracerebroventricular (ICV) injection of gastrin-releasing peptide
(GRP), a member of the bombesin family, caused primary anorexigenic effect in
broiler chicks. Currently the central mechanisms behind the anorexigenic effect of
GRP are poorly understood. We hypothesized that this effect is likely mediated
through similar neurotransmitters that are known to be involved in hypothalamic
appetite regulation. c-Fos immunoreactivity was used in order to visualize changes in
neuronal activity in regions primarily associated with satiety and appetite regulation
as a result of ICV administration of GRP. We are currently in the process of further
studying the central mechanisms of GRP via real-time PCR Further investigation
into the central mechanisms of GRP-induced anorexia could have significant
implications in the management of obesity if pharmaceutical applications are
pursued.

OPIOID AND HIV-1 ASSOCIATED NEURODEGENERATION: POSSIBLE
REGULATION BY THE P2X 4 RECEPTOR IN PRIMARY MOUSE STRIATAL
CELLS. ME Sorrell , S. Zou, P.E. Knapp, and K.F. Hauser, Department of
Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, VA
23298. HIV-I-associated neurocogmtive disorders (HAND) are seen in up to 50%
of AIDS patients and individuals who abuse opiates can have an increased incidence
of HAND with more severe symptoms. Microglia play a role in HIV-1
neuropathogenesis since activation produces cytokines, chemokines, and other
inflammatory molecules that can lead to neuronal injury and death. Recently,
morphine has been reported to increase microglial motility by modifying P2X 4
signaling. This is important because mu opioid receptor agonists can increase HIV-1
replication, potentiate the release of oxyradicals and glutamate, and increase cytokine
production in HIV-1 Tat-exposed microglia. To examine whether HIV-1 and/or

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opioid-induced microglial activation and neurotoxicity are mediated via purinergic
signaling, co-cultures of primary neurons and mixed glia from mouse striatum were
treated with vehicle. Tat and/or morphine ± TNP-ATP, which blocks P2X 1 _ 7
receptors. Individual neurons were repeatedly tracked for 72 hours and neuron
survival vs. time was assessed. In neurons co-exposed to Tat ± morphine, treatment
with TNP-ATP significantly blocked Tat-mediated neurotoxicity. Further neuron
survival studies showed drat TNP-ATP dose dependently prevented Tat and
morphine toxicity, both when treated in combination and separately. Our data
implicates that the activation of P2X receptors are critical in opiate and HIV
neurotoxicity, while further work needs to be done to verify the role of the P2X 4
receptor in our HIV and opioid model.

THE IMPACT OF ADOLESCENT NICOTINE EXPOSURE ON DRUG
DEPENDENCE IN ADULTHOOD. Mai Alajaji, M S. & M. Tmad Damaj, Ph D
Department of Pharmacology & Toxicology Virginia Commonwealth University,
Richmond, VA 23298. Smoking among adolescents is a strong predictor of future
drug abuse and dependence in adulthood. A number of studies have suggested that
adolescents pre-exposed to nicotine may suffer permanent disruption of the brain’s
reward systems through changes in dopamine receptor function. We hypothesize that
nicotine exposure during adolescence causes long lasting neurobiological alterations
that increase the likelihood of cocaine use in adulthood. Conditioned place preference
data showed that a 7-day exposure to 0.5 mg/kg of nicotine altered cocaine-induced
responses. In contrast, neither 1 day exposure nor a low dose of nicotine (0.1 mg/kg)
elicited this effect. A follow-up study was undertaken to determine if this
enhancement generally applies to other drugs of abuse. Pre-exposure to 0.5 mg/kg
nicotine during early adolescence demonstrated significant enhancement to
d-amphetamine and morphine preference in a CPP model. Similar to the effects seen
with reward, exposure of early adolescent mice to nicotine also enhanced acute
locomotor activity and locomotor sensitization to cocaine in adulthood. Our data
strongly suggest that nicotine intake during adolescence may act to cross-sensitize the
brain to cocaine’s long-term changes in the brain. Further research will be required in
order to more fully examine the mechanisms of action for the observed changes in
cocaine rewards.

CHANGES IN GENE EXPRESSION DURING IMCD3 CELL
DIFFERENTIATION. Brittany N, Abbatiello & Deborah L. Zies, Dept, of Biol.
Sci., Umv. of Mary Washington, Fredericksburg, VA. Renal Cell Carcinoma (RCC)
accounts for 90% of kidney cancer cases and is extremely difficult to treat.
Understanding the mechanisms by which renal cells become de-differentiated during
the transformation process is an important step in understanding this deadly disease.
Identification of the genes involved in kidney cell differentiation may lead to a better
understanding of the de-differentiation process. Therefore, the goal of this study is
to identify genes involved in the differentiation of kidney cells. We determined
growth conditions for undifferentiated and differentiated IMCD3 cells. Cells were
plated into 6 well plates and duplicate wells were lysed each day for 9 days. RNA
was collected, converted to cDNA, and QPCR was performed for Glyceraldehyde
3-Phosphate Dehydrogenase (GAPDH) and Epithelial sodium channel subunit alpha

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61

(ENaCa). GAPDH expression did not change over time. IMCD3 cells were
determined to be differentiated when QPCR results correlated with expected increase
in expression of ENaCa. The RNA from undifferentiated samples (day 4) and
differentiated samples (day 7) were compared in a microarray experunent.
Bioinformatic analysis of the results identified four candidate genes, neuronal
guanine nucleotide exchange factor (NGEF), roundabout homolog 1 (ROBOl),
transforming growth factor 1 beta (TGFB1), and brain derived neurotropic factor
(BDNF). These genes differ in expression between undifferentiated and
differentiated cells and have functions blown to be associated with development or
differentiation. Future studies will characterize these genes to determine if they play
a role in cell differentiation or cancer cell de-differentiation.

ASPECTS OF ETHANOL RESPONSE IN C. ELEGANS. J. Alaimo . A. Davies & J.
Bettinger, Pharm/Tox, VCU, Richmond VA 23298. Alcohol abuse is a disorder with
a poorly understood etiology that includes both genetic and environmental influences.
One factor found to influence drinking behavior is variation in genes encoding
ethanol metabolism machinery. Alcohol dehydrogenase (ADH) and aldehyde
dehydrogenase (ALDH) are two major enzymes involved in the ethanol metabolic
pathway. We directly tested the effects of altering the function of these enzymes on
ethanol responsive behaviors in the nematode, C. elegans. Through homology
searches we have identified two ADH-like enzymes that are encoded by sodh-1 and
H24K24.3. We have found that at 200mM and 400mM exogenous ethanol, worms
lacking SODH-1 are hypersensitive to the initial effects of ethanol relative to wild
type, but knockdown of H24K24.3 caused no significant behavioral changes at either
dose. We tested if this sensitivity to ethanol reflected an increase in internal ethanol
concentrations and found that tissue concentration is increased in sodh-1 mutants, but
not in H24K24.3 mutants. Moreover, sodh-1 and H24K24.3 mutants develop acute
tolerance to ethanol suggesting that these components of the alcohol metabolism
machinery' are dispensable for this process The C. elegans genome encodes 13
ALDH enzymes that are highly conserved between worms and humans. We have
found that knockdown of either alh-6 or alh-13 results in altered ethanol sensitivity at
400mM exogenous ethanol, but internal tissue concentrations trend towards an
increase relative to wild type. This suggests that the lack of ALDH function may
cause a buildup of acetaldehyde, which would be converted by ADH into ethanol.
Collectively, we find that altered ethanol metabolism in worms results in a mild effect
on ethanol response behaviors.

GENE NETWORKS OF ALCOHOL DEPENDENCE. S.P. Farris . K.L. Shelton, &
M.F. Miles. Department of Pharmacology and Toxicology, Virginia Commonwealth
University', Richmond VA 23298. The molecular mechanisms underlying alcohol
abuse and alcoholism are largely unknown, however, divergent patterns in gene
expression are critical molecular endophenotypes regulating the risk of developing
alcohol dependence. Using a mouse model of intermittent ethanol self-administration
we have conducted Weighted Gene Co-expression Network Analysis (WGCNA) of
medial prefrontal cortex (PFC) to identify gene expression networks associated with
ethanol intake. WGNA revealed a significant correlation of ethanol intake to
myelin-associated gene expression. Prior work from our laboratory and others’ have

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identified decreased expression of inyelin in the PFC of alcoholics. Additionally, our
laboratory has shown myelin gene expression may be a latent factor influencing acute
ethanol sensitivity winch influences long-term drinking behavior. Therefore, we
hypothesized a myelin network functions in acute ethanol behavioral sensitivity.
Using the cuprizone model of demyelination in C57BL/6J male mice we determined
that differential expression of a myelin gene network impacts the duration for the loss
of righting reflex, a phenotypic measure of acute ethanol behavioral sensitivity.
Cuprizone treated mice and controls exhibited no difference in blood ethanol
concentrations, suggesting the phenotypic impact of ethanol administration is a
pharmacodynamic effect of myelin and not due to non-specific effects on the
pharmacokinetics of ethanol. Our results suggest myelin may be a molecular
endophenotype influencing acute behavioral sensitivity and the risk of developing
alcohol dependence. Supported by NIAAA grants U01 AAO16667 to MFM and F31
AA018615 to SPF.

GENE EXPRESSION OF GASTRIC CYTOKINES AND HORMONES IN
OBESITY ASSOCIATED LIVER INFLAMMATION. R. Mehta :[ A A. Birerdinc 1,2 ,
L. Wang 1,2 , N. Hossain 2,3 , A. Afendrty V. Chandhoke 12 , Z. Younossi 1,2,3 & A.
Baranova 12 'Molecular and Microbiology Department and Center for the Study of
Genomics in Liver Diseases, George Mason University, Fairfax, VA, USA.
2 Translational Research Institute, lnova Health System, Falls Church, VA, USA.
^Center for Liver Diseases, Inova Fairfax Hospital, Falls Church, VA, USA.
Obesity is one of fastest growing global epidemics. Obesity is a complex multisystem
disease resulting from a failure in the normal homeostatic mechanisms which
regulates food intake, fat storage, and energy utilization. Further in obesity, there is
interplay of substances produced by various organs, including visceral adipose, liver,
muscle and gastric tissues. Obesity is strongly associated with numerous
degenerative conditions, particularly non-alcoholic fatty liver disease (NAFLD)
Consequently, deregulation of genes involved in energy homeostasis and expenditure
may play an important role in not only obesity, but also the pathogenesis of the
diseases stemming from this condition. Soluble molecules produced by gastric tissues
and their receptors may play a role in the pathogenesis of obesity-related NAFLD In
this study, we assessed the association of gastric tissue gene expression of obesity
related genes and inflammatory cytokines with NAFLD in obese patients. We
demonstrate a correlation of gastric CNR1 and cytokines with hepatic inflammation.
This data indicates that gastric tissue may play a previously unexplored role in the
pathogenesis of obesity associated NAFLD. With further validation in a larger
cohort, the differentially expressed genes detected m this study may provide novel
mechanistic insights mto the pathology of NAFLD.

DYNAMICS OF SUBSTRATE INTERACTIONS IN tRNA (m 1 G37)
METHYLTRANSFERASE: IMPLICATIONS FOR DRUG DISCOVERY. M, K.
Palesis 1 . J. C. Hackett 2 , & W.M. Holmes 3 , Departments of 'Pharmacology and
Toxicology, 2 Medicinal Chemistry, and 3 Microbiology, Virginia Commonwealth
University, Richmond, VA 23298. The bacterial enzyme t-RNA (m , G37)
methyltransferase (TrinD), a potential antimicrobial drug target, methylates the G37
nucleotide of tRNA using S-adenosyl methionine (SAM) as the methyl donor. This

ABSTRACTS

63

methylation reduces the occurrence of frame shift mutations during protein
translation. Molecular dynamics simulations of TrmD implicate secondary structural
elements which may contribute to active site accessibility. Substrate-enzyme
interactions were studied using Isothermal Titration Calorimetry (ITC). Molecular
dynamics simulations together with ITC data suggest cooperativity between two
active sites on the enzyme. To further investigate this cooperativity, a series of
experiments have been initiated. Because SAM is a ubiquitous substrate for many
different methyltransferases in the cell, analogs of this ligand may not be suitable for
drug development. It is therefore important to understand the structural elements
involved in the mechanism of action which will subsequently enable a more targeted
approach to the development of new antimicrobial drugs.

COMBINING CANNABINOIDS AND RADIATION THERAPY IN BREAST
CANCER. S.M. Emery . E, Sumner, A. Lichtman & D.A. Gewirtz, Department of
Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, VA
23298. Numerous reports have shown that in several preclinical models
cannabinoids have the potential for improving the effectiveness of cancer treatment.
Given that cannabinoids are already used as a palliative treatment during cancer
therapy, we investigated whether select agents could enhance the effectiveness of
radiation, which is used in treatment of breast cancer. We assessed the impact of
Win55, 212-2 (Win2), a full efficacy cannabinoid agonist, as well as Cannabidiol
(CBD) and A9- Tetrahydrocannabinol (THC) in combination with radiation in
MCF-7 (human p53 wild type), MDA-MB-231 (human p53 mutant), and 4T1 (mouse
p53 null) breast tumor cell lines as well as in MCE-10a cells, a human
non-transformed cell line. Win2 significantly enhanced the antiproliferative
actions of radiation (2 Gy) in MCF-7, MDA-MB-231 cells as well as 4T1 cells but
not in MCF-lOa cells. Unexpectedly, CBD and THC did not produce the same
effect as Win2, suggesting differences between the compound classes. We also
were able to show in MCF-7 cells that when using Win55,212-3 the inactive
enantiomer of Win2 there was no effect alone as well as no enhancement with the
combination. Our data indicates that Win2 (but not THC) has the potential to enhance
current radiation therapy used against breast cancer.

miRNAs AND THEIR TARGET GENE EXPRESSION IN THE VISCERAL
ADIPOSE TISSUE OF NON-ALCOHOLIC FATTY LIVER DISEASE (NAFLD)
PATIENTS. H. Sharma 1 y M. Estep 1 , N. Hossain 1 , H. Elarainy 1 , Z. Goodman 23 , V.
Chandhoke 2 , A. Baranova 2,3 , Z. Younossi 1 2 ’ 3 , 'Betty and Guy Beatty Center for
Integrated Research, Inova Health System, Falls Church VA, 2 School of Systems
Biology, George Mason University, Manassas, VA and ’Center for Liver Diseases,
Inova Fairfax Hospital, Falls Church , VA There are many biological molecules
involved in the pathogenesis of Non-Alcoholic Steatohepatitis (NASH) and its
progression from NAFLD. miRNAs are known to inhibit the translation of their
target mRNA. At least 113 species of miRNAs are differentially expressed in patients
with NASH within visceral adipose. In this study, we set out to measure the
differential levels of expression for six miRNA target genes and the components of
the miRNA processing apparatus in patients with simple steatosis vs NASH We
discovered that while in NASH patients expression levels increase for only two

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pre-processed miRNAs, for miRNA processing enzymes mRNA level increase
throughout the pathway. When the expression levels of several NASH-related genes
were studied, we observed concerted changes in the expression levels of the target
genes (increased) and levels of miRNA responsible for regulation of these genes
(decreased). This observation suggests that regulation by miRNA may be involved in
the pathogenesis of NASH. These results warrant additional research in order to
establish the precise role of the miRNA regulated genes and their miRNA regulators.

