The Ohio Journal of - The Ohio Academy of Science

Vol. 112(1)
average zone of 32 mm for Staphylococcus aureus, 35 mm for
Staphylococcus epidermidis, and 26 mm for Lactobacillus
acidophilus. The Thieves’ oil had an average zone of 15
mm for Staphylococcus aureus, an average of 12 mm for
Staphylococcus epidermidis, and no zone for Lactobacillus
acidophilus. The colloidal silver and garlic oil had no zone
for each trial and were therefore ineffective. This study
demonstrates that alternative remedies may have efficacy
in treating some antibiotic resistant infections.
Poster Board No. 057 THE KEY ROLE OF ADRA1D IN MAKING
B-BLOCKERS MORE EFFECTIVE. Peeyush Shrivastava, 6555
Glenstone Way, Mason OH 45040 (Mason HS).
The adrenergic receptors mediate the actions of
norepinephrine and epinephrine throughout the body;
stimulation of an adrenergic receptor occurs by epinephrine
binding to an agonist. The myocardial response is a fatal
increase in heart rate, known as a ventricular tachycardia
(VT). The objective of this study is to identify the cause for
the failure in beta-blocker treatments, and identify how to
overcome the problem. ADRA1D was the variable of gene
knockdown in this study because ADRA1D was the most
unique receptor subtype, showing no fluctuations in cAMP,
unlike other receptors. The efficiency of ADRA1D gene
silencing was determined using Western Blot analysis
in which cell lysate samples were loaded onto 12% SDS
gels and then transferred onto nitrocellulose membrane.
For the average of the three trials conducted for Sotalol,
it appears that the control heart rate of the cardiac
myocytes was reduced from the initial contraction rate
of 131.67 bpm to 103 bpm with the applied non-selective
beta-blocker. These data are consistent with the original
purpose of experimentation considering the non-selective
beta-blocker did not effectively reduce heart rate to below
100 bpm. Furthermore, once ADRA1D was silenced
through N-terminal encoded siRNA transfection, heart
rate surmounted from the initial 131.67 bpm to 157.33
bpm; the fact that the blocker was applied simultaneously
as ADRA1D was silenced and heart rate still increased in
the cardiac myocytes supports the hypothesis that the 30%
of failure of beta-blockers to reduce heart rate in cardiac
myocytes can be traced to the key role of ADRA1D.
Poster Board No. 058 ATTACHING MOLECULES TO COWPEA
MOSAIC VIRUS FOR APPLICATIONS IN DRUG DELIVERY.
Aparna Narendrula aparna.n.14@gmail.com, (Nicole F.
Steinmetz, nfs11@case.edu) 34240 Seminole Way, Solon OH
44139 (Hathaway Brown School)
Cancer is the second leading cause of death in America.
Chemotherapy, the leading cancer treatment, treats
cancer with chemicals destroying cancer cells. It does not
differentiate between cancer and healthy cells, therefore
results in harmful side effects. Through nanomedicine,
nanoparticles deliver therapeutics to cancer while avoiding
healthy tissues, increasing therapy and decreasing side
effects. The goal of this research is to see if organic and
benign plant viral nanoparticle cowpea mosaic virus (CPMV),
can be used as a vehicle to carry drugs to target areas. The
hypothesis is CPMV will bind cancer specific targeting
proteins. To test this hypothesis, a sample of CPMV was
purified and modified by attaching Oregon Green 488®,
which was used to image and detect the particles in cells, and
biotin, which served as a molecular handle to bind cancer
specific targeting proteins. These CPMV nanoparticles were
produced in cowpea plants and isolated from infected leaves
through centrifugation and chemistry; concentration was
determined using UV-Vis spectroscropy: a peak at protein’s
absorbance indicated CPMV’s presence (0.401 absorbance).
Once purified, these nanoparticles were synthesized with the
dye; agarose and SDS gels and UV-Vis spectroscopy affirmed
the presence of Oregon Green 488® (there was a peak (0.021)
at the dye’s absorbance spectrum). There were 51 and 45
dyes per particle for the samples. Biotin was attached and
confirmed with a stained and unstained agarose gel and a dot
blot. This unit was used to target cancer cells. This technology
has the potential to replace chemotherapy in treating cancer.
