The Ohio Journal of - The Ohio Academy of Science

Vol. 112(1)
by a comparison to a ouabain standard curve where the IC 50
was 2.3 μg/ml. Comparison of the plant extracts with the
ouabain standard curve showed that 1.73 g of Convallaria
majalis and 2.16 g Verbascum phlomoides contained 1 mg
of ouabain-like activity. The remaining extracts contained
no detectable levels of oubain-like activity. These results
demonstrate digitalis-like factors are not restricted to
the figwort family, although they are present at lower
concentrations in related species.
Poster Board No. 008 ARTIFICIALLY-INDUCED
OLIGODENDROCYTE APOPTOSIS AS A MODEL FOR MULTIPLE
SCLEROSIS. Ingrid N. zippe. ingridzippe@gmail.com.
17370 South Park Boulevard, Shaker Heights, Ohio 44120.
(Hathaway Brown School)
Multiple sclerosis (MS) is an autoimmune disorder resulting
from myelin degeneration in the central nervous system
(CNS). A growing body of research suggests that an early
stage of aberrant oligodendrocyte apoptosis precedes
T-lymphocyte infiltration and myelin deterioration in
MS. An experiment was designed to study the effects of
oligodendrocyte apoptosis within the CNS. Caspase 9 is an
enzyme that begins cellular apoptosis. A line of transgenic
mice was engineered in which the MBP (myelin basic
protein) promoter unique to oligodendrocytes promotes both
an inducible version of Caspase 9 (iCP9) and a fluorescent
marker (DsRed). Systemic exposure to an iCP9 crosslinking
agent (CID) induces the oligodendrocyte apoptosis
cascade. The dorsal column region surrounding the spinal
cord of wild-type and transgenic mice were injected with
either saline or CID at postnatal day-4. The animals were
sacrificed, and the spinal cord tissue was fixed at postnatal
day-7. Immunohistochemical staining in transgenic mice
revealed co-labeling of MBP and DsRed, as well as exclusivity
of DsRed and other neural cell markers, substantiating
the oligo-specificity of the model. Staining for CC1, an
oligodendrocyte marker, demonstrated a 43% decrease in
CC1+ cells between wild-type and CID-injected transgenic
mice, and staining for DsRed demonstrated an 80% decrease
in DsRed+ cells between saline-injected transgenic mice
and CID-injected transgenic mice. MBP and neurofilament
staining of experimental tissue revealed scattered myelin
sheaths and axons, similar the typical phenotype of
late-stage MS tissue. We conclude that oligodendrocyte
apoptosis may contribute to myelin disorganization and
morphological change in axonal arrangement.
Poster Board No. 010 INVESTIGATION OF DEXAMETHASONE
LOADING IN SYNTHETIC PLATELETS. Serena W. Chang,
schang13@hb.edu, Ranti Ositelu, rositelu@gmail.com,
Andrew Shoffstall, andrew.shoffstall@case.edu, Erin Lavik
Sc.D., erin.lavik@case.edu, Case Western Reserve University,
3418 Thomson Circle, Rocky River OH 44116. (Hathaway
Brown School)
Uncontrolled hemorrhage is a prevalent cause of death in
military and civilian trauma patients. Synthetic platelets
in the form of functionalized nanoparticles have been
recently developed to help staunch internal hemorrhage.
This project aims to extend these synthetic platelets’
functionality even further by investigating their loading
with the corticosteroid dexamethasone to address the
subsequent inflammation that develops after trauma.
Since the synthetic platelets collect at an injury site, they
have the potential to address this secondary issue locally.
Particle size is an important factor that determines the
drug release profile as well as determines the safety for
intravenous injection; particles must be larger than ~50
nm so as not to be filtered by the kidneys and substantially
smaller than the ~5 micron capillaries through which they
are proposed to circulate. This study investigated particle
diameter after loading them with dexamethasone at 10%,
20%, and 40%, weight/weight. Size was characterized with
dynamic light scattering and scanning electron microscopy
and was distributed between 400 and 600 nanometers,
with 20% loading having the smallest diameters. These
results are important because they show that drug loading
The Ohio Journal of Science A-39
with dexamethasone is possible and maintains a viable
size distribution for intravenous injection.
