Poster Presentations
SESSION 1: MICROBIOLOGY, MOLECULAR AND CELLULAR BIOLOGY I
FIRST PLACE
Ancar, Rachel. Phi Rho, University of Colorado/Denver. Activation of Src Tyrosine Kinase by
Lipid Phosphatidic Acid (PA) in Lipid Rafts.
During fertilization, the signaling pathway initiated by the binding of the sperm to the
egg begins at the plasma membrane of the egg. Lipid rafts are required for fertilization
(Belton Jr et al., 2001; Sato et al., 2002). A measure of rafts are detergent resistant
membranes (DRMs), which have a liquid-ordered state, high levels of sphingomyelin,
and high levels of cholesterol (Luria et al., 2002). We isolated Xenopus egg DRM and
non-DRM fractions with 1% Triton-X-100 and density gradient centrifugation using
Optiprep. A light scattering band near the top of the centrifuge tube contains the DRMs.
One mL fractions were collected from the top; rafts were in fractions #3 and #4, whereas
the non-raft membrane in fractions #9 and #10. Sample buffer was then added to each
density gradient fraction, and Western detection was performed for caveolin 1 (a raft
marker), Src and PLCγ. In Xenopus, Src is present in DRM before fertilization whereas
PLCγ translocates to DRM only after sperm addition (Luria et al., 2002; Sato et al., 2002;
Sato et al., 2003). As noted, rafts are required for fertilization, and Src is located in egg
DRM before fertilization whereas PLCγ translocates to DRM after fertilization (Sato et
al., 2002; Sato et al., 2003). Five minutes after addition of dPA, the amount of Src in
DRM decreased by ~30%, but levels returned to control levels by 10 minutes and then
doubled by 30 minutes. Based on this and the cellular phosphoSrc data, PA may activate
Src rapidly but this activation could lead to lowered levels of Src in rafts. In contrast to
Src, dPA induces a 2.2-fold increase in PLCγ in DRM at 5 minutes and levels return to
basal by 15 minutes. As opposed to Src, which is already in rafts before fertilization,
dPA induced PLCγ translocation to DRM to mimic fertilization.
SECOND PLACE
Ignatz, Christopher. Xi Psi, Duquesne University. Pain, Depression, and Anxiety Modulation
Through a Novel CNS Active Marine Cyanobacterial Compound.
Comorbid pain and depression affects millions of Americans. Most pharmaceuticals treat
either pain or depression, but rarely both. Those suffering from comorbidity take multiple
drugs, sometimes eliciting negative side effects. This project focuses on the antidepressive, analgesic, and anti-anxiety capabilities of a novel compound from a marine
cyanobacterium. Fraction 2064h was extracted and purified from a marine
cyanobacterium using chromatography and analyzed using NMR and HPLC-MS.
Fractions were pre-screened for activity of the CNS receptors. Fraction 2064h was found
to bind to 5-HT2C receptors. It was tested in vivo through administration to the CNS
utilizing intracerebroventricular cannula injections in C57Bl/6J mice. Analgesic qualities
of 2064h were tested in vivo using the spontaneous formalin test. We then performed the
tail suspension test and forced swim test, widely used depression assays, to test the antidepressant potentials of this compounds. Lastly, anti-anxiety properties of 2064h were
assessed through the open field test. Biological results to date will be presented.
THIRD PLACE
Singh, Shivali D. Theta Omega, Gannon University. Relative expression of CMADS2,
CMADS3, and CMADS6 in the Fern Ceratopteris richardii
MADS box-genes are a family of transcriptional regulators in eukaryotic development.
In plants, MADS genes are involved in processes throughout the plant life cycle,
including both gametophyte and sporophyte stages. In the model system Ceratopteris
richardii, a homosporous fern, a small set of MADS genes including CMADS2, CMADS3
and CMADS6 have been shown to be expressed. Here we develop a technique for
determining the relative expression of CMADS2, CMADS3 and CMADS6 in
hermaphrodite and male gametophytes as well as sporophyte tissue using RT-qPCR.
RNA was isolated from three-week old hermaphrodite and male gametophytes, exposed
to DNase, and converted to cDNA. RNA was also isolated and processed from the initial
leaves produced by young sporophytes. Primers were designed to recognize each unique
member of the CMADS gene family. Sequence analysis verified that the amplicon
produced was the same for each CMADS gene in hermaphrodite, male, and sporophyte
tissue, but distinct for each CMADS gene. CMADS2, CMADS2 and CMADS6 are
expressed in hermaphrodites, males, and sporophytes, though at different levels.
HONORABLE MENTION
Blum, Samuel. Lambda Nu, Frostburg State University. Effects of The Hessian Fly Responsive
Gene on Tobacco Hornworm Development.