THE ROLE OF CCR5 IN HIV-1 TAT AND MORPHINE MEDIATED
NEURODEGENERATION. E M Podhaizer '. Y. Zhang 2 , P.E. Knapp 13 , & K.F.
Hauser 1 , Depts. of 'Pharmacology & Toxicology, 2 Medicinal Chemistry, ’Anatomy
& Neurobiology, Virginia Commonwealth University, Richmond VA, 23298.
Injection drug use, such as heroin abuse, is closely linked to both the spread and
pathogenesis of HIV-1. The effects of HIV-1 Tat, in models of HIV-1 infection, have
been shown to be worsened with the addition of morphine and include measures of
neurotoxicity, dendritic pathology, and astrocyte calcium and cytokine/chemokine
release. CCR5 has been suggested to be involved in Tat-induced signaling through
Tat's effects on pro-inflammatory CCR5 ligand secretion. Additionally, morphine
and the mu opioid receptor have been shown to regulate CCR5 ligand secretion and
both receptor expression and protein levels. Thus, it was hypothesized that inhibition
of CCR5 would be protective against neurodegeneration and glial inflammatory
signaling. Neurotoxicity was examined in neuronal-mixed glial co-cultures treated
with morphine (500 nM), Tat (100 nM), or the combination, in the presence or
absence of the CCR5 antagonist maraviroc (50 nM). Maraviroc pretreatment
prevented the synergistic neurotoxic effects of collective Tat and morphine treatment.
CCR5 inhibition in cultures heavily enriched in astrocytes suppressed both RANTES
and MCP-1 chemokine production, but had no effect on TNF-a. This data suggests
that CCR5 is a critical convergence point of Tat and morphine signaling. Supported
by NIHP01 DA019398 & T32 DA007027.

NADPH OXIDASE-MEDIATED INFLAMMASOME ACTIVATION BY
ELEVATED HOMOCYSTEINE IN MOUSE PODOCYTES AND GLOMERULI
J. M. Abais . C. Zhang, M Xia, K Boini, E A Laperle, A. M, Thacker & P-L. Li,
Department of Pharmacology & Toxicology, Virginia Commonwealth University,
Richmond VA 23298. Hyperhomocysteinemia (hHcys) is an important pathogenic
factor contributing to the progression of end-stage renal disease. Recent studies in
our laboratory have found that Hcys can activate the mflammasome, potentially
turning on the inflammatory response. Hcys is also known to activate nicotinamide
adenine dinucleotide phosphate (NADPH) oxidase, and this present study was
designed to test whether NADPH oxidase-mediated redox signaling contributes to
Hcys-mduced inflammasome activation m podocytes. Confocal microscopy showed
that Hcys caused a 2.5-fold increase in colocalization between the 3 major
inflammasome components - NALP3, apoptosis-associated speck-like protein
(ASC), and caspase-1. Furthermore, Hcys increased caspase-1 activity, IL-1(3, and
superoxide production, and all of these changes in podocytes were significantly
blocked by inhibition of NADPH oxidase using apocynin, gp91 /,/,ov peptide, and
gp91 phox siRNA. In vivo , gp9 l phox knockout mice protected against inflammasome

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65

formation and dysfunction induced by hHcys. These results suggest that
Hcys-induced NADPH oxidase activation is importantly involved in turning on
NALP3 inflammasomes, making both NADPH oxidase and the inflammasome
potential therapeutic targets for preventing Hcys-induced glomerular sclerosis.

ROLE OF ALPHA7 NTCOTINIC ACETYLCHOLINE RECEPTOR IN A MURINE
DEXTRAN SULFATE SODIUM-INDUCED COLITIS MODEL. S. AlSharan . H.
Akbarali, & M.I. Darnaj. Department of Pharmacology & Toxicology, Virginia
Commonwealth University, Richmond, VA 23298. Substantial evidence suggests an
association between cigarette smoking and the incidence and severity of ulcerative
colitis (UC), a common human inflammatory bowel disease (1BD). Nicotine seems to
be a key mediator of this response as has been demonstrated by the use of transdermal
patches and nicotine enemas The mechanism through which nicotine acts is not well
understood. Identification of nAChR subtypes and their role in UC would lead to
better understanding of the disease and the development of better pharmacological
treatment of UC. We investigated the role of a5, 0 . 6 , al and (32 nAChR subtypes in a
murine model of UC in which dextran sulphate sodium (DSS) to induce colitis in C57
mice and measured a Disease Activity Index (DAI). In addition, al KO mice
displayed a significantly increased in DAI value (diarrhea, irritation around the anal
area, hematochezia, and loss of body weight) compared to their WT litter mates
Furthermore, treatment with PHA-543613, a selective al agonist, significantly
reduced DAI signs in DSS-treated mice compare with vehicle DSS-treated mice
Moreover, we use PNU-120596, a positive allosteric modulator, and Choline
chloride, an al nAChR full agomst, were both reduced DAI signs in DSS-treated
mice. These results suggest that al nAChR has a protective role in colitis.

THE ROLE OF 2-AG ENDOCANNABINOID NEUROTRANSMISSION IN
NICOTINE REWARD AND WITHDRAWAL. P. Muldoon . A. Lichtman, & M.I.
Damaj, Ph.D. Department of Pharmacology and Toxicology, Virginia
Commonwealth University, Richmond, VA 23298. Nicotine is the main addictive
component of tobacco that plays a major role in dependence. Emerging evidence
suggest that the endogenous endocannabinoid system may modulate these effects.
2-AG is the most abundant endocannabinoid in the brain, required for retrograde
transmission and exerts its action via CB1 receptors. We hypothesize that 2-AG
neurotransmission is altered during Nicotine Dependence (ND). We first evaluated
2-AG levels in relevant brain regions after acute administration of nicotine (1,
2mg/kg s.c.) using liquid chromatography-mass spectrometric (LC-MS) method.
Next, assessed the effect of repeated nicotine injections (2 times a day for 7 days)
on 2-AG levels in LC-MS. Unfortunately, in either treatment group we did not see
an effect of nicotine on 2-AG neurotransmission. However, this effect could be due
to the sensitivity of the assay. Secondly, there could be a dilution of effects since we
take whole tissue. We then wanted to measure the effect of enhanced 2-AG via
MAGL inhibition by JZL184 in ND. JZL184 is a potent and selective inhibitor of
MAGL. Nicotine reward in the mouse was evaluated in an unbiased conditioned
place preference paradigm (CPP) in induction. Our results showed that degradation
of MAGL dose-dependently decreased nicotine preference compared to nicotine
control in our CPP paradigm. Finally, we assessed JZL184's effect on nicotine

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withdrawal. Our results verified that JZL184 dose-dependently decreased somatic
signs. Our results suggest that MAGL inhibition may be a therapeutic target in
ND. P50DA005274

DETERMINING RAI1 BINDING SITES WITHIN CLOCK. C.A. Bax 1 , S.
Williams 2 , S. Elsea 2 , & D.L. Zies 1 , Department of Biological Sciences, University of
Mary Washington, Fredericksburg, VA 22401, "Virginia Commonwealth University,
Richmond, VA 23284. Smith-Magenis Syndrome (SMS) is a neurobehavioral
disorder characterized by mental retardation, sleep disturbances, obesity and self
injurious / attention seeking behavior. The major symptoms of SMS are caused by
haploinsufficiency of the retinoic acid-induced 1 (Kill) gene, which is thought to be
a transcription factor involved in the regulation of many other genes. A Chromatin
Immunoprecipitation-microarray chip (ChlP-chip) experiment was conducted to
determine genes regulated by RAH. One of the genes identified was Circadian
Locomotor Output Cycles Kaput (CLOCK). The purpose of this project was to
confirm RAI1 as a regulator of CLOCK and identify specific binding sites within the
gene. Transient transfection and luciferase assays were performed. A statistically
significant increase in transcription of luciferase was seen when RAI1 was
cotransfected with the regulatory region of CLOCK. No significant drop in
luciferase levels was seen when the shortened CLOCK construct was cotransfected
with RAI1. Therefore, the binding site is located within the 227 bases that were not
removed by the deletion. The identification of a specific RA11 binding site would
enable researchers to identify other genes regulated by RA11. Knowing the cohort
of genes regulated by RAI1 would provide a better understanding of the specific
function of RA11 and the downstream effects that lead to the symptoms of SMS.

BOTH POLYCYSTIC OVARY SYNDROME (PCOS) AND NON-ALCOHOLIC
FATTY LIVER DISEASE (NAFLD) ARE DISEASES OF METABOLIC
SYNDROME SPECTRUM. T P. Tran 1 - 2 . A. Baranova 1 ’ 2 , A. Birerdinc 12 & Z.M.
Younossi"' , Betty and Guy Betty Center for Integrate Research, Iona Health System,
Falls Church VA 22042, 2 Center for the Study of Genomics in Liver Diseases,
Molecular and Microbiology Department, George Mason University, Fairfax VA
22030, 3 Center for Liver Diseases, Inova Fairfax Hospital, Falls Church VA 22042
Obesity has been widely known to cause non-alcholic fatty liver disease (NAFLD)
and aggravate polycystic ovary syndrome (PCOS). Insulin resistance (IR), a
hallmark of metabolic syndrome is observed in 50-80% of women with PCOS and
patients with NAFLD. Here, we performed a systematic review using
PubMed-search for peer-reviewed articles related to PCOS and NAFLD Based on
the association of PCOS and other metabolic abnormalities, such as IR,
hyperandrogenism, obesity and NAFLD, the candidate genes have been proposed
for PCOS. Closer scrutiny of these genes places most of their proteins at the
crossroads of three highly inter-related conditions: metabolic syndrome, obesity and
NAFLD. We showed that the prevalence of both PCOS and NAFLD rises
proportionally to the degree of IR and increases in the mass of adipose tissue. These
conditions often co-exist and may respond to similar therapeutic strategies. We
suggest that it is appropriate to consider PCOS as the ovarian manifestation of

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67

metabolic syndrome, similarly to NAFLD that was earlier proposed as the hepatic
manifestation of metabolic syndrome.

THE ROLE OF NEURONAL NICOTINIC ACETYLCHOLINE RECEPTORS IN
MORPHINE DEPENDENCE. Lindsay M. Kondo & M. Imad Damaj, Dept, of
Pharmacology and Toxicology, Virginia Commonwealth University, Richmond
VA 23298. Opiate addiction is a growing concern in today’s society. Opiate users
show little success in maintenance treatments and improved clinical therapies are
needed. Evidence suggests that neuronal nicotinic acetylcholine receptors
(nAChRs), which contribute to nicotine dependence, may also influence morphine
dependence. We used two complementary approaches, genetically modified mice
and pharmacological ligands, to study the involvement of 06 * nAChRs (* denotes
additional subunits). In the first experiment, a 6 * knockout (KO) mice and wild type
(WT) littermates were injected twice a day with ascending doses of morphine (20,40,
80,100 mg/kg morphine, s.c.) for 8 days. On day 9, mice received a pretreatment of
morphine two hours before naloxone challenge (2 mg/kg, s.c.) and abstinence signs
were observed for 30 minutes including jumps, paw tremors, head shakes, ptosis,
backing, and diarrhea. We observed that the KO mice showed an increased intensity
of withdrawal signs in comparison to the WT controls. To complement our
transgenic approach, we used a peptide-like a6|32* selective antagonist, a-conotoxin
MII-[H9A;L15A], In this study, mice were injected three times per day for three
days with ascending doses of morphine (50, 75 and 100 mg/kg morphine, s.c.). On
day 4, mice received a morphine pretreatment followed by an injection of
a-conotoxin MI1-[H9A;L15A] (12 pmol, i.c.v.) 5 minutes before naloxone.
MII-[H9A;L15A] treatment enhanced the intensity of global somatic withdrawal
signs (pO.05). Taken together, these results suggest that a 6 * nAChRs offer a novel
target for the treatment of physical morphine dependence.

AUTOLOGOUS PLATELET-RICH PLASMA (PLATELET GEL) ENHANCES
BLOOD FLOW IN LARGE SOFT TISSUE WOUNDS IN VIVO AND REDUCES
BACTERIAL GROWTH IN VITRO. B F, Host . F Li & B. Hargrave, Center for
Bioelectrics, Old Dominion University, Norfolk VA 23508, Platelet-rich plasma
(PrP) enhances the process of wound healing in soft tissues. Platelets in whole blood
were concentrated 7-10 times using a Harvest Technologies Concentrator.
Concentrated platelets were then placed into a sterile pulsing cuvette and activated by
exposure to a 30-kV/cm electric field for 5 pulses, with each pulse having a duration
of 300 ns. With the annual under a surgical plane of anesthesia, a 3x8 cm surgical
wound was created on the dorsal surface of each animal, and the autologous
platelet-rich plasma (formed from rabbit blood) was applied directly to the wound
site. To assess the return of blood flow to the wounded area, the wound was imaged
using a Moor Instruments Laser Doppler Imager just prior to the creation of the
wound, and also post-operatively on days 0, 3, 7, 14, and 21. The wounds of 4
animals were treated with saline and these animals served as our controls. The
wounds of 6 animals were treated with PrP. Results are reported as the mean ±
standard deviation. Our current data suggest that the return of blood flow to the distal
region of the wound was significantly greater in the PrP-treated animals than in the
control animals treated with saline.

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IDENTIFICATION OF GENES TEAT MEDIATE ETHANOL-INDUCED ACUTE
FUNCTIONAL TOLERANCE IN C. ELEGANS. Ka-Po Leung . M. Bolling, J.
Gardner, A. Davies & J. Bettinger, Deptartment of Pharmacology and Toxicology,
VCU, Richmond, VA 23298. Alcohol abuse and alcoholism are prevalent diseases
in our society. There are few adequate treatments available, in part because the
molecular mechanisms behind the development of alcoholism are still unclear. We
know from human genetics studies that there is a significant genetic component that
influences disease susceptibility, and that an individual's development of acute
functional tolerance (AFT) after alcohol consumption are strong predictors of
lifetime development of addiction. We have taken a genetic approach to study the
development of tolerance to ethanol. We used C. elegans as a behavioral model for
the study of alcohol responses because of its simple nervous system and conserved
neurobiology. We performed a genetic screen looking for mutations in genes that are
required for the development of AFT to ethanol. Previous studies have shown that the
gene NPR-1, a G protein-coupled neuropeptide receptor in C. elegans that is
homologous to the mammalian neuropeptide Y (NPY) receptor, negatively regulates
the development of AFT; animals that lack NPR-1 function develop tolerance to
ethanol at a faster rate relative to animals with intact NPR-1. We used an inactivated
npr-1 gene strain, npr-1 (0), as a sensitized background for a phenotypic screen to
identify animals that are unable to develop AFT. We have identified a mutation,
bet 11 , that causes mutant animals to be defective in the development of AFT.
Identifying the genes that are responsible for alcohol-induced development of AFT
will provide a better understanding of the mechanisms that cause alcohol abuse and
alcoholism.