Poster Board No. 011 COMBATING BIOTERRORISM:
DETERMINATION OF MINIMUM PATHOGEN DNA TEMPLATE
CONCENTRATION (MPC) FOR RAPID, ACCURATE, AND
EFFICIENT DETECTION OF BIOTERROR AGENTS!. Anamika
Veeramani, malar44133@yahoo.com, 9388 Chesapeake Dr.,
North Royalton OH 44133. (Laurel School)
This project estimates Minimum Pathogen DNA template
Concentration (MPC) for select pathogens to ascertain the
feasibility of predetermining and publishing MPC data for
bioterror agents. MPC is defined as the lowest concentration
of the pathogen DNA template in the PCR assay that
results in a clear gel band indicating the presence of the
source pathogen. pUC plasmid extract and DNA extracts of
Escherichia coli, and Bacillus subtilis were chosen as DNA
templates. DNA template concentrations were prepared
using serial dilution method. illustra puReTaq Ready-To-
Go PCR beads were used with primers, Eub16S1 and
Eub16S2 for E.coli, and B.subtilis, and primers M13F
and M13R for pUC plasmid PCR assays. Agarose gel
electrophoresis molecular technique was used to detect the
presence of source pathogens and establish MPC. Presence
of gel bands on computer-generated PCR gel images
was visually inspected to identify the presence of source
pathogen. PCR assays were prepared for E.coli(65µg/
µl, 6.5µg/µl), B. subtilis(10µg/µl, 1µg/µl), and pUC(30µg/
µl, 3µg/µl) at two different DNA template concentrations
and PCR cycles were carried out. Gel run was conducted
for each of the assays. Separate negative controls for the
bacterial DNA extracts and pUC were included in the gel
run, along with 1KB ladder DNA standard size marker and
Lambda Hind III standard molecular weight marker. The
concentration for which the gel band was clearly visible was
recorded for each of the pathogen DNA templates. Another
gel run of the PCR assays for E.coli (6.5µg/µl, 0.65µg/µl,
0.065µg/µl, 0.0065µg/µl, 0.00065µg/µl), B.subtilis(1µg/µl,
0.1µg/µl, 0.01µg/µl, 0.001µg/µl, 0.0001µg/µl), and pUC(µl,
0.3µg/µl, 0.03µg/µl, 0.003µg/µl, 0.0003µg/µl) was repeated
at five different DNA template concentrations to zoom
in on the minimum concentration that yielded a clear
gel band. This minimum DNA template concentration at
which the source pathogen’s presence was detectable was
documented as the MPC value(0.03µg/µl for pUC; 0.01µg/
µl for B. subtilis; 0.065µg/µl for E. coli). The above study
demonstrates that MPC thresholds can be predetermined
and catalogued for bioterror agents. Publishing MPC data
may help with optimal PCR assay design for faster and
more accurate detection of the agents in case of a bioterror
outbreak when time to detect becomes crucial.
Schar Room 203
Poster Board No. 017 THE EFFECT OF LIGHT TYPE AND
SPECTRUM ON FIR (Abies bAlsAmeA) NEEDLE ABSCISSION.
Samantha P. Rowland, carrie.rowland@wright.edu, 747 Hawk
Avenue, Tipp City OH 45371. (Tippecanoe Middle School)
Natural production of ethylene gas, coupled with heat,
likely trigger fir (Abies balsamea) needle abscission in
Christmas trees. While research has been conducted
to limit the effect of ethylene gas on needle abscission,
methods rely heavily on chemical treatments. This
experiment compared two different decorative holiday
light types; light emitting diode (LED) and incandescent,
and three different spectrums of light; red, white, and blue
on the rate of fir needle abscission as an alternative to
chemical treatment. It was hypothesized that white lights,
being most similar to full spectrum sunlight, in the cooler
LED treatment would result in the least needle abscission.
A total of 14, three foot tall, 15 year old fir trees were
exposed for 21 days to red, white, or blue lights in both
LED and incandescent forms (N=2 trees per treatment).
Overall, the fir trees exposed to LED light exhibited the
least needle loss, however, color also seemed to make a
difference. Trees exposed for 21 days to red LED light lost