Insect pests are becoming an ever increasing problem in food production, especially as
world populations continue to grow. One way to combat these insects is to engineer
plants to resist them using genes from various species. Transgenic tomato plants were
engineered using 1 of 3 genes from wheat and used in experiments to see if they are more
or less resistant to Tobacco Horn Worm. Caterpillars that fed on plants with the Hfr-1
gene did have a significantly higher mortality rate. In addition, plants containing Hfr-1 or
Hfr-3 inhibited caterpillar growth during the later stages of development. These results
indicate that plant genes from various species of plants can be used to engineer plants that
are more resistant to insect damage, thus providing more food for growing world
populations.
SESSION 2: MICROBIOLOGY, MOLECULAR AND CELLULAR BIOLOGY II
FIRST PLACE
Tepper, Sarah, Denis Avey, and Fanxiu Zhu. Sigma Tau, Florida State University.
Characterization of the Processivity Factor PF-8 in Kaposi’s Sarcoma-associated
Herpesvirus.
Kaposi’s sarcoma-associated herpesvirus (KSHV) is an oncogenic DNA virus and the
etiological agent of three human cancers: Kaposi’s sarcoma, primary effusion lymphoma,
and multicentric Castleman’s disease. To ensure its propagation, KSHV encodes a DNA
polymerase designed to replicate the viral genome. Achieving high replication efficiency
requires a processivity factor, known in KSHV as PF-8, which prevents polymerase
dissociation from the template DNA strand. After infection, KSHV exists in a dormant,
non-replicative state called latency. To maintain this state, the virus uses host cell
histones to package the genome and repress transcription. KSHV must transition from
latency and enter the lytic cycle to actively replicate its genome and produce progeny
virions, necessitating regulatory proteins able to manipulate histones and remodel the
chromatin. We recently discovered that PF-8 co-purifies with histones in KSHV
lytically-infected cells. Interesting, we also found that PF-8 undergoes a post-translational
modification called biotinylation and that the modified form is present in the histone
extracts. Further examination of PF-8 is necessary to determine the significance of its
biotinylation and association with cellular chromatin to KSHV lytic replication and
pathogenesis. The targeted inhibition of the essential functions of PF-8 may represent
novel therapeutic strategies to treat or prevent KSHV-associated diseases.
SECOND PLACE
McGuire, Joshua Tanner and Dr. Edith Osborne. Epsilon Sigma, Angelo State University.
Hofmeister Effects on the Structure and Fluorescence of Green Fluorescent Protein.
The Hofmeister series describes a set of cations and anions that have been arranged in
order of their ability to affect the physical properties of solutions and proteins. Salts in
the Hofmeister series will be used to investigate the structural properties of Green
Fluorescent Protein, GFP. The goal is to detect structural intermediates induced by the
Hofmeister salts prior to the major unfolding transition. Structural changes will be
detected with circular dichroism spectropolarimetry and compared to changes in
fluorescence and fluorescence quenching induced by Hofmeister salts. These
experiments will contribute to the understanding of Hofmeister effects and help
characterize how subtle structural changes affect the fluorescence of GFP. Data
collection is in the final stages.
THIRD PLACE
Mundy, Paige, Matthew Williams, Zachary Sterner, and Irene M. Wolf, Ph.D. Upsilon Beta,
Saint Francis University. Analysis of Various Response Mechanisms to Cellular Stress
Using Plant Models Elodea densa and Arabidopsis thaliana
This project aims to assess the cellular stress on plant models. One of the main avenues used is
heat shock protein 70. The submergible plant model Elodea densa has been subjected to varying
temperatures to measure HSP70 expression. This was done with intentions of discovering the
temperature at which HSP70 is induced. It is known that plants are more prone to methylation
under stressful conditions in order to regulate gene expression. Generally, the more methylation
occurring on the DNA of the promoter region for the gene, the more repression of gene
transcription occurs. We seek to determine whether changes in methylation are occurring on the
HSC70-2 gene in response to heat stress. In order to asses this methylation, a hydroponic growth
system was created for Arabidopsis growth. Methylation sensitive restriction enzymes were used
to assess methylation occurring in the promoter region of HSC70-2 and Rubisco genes. In order
to explore different physiological responses to stress we are currently evaluating cell viability
and reactive oxygen species in elodea and arabidopsis using florescent staining.
SESSION 3: ECOLOGY
FIRST PLACE
Blatzheim, Lauren 1, Corey D. Bower2, Trimelle Polk3, Amhed Karahan4, Dilan Ikizoglu4,
Brianna Levinson5, Nazmiye Guneş4, Ibrahim Çakmak4, Harrington Wells6, and John M.
Hranitz2 1Southwestern Oklahoma State University, 2Bloomsburg University, 3Southern
Nazarene University, 4Uludağ University, Bursa, Turkey, 5University of California, San
Diego, 6University of Tulsa. Delta Sigma, Southwestern Oklahoma University.