THE GAL ACT OLIPID SULFATIDE IS A NEGATIVE REGULATOR OF CELL
PROLIFERATION. S.A Freeman, A.D. Pomicter & J.L. Dupree, Department of
Anatomy and Neurobiology, Virginia Commonwealth University, School of
Medicine, Richmond VA, 23284. Myelin is an electrically insulating sheath that
oligodendrocytes wrap around neurons, greatly increasing the speed and efficiency of
electrical signaling between neurons. Unlike other membranes, myelin is lipid rich
and composed of approximately 70% lipid. Myelin is also unique in that it contains
an abundance of galactolipids. The two most prominent myelin galactolipids are
galactocerebroside and its sulfated derivative sulfatide, made by the enzyme
cerebroside sulfotransferase (CST). The CST null mouse exhibits an increased
population of oligodendrocytes due to enhanced proliferation and decreased cell
death in the spinal cord, but the mechanism of these changes is not known. The goal
of this project is to determine if the inhibition of sulfatide results in increased
proliferation in Madin Darby canine kidney (MDCK) cells, a sulfatide expressing,
immortal cell line. In separate experiments, we will utilize sodium chlorate to block
sulfatide production The sulfatide antibody 04 and bromodeoxyuridine (BrdU) will
be used to examine the amounts of sulfatide and proliferation, respectively.
Preliminary results suggest that MDCK cells treated with sodium chlorate exhibit an
elevated cell population via a mechanism favoring increased proliferation, and
additionally, sodium chlorate treated cells show decreased morphologic complexity,
which is consistent with findings in the CST null mouse. Understanding the complex

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69

factors that regulate oligodendrocyte proliferation is critical for future production of
therapeutics for multiple sclerosis patients.

THE RELATIONSHIP BETWEEN PHYSICIAN SPECIALIZATION AND
COMMUNICATION EFFICACY. Amber Collier 1 , Matthew Cronin 1 , School of
Management, George Mason University, Fairfax, VA 22030. A central challenge in
being an effective physician is communication; patients need reassurance and
emotional support, and also need to be informed about risks and requirements of
treatment. The challenge is for the physician to be compassionate and communicate
complex information in away that the patient can understand. As physicians become
more specialized, these functions may become more difficult to perform. This
presents a challenge as the patients who go see specialists often do so because their
health conditions are more serious. If specialists are having greater difficulty with
communication this could have significant effects on the patient’s health outcome.
Currently, there is no existing research that explores this area of communication and
its corresponding relationships. Thus, this project aims to discover whether and how
increased specialization negatively affects the quality of communication, and
ultimately the patient’s health. We plan to achieve this through survey scales that
gauge different dimensions of communication and objective/subject health measures.
If we find it does, we hope to be able to suggest mitigation measures.

COMPONENTS OF THE METABOLIC SYNDROME AND THEIR RELATION
TO NAFLD IN MORBIDLY OBESE PATIENTS. C.T. Nguvenngo 1 .
A.Baranova 2 Z.Younossi’, M.J. Estep 1 T 'Biology Department College of Science,
George Mason University, Fairfax, VA 22030, 2 School of Systems Biology, College
of Science, George Mason University, Fairfax, VA 22030, and 'Center for Liver
Diseases, Inova Fairfax Hospital, Falls Church, VA 22042. Non-alcoholic fatty
liver disease (NAFLD) and its most severe form non-alcoholic steatohepatitis
(NASH) are often referred to as the hepatic manifestations of metabolic syndrome.
However, firm connections between individual components of metabolic syndrome
and the progression of NAFLD have yet to be conclusively established. The aim of
this study was to examine the relationship between lipid profiles and NAFLD
progression in morbidly obese patients Circulating concentrations of fasting glucose,
HDL, LDL, Total cholesterol, and triglycerides was obtained using Cholestech-GLU
kits. Descriptive statistics, Mann-Whitney U-tests, and Spearman correlations were
calculated. Of the 416 NAFLD patients, 245 had progressed to NASH. Significant
differences were seen between NASH and non-NASH NAFLD in circulating
concentrations of HDL, Triglycerides, and Glucose. Diagnosis of NASH and
circulating concentrations of HDL were negatively correlated, while NASH and
Triglyceride levels and NASH and glucose levels were positively correlated. The
individual components of metabolic syndrome are associated with stage of NAFLD
progression to NASH.

LIVER FUNCTION IN HCV PATIENTS WITH DIFFERENT IL28B ALLELIC
GROETPS, R. Guragain 1 . A. Baranova 2,3 , Z Younossi 3 , & M Estep 1,3 , 'Bio, Dept.,
2 School of Sys Bio., George Mason University, Fairfax VA, and 3 Center for Liver
Diseases, Tnova Fairfax Hospital, Falls Church VA. IL28B genotype is an important

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host factor in treatment of Hepatitis C virus (HCV) infection. HCV patients could be
either homozygous for “C” allele (CC-group), or homozygous for “TV heterozygous
(non-CC-group). The success rate of achieving Sustained Viral response (SVR) in the
CC-group is significantly higher. However, other host-related factors that affect liver
function may also affect rates of SVR achievement. We examined distinctions in
clinical parameters between CC-group and non-CC-group HCV patients. Blood
samples and laboratory data were obtained for 56 patients who had undergone
standard HCV treatment. DNA was extracted using QIAmp Minikit (Qiagen). Allelic
discrimination was performed by tetra-primer PCR and electrophoresis. Spearmen
correlations and Mann-Whitney tests were performed. Of the 23 patients that
comprised the CC group, 15 had achieved SVR. Of the 33 non-CC-group patients,
only 8 had achieved SVR. SVR negatively correlated with occurrence of significant
steatosis (r= -0.31, P=0.03). Interestingly, among the patients that achieved SVR,
those with CC genotype had significantly increased liver enzymes compared to
non-CC-group (average ALT levels 59.75U/1 and 35.4U/1, respectively (P=0.007)).
Similar trends were observed for AST levels (86.91U/1 vs. 63.8U/1 (P=0.02)).
Additionally, urine nitrogen was also significantly higher in the CC-group that
achieved SVR (18.5mg/dl vs. 8mg/dl, (PO.OOl)). The results support the hypothesis
that factors affecting liver function such as steatosis may have a more significant
impact on the achievement of SVR in the non-CC-group.

HOMA CALCULATOR TO CALCULATE RESISTANCE SCORE USING
HOMEOSTASIS MODEL ASSESSMENT. L. Alomair 1 . L. Wang 1 , A. Birerdmc 1 ’ 2 ,
J.M. Estep 2 J & A Baranova 12 J , ’School of Systems Biology, College of Science,
George Mason University, Fairfax, VA, USA, 2 Betty and Guy Beatty' Center for
Integrated Research, Inova Health System, Falls Church, VA, USA, ’Center for Liver
Diseases, Inova Fairfax Hospital, Falls Church, VA, USA. Insulin is a protein that
plays a major role in glucose metabolism. Insulin may be quantified in the serum as
whole insulin or as C-peptide, where C-peptide is more stable to degradation in
during sample processing and storage. C-peptide is a byproduct created when the
hormone insulin is produced. Eqmmolar amounts of C-peptide and insulin are then
stored m secretory granules of the pancreatic beta cells and both are eventually
released to the portal circulation. Homeostasis Model Assessment (HOMA) scores
are a combination of fasting insulin and glucose measurements m the serum. HOMA
indices are used to assess insulin resistance (lR). Currently, there no publically
available software to calculate HOMA scores using C-peptide data. Here we present a
software tool capable of calculation of Homeostasis Model Assessment (HOMA)
scores based on both fasting C-peptide and fasting insulin levels. Use of this tool will
allow researchers to optimize their analysis of insulin resistance in human subjects.

THE DISCRIMINATIVE STIMULUS EFFECTS OF NITROUS OXIDE IN MICE.
Kellianne J. Richardson & Keith L. Shelton. Deptartment of Pharmacology &
Toxicology, Virginia Commonwealth University, Richmond, VA 23298. Despite
the high prevalence of clinical and illicit use, the in vivo actions of nitrous oxide are
unclear. In vitro studies have shown that nitrous oxide alters the function of NMDA,
GABA a , and nicotinic acetylcholine receptors, among others. However, the receptor

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71

system or systems responsible for the intoxicating, subjective effects of nitrous oxide
are unknown. Our objective was to use drug discrimination in mice to assess the
neurotransmitter systems responsible for producing the subjective stimulus effects of
nitrous oxide. Sixteen male B6SJLF1/J mice are being trained to discrimmate 10 min
of exposure to 60% mhaled nitrous oxide from 100% oxygen. Discrimination training
under conditions in which training and test sessions were done in room air after
cessation of nitrous oxide exposure resulted in lengthy training requirements and
poor stimulus control. As a result tire procedure was modified to conduct
discrimination training and testing while being concurrently exposed to nitrous oxide.
Thirteen mice thus far have acquired tire discrimination and begun substitution
testing with increasing nitrous oxide concentrations. The results indicate that under
proper training conditions nitrous oxide can serve as a discriminative stimulus.
Further testing will be conducted using site-selective test drugs to determine tire
neurotransmitter systems responsible for the discriminative stimulus effects of
nitrous oxide.

CHARACTERIZATION AND OPTIMIZATION OF THE
SYNAPTONEUROSOME PREPARATION IN ORDER TO
INVESTIGATE ETHANOL INDUCED ALTERATIONS TO THE SYNAPTIC
TRANSCRIPTOME. Megan A. O’Brien & Michael F. Miles, Department of
Pharmacology and Toxicology, Virginia Commonwealth University', Richmond VA
23298. It is well established that nrRNA can be transported to neuronal distal
processes, where it can undergo localized translation regulated in a spatially restricted
manner in response to stimulation, potentially playing a role in synaptic plasticity that
results in long-term adaptive brain responses. In order to investigate our hypothesis
that ethanol behavioral sensitization results, at least in part, from alterations in the
trafficking of mRNAs to distal processes, we have worked to optimize a protocol for
obtaining synaptoneurosomes from the frontal pole of mice treated with repeated
ethanol In the protocol, fresh tissue undergoes homogenization and fractionation
resulting in a pelleted fraction (P2) that should be enriched with vesicularized pre-
and post-synaptic elements from which RNA and protein can be extracted.
Characterization of the preparation through electron microscopy, western blotting,
and quantitative PCR indicate synaptic enrichment of the putative synaptoneurosome
fraction. Attempts to further isolate synaptoneurosomes were undertaken through an
immunoaffinity purification scheme targeting the synaptic transmembrane protein,
Neuroligin. We conclude that the synaptoneurosome preparation will provide us
with samples enriched in synaptically localized mRNAs and proteins that will aid
our investigation into the underlying molecular alterations that contribute to
behavioral sensitization in response to repeated ethanol.

CHRONIC INHIBITION OF MONOACYLGLYCEROL LIPASE REDUCES THE
INTENSITY OF PRECIPITATED WITHDRAWAL SIGNS IN OPIOID
DEPENDENT MICE. R A. Owens 1 . D. Ramesh 1 , B.F. Cravatt 2 & A H. Lichtman 1 ,
‘Department of Pharmacology and Toxicology, Virginia Commonwealth
University, Richmond, VA, 23298, 2 Scripps Research Institute, La Jolla, California,
92037. Chronic use of opioids can lead to dependence that results in a clinically
significant withdrawal syndrome upon drug cessation. It has been shown that

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A '-tetrahydrocamiabmol (THC) reduces opioid withdrawal signs in rodents via CEb
receptors. The endocannabinoids, anandamide (AEA) and 2-arachidonyl glycerol
(2-AG), also activate CBj receptors, but they are rapidly metabolized by their
respective enzymes, fatty acid amide hydrolase (FAAH) and monoacylglycerol
lipase (MAGL). Research from our laboratory suggests that acute administration of
these catabolic enzyme inhibitors reduces naloxone-precipitated withdrawal signs in
morphine-dependent mice. In the present study, we investigated whether the
MAGL inhibitor JZL184 would retain its anti-withdrawal effects following repeated
administration. ICR mice were Implanted with 75 mg morphine pellets, and
challenged 72 h later with the opioid receptor antagonist naloxone to precipitate
withdrawal. The presence of somatic withdrawal signs such as such as jumps,
front paw flutters, weight loss, and the occurrence of diarrhea were measured. As
previously reported, acute treatment of THC (lOmg/kg) significantly reduced the
occurrence of jumps and diarrhea. Repeated treatment with high dose of JZL184
(40 mg/kg), but not the low dose (4 mg/kg), reduced all measured signs of opioid
withdrawal. The results of the present study indicate that inhibitors of MAGL
offer a promising target to treat opioid dependence.

LYSOPHOSPHATIDIC ACID UPREGULATES EXPRESSION OF
PEROXISOME PROLIFERA T OR-ACTTV ATED RECEPTOR GAMMA IN
OVARIAN CANCER CELLS. J.K. Ngwammbi. A. Mukherjee & F. Fang,
Department of Biochemistry and Molecular Biology, Virgmia Commonwealth
University, Richmond, VA 23298. Ovarian cancer is the fifth most common
cancer among women in the United States and it is the leading cause of death
among all gynecological cancers. Most early stage ovarian cancers are detected
incidentally on routine pelvic examinations, and five year survival rates of over
90% can be achieved for localized disease. Lysophosphatidic acid (LPA), is a
soluble phospholipid made up of a single fatty acyl chain, a glycerol back bone and
a free phosphate group. It stimulates proliferation, survival, migration and invasion
of ovarian tumor cells by acting on its cognate G protein-coupled receptors.
Previous studies by others suggest that LPA binds and activates peroxisome
proliferator-activated receptor gamma (PPARy), a nuclear transcription factor
important in regulation of fat and energy metabolism. In the current study, we did not
observe any evidence for direct interactions between LPA and PPARy instead, we
demonstrated that LPA induces the expression of PPARy in ovarian cancer cells
leading to an increase in its transcriptional activity. This work points to a novel
mechanism for activation of PPARy by LPA in mammalian cells.