The Ohio Journal of Science A-47
Poster Board No. 059 IMMUNOHISTOCHEMICAL
IDENTIFICATION OF CELLS IN THE RESPIRATORY RHYTHM-
GENERATING CENTER OF THE DEVELOPING RAT BRAIN.
Jennifer L. Jones 1 , jlhbjones@sbcglobal.net, Catherine A.
Mayer 2 , caa4@case.edu, Christopher G. Wilson 2 , cgw5@
case.edu, 1 2201 Chatfield Dr., Cleveland Heights OH 44106
(Hathaway Brown School) and 2 Dept. of Pediatrics, Case
Western Reserve Univ., Cleveland OH 44106.
Episodes of apnea that can lead to long-term respiratory
morbidities are one of the major problems faced by
premature infants in neonatal intensive care units.
This project investigated the developmental changes in
expression of the underlying neurotransmitters and neural
network using immunohistochemistry. The regions of the
brain responsible for respiration have been extensively
studied in the rat. The regulation of mammalian respiration
by the inhibitory neurotransmitter adenosine evolves
during early brain development. Activating adenosine A 2A
receptors can induce apnea in young rats, but not in rats
older than postnatal day 21 (P21). Sprague Dawley rats
were studied at ages P0, P4, P7, P14, and P21. Brainstem
tissue sections (20 µm) were taken and immunostained for
neural and astrocytic marker proteins (NK1, NeuN, A 2A,
GAD, and S100β) taking 5 samples of each age/protein
combination. Commercially manufactured primary
antibodies were used to characterize cell types (Millipore
AB5060, Millipore MAB377, Genway GWB-32D8EE,
Millipore MAB5406, and Abcam AB4066 respectively).
The stained sections were viewed by a camera attached
to a light microscope, and the images analyzed by
Stereology Resource Center unbiased stereology software
which counted the numbers of stained cells. This
allowed neurons, A 2A receptors, GABAergic neurons, and
astrocytes in the respiratory rhythm-generating center,
the preBötzinger Complex (pBC), to be localized and cell
densities to be calculated. Neuron density (cells/10 6 µm 3 )
decreased 53% from 168 at P0 to 79 at P7. A 2A receptor
density (receptors/10 6 µm 3 ) decreased 87% from 163 at P0 to
21 at P7. GABAergic neuron density was relatively stable
over P0 to P21, with 9% variation, while astrocyte density
rose 27% over P0 to P21. The observed rapid drop in A 2A
expression in the pBC of rat pups during the first week
of life suggests that the reduced susceptibility to apnea
observed in maturing neonates may be a consequence
of the declining role of the inhibitory neurotransmitter
adenosine in the neural network controlling respiration.
Poster Board No. 060 PATHOLOGICAL EFFECTS OF TBI IN A
MOUSE MODEL OF ALzHEIMER’S DISEASE. Lena J Ransohoff,
LRansohoff13@gmail.com, Hathaway Brown School, 19850
Marchmont Rd., Shaker Hts. OH 44122, Olga N Kokiko-
Cochran, Kokikoo@ccf.org, Bruce Lamb, Lambb@ccf.org
Alzheimer’s Disease (AD) is the most common cause for
dementia. Post-mortem AD brain tissues show betaamyloid
(Aβ) plaques, intracellular neurofibrillary tangles
(NFT), neuroinflammation and neuronal loss. Traumatic
brain-injury (TBI) is a significant risk factor for AD. At
post-mortem, TBI brain tissues can exhibit AD-like
neuropathological changes and patients can show memory
impairment during life. This project studied a mouse
model for the interaction between Aβ-deposition and TBI,
using R1.40 mice, in which a transgene contains a genomic
copy of the human amyloid precursor protein (APP)
with a familial AD mutation. Previous studies showed
neuroinflammation at 6 months of age, with Aβ plaques,
learning and memory impairment after 12 months. Strainmatched
wild-type (WT) mice were used as controls. At 2
months of age, WT and R1.40 mice (N=4/group) received
fluid-percussion brain injury. Behavioral testing, sacrifice
and pathological examination were all done 3 months later.
Rotorod (motor), Y-maze (memory) and Morris water-maze
(learning) were tested to assess behavior. Histochemistry
characterized lesion size and cellular pathology. Rotarod
testing showed equivalent deficits in percussion-injured
WT and R1.40 mice. WT and R1.40 sham mice showed
average of latency (in seconds) which was borderline