Neonicotic Pesticide Thiamethoxem Affects Motor Responses and Foraging Behavior of
Honey Bees.
Neonicotinoid compounds (e.g., thiamethoxam) are the next generation of pesticides to
target nicotinic-Ach receptors to be broadly used in agriculture. Neonicotinoids have
been have been implicated as causal agents in honey bee colony collapse disorder (CCD),
since the onset CCD coincided with the widespread use of neonicotinoids. The doses that
bees encounter in agriculture are well below the LD50 and considered "safe" but
sublethal effects may be important. Our goal was to study the sublethal effects of
thiamethoxam on the motor coordination of captive bees and foraging behavior of freeflying bees. We tested doses at 1/5 to 1/500 of the LD50 for motor responses after 4 h of
administration for harnessed bees and 60-150 minutes after administration in free flying
bees. At 4 h post-administration, motor coordination was assessed for antennal
movement, proboscis extension reflex, leg, and abdomen movement. Motor coordination
of bees treated with 1/5 LD50 was lower than motor coordination of control bees. We
assessed the visitation rate and foraging abilities of free-flying bees on an artificial flower
patch. The return rate was significantly lower for pesticide-treated bees at doses as small
as 1/10 LD50 and the ability of foragers to distinguish between high and low sucrose
nectar was impaired at doses higher than 1/50 LD50. Our results show significant
sublethal effects of thiamethoxam, impairing basic motor coordination and foraging
performance of honey bees at doses substantially lower than the LD50.
SECOND PLACE
Peterson, Kendra, J. A. D. Parrish, and Travis E. Wilcoxen. Iota Epsilon, Millikin University. Effect
of Herbivory on Photosynthetic Rates in Soybean (Glycine max) Plants of Different Ages.
We explored the effects of herbivory on photosynthetic rates of different aged soybean
(Glycine max) plants. We planted soybeans on different dates to get four different age groups
of plants aged 40, 45, 58, and 71 days old. We used 60 total plants from each age for three
separate experimental groups, one with no damage for the control group (n = 20), a group
subjected to mechanical damage on 60% of the center leaflet (n = 20), and another group
subjected to feeding by five-day old painted lady caterpillars (Vanessa cardui) for ten hours
(n = 20). We measured photosynthetic rates of the leaflet adjacent to the damaged one on the
third trifoliate and the middle leaflet of the leaf above the damage (4 th trifoliate) every day for
five days following damage. The 58 day-old soybean plants showed the greatest increase in
photosynthetic rates after caterpillars herbivory. Compared to the control plants, there was a
significant difference among the treatments, age of the plants, and day tested. Overall, the
photosynthetic rates were significantly higher for the caterpillar and mechanical damage
treatments, showing a compensatory increase in photosynthetic rate in the remaining leaf
tissue. Although the control photosynthetic rates did not change significantly over time,
leaves fed on by caterpillars had the highest photosynthetic rates at all ages. Leaflets above
the treated leaflet followed the same pattern of rate increases, with the newer, higher leaves
having significantly higher rates than compared leaflets. This study provides evidence that
moderate damage to soybeans by insects may result in increased yields.
THIRD PLACE
Fuller, Brian, Tyler Watson, Ellen Butts, Dr. Greg Andraso, and Dr. Kelly Grant. Theta Omega,
Gannon University, Erie Pennsylvania Detecting Invasive Species in the Feces of
Predatory Fish
The ecology of Lake Erie is disrupted by non-native species, such as the tubenose goby
(Proterorhinus semilunaris), recently discovered in Presque Isle Bay. Its impacts on the
food web are relatively unstudied; we are interested in discovering what fish prey upon
the tubenose goby; to extend the utility of our study, we included another invasive
species, the round goby (Neogobious melanostomus), even though predation on it is
better studied. Our goal is to specifically detect the DNA of these invasive species in the
feces of predatory fish. This approach offers advantages compared to traditional gut
content analysis; it allows us to examine many more samples without sacrificing the fish.
In order for this assay to be successful, we must be able to specifically identify DNA
from tubenose and round gobies. During the initial phase of this project, we optimized
primers to specifically amplify segments of cytochrome oxidase I (COI) from tubenose
and round gobies. We tested the specificity of our primers against a panel of DNA
purified from different fish species. Currently, we are testing the effectiveness of our
primers when used with DNA extracted from fecal samples. These results will allow us to
determine whether the predator recently consumed either tubenose goby or round goby.