Natural History & Biodiversity

TWELVE YEARS LATER: ECOLOGICAL COMMUNITIES OF THE ABRAMS
CREEK WETLANDS IN WINCHESTER AND FREDERICK COUNTY,
VIRGINIA. Briana L, Barron , James E. Bisset, Linden E. Lewis, Daniel A. Milhon,
Cory M. Miller, Benjamin S. Sawyer, Amy L. Smith, John V. Stevens & Woodward
S. Bousquet, Environmental Studies Department, Shenandoah University,

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73

Winchester VA 22601. Located in northwestern Virginia, the Abrams Creek
Wetlands support 18 vascular plant species and two community types (calcareous
muck fen, wet-mesic prairie) considered rare in the state. A 1998 study by
Shenandoah University (SU) supplemented investigations by Virginia's Natural
Heritage Program in documenting the property's ecological significance. In the
spring and fall of 2010, SU researchers resurveyed three communities and studied
one new community at this site. Using the releve (Braun-Blanquet) method,
community plots were described by their vegetation and physical characteristics. The
new plot includes Car ex lasiocarpa , which is listed as critically imperiled (SI) in
Virginia. Percent similarity comparisons of the three resurveyed plots show
substantial change (PS = 15.5%, 17%) in two but little change (93.8%) in the third
since 1998. Recommendations for managing the Abrams Creek Wetlands include
removing invasive species and minimizing future disturbances from the area’s
planned housing developments and walking-biking paths.

REPRODUCTIVE BEHAVIOR OF THE POTOMAC SCULPIN (COTTUS
GIRARDI). Kimberly J. Bolyard, Dept, of Biology, Bridgewater College,
Bridgewater, VA 22812. Potomac sculpin (Cottus girardi) fish from Dry River,
Virginia were brought into the laboratory to study their reproductive behavior and to
evaluate if these fish are subject to sexual selection pressures. Reproductively
mature males vaiy in standard length I tested the hypothesis that females should
prefer to mate with larger males because they should be able to defend the nest and
eggs from predators better than smaller males. Studies were conducted during the
breeding season, February-March, 2010 and 2011. Observations were made of
groups consisting of two males, one large and one small, and one female. Male
reproductive behavior included pectoral fin waving and body shaking associated with
darkening the body color. An actively courting male would approach the female and
bite her in the head region while turning her towards his nest cavity. If she entered
the nest, the male would rest on her while shaking intermittently Occasionally, the
male would leave the female to chase the other male away from the nest area. More
females, six of seven, associated with or spawned with the larger of the two males.
While not statistically significant, the observations suggest that female mate choice
may play a role in the reproductive ecology of these fish In addition, Potomac
sculpins may be subject to male-male competition for nest sites and for females;
hence, intrasexual selection will be the focus of future studies.

INVESTIGATION OF PRE- AND POST-ZYGOTIC REPRODUCTIVE
BARRIERS BETWEEN TWO HOST-COMPLEX RACES OF A PARASITIC
WASP. Justin P. Bredlau & Karen M. Kester, Dept, of Biology, Virginia
Commonwealth University, Richmond VA 23284. Understanding the mechanisms of
speciation and reproductive isolation has important implications for studies in
biodiversity, evolution, and ecology Insect parasitoids in particular display rapid
speciation that may be associated with chemically diverse host plants. Investigations
of host-associated differentiation of parasitoids have largely focused on the degree of
molecular genetic differentiation, but a true test of species status must consider
whether differentiated populations are capable of interbreeding and producing viable
offspring. We examined possible mechanisms of isolation between two genetically

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distinct (2% divergence in the mtDNA COT locus) host-plant complex races of the
parasitoid, Cotesia congregata (Hymenoptera: Braconidae) originating from
Manduca sexta on tobacco and Ceratomia catalpae on catalpa. We compared male
responses to female pheromones, elements of male acoustic courtship signals, and
mating success between the two groups. Males responded to female pheromones of
either race and male courtship songs showed only subtle differences, suggesting that
factors other than courtship behavior may be involved in pre-zygotic isolation of the
two races. Wasps from the two sources mated and produced offspring; however, 90%
of females from one hybrid cross failed to produce offspring, leading to post-zygotic
isolation. Development time, emerged brood size, and sex ratios among the wasp
races also differed. Results suggest that the two host-complex races represent distinct
sibling species.

MOLECULAR AND MORPHOLOGICAL VARIATION IN THE BARNACLE
PREDATOR NEMERTOPSIS BEIT ATT A (NEMERTEA, HOPLONEMERTEA).
Serena A. Caplins'. Jon. L. Norenburg 2 , & Clint Turbeville 1 , 'Dept, of Biol., Virginia
Commonwealth University; Richmond Va. 23284, 2 National Museum of Natural
History, Smithsonian Institution, Washington D.C. The nemertean worm
Nemertopsis bivitatta , is a member of the hard-bottom intertidal community,
occurring along the coasts of Europe, Brazil, and the Southeastern United States.
Our laboratory observations indicate that this nemertean is a suctorial feeder of
barnacle crustaceans. Individuals of N. bivitatta are typically pale yellow to whitish
in color with a pair of dorsally-situated dark greenish brown pigment bands that
extend nearly the entire length of the worm. In most individuals these stripes are
separated anteriorly, but in some individuals they are fused anteriorly. Variation in
stripe configuration has been considered to reflect intraspecific variation. We
describe variation in stylet basis morphology and assess genetic variation, which may
be correlated with stripe configuration. Sequences of the mitochondrial
cytochrome-oxidase one gene (COl, 500-650 bps) were obtained for 8 individuals
exhibiting the anterior fusion of pigment bands and for 14 individuals without this
feature. A pair-wise comparison revealed a 14.8% difference exists between the
morphs. This lies within the range of variation observed between species in
Hoplonemertea. A tree-based approach will be used to further test whether these
morphs represent different species.

TADPOLE DENSITY CHANGES THE RELATIONSHIP OF RED-EYED
TREEFROG MORPHOLOGY AND JUMPING PERFORMANCE.
J.FCharbonmer 1 . T.Landberg 2 & J.Vonesh 1 , 'Department of Biology, Virginia
Commonwealth University, Richmond VA 23298 and "Department of Biology,
Boston University, Boston MA 02215. As organisms develop, increased body size
is often accompanied by shape changes that alter the morphology-performance
relationship. Animals with different growth histories may also have different shapes
at similar body sizes. To investigate how larval growth history affects the
morphology-performance relationship, we raised red-eyed treefrog tadpoles
(Agalychnis calhdiyas ) at three densities (5, 25 and 50 tadpoles per 400 L tank) and
measured jump distance during metamorphosis. We predicted that tadpoles grown at
low density would metamorphose into larger frogs with relatively longer legs than

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those grown at higher densities. We also expected low density frogs to jump further -
both absolutely, because of their larger body size and relative to their size if they had
longer legs. Frogs from low density had longer snout-vent lengths (SVL) than those
from medium and high densities and longer tibiafibula lengths and greater masses
relative to their SVL. Jump distance was strongly correlated with tibiafibula length;
however, there was a significant density*tibiafibula interaction. While longer
tibiafibulas in high and medium density frogs were correlated with longer jump
distances, there was no such correlation in the frogs from low density tanks. We
interpret these results as a consequence of the greater relative mass of tine frogs from
low density tanks. The relatively long legs of these frogs may not fully compensate
for their disproportionately higher mass. This study demonstrates that different larval
densities can change not only red-eyed treefrog morphology, but also the
morphology-performance relationship. It suggests a trade-off where low larval
density can increase body size and presumably fat reserves, but the latter may
decrease jumping performance.

FLEAS (SIPHONAPTERA) FROM A SMALL MAMMAL COMMUNITY IN A
GUATEMALAN CLOUD FOREST. R. P. Eckerlin 1 . W. Bulmer 1 , H. Lanier, J. O.
Matson’ & N. Woodman 4 , 'Natural Sciences Div., Northern Virginia Community
College, Annandale, VA 22003, : Dept. Biological Science, University of
Alaska-Fairbanks, AK 99775, ’Dept, of Biological Sciences, San Jose State
University, San Jose, CA, 95192, 4 USGS Patuxent Wildlife Research Center, U.S.
National Museum, Smithsonian Institution, P.O.Box 37012, Washington, DC 20013.
The flea fauna of a mammal community at a specific site has never been recorded for
Guatemala. We collected 104 small mammals (6 spp. shrews & rodents) and 323
fleas from remnant cloud forest at 2950-3160m in El Retiro, Huehuetenango,
Guatemala. Fleas were brushed off the hosts, preserved in alcohol and later mounted
on slides for identification. Plusaetis vermiformis , previously known from a single
specimen in Guatemala, was the most common flea but found only on Peromyscus
beatae and P guatemalensis, An undescribed species of Ctenophthalmus was present
on Sorex scmssurei. Plusaetis mathesoni , found on several rodent host species, is a
new country record for Guatemala. A single specimen of an Atyphloceras sp from P
beatae extends the range of this genus south of central Mexico. Other species present
on rodents were Ctenophthalmus sanborni , Baculomeris schmidti, Kohlsia osgoodi,
Jellisonia painter 7 , and Strepsylla sp. Most of the 3 species of Reithrodontomys that
we examined had no fleas. Although the mammal diversity was judged to be low the
flea fauna was robust and diverse.

TROPHIC DYNAMICS OF CHESAPEAKE BAY PLANKTON Todd A
Egerton , Matthew R. Semcheski & Harold G. Marshall, Department of Biological
Sciences, Old Dominion University, Norfolk VA 23529. Examining the relationship
between diversity and ecosystem processes has become an important topic in ecology
given the increased rate of species extinctions. Previous work has shown that
stations in lower Chesapeake Bay with increased nutrient concentrations have higher
productivity and algal abundances but lower phytoplankton diversity. However, the
potential effect of algal diversity on higher trophic levels has not been examined.
Using a 16 year dataset (1986-2001) of phytoplankton and zooplankton community

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composition, water quality parameters, and commercial fisheries landings, several
relationships were identified. Zooplankton diversity is positively correlated with
phytoplankton diversity and negatively correlated with total phytoplankton
abundance. Long-term trend analysis indicates a significant decline in zooplankton
abundances, as well as a significant negative correlation between total nitrogen
concentrations and zooplankton species richness. Furthermore, there has been a
significant decline in the average annual landing of Virginia commercial fisheries.
Fishing yield is positively correlated with zooplankton abundance and negatively
correlated with algal productivity. This suggests that bottom up factors are driving
the system, where increased nutrient levels lead to more productive less diverse algal
communities capable of supporting few'er, less diverse grazers and an overall decline
in fish populations. Supported by VADEQ.

EFFECT OF PHOTOPERIOD ON IMMATURE DEVELOPMENT IN THE TIGER
MOSQUITO, AEDES ALBOPICTUS. Allen Frisa & Deborah Waller, Dept, of
Biol., Old Dominion Univ., Norfolk, VA 23529. Tiger mosquitoes are active from
spring through fall and overwinter in the egg stage. We examined whether mosquito
development varied depending on the photoperiod the immatures experienced.
Late-instar larvae were collected from an incubator with a summertime photoperiod
(16L;8D), placed in individual cups with 20 ml deionized water and fish food, and
incubated in the following photoperiods: constant dark (DD), spring/fall (12L:12D),
summer (16L:8D), reverse summer (8D: 16L) and constant light (EL). There were ten
replicates per photoperiod. Time from pupal formation to adult eclosion was recorded
along with the gender of the adults. Under constant temperatures with a mean of
26°C, pupal durations were 2-3 days for both male and female with no differences
related to photoperiod. In a second experiment in which spring/fall and constant light
incubators were kept at a mean of 23°C, pupal durations were 3-4 days for both males
and females. Therefore temperature influenced developmental rates but there was no
influence of photoperiod on pupal duration

A SURVEY OF MUSKRAT INTESTINAL PARASITES IN VIRGINIA.
Jeronimo G. Da Silva Neto , Elizabeth R. Hepner, Allison E. Salinger, Emily A.
Kimminau & Lisa K. Belden, Department of Biological Sciences, Virginia Tech,
Blacksburg, VA 24061. Infectious diseases of humans and wildlife have increased
dramatically in recent decades. These diseases can have important impacts on
global health, agriculture and biodiversity Despite these increases, we still know'
relatively little about disease in natural wildlife populations. In many cases, we lack
even basic information about what parasites and pathogens are present in wildlife
species. This prompted us to conduct a survey on local populations of muskrats, a
common semi-aquatic rodent found in the United States. We conducted necropsies on
donated muskrat carcasses collected from across Virginia, and began to establish a
baseline database on parasites of muskrats, with a focus on the macroparasitic worms
in the gastrointestinal tract (stomach, small intestine, large intestine and cecum).
Since 2008, we have necropsied over 100 individuals. Three genera of parasitic
flatworms (trematodes) dominated the samples - Echinostoma, Ouinqueserialis and
Wardius. As these parasites have complex life cycles involving multiple hosts in
addition to muskrats, we can potentially use the presence of these parasites to infer

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something about the ecological communities of the sites at which the muskrats were
collected. We have also completed some analyses regarding the distribution of the
parasites among hosts. We have not seen any significant relationship between total
worm burden and either sex or mass of the host. Our research provides an important
baseline for understanding parasite dynamics in natural muskrat populations. This
allows a better understanding of potentially emerging diseases and can improve how
we manage wildlife disease.

ALLEGHENY WOODRATS (. NEOTOMA MAGISTER) IN VIRGINIA:
COMPARISONS OF OCCUPANCY OVER TIME. Jennifer M, Kamne. Michael
T. Mengak & Steven B. Castleberry, Warnell School of Forestry and Natural
Resources, LTniversity of Georgia, Athens, Georgia 30602. The Allegheny woodrat
(Neotoma magister) is a cryptic small mammal that occupies rocky outcrop and cave
habitats throughout the Appalachian Mountains. Populations have been
documented as declining throughout much of its historic range. Population declines
have been attributed to a suite of effects occurring in the habitat including interactions
with raccoon roundworms, loss of American chestnut, habitat reduction, and
fragmentation. We surveyed 232 sites in Virginia in 2008-10 for woodrat
occupancy and compared our results to results of surveys conducted from 1990-1999
to assess changes in woodrat occupancy over time. We also examined if our
trapping protocol was adequate to determine occupancy through a combination of
extending the number of nights trapped and through the use of remote cameras.
Naive occupancy during the 1990s was 67%, while only 49% of sites were occupied
from 2000-2010. Woodrat populations exist as metapopulations, where sites consist
of spatially separated populations that interact. Metapopulations systems have some
groups that may be extirpated in some years which are followed by re-colonization in
subsequent years. We have documented 38% of our sites having variable occupancy
over time. Results of our camera surveys indicate that our current monitoring
protocol is effective at determining occupancy 85% of the time.

A SKULL OF THE EXTINCT RIGHT WHALE BALAENULA FROM THE FATE
PLIOCENE OF NORTH CAROLINA. Laura Kellam, Alton C. Dooley, Jr., &
Vince Schneider. Roanoke College, Salem, VA 24153, Virginia Museum of Natural
History, Martinsville, VA 24112, North Carolina Museum of Natural Science,
Raleigh, NC 27601 A nearly complete balaemd skull was collected in May 2008
from the Bear Bluff Formation in Lake Waccamaw, North Carolina. The cranium as
preserved is 165 cm in length. The tight closure of most of the cranial sutures
suggests that this was a mature animal at the time of death. It has extreme telescoping,
arched rostral bones, dorsoventrally compressed tympanic bullae, and the dentary has
an anterior torsion and lacks a distinct coronoid process, all features that are typical of
balaenids. Features shared with the genus Balaenula include: 1) small overall size; 2)
large, block-like nasals; 3) approximately right angle between posterior and anterior
processes of the petrosal; 4) anteriorly oriented squamosal; 5) right angle formed
between supraorbital processes of frontals when viewed anteriorly However, it
differs from B. astensis from Italy and B. balaenopsis from Belgium in a number of
characters. This is the first report of Balaenula from the Bear Bluff Formation, which
has an approximate age of 2.75 Ma All other reported specimens of Balaenula from

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the western Atlantic are from the lower Yorktown Formation, with an approximate
age of 4.5 Ma. Balaenula balaenopsis from Belgium and undescribed Balaenula
specimens from Japan and California are Pliocene in age, but their ages are poorly
constrained, while B. astensis from Italy has an approximate age of 3.5 Ma. The Lake
Waccamaw specimen is thus possibly the youngest known occurrence of Balaenula.

PRIMARY FEATHER PIGMENTATION IS CONSTRAINED BY FUNCTIONAL
NECESSITY. Joshua D. Kreitzer & Lisa Horth Dept, of Biology, Old Dominion
University, Norfolk, VA 23529. Primary feathers are critical for powered flight.
High exposure of these feathers to the wind and associated abrasive particles
necessitates that they be strong relative to other, less exposed feathers. The black
pigment, melanin, has been associated with increased feather strength via an increase
in keratin thickness near the melanin and via the added strength provided by the
melanin granules themselves. Furthermore, melanin provides protection from the
damaging affects of ultraviolet radiation in comparison to paler feathers. We tested
the hypothesis that primary feathers would be more frequently pigmented with
melanin than a flight-neutral feather patch, the upper back. A survey of4006 species
of non-passerine and 1823 species of passerine birds showed that 63.4% had black in
the primaries, 35.4% had other color as the darkest color, and 1.2% had wholly white
primaries. The upper back feathers showed a different pattern, with 29.0% of species
having black, 69 2% having other, and 2.0% having white. Chi-square analysis
showed that the pigmentation of the primary feathers was significantly different from
the expected color distribution of the more flight-neutral upper back feathers (/?-value
<0.0001). These results may be understood as evidence supporting the hypothesis
that primaries are more constrained to be black than the upper back feathers

RESPONSE OF IMMATURE MOSQUITOES TO PREDATOR PRESENCE.
Jackie McKee & Deborah Waller, Dept, of Biol., Old Dominion Univ., Norfolk, VA
23529. Mosquito immatures are consumed by a variety of aquatic predators. We
tested responses of mosquito larvae and pupae to the presence of dragonfly nymphs,
backswimmers and whirligig beetles hi the laboratory. Experimental units consisted
of 14 cm diam petn dishes with a grid of 2 cm diam squares etched on the bottom.
Petri dishes were filled with 80 ml deionized water and set m sleeves that were either
black, white or half blaCk/half white. Previous experiments demonstrated that first
instar larvae prefer white backgrounds, later instars show no preference, and pupae
prefer black backgrounds, and mosquito activity declines with age. A predator was
placed in the petn dish and then either a larva or pupa was introduced to the center of
the dish. Positions of both the mosquito and the predator were noted every 30 seconds
for five minutes. Five different mosquitoes were tested in each experiment. Predators
were replaced with a new predator of the same species if it consumed the mosquito.
All three predator species ate mosquito larvae and pupae on occasion. There was no
response by mosquito immatures to predator presence. Larvae were more active than
pupae and pupae moved less frequently on black than on white backgrounds. These
patterns are similar to mosquito behavior in the absence of predators, where larvae
move frequently to find food and the non-feeding pupal stage seeks out dark areas.

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HIGH SPEED EXAMINATION OF PECTORAL STRIDULATION SOUND
GENERATION IN BLUE CATFISH ICTALURUS FURCATUS. Y J. Mohaier ,
M. L. Fine & Z. N. Ghahramani, Department of Biology, Virginia Commonwealth
University, Richmond VA 23284-2012. The blue catfish lctalurus furcatus is an
invasive species whose population numbers have exploded in Chesapeake Bay
estuaries. Catfishes produce stridulation sounds by rubbing ridges on the dorsal
process of the pectoral spine against a rough surface on the cleithrum to produce a
series of pulses during abduction. We studied the mechanism of sound generation by
synchronizing audio recordings with a high speed camera at 2000 frames per second.
Unlike channel catfish that typically keep their pectoral fins retracted (adducted),
blue catfish tend to maintain them in a forward position. Therefore sound production
requires the fish to adduct the spine before forward stridulatory movement.
Adduction movements were about a third as long as abductions (means of 49 and 152
ms respectively ). The amplitude of abduction and adduction movements was similar
at 25.9 and 26.5 degrees. Stridulatory abductions consisted of a series of 3-17 short
rapid movements (median of 13) with an average rotation of 3.1 degree over 1.5 ms.
Movement was followed by an interpulse period of 13.2 ms in which the spine was
stationary. Sound amplitude was low during movement and increased during the
stationary period suggesting time was required to activate vibrations in the pectoral
girdle. Sounds are produced by a slip-stick mechanism similar to a bow over a violin
string.

SYSTEMATIC REVISION OF THE NORTHERN SHORT-TAILED SHREW,
BLARINA BREVICAUDA (SAY). N. D. Moncnef 1 . Wm. D. Webster, J. R. Choate 3
(deceased) & H. H. Genoways 4 , *VA Museum of Natural History, Martinsville, VA
24112, : Dept. Biology, Univ, of North Carolina Wilmington, Wilmington, NC,
28403, 3 Sternberg Museum of Natural History, Fort Hays State University, Hays, KS
67601, and 4 University of Nebraska State Museum, Lincoln, NE 68588. The
northern short-tailed shrew, Blarina brevicauda (Say, 1823) occurs from southern
Canada southward to the central Great Plains, in the Appalachian Mountains south to
Georgia and Alabama, and along the East Coast as far south as southeastern North
Carolina We analyzed 9 cranial and mandibular measurements of 2,736 specimens
using a single classification ANOVA and a principal component analysis. We
conclude that B. brevicauda consists of 7 well-defined subspecies Two subspecies
( B. b. brevicauda and B. b. talpoides ) occupy almost the entire geographic range of
the species. The other 5 subspecies (including 3 that were previously undescribed)
occupy small-to-modest geographic ranges at the periphery of the species’ range and
in isolated geographic areas. We recognize these 7 subspecies because we believe
that each has begun to follow its own evolutionary path. Mitochondrial DNA data and
our morphological data indicate that most or all of the peripheral subspecies are
derived from B. b. talpoides. These data also indicate that B. b. brevicauda and B. b.
talpoides are semi-species.

A COMPARISON OF TOOTH BREAKAGE RATES IN MIOCENE
CARCHARHINIFORM SHARKS FROM THE CARMEL CHURCH QUARRY,
CAROLINE COUNTY, VIRGINIA. Brandi Neifert, Roanoke College, Salem, VA
24153 & Alton C. Dooley, Jr., Virginia Museum of Natural History, Martinsville, VA

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24112. Excavation of marine sediments of the middle Miocene Calvert Formation at
the Carmel Church Quarry has resulted in the collection of thousands of shark and ray
teeth. Unlike most large collections of shark teeth, the Carmel Church specimens
were all collected in situ from a single bed with a maximum thickness of less than 1
m. While tire majority of teeth from Carmel Church represent clearly reworked
specimens, a substantial portion show no evidence of reworking and provide a sample
that may represent the local population of sharks over a relatively short period of
time. Multiple shark taxa with similar tooth morphologies and body sizes may have
coexisted through niche partitioning. This could have been achieved through
behavioral variations such as temporal segregation (diurnal or nocturnal habits, or
seasonal movements) or through variations in dietary preferences. In order to test for
evidence of variation in dietaiy preferences, tooth breakage frequency and height-
thickness ratios were examined in four shark genera that are broadly similar to each
other in size and tooth morphology - Hemipristis , Galeocerdo , Physogaleus and
Carcharhinus. Preliminary results show similar overall breakage rates among the
four genera, but a higher rate of catastrophic breakage in Carcharhinus and
Galeocerdo.

MODELING THE PERSISTENCE OF RARE GENOTYPE IN NATURE.
Iordanka N. Panayotova, Department of Mathematics and Statistics & Lisa Horth,
Department of Biological Sciences, Old Dominion University, Norfolk, VA 23529.
Major theoretical constructs explaining the maintenance of rare genotypes include
mutation-selection balance and balancing selection. Empirical work demonstrates
that mutations arise on the order of -lO'Vgene/generation (± an order of magnitude
contingent upon data used). Thus, novel mutants that arise from one or two single
nucleotide polymorphisms and that persist at a frequency exceeding the mutation
rate (by three or four orders of magnitude) are not trivially explained by
mutation-selection balance and are extremely understudied. Empirical
demonstrations of negative frequency dependent selection tend to only address the
maintenance of polymorphisms that oscillate around a frequency far in excess of
~10' 2 Thus, what maintains the rare genotypes that persist at frequencies of
10 2 -l0 4 ? Here, we present a model that incorporates empirical life-history data on
mosquitofish from 15 years of field-work. Our model simulates the maintenance of
a stable polymorphism where the rare genotype persists at a frequency of ~ 10 " (the
actual frequency of the melanic pigmented genotype in nature) Our results present
a novel and intriguing find by providing a robust mechanism for the maintenance of
a rare (but not too rare) genotype in a stable polymorphism

POPULATION GENETICS OF TUNDRA ( CYGNUS COLUMBIANUS
COLUMB1ANUS) AND TRUMPETER ( CYGNUS BUCCINATOR ) SWANS: THE
IMPACT OF CLIMATE CHANGE ON THE BREAKDOWN OF PRE-ZYGOTIC
ISOLATING MECHANISMS. Lauren E Wilson, Department of Environmental
Science and Policy, George Mason Univ., Fairfax, VA 220304444. The Tundra
(Cygnus cohimbianus cohimbiamis) and Trumpeter (C. buccinator) Swans are two
migratory swan species with significant breeding populations in Alaska.
Historically, breeding ranges have remained sympatric; the Trumpeter nests on
boreal forest wetlands while the Tundra nests near tundra wetlands. However,

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81

increased population size of both species and expansion of the boreal forest, a likely
result of global climate change, have caused areas of geographic overlap to occur.
The Trumpeter and Tundra Swans are known to be reproductively compatible, as a
viable hybrid population exists in Virginia at the Airlie Center’s Swan Research
Program (SRP). Further, accounts from local wildlife managers indicate the
presence of fertile hybrids in overlapping breeding areas in Alaska. This research
seeks to confirm tire presence of viable wild hybrids and to elucidate tire genetic
structure and relatedness of these interbreedmg populations by comparison of
phylogenetically relevant genetic loci of wild individuals to known hybrids from the
SRP. We will also incorporate “historic” samples collected in 1920 (and prior) to
confirm that this interbreeding is a recent phenomenon and likely due to habitat
disturbance from climate change.

EFFECTS OF PREDATION, COMPETITION, AND ISOLATION ON THE RATE
OF SHELL SELECTION IN THE HERMIT CRAB PAGURUS LONGlCARPUS.
Amber L. King & Deirdre Gonsalves-Jackson, Biology Department, Virginia
Wesleyan College. Hermit crabs are most notably known for their use of gastropod
shells to protect their soft bodies, which lack calcification on their abdominal
exoskeleton Because of the necessity for a gastropod shell, hermit crabs will often
fight amongst each other for shells due to the scarcity of available shells because
without a shell to provide adequate protection, they risk exposure. Pagurus
longicarpus was used in this study to determine the effect of various factors on the
rate of shell switching, specifically the effect of isolation, conspecifics, and
predation. It was proposed that more hermit crabs would switch shells and the rate of
shell switching would increase when isolated, rather than when in the presence of
conspecifics or a predator. Three different treatments were set up and based on the
results, the two hypotheses were supported because more hermit crabs switched
shells and at a faster rate in the isolation treatment, rather than in the other two
treatments, suggesting that predation may have posed as significant a threat in my
study because the hermit crabs failed to switch shells.

Psychology

SEX ANXIETY AND SEX GUILT FROM 1980 TO 2010 Yang Deng & Louis H
Janda, Department of Psychology, Old Dominion University, Norfolk VA, 23529.
The present study examined perception of social desirability regarding sex anxiety,
perceived norms regarding sex guilt, and attitude changes in sex anxiety and sex guilt
over the past 30 years Undergraduate students completed the Sex Anxiety Inventory
(SAI) honestly, attempting to present a favorable impression and attempting to
present an unfavorable impression. They also completed the sex guilt subscale of
Mosher Forced-Choice Guilt Inventory (MFCG1) by describing their own feelings
and estimating their same-sex peer responses Both sex anxiety scores and sex guilt
scores were compared among different instructions. The self-descriptive scores on
sex anxiety and sex guilt were compared with those in 1979/1980, The results
indicated that the favorable-impression scores on sex anxiety were significantly
higher than the unfavorable-impression scores and the self-descriptive scores; the

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self-descriptive scores on sex guilt were significantly higher than the estimated,
same-sex peer scores, consistent with the concept of pluralistic ignorance ; and the
self-descriptive scores on sex guilt in 2010 were significantly higher than those in
1979. The results suggest that people are holding a conservative standard for sex but a
permissive perception of sex norm. Changes in sex guilt and sex anxiety between
1979/1980 and 2010 are discussed.

THE EFFECT OF A CONTEXTUAL AID DURING A VISUAL SEARCH TASK.
Molly Liechtv & Poornima Madhavan, Old Dominion University, Norfolk, VA
23529. Eidividuals utilize contextual cues (familiar objects within an environment)
to simplify their world and anticipate where items will be. The goal of this study was
to simulate the way in which airport security screeners utilize contextual cues while
searching for threat objects. Twenty undergraduates from Old Dominion University
performed a simulated luggage-screening task during which they were connected to
an eye tracker. Participants screened 375 bags, 75% of the images contained a target
and/or a distractor (an item in the visual scene semantically unrelated to the threat
object, i.e., iPod). Results revealed that when the distractor was present, fixation
durations increased while fixation counts, dwell times, and saccade counts decreased,
demonstrating an aggressive search pattern and an expectation of target presence. We
contend that participants formed an implicit association between the distractor and
the target and used this association to improve the efficiency of their visual search.
This implicit association of objects demonstrates the potential that exists to bias
airport security screeners and could aid in training security screeners.