SESSION 4: CELL, ORGANISMAL AND DEVELOPMENTAL BIOLOGY
FIRST PLACE
Nick Sheehan. Lambda Theta, Mount Saint Mary College. The Effects of Roundup Herbicide on
Head Regeneration in Dugesia tigrina, Brown Planaria, and Dugesia dorotocephala, Black
Planaria
Though ecological toxicity from the use of Roundup herbicide has been reported, little is
known about its effects on invertebrates. The purpose of this study was to determine its
toxicity to head regeneration in Dugesia tigrina, brown planaria, and Dugesia dorotocephala,
black planaria. Commercially available Roundup Herbicide was exposed to decapitated
brown (5-10 ppm) and black flatworms (10-15ppm), cultured under artificial conditions ( 23
C and L:D cycle of 14:10hr) . Though high concentrations of herbicide impede the head
regeneration process, low concentrations did not adversely affect regeneration of either
species. In addition, the negative phototropic response was observed during the various
stages of head regeneration. Though the herbicide prolonged (5ppm) or blocked (10ppm) the
negative phototropic response in brown planaria, it only retarded (at 15ppm and 10ppm) this
response in black planaria throughout head regeneration. Though the toxic effect of Roundup
(10ppm) on head regeneration appears attenuated, it still prolongs the negative phototropic
response in brown planaria. Analysis of the total proteins of the regenerates suggests that
exposure may alter the timing of protein expression in herbicide-exposed black planaria.
Though reports have praised the use of this herbicide for crop productivity, it is important to
evaluate the impact of Roundup on the larger ecosystem.
SECOND PLACE
Acquaviva, Joseph T. III, Cody F. Bahavar, Fefian Zhou, and Wei R. Chen. Psi Mu, University
of Central Oklahoma. Anti-tumor Response Induced by Immunologically Modified
Carbon Nanotubes and Phototherapy Using a Rat Mammary Tumor Model
Since ancient times, cancer has plagued society. So far, effective treatment modalities for
metastatic cancers have been elusive. Previously, an innovative cancer treatment, laser
immunotherapy (LIT), has induced effective immunological anti-tumor responses in
patients with late stage metastatic cancer. Paramount to the success of LIT is the antitumor immune response induced by local laser irradiation and immunological
stimulation. Recently, we conjugated single-walled carbon nanotubes (SWNTs) with the
immunostimulant glycated chitosan (GC), creating the novel immunologically modified
carbon nanotube (SWNT-GC) for the treatment of cancers. First, DMBA-4 mammary
cancer cells (1.0 x 105) in a 0.1 ml solution were injected into the flank region of female
Wistar Furth Rats. The primary tumors of the rats, injected with SWNT-GC, received
local phototherapy with a power density of 1W/cm2 at 980nm. We hypothesized
Laser+SWNT-GC with a longer duration of laser irradiation (phototherapy) would be
more effective at inducing an anti-tumor response in rats. After treatment, the sizes of the
primary and subsequent metastatic tumors were measured. Laser+SWNT-GC with a
longer irradiation time (10 minutes) proved to be more effective at causing regression of
primary and metastatic tumors. Laser+SWNT-GC for a longer irradiation time (10
minutes) produced a higher survival rate, 50% n=10. While Laser+SWNT-GC with a
shorter irradiation time (5 minutes) did not produce a survival rate, 0% n=8. This novel
nanotechnology-based laser immunotherapy could lead to an effective therapy for
metastatic cancers.
THIRD PLACE
Hylton, Andrew. Mu Epsilon, Troy University. Synergistic behavioral, neuronal, and cortisol
alterations in zebrafish (Danio rerio) following exposure to fluoxetine and ketamine.
This pilot study will investigate the efficacy and mechanisms-of-action of acute subanesthetic doses of ketamine and fluoxetine for the treatment of severe depression; using
an established animal model, zebrafish (Danio rerio), in Dr. Pittman’s laboratory. The
overall aim of this project is to evaluate the role that several neurotransmitters and glia
play in major depression and possibly suicide. The specific aims are to: (1) estimate
quantitatively the changes in morphology and distribution of neurons from a number of
brain nuclei from pharmacologically induced depressed zebrafish, (2) explore the
efficacy of ketamine as compared to a selective serotonin reuptake inhibitor (SSRI),
fluoxetine, on the depression-related behaviors of zebrafish using behavioral assays and a
videotracking system developed in Dr. Pittman’s laboratory. The project is innovative in
that it directly tests the importance of a wide spectrum of neurotransmitters, and aims to
elucidate the circuits of most importance. The results of the project should greatly aid in
understanding the neuroanatomical substrates of depression and offer insight at the
molecular level of ketamine’s mechanism-of-action following an acute exposure. It is
anticipated that the pilot study findings will lead to explanations of how ketamine and
fluoxetine may synergistically contribute not only to the rapid means of alleviating severe
symptoms, but more importantly identify new drug targets with perhaps even greater
efficacy, which is desperately needed in emergency situations involving major depressive
disorder and suicide.