WHEN FEAR AND SADNESS ARE GOOD: THE EFFECTS OF ANGER, FEAR,
AND SADNESS ON THREAT RECOGNITION DURING LUGGAGE
SCREENING. Ray C. Heikens & Kimberly E. Culley, Dept, of Psychology, Old
Dominion University, Norfolk VA, 23529-0001. Research involving the impact of
negative emotions on decision-making shows that they can have an effect on one’s
perception of risk. Anger has a significant reduction in the amount of perceived risk
and is able to increase hopefulness for the future. Fear has the opposite effect,
increasing perceived risk and reducing optimism. Sadness has a limited history of
examination, but can drive one’s risk assessment more logically. Current standards of
luggage security screening allow personnel to view bags being x-rayed for 4 seconds.
The purpose of this study was to investigate the effect of anger, fear, and sadness as
well as tire effect of stimulus duration on luggage screenmg accuracy Participants (n
= 228) viewed linages selected to induce one of the three test emotions, then
attempted to locate threatening objects in x-rays of luggage after either 2, 4, or 6
seconds of viewing. Results showed that 4 seconds of viewing enable the highest
percentage of hits when participants were angry or afraid. Sad participants had a
higher hit rate when viewing the x-ray for 6 seconds and 6 seconds clamred the lowest
percentage of false alarms overall. The implications of this research could aid in
improving training and optimizing decision-making for security personnel.

PARENTAL ALCOHOL ABUSE, NEGLECT, PSYCHOLOGICAL ABUSE, AND
PHYSICAL ABUSE: A BRIEF LOOK AT INTIMATE PARTNER VIOLENCE.
Emily M. Muschek, Old Dominion Univ., Norfolk, VA. Adverse childhood

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experiences (parental alcohol abuse, neglect, psychological abuse, and physical
abuse) have long-term negative consequences. This study examines the relationship
between these 4 types of adverse childhood experiences and emotional and physical
abuse in college dating relationships. 426 college students completed 4 measures
that assessed their adverse childhood experiences, exposure to parental alcohol abuse,
emotional abuse, and physical aggression. Overall, ACOAs were more likely to be
in a relationship in which they are bemg emotionally abused in all four dimensions.
ACOA status also predicted restrictive engulfment behaviors as well as behaviors
resulting in injury toward their partners. ACOAs were more likely to be in a
relationship in which their partner was psychologically aggressive. Participants who
were exposed to parental alcohol abuse were more likely to express behaviors that
would injure their partner. Participants who experienced childhood psychological
abuse were more likely to exhibit behaviors of hostile withdrawal and
dominance/intimidation toward their partners. They were also more likely to engage
in acts of assault leading to possible injury towards their partner. Participants who
experienced physical abuse were more likely to exhibit behaviors of restrictive
engulfment, denigration, and hostile withdrawal, and to be more aggressive and
engage in acts of assault and be psychologically aggressive towards their partner.
Neglect, psychological abuse, and physical abuse were not found to be predictors of
participant perpetration in any of the 4 dimensions of emotional abuse in dating
relationships.

CONCORDANCE OF ALCOHOL USE AMONG UNDERGRADUATE WOMEN
IN DATING RELATIONSHIPS. Michelle K. Crewe & Michelle L. Kelley,
Department of Psychology, Old Dominion University, Norfolk, VA 23529-0267
This study exammed concordant drinking among 315 undergraduate women between
the ages of 18 and 30 and their dating partners. The Substance Abuse Subtle
Screening Inventory -3 was used to assess their own and their reports of partner’s
alcohol use. Four types of couples were expected: alcohol-abusing participants with
alcohol-abusing partners (i.e., abuse-concordant couples, n = 38), alcohol-abusing
participants with a non-alcohol-abusing partner (n = 20), non-abusing participants
with an alcohol-abusing partner (n = 46), and non-alcohol-abusing women and men
(.n = 211). Abuse-concordant couples were compared to other couple types to
examine potential differences in relationship satisfaction as measured by the Dyadic
Adjustment Scale, quantity of alcohol use as measured by the Daily Drinking
Questionnaire, and alcohol consequences as measured by the Rutgers Alcohol
Problem Index. As hypothesized, participants who met criteria for alcohol abuse on
the SASSI-3 were significantly more likely than expected by chance to report that
their dating partner also abused alcohol. Participants in alcohol-concordant couples
reported significantly higher alcohol use than participants in other couple types,
indicated more alcohol consequences than other couples types except
alcohol-abusing women with non-abusing men, and had higher relationship
satisfaction than non-abusing women with alcohol-abusing men.

DON’T BE AFRAID TO EXAGGERATE: A STUDY OF EMOTIONAL
INTELLIGENCE & PHYSIOLOGICAL MANAGEMENT. Daniel T. Bice .
Alexander G Morris . Victoria Toadvine . Joshua K. Vickstrom . & Hilary E.

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Stebbins., Dept, of Psych., Univ. of Mary Washington, Fredericksburg VA 22401.
This study investigated the effect of emotional intelligence (ET) and exaggeration of
emotional responses on the ability to recover from an aroused state. We
hypothesized that participants high in El would recover faster regardless of
condition and that exaggeration would produce a greater discrepancy between the
minimum and maximum arousal levels. After completing the Wong-Law Emotional
Intelligence Scale, 47 participants (18 men) were randomly assigned to an
exaggerate or natural condition and exposed to a 3-minute video clip from 1 Know
What You Did Last Summer (1997). Both, physiological arousal and management
ability were measured via Galvanic Skin Response. A statistically significant
interaction between condition and El on baseline recovery rate was found such that
those high in El did recover faster that low El, but only in the non-exaggerate
condition. Arguably, exaggeration allows low El individuals to be more aware of
physiological changes and better manage them, but interferes with high El
individuals’ natural awareness and management abilities. This finding illustrates
that there are potential techniques, such as exaggeration, that can be taught or used
to increase emotional intelligence.

BULLYING IN COLLEGE AND ITS EFFECT ON ACADEMIC
ACHIEVEMENT Calesha W. Haves & Barbara Winstead, Old Dominion
University, Norfolk, VA. Bullying and its effects on academic achievement were
examined. It was hypothesized that engaging in bullying would negatively correlate
with academic achievement in high school and college and that being bullied would
negatively correlate with academic achievement in high school and college.
Undergraduate Old Dominion University students (A-190) were the participants.
Participants were presented with a definition of bullying and given an anonymous
self-paced questionnaire about academic achievement and bullying others and being
bullied in high school and college. The correlation between bullying others in both
high school and college with academic achievement was not significant. The
correlation between being bullied in high school and college with academic
achievement was not significant A curvilinear relationship was found between the
college victim and bully scores and GPA The medium GPA group was found to have
significantly higher bullying and victim scores than the high GPA group, while the
low GPA group did not differ significantly from either group. A revised questionnaire
or definition of bullying that includes lower levels of bullying and victimization is
discussed.

THE INFLUENCE OF SIMULATION STYLE ON PERFORMANCE. Chelsea D
Picot, Rachel R. Phillips, & Poomima Madhavan, Department of Psychology, Old
Dominion University, Norfolk VA 23529. Despite tire fact that luggage screening
involves a dynamic presentation of images, most luggage screening research is
conducted with static image presentation. Although some research has suggested that
performance with static and dynamic images is equivalent, this has not been
addressed in the context of luggage screening. To address this gap, we conducted an
experiment to compare luggage screening performance between static and dynamic
conditions. Fifty-six students participated for a partial fulfillment of course
requirements and were told that they would be playing the role of airline luggage

ABSTRACTS

85

screeners looking for knives in x-ray images of luggage. The experiment began with a
200 image training session during which participants viewed each image in the center
of the screen for 3 seconds. Participants had to indicate if they believed a knife was
present before the mrage timed out. After a short break, participants completed a
transfer block of200 images in one of three conditions: 3 seconds centered, 9 seconds
centered, or scrolling. Results revealed that participants had a higher hit rate and a
lower false alarm rate in the transfer block than in the training block indicating that
their performance unproved between training and transfer. In tire transfer conditions,
participants in the scrolling condition had a significantly lower hit rate than those in
the 9 second centered condition. These results indicate that performance with a
dynamic versus static image display may differ when it comes to a complex visual
search task such as luggage screening and that simulation style may influence results
in a meaningful way.

METHODOLOGICAL CONSIDERATIONS LOR USE OF A MULTIMODAL
STROOP-LIKE TASK. J. Christopher Brill & Elizabeth L. Ferguson, Department
of Psychology, Old Dominion University, Norfolk, VA 23529. Development and
usage of a multi-modal Stroop-like task for assessing attentional processing requires
specific methodological considerations. A task requiring the learning and use of a
spatial translation code that supersedes stimulus-response compatibility may
facilitate tapping into the same psychological processes involved in performing the
classic Stroop task. Information is presented visually, aurally or tactually in a spatial
array, and participants respond manually using a spatially congruent response box.
Special considerations, however, are required to fairly compare performance for a
tri-modal Stroop-like task. Among the considerations are creating highly
discernable signals, equating perceptual loudness across sensory modalities,
considering response mode, custom fitting the vibrotactile array to each participant,
and use of hardware that facilitates accurate measurement of response time. Current
research explores the use of this task as a means for assessing differences in
processing for multimodal signals. Future research will explore the use of
redundantly-coded and spatially incongruent signals.

SELF-REPORTED AGGRESSIVE DRIVING BEHAVIOR AMONG COLLEGE
STUDENTS J. Christopher Brill, Lindsey C. Chase, Christina M. Rieger, &
Rebecca L. Tiefenback, Department of Psychology, Old Dominion University,
Norfolk, VA 23529. The National Highway Transportation Safety' Administration
identified aggressive driving behavior as a serious hazard to driving safety. We
sought to identify potential patterns of aggressive driving behavior among college
students usmg a recently constructed questionnaire: The Aggressive Driving
Behavior Questionnaire .The ADBQ is a 20-item self-report questionnaire in which
participants rate on a six point Likert-type scale the extent to which they engage in a
series of behaviors while drivmg. Participants completed a basic demographics
form and driving history questionnaire, as well as the ADBQ. The results suggest
the majority of participants endorse feeling angry at other drivers’ behavior, but
substantially fewer reported engaging in aggressive acts (e.g., flashing their bright

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lights at other drivers, slowing down in front of drivers who are tailgating, and cutting
off other drivers). Most participants reported engaging in more passive behaviors,
such as shaking their heads at other drivers.

Statistics

FINDING AN OPTIMAL DESIGN USING PSEUDOFACTORS. Mary A.
Marion, Department of Statistics, Virginia Polytechnic Institute and State University.
This paper was written as a result of an Industrial Systems Engineering project
performed at Virginia Tech. This paper reflects an evolving procedure to design an
industrial experiment utilizing optimality criteria, AIC statistic and the usual
regression/ANOVA model statistics, Discrete factor settings were coded as
continuous to utilize response surface methods to find the best settings to reach a
specified target. While the industrial example is trivial the characteristics of the
project lend themselves to illustrate the complexity of real life applications

AN INCREMENTAL FORWARD ST AGE WISE REGRESSION ALGORITHM
FOR DICHOTOMOUS RESPONSE VARIABLES. Adam Sima, Department of
Biostatistics, Virginia Commonwealth University. The Incremental Forward
Stagewise Regression (TFSR) procedure was developed by Hastie, et a). (2001) as a
flexible estimation procedure for fitting penalized linear models. To generalize this
procedure, the IFSR estimation method was extended for use with a dichotomous
response variable. In particular, a simulation study was used compare both the
accuracy in prediction and model fit to similar algorithms that snnultaneously fit a
model and estimate parameters. The results show that this method is comparable to
some commonly used algorithms.

INTRODUCTION TO DISCRETE CHOICE MODELS. Bhaskara S. Ravi and N.
Rao Chaganty, Department of Mathematics and Statistics, Old Dominion University.
We often encounter with decisions that involve choosing between alternatives or
choices such as “which phone to buy” or “which minute plan” to choose or “which
brand of shampoo to buy” etc. Interestingly, these decisions not only depend on
individual characteristics but heavily on alternatives available. Discrete choice
models analyze such choice behavior and these are very popular in economics. This
talk aims at introducing very famous McFadden’s conditional logit model and the
importance of ITA (Independence of irrelevant attributes) assumption. Also, a
review of current trends and challenges in this popular research area are presented.

Structural Biology, Biochemistry and Biophysics

MECHANISM OF ACTION OF UDP-GALACTOPYRANOSE MUTASE FROM
TRYPANOSOMA CRUZI. Michelle Qppenheimer . Ana L. Valenciano, Jun Qi, &
Pablo Sobrado, Department of Biochemistry, Virginia Tech, Blacksburg, VA
24061. Trypanosoma cruzi (T. cruzi) is the causative agent of Chagas’ disease,
which if untreated leads to chronic inflammation of the heart. UDP-galactopyranose

ABSTRACTS

87

mutase (UGM) is a flavoenzyme that catalyzes the conversion of
UDP-galactopyranose (UDP-Galty) to UDP-galactofuranose (UDP-Gal/). UGM are
important in parasitic pathogenesis and are absent in mammals, making UGM an
attractive drug target. The chemical mechanism of UGM is not well understood. We
have used steady-state kinetics, rapid-reaction kinetics, and trapping intermediates to
better understand the mechanism of function of UGM. A k cat value of 8.1 ± 0.3 and a
K m value of 43 ± 7 pM were determined for T. cruzi UGM (TcUGM) using
steady-state kmetics. Spectral change indicative of a predicted flavin iminium ion
were detected by rapid reaction kinetics analyses, consistant with a postulated flavin
galactose adduct. The intermediate forms at a rate of 310 ± 40 s' 1 , suggesting this is
not the rate limiting step. We have trapped this intermediate and have been able to
isolate the adduct by both the HPLC and identify it by mass spectrometry. Lastly, We
sought to determine the redox partner for TcUGM. We show that TcUGM can be
reduced by NAD(P)H and maintained the reduced state for several catalytic cycles.
This activity is unique to eukaryotic UGMs as prokaryotic LTGMs do not react with
NAD(P)H. Supported by NIH grants GM094469 and AI082542 and the American
Heart Association.

PROBING THE MECHANISM OF HIGH-FIDELITY DNA SYNTHESIS USING
X-RAY CRYSTALLOGRAPHY. Eugene Wu, Department of Biology, University
of Richmond, Richmond, VA. DNA polymerases replicate DNA with higher
fidelity than would be expected from free energy differences between
complementary and mismatched base pairs. One approach to studying replication
fidelity is to determine X-ray crystal structures of reaction intermediates and
polymerase complexes with mismatched base pairs. A crystal structure of a
guanosine;thymidine triphosphate mismatch in the active site of DNA polymerase I
from Bacillus stearothermophilus revealed a key intermediate conformation
between the “open” and “closed” conformations, The previously unknown “ajar”
conformation allows the template to interact with the incoming nucleoside
triphosphate and position it relative to the polymerase active site. Complementary
dNTPs advance past this conformation to a correctly aligned closed conformation
for catalysis, while mismatches are misaligned, leading to substrate release Other
DNA polymerase I family enzymes share features important for the three-state
sorting mechanism and are expected to use thia mechanism. One homologue with
an altered active site sequence may show a preference for the ajar conformation.
This homologue’s crystal structure has been solved by multiwavelength anomalous
dispersion and may lead to confirmation of the three-state or new insights into
nucleotide selection by DNA polymerases.

INCORPORATION OF TAUTOMERISM INTO MOLECULAR MODELING;
PROGRESS IN DEVELOPING PYRROLE-BASED ANTI-TUBULIN AGENTS.
C. Da, 1 G. E. Kellogg, 1 N. Telang 2 & J. Gupton 2 , 'Virginia Commonwealth
University, Richmond VA 23298-0540 2 University of Richmond, Richmond VA
23173. Tautomerism is a commonly observed chemical phenomenon that involves
readily available structural changes of the positions of protons and double bonds.
However, alternative tautomeric forms are often ignored in many molecular
modeling applications. In docking, inaccuracy in predicting binding affinities can be

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related to the failure to consider all possible states including tautomeric states. We are
working to incorporate tautomerism into modeling tools based around HINT
(Hydropathic INTeractions). A preliminary version of the program has been
developed to apply a simple, straightforward algorithm to identify and enumerate
tautomeric forms. It is being used in an ongoing project of designing and developing
pyrrole-based anti-tubulin agents, to identify and evaluate potential tautomers, and to
build quantitative predictive models.

HIGH-THROUGHPUT ASSAY TO IDENTIFY INHIBITORS AGAINST
UDP-GALACTOPYRANOSE MUTASE (UGM) FROM ASPERGILLUS
FUMIGATUS. Jun Qi, Michelle Oppenheimer, & Pablo Sobrado, Department of
Biochemistry, Virginia Tech, Blacksburg, VA 24061. The flavoenzyme
UDP-galactopyranose mutase (UGM) catalyzes the isomerization of
UDP-galactopyranose to UDP-galactofuranose, the biosynthetic precursor of
galactofuranose (Gal/). Gal/ residues are essential components in the cell wall of
pathogens and play vital roles for their virulence. Thus inhibitors of LTGM that block
the biosynthesis of Gal/ could lead to novel therapeutics. To date, no eukaryotic
UGM inhibitors and only a few prokaryotic L1GM inhibitors have been reported. We
present the development of a high-throughput fluorescence polarization (FP) assay to
identify specific inhibitors of eukaryotic UGM from human pathogenic fungus
Aspergillus fumigatus. Our FP binding assay demonstrates that specific binding to
eukaryotic 4/UGM were only obtained from UDP-TAMRA chromophores, and a
UDP-TAMRA chromophore with K d value of 2.6±0.2 pM was selected as the
fluorescent probe in the high-throughput FP assay. The competitive binding assay
indicates that this UDP-TAMRA chromophore shares the same binding site with
UDP, a known UGM ligand that binds to the active site of UGM The FP assay was
evaluated and displayed excellent Z’ factor (0.79±0.01) and good tolerance to DSMO
Nine compounds were screened in this system, and one compound was identified as
4/UGM ligand. This compound was further confirmed to inhibit the activity of
4/UGM in 4/UGM activity assay, indicating that the compounds identified by our FP
high-throughput screening system are inhibitors of 4/UGM. Supported by a NIH
grant RO1-AT082542 (R Tarleton PI).

HOMOLOGY MODELS OF C-C CHEMOKINE TYPE-5 RECEPTORS. ARE
BOUND WATERS IMPORTANT IN BINDING SITES OF MEMBRANE BOUND
RECEPTORS? Saheem A. Zaidi . Philip D. Mosier, Yan Zhang & Glen E. Kellogg,
Department of Medicinal Chemistry, Virgmia Commonwealth University,
Richmond, VA 23298 HIV-1 uses two plasma membrane receptors, CD4 and a
co-receptor, to facilitate its entry into the target plasma membrane. Dependmg upon
the type of virus, this co-receptor can either be C-C chemokine receptor type-5
(CCR5) or C-X-C chemokine receptor type 4 (CXCR4). Studies suggest that CCR5
plays a dominant role in early stages of infection. Many allosteric antagonists of
CCR5 are known to inhibit fusion/entry process and subsequent infection of HIV-1.
In the present study we explored an allosteric binding site of CCR5 and the possible
role protein-bound water molecules may play in the interaction. A homology model
of CCR5 was built using recently crystallized CXCR4 as a template, followed by
virtual docking of maraviroc, a known allosteric antagonist. Mutagenesis studies

ABSTRACTS

89

from the literature agreed with the docking pose obtained. Possible protein-bound
water sites were generated and the antagonist was re-docked. According to our model
there are at least two important protein-bound waters. One of the bound waters forms
a hydrogen bond bridge between the ligand and the protein while the second bound
water is displaced due to ionic-bridge formation between die ligand and the protein,
and thus increasing the binding affinity due entropic gain resulting from disruption of
water network in the unliganded receptor.

ANALYSIS OF POST-TRANSLATIONAL MODIFICATIONS OF SIKE
FOLLOWING DOUBLE-STRANDED RNA STIMULATION. Charlotte F.
Roberts, James D. Marion, R. Jason Call and Jessica K. Bell, Department of
Biochemistry and Molecular Biology, Virginia Commonwealth University,
Richmond, VA 23298. The innate immune system is the body’s first line of defense
against infectious agents. Essential to this response are cellular mechanisms that
recognize, sequester and eradicate these invading organisms. Toll-like receptor 3
(TLR3), a pathogen recognition receptor, is stimulated by the viral genomic material
double stranded RNA (dsRNA). TLR3 stimulation initiates a signaling cascade that
leads to the production of type 1 interferon. Critical to this signaling pathway is a
kinase complex, NAP1 (NAK associated protein 1)-TBK1 (TANK Binding Kinase
l)-IKKe (IkB kinase epsilon), which leads to the phosphorylation of IRF3 (interferon
regulatory factor) and IRF7 and production of LFNp. To control this activity, SIKE
(Suppressor of IKKe) acts as a physiological inhibitor of TKKs and TBK1 activity
through an undefined mechanism. The role of post-translation modifications to
control SIKE function was examined. FLAG-tagged SIKE DNA was transiently
transfected into HEK293 followed by dsRNA stimulation for 24 h. Cell lysates were
harvested, SIKE unmunoprecipitated using anti-FLAG antibody agarose beads.
Immunoblot analysis showed serine phosphorylation state following dsRNA
stimulation. Usmg in vitro kinetic assays, TBK1 mediated SIKE phosphorylation
These results suggest that upon pathway stimulation and TBK1 activation, TBK1
phosphorylates SIKE signaling SIKE’s release from the kinase complex. Funding
provided in part by the American Cancer Society.

SUPPRESSOR OF IKK-EPSILON IS A MIXED TYPE INHIBITOR OF THE
TYPE I INTERFERON RESPONSE. James D. Marion, Charlotte F. Roberts, R.
Jason Call, and Jessica K. Bell, Department of Biochemistry and Molecular Biology',
Virginia Commonwealth University, Richmond, VA 23298. Kinases act in
signaling pathways to propagate the cellular response by phosphory lation of specific
targets that trigger downstream events that alter the cell’s transcriptional program. To
control kinase activity, the cell has developed an intricate series of protein
interactions that are required for activation or mediate inhibition. In innate immunity,
viral-derived ligands activate innate immune receptors to initiate an anti-response
including production of type 1 interferons. Downstream of these receptors, a critical
kinase complex, NAP1 (NAK associated protein 1)-TBK1 (TANK Binding Kinase
l)-IKKe (IkB kinase epsilon) phosphorylates of the transcription factor, 1RF3
(interferon regulatory factor) and IRF7, which leads to the production of IFN-beta. To
control this activity, SIKE (Suppressor of IKK-epsilon) acts as a physiological
inhibitor the kinase activity through an undefined mechanism. Our goal is to define

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the inhibitory mechanism of SIKE in the kinase reaction. Using an in vitro kinase
assay, the Michael is constant for TRF3 phosphorylation was determined (5
micromolar). Upon addition of increasing concentrations of SIKE, a decrease in
Vmax for IRF3 phosphorylation was observed with minimal change in Km. Further
analysis showed that SIKE functioned as a mixed type inhibitor with an approximate
IC50 of 450 nM. The effect of SIKE phosphorylation on inhibitory function is
currently under study. Funding provided in part by the American Cancer Society.

STRUCTURE BASED PREDICTIVE MODELS FOR PROTEIN
POST-TRANSLATIONAL MODIFICATIONS. M. Zhang' , V. A. Yakovlev 2 , R.
B. Mikkelsen 2 & G. E. Kellogg 1 , 'Department of Medicinal Chemistry and Institute
for Structural Biology and Drug Discovery, Virginia Commonwealth University,
Richmond, VA 23298 department of Radiation Oncology, Massey Cancer Center,
Virginia Commonwealth University, Richmond, VA 23298. Nitric oxide (NO) has
long been identified as a diffusible signaling molecule, but only until recent years had
gained prominence as an inducer of redox-based protein post-translational
modifications (PTMs). NO-dependent PTM is achieved, in a large part, through
cysteine S-nitrosylation, the covalent addition of the NO moiety to a reactive cysteine
thiol (other modes of action also exist, such as tyrosine nitration). Over the last
decade, the number of reported substrates for S-nitrosylation has grown
exponentially, and are shown to participate in a wide range of biological processes,
however, the precise mechanism of S-mtrosylation is still elusive, the only certainty
is that targeting of S-nitrosylation between proteins, and especially between Cys
residues within substrate proteins reflects the interactions of a number of
determinants of specificity. In an attempt to untangle these determinants, we have
developed computational methods to build models of protein post-translational
modifications based on 3D structural features, which have been successfully applied
to predict tyrosine nitration. In the current study, we are developing models that we
hope can accurately predict Cys S-nitrosylation. Preliminary analysis with 821
cysteines has proven to be promising, some interesting interactions between
structural features have been identified with a logistic regression model, and a much
simpler but more accurate model was built with a novel ant-colony algorithm.

VJAS AWARDS

91

VIRGINIA JUNIOR ACADEMY OF SCIENCE
MAY 25, 2011, 70TH ANNUAL MEETING
UNIVERSITY OF RICHMOND
RICHMOND, VIRGINIA

Agriculture and Animal Science

Honorable Mention:

Kintora Chapman

Central Virginia Governor's School

Honorable Mention:

Haven Headley

Chesapeake Bay Governor's School

Honorable Mention:

Mallory Banton

Hanover High School

Third Place:

Ryan Boles

Chesapeake Bay Governor's School

Second Place:

Clayton Geipel and Jason McAleese

SMT at Godwin High School

First Place:

Jennifer Heyward

Shenandoah Valley Governor's School

Animal Behavior

Honorable Mention:

Thomas Coleman, Kevin Gilbert and Rahim Islam
Chesapeake Bay Governor's School

Honorable Mention:

Stuart Geipel

Deep Run High School

Honorable Mention:

Sarah Graham and T.A. Njuyen

Thomas JeffersonHigh School for Science and Technology

Third Place:

Ashley Wayne

Central Virginia Governor's School

Second Place:

Natalie Hahn

SMT at Godwin High School

First Place:

Hannah Heimer

Central Virginia Governor's School

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VIRGINIA JOURNAL OF SCIENCE

Botany

Honorable Mention:

Adam Grim

George H. Moody Middle School

Honorable Mention:

William O'Brochta

Roanoke Valley Governor's School

Honorable Mention:

Jagannath Kadiyala

Godwin High School

Third Place:

Tyler Laredo

Washington-Lee High School

Second Place:

Brooke Baker and Amber Brown

Chesapeake Bay Governor's School

First Place:

Erik Zom

Roanoke Valley Governor's School

Chemistry A

Honorable Mention:

Corimie Moorefield

Ocean Lakes High School

Honorable Mention:

Kaixin Chen

Godwin High School

Honorable Mention:

Jee In Seo

Thomas Jefferson High School for Science and Technology

Third Place:

Joel Goodloe

Hanover High School

Second Place:

Arjun Jaini

Lee-Davis High School

First Place:

Prasana Joshi

SMT at Godwin High School

Chemistry B

Honorable Mention: Victoria Nguyen

Bishop O'Connell High School

VJAS AWARDS

93

Honorable Mention:

Sathya Areti and Garrick Sin

Henrico High School

Honorable Mention:

Kevin Zhan

George H. Moody Middle School

Third Place:

Ashley Canaan

George H. Moody Middle School

Second Place:

Priya Sarkar

Henrico High School

First Place:

Soyun Kim

Central Virginia Governor's School

Computer Science

Honorable Mention:

Kelvin Chen

Deep Run High School

Honorable Mention:

Kyla Bouldin

Washington-Lee High School

Honorable Mention:

Zuahear Mansiv

Washington-Lee High School

Third Place:

Bradley Lyman

Mountain Vista Governor's School

Second Place:

Christian Dolliff

Washington-Lee High School

First Place:

Luke Faraone

Yorktown High School

Consumer Science A

Honorable Mention:

Jessica Hawkins

Shenandoah Valley Governor's School

Honorable Mention:

Jessica Evans

Shenandoah Valley Governor's School

Honorable Mention:

Eileen Breslin

Washington-Lee High School

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VIRGINIA JOURNAL OF SCIENCE

Third Place:

William Joyce IV

Mountain Vista Governor's School

Second Place:

Tanner Westmoreland

Mountain Vista Governor's School

First Place:

Amna Adrees

Shenandoah Valley Governor's School

Consumer Science B

Honorable Mention:

Michelle Heath

Central Virginia Governor's School

Honorable Mention:

Mark Feinberg

Thomas Jefferson Middle School

Honorable Mention:

Frank Gaskins and Mischa Turner
Chesapeake Bay Governor's School

Third Place:

Ciara King and Mariah Martin
Chesapeake Bay Governor's School

Second Place:

Kiersten Garcia

Central Virginia Governor's School

Earth and Space Science

Honorable Mention:

Purnima Ghosh

George H. Moody Middle School

Honorable Mention:

Emily Hastings

Hanover High School

Honorable Mention:

Alan Booth

Hampton High School

Third Place:

Ramsey Natour

Central Virginia Governor's School

Second Place:

Benjamin Wong

George H. Moody Middle School

First Place:

Zoey Dorman

Central Virginia Governor's School

VJAS AWARDS

95

Honorable Mention:

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Engineering

Paul DiMartino

George H. Moody Middle School

Samantha Marquez

Maggie L. Walker Governor's School

Nick Ladocsi

Mathematics and Science High School at Clover Hill
Liz Carpenter

Central Virginia Governor's School
Akshay Deverakonda

Thomas Jefferson High School for Science and Technology

Andrew Gatow, Sherwood Lin and Robert Shock
Blacksburg High School

Environmental Science A

Ariel Bobbet

Washington-Lee High School

Sarah Goodman
Washington-Lee High School

Angela Mestre and Emilia Zevallos
Washington-Lee High School

George Maurakis

Mathematics and Science High School at Clover Hill
Casey Cooke

Mountain Vista Governor's School
Darwin Li

Thomas Jefferson High School for Science and Technology

Environmental Science B

Akshita Sharma

George H. Moody Middle School

Jordan Word
Robious Middle School

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VIRGINIA JOURNAL OF SCIENCE

Honorable Mention:

Hannah Patrick

Mountain Vista Governor's School

Third Place:

Aolin Zhang

JR Tucker High School

Second Place:

Mark Pfeifer

Mountain Vista Governor's School

First Place:

John-Reid Ryan

Hanover High School

Environmental Science C

Honorable Mention:

Margaret Anne Seif

Chesapeake Bay Governor's School

Honorable Mention:

Samantha Schwartzkopf

George H. Moody Middle School

Honorable Mention:

Sarah Sisson

Chesapeake Bay Governor's School

Third Place:

Connor Wood

George H. Moody Middle School

Second Place:

Emma Hollowell

Chesapeake Bay Governor's School

First Place:

Kendall Trivett

Chesapeake Bay Governor's School

Environmental Science D

Honorable Mention:

Seth Diggs

Chesapeake Bay Governor's School

Honorable Mention:

Hannah Wallace

Chesapeake Bay Governor's School

Third Place:

Rebecca Walker

Chesapeake Bay Governor's School

Third Place:

Andrea Green and Catherine Mitchell
HB Woodlawn High School

VJAS AWARDS

97

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Honorable Mention:

Third Place:

First Place:

Valentina Lohr
Williamsburg Middle School

Rachel Corrigan

Chesapeake Bay Governor’s School

Genetics and Cellular Biology

Reshini Premaratne

George H. Moody Middle School

Rhiannon Edwards
Gildersleeve Middle School

Anant Kharkar

George H. Moody Middle School

Radha Venkatesan
York High School

Nadine Nikolova
Deep Run High School

Mathematics

Yeji Kim

Deep Run High School
Agustey Mongia

George H. Moody Middle School

Medicine and Health A

Diana Otaya

Mountain Vista Governor's School

Paul Girgis
Godwin High School

Sayantanee Das
Godwin High School

Yanni Hedjar

Shenandoah Valley Governor's School
Harsh Patolia

Roanoke Valley Governor's School

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VIRGINIA JOURNAL OF SCIENCE

Medicine and Health B

Honorable Mention:

Aejoo Hong

Central Virginia Governor's School

Honorable Mention:

Michael Bonini

Bishop O'Connell High School

Honorable Mention:

Elena Anderson

Southwest Virginia Governor's School

Third Place:

Elizabeth Hilbert

Appomattox Regional Governor's School

Second Place:

Jason Cui

Langley High School

First Place:

Riley Ennis

Thomas Jefferson High School for Science and Technology

Medicine and Health C

Honorable Mention:

Avery Mahaney

Southwest Virginia Governor's School

Honorable Mention:

Elizabeth Levit

George H. Moody Middle School

Honorable Mention:

Sehaj Khara

JR Tucker High School

Third Place:

Nathan Lin

George H. Moody Middle School

Second Place:

Sreemoyee Som

Roanoke Valley Governor's School

First Place:

S ameer Sarlcar

Maggie L. Walker Governor's School

Microbiology A

Honorable Mention:

Angela Huckstep

Hanover High School

VJAS AWARDS

99

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Honorable Mention:

Ao Zhang

SMT at Godwin High School

Sarah Jensen
Yorktown High School

Ashley Gartin

Shenandoah Valley Governor's School
Jacob Melton

Math and Science High School at Clover Hill
Levi Helm

Roanoke Valley Governor's School

Microbiology B

Sruthi Jayaraman
Robert Frost Middle School

David Lanman

Thomas Jefferson Middle School
Emily Lodge

Bishop O'Connell High School
Monika Payerhin

Central Virginia Governor's School

Leigh Anna Morris and Marian Walker
Chesapeake Bay Governor's School

Coly Elhai

George H. Moody Middle School
Physical Science A
Kayla Sell

George H. Moody Middle School

Rebecca Merriman-Goldring
Swanson Middle School

Jacqueline Anders
Swanson Middle School

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VIRGINIA JOURNAL OF SCIENCE

Third Place:

Ryan Lucia

Stonewall Jackson Middle School

Second Place:

Claire Spaulding

Swanson Middle School

First Place:

Seth Peacock

Stonewall Jackson Middle School

Physical Science B

Honorable Mention:

Jake Cui

Kilmer Middle School

Honorable Mention:

Stephen Russell

George H. Moody Middle School

Honorable Mention:

William Deng

George H. Moody Middle School

Third Place:

Myra Ahmad

George H. Moody Middle School

Second Place:

Mihir Paithane

George H. Moody Middle School

First Place:

Caroline Fudala

James River High School

Physics A

Honorable Mention:

Molly Huss

Shenandoah Valley Governor's School

Honorable Mention:

Juliana Bain

Swanson Middle School

Third Place:

Thomas Varley

Yorktown High School

Second Place:

Sarah Thibodeau

Shenandoah Valley Governor's School

First Place:

Ellen Korcovelos

SMT at Godwin High School

VJAS AWARDS

101

Physics B

Honorable Mention:

Austin Holmes

Central Virginia Governor's School

Honorable Mention:

Melissa Havel

Ocean Lakes High School

Honorable Mention:

David Carson

Roanoke Valley Governor's School

Third Place:

Conner Peters

Central Virginia Governor's School

Second Place:

Thomas Sheehy

Thomas Jefferson Middle School

First Place:

Swapnil Pande

George H. Moody Middle School

Psychology-General

Honorable Mention:

Maya Ravindran

Douglas S. Freeman High School

Honorable Mention:

Supreeti Sharma

Mountain Vista Governor's School

Third Place:

Tori Auerhan

HB Woodlawn High School

Second Place:

Megan Geboski

Mountain Vista Governor's School

First Place:

Lakshmi Bodapati

JR Tucker High School

Psychology-Learning and Perception A

Honorable Mention:

Evelyn Strope

Roanoke Valley Governor's School

Honorable Mention:

Megan Wentworth

Roanoke Valley Governor's School

Honorable Mention:

Arm Marie May

Gildersleeve Middle School

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VIRGINIA JOURNAL OF SCIENCE

Third Place:

Amanda Hinckle and Alicia Talley

Deep Run High School

Second Place:

Elizabeth McDonald

Yorktown High School

First Place:

Michael Nguyen and Kevin Zhou

Thomas Jefferson High School for Science and Technology

Psychology-Learning and Perception B

Honorable Mention:

Diamond Pollard

Central Virginia Governor's School

Honorable Mention:

Bria Atkins

Central Virginia Governor's School

Honorable Mention:

Preston Rogers

Central Virginia Governor's School

Third Place:

Eugene Jeong and Peter Pak

Deep Run High School

Second Place:

Drew Precious

Central Virginia Governor's School

First Place:

Sarah Murphy

Douglas S. Freeman High School

Psychology-Social

Honorable Mention:

Katherine Mireles

Ocean Lakes High School

Honorable Mention:

Kelly Johnson

Shenandoah Valley Governor's School

Honorable Mention:

Lara Phillips

Yorktown High School

Third Place:

Leah Dillard

George H. Moody Middle School

Second Place:

Alison Eddins

George H. Moody Middle School

VJAS AWARDS

103

First Place:

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Honorable Mention:

Honorable Mention:

Honorable Mention:

Third Place:

Second Place:

First Place:

Margaret Doyle
Williamsburg Middle School

Statistics

Kem Anderson
Goochland High School

Brittany Moncure
Deep Run High School

Alison Hauslee

SMT at Godwin High School

Eddie Hanks
Pulaski High School

Nitin Iyer

Ocean Lakes High School
Zoology
Climothy Morris

Chesapeake Bay Governor's School

Alexander Schmitt

Mountain Vista Governor's School

Abhya Panya Vij and Jennifer Wang

Thomas Jefferson High School for Science and Technology

Andrew Leffler

George H. Moody Middle School

Andre Desruelle
HB Woodlawn High School

Natasha Sheybani

Appomattox Regional Governor's School

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VIRGINIA JOURNAL OF SCIENCE

Special Awards

Botany Section Award

Botany Section Award, given by the Botany Section of the VAS, to the best paper on
a botanical subject.

Erik Zorn

Roanoke Valley Governor's School

Mathematics Award

Mathematics Award for the paper that evidences the most significant contribution in
the field of Mathematics.

Agustey Mongia

George H. Moody Middle School

Statistics Award

Statistics Award for the paper that evidences the most significant contribution in the
field of Statistics.

Nitin Iyer

Ocean Lakes High School

Speleological Society Award

Given by the Richmond Area Speleological Society for outstanding research addressing
karst or topics related to speleology (bats, caves, carbonate geology,
paleoenvironments, limestone fossils, sinkholes, etc.

Rachel Marzen

Thomas Jefferson High School for Science and Technology

Smith Shadomy Infectious Disease Award

Given to commemorate Dr. Smith Shadomy by the Virginia Chapter of the National
Foundation for Infectious Diseases to the paper that evidences outstanding research in
the field of infectious diseases.

Prasana Joshi

SMT at Godwin High School

Roscoe Hughes Award

Roscoe Hughes Award for the best paper in the field of Genetics.

Rhiannon Edwards
Gildersleeve Middle School

Rodney C. Berry Chemistry Award

Rodney C. Berry Chemistry Award for the paper that evidences the most significant
contribution in the field of chemistry.

Prasana Joshi

SMT at Godwin High School

VJAS AWARDS

105

Dr. and Mrs. Preston H. Leake Award in Applied Chemistry

The Dr. and Mrs. Preston H. Leake Award in Applied Chemistry will be given to the
author of a research paper which best exemplifies how chemicals, chemical principles,
or chemistry have been used, are used, or might be used to enhance or even to save life.

Riley Ennis

Thomas Jefferson High School for Science and Technology

Catesby Jones - Russell J. Rowlett Award

Catesby Jones - Russell J. Rowlett Award for the Best Research Paper of the Year.

Michael Nguyen and Kevin Zhou

Thomas Jefferson High School for Science and Technology

Virginia Sea Grant College Program

Virginia Sea Grant College Program Award is given by the Virginia Sea Grant College
Program for outstanding marine or coastal research.

Rachel Corrigan

Chesapeake Bay Governor's School

American Cancer Society Award

American Cancer Society Award - This award is to recognize outstanding science
papers related to cancer research. These awards are provided by the American Cancer
Society.

Riley Ennis

Thomas Jefferson High School for Science and Technology

Sayantanee Das
Godwin High School

Radha Venkatesan
York High School

Gamma Sigma Delta Award (Agriculture)

The Gamma Sigma Delta Award (Agriculture). Presented by the VPI & SU Chapter of
the Honor Society of Agriculture. This award is presented in recognition of excellence
in research dealing with application of new technologies and/or concepts in agriculture
forestry, or veterinary medicine.

Jennifer Heyward

Shenandoah Valley Governor's School

Do mi nion - W.W. Berry Award

Dominion - W.W. Berry Award. This award is given by Dominion Virginia Power in
honor of Mr. W. W. Berry who was a past Chairman of the Board of VA Power.

Andrew Gatow, Sherwood Lin and Robert Shock
Blacksburg High School

Joyce K. Peterson Award

The Joyce K. Peterson Award is presented for the outstanding paper by a middle school

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VIRGINIA JOURNAL OF SCIENCE

student. It is presented in honor of Mrs. Joyce K. Peterson who has been an outstanding
teacher in the Arlington County Schools.

Valentina Lohr
Williamsburg Middle School

Ann M. Hancock Award

The Ann M. Hancock Award - This award is given to the best paper in cellular biology
and is given in memory of Anne Hancock who retired from Patrick Hen 17 High School
in Hanover County and who gave many years of service to the Jr. Academy not only
by teaching but also serving on the Jr. Academy Committee.

Nadine Nikolova
Deep Run High School

Dorothy Knowlton Award

Dorothy Knowlton Award - This award is given to the best paper in the Consumer
Science section(s) and is given in honor of Dorothy Knowlton, former Science
Coordinator of Arlington County Schools.

Lukas Mohnach

Chesapeake Bay Governor's School

VABE Award

outstanding research in Zoology VABE Award - This award is presented by the
Virginia Association of Biology Educators and is given for outstanding research in the
Zoology section.

Natasha Sheybani

Appomattox Regional Governor's School

Virginia Museum of Natural History Award

Virginia Museum of Natural History Award - Presented by the Friends of the Virginia
Museum of Natural History in recognition of significant contribution in the study and
interpretation of Virginia's Natural Heritage.

Rachel Corrigan

Chesapeake Bay Governor's School

AAAS Aw ard and Honorary Membership

The AAAS Award is given for presenting an outstanding paper at this symposium. This
student will also receive an honorary membership to AAAS and a cash award of $200.

Prasana Joshi

SMT at Godwin High School

VAS Honorary Membership

One year's membership, without cost, to the Virginia Academy of Science including
a subscription to the Virginia Journal of Science will be awarded.

Ana O'Harrow
Yorktown High School

VJAS AWARDS

107

Bethel High School Scholarship

Bethel High School Scholarship - This $1,000 Scholarship Award comes from the
interest earned from a $ 10,000 endowment contributed by the students of Bethel High
School, Hampton, Va., over a two year period. This award is based on both the
student's presentation and paper.

Rachel Corrigan

Chesapeake Bay Governor's School

Henry MacKenzie Environmental Scholarship (VEE)

Henry MacKenzie Environmental Scholarship - This $5,000 scholarship will be
awarded to the student whose paper evidences the most significant contribution in the
field of Environmental Science dealing with the James River Basin and Chesapeake
Bay. The Virginia Endowment and VJAS offer this scholarship in tribute to the
outstanding and generous services of Judge Henry W. MacKenzie, Jr., one of the
founding directors who has a great interest in the James River and the Chesapeake Bay.

Alan Booth
Hampton High School

Frances and Sydney Lewis Environmental Scholarship (VEE)

Frances and Sydney Lewis Environmental Scholarship: A $ 14,000 scholarship ($3,500
per year for four years) for the best effort by a student in grades 9 to 12 in the field of
environmental science. This scholarship is in the name of Frances and Sydney Lewis
and is given by the Virginia Environmental Endowment.

Sarah Sisson

Chesapeake Bay Governor's School

E.C.L. Miller Science Teacher of the Year Award

E.C.L. Miller Science Teacher of the Year Award is given to an outstanding science
teacher.

Tim Criner

Gildersleeve Middle School

Distinguished Service Award

VJAS Distinguished Service Award, most prestigious award given by the VJAS, is
presented to a person for exceptionally outstanding service to the VJAS.

Michael Lovrencic
Yorktown High School

NOTES