Kentucky Junior Academy of Science 2000 Symposium Abstracts
Effect of Dopamine Depleting Agent Tetrabenazine on Malonate-Induced
Lesion. MINDY SIMONS Nicholas County High School, 103 School Drive, Carlisle,
KY 40311-9124.
Huntington's Disease is a fatal autosomal
dominantly inherited neurodengenerative disorder which affects approximately
five to ten out of 100,000 people. In 1993, a mutated gene on chromosome
4 consisting of an expanded number of CAG tri-nucleotide repeats was discovered.
Neuropathologically HD causes a progression of neurodegeneration of neurons
in the corpus striatum which appears to be a type of cell death called
apoptosis. One factor that appears to be related to this cell death
is the sensitivity of the striatum to the presence of dopamine. Dopamine
in high concentrations is neurotoxic. Moreover, removal of striatal
dopamine has been shown to confer protection in several models of cell
death. In order to evaluate this finding a suitable model is required.
Intrastriatal injections of malonate a mitochindrial inhibitor, produces
lesions in rats which mimics the neuropathology of HD. In previous
studies dopamine levels were reduced by surgical ablation of dopaminergic
fiber bundles. These lower levels of dopamine resulted in reduction
in size of malonate-induced lesions. Surgical manipulation would
not be treatment of choice so a pharmacological approach is desired.
This study was designed to see if a compound TBZ, which is used to
treat Huntington's Disease, works by reducing lesion volume in rats.
Lesions were induced by injecting malonate
into striatum of animals treated with i.p. injection of TBZ and control
animals untreated. Sections of striatal tissue were analyzed for
lesion volume using computer-based image analysis. Results revealed
that there was no significant differences in lesion volume between the
TBZ treated animals and controls. While lesion reduction was the
same, the lesions were much smaller by approximately 50%. It was
also noted that the animals did respond to the TBZ treatment due to apparent
lack of movement. This indicates dopamine was depleted.
This study suggests that further research
is warranted and that different dosages amounts or different dosage regimes
of TBZ should be employed.
Asthma and Allergies. ANNE M. CHMILEWSKI, duPont Manual High
School, 120 W.
Lee St., Louisville, KY 40208.
Louisville, KY, has long been considered a
worse city than many for allergies and asthma
as result of several factors, including its location in a valley, long
pollen season, and diverse plant population. A survey was designed
to test the validity of considering Louisville to be an unhealthy residence
for allergy and asthma sufferers. The survey, which asked questions
concerning severity, longevity, and occurrence of allergies and asthma,
was completed and returned by 52 15- and 16-year old students at duPont
Manual High School. The results of this survey presented several
interesting facts. It was found that 63% of those surveyed had some
form of allergies, compared to the national rate for the entire population
which is only about 20%. Asthma sufferers accounted for 19% of the
survey population, but only makes up 7% of the US population under age
18. It is also important to note that 56%, or more than half, of
the students claimed to be sensitive to tobacco smoke to some degree.
Finally, non-native students (students not born in Louisville) had only
a 53% allergy rate, compared to a 70% allergy rate for native Louisville
residents. The experiment was greatly hampered by its limited size
and range of survey participants, but the results can be seen as evidence
of the need for greater study of allergies and asthma in all ages of Louisville
residents.
The effects of vitamin C on runners. LISA MUDD, duPont Manual
High School, 120 W.
Lee Street, Louisville, KY 40208.
Many runners spend several days recovering
after hard races and practices. These recovery days" often infringe
on vital workouts, causing the runner to lose ground. By giving runners
500-milligrams of vitamin C, I hope to determine whether vitamin C has
an effect on the recovery time in the muscles of runners.
Each runner was given one 500-milligram vitamin
C capsule each day for five days. Each day their flexibility would
be measured before they participated in a track workout. They would
take the weekend off, and then they would do the same with a placebo.
Females, being more flexible than males noticed
the smallest change in flexibility (2-4 inches). Males noticed a
larger change in flexibility (2-6 inches). The females also experienced
a smaller decrease in flexibility (2-4 inches), while the males experienced
a larger decrease (3-6 inches).
Methods to Improve Body Composition. JEFF KARP and MATT ROBERTS.
duPont Manual High School, 120 West Lee Street, Louisville, Kentucky 40208.
This project examines the
impact of weight training and running on subjects' body fat percentage
(BFP) and maximum weight lifts. Similar tests have been conducted
before, but none dealing with this combination.
To qualify for the tests, participants had to be between the ages of
14-18, and must adhere to the schedule set out by the researchers.
The schedule required the subjects to complete a running program and a
weight training session four times a week, for four weeks. The running
program lasted approximately one hour per day and consisted of form running,
agility drills, distance running, and traditional sprints. The weight-training
program also lasted approximately one hour, and consisted of exercises
that focused on the chest and lower body. The bench press, leg press,
and squat were the three lifts used for supporting evidence.
Once all of the information was gathered,
it was analyzed and graphed. After running t-tests on the data we
could see that there was a significant difference in the body fat percentage
and the maximum weight lifts. This can be attributed to one reason:
more than half of the subjects were exposed to this type of rigorous conditioning
for the first time, therefore, extensively improving their BFP and maximum
weight lifts.
The Involvement of the Pro-Apoptotic Protein in Prostate Apoptosis
Response-4 in Amyotrophic Lateral Sclerosis. CRYSTAL PRYOR. Nicholas County
High School, 103 School Dr., Carlisle, KY 40311-9124.
Amyotrophic lateral sclerosis (ALS) is caused
by degeneration of motor neurons in the spinal cord and brainstem.
Familial type, which represents about 20% of all ALS cases, is caused by
a gene mutation in Cu/Zn superoxide dismutate. This mutation results
in an increase in oxidative stress.
Recent studies have noted that the oxidative stress in familial ALS
cases result is higher levels of lipid peroxidation products, which is
thought to cause a type of cell death called apoptosis.
A new method of detecting neuronal cell death is in measuring levels
of a protein called prostate apoptosis response-4 (Par-4).
This research was involved in determining
if an increase in Par-4 could be detected using transgenic murine spinal
cord cells containing the Cu/Zn SOD mutation: and also attempting to measure
Par-4 in NSC-19 cells transfected with the mutation after treatment with
lipid peroxidation products.
It was expected that both cell types would
demonstrate an increase in Par-4. Western blot analysis was used.
Results determined that cells from transgenic murine spinal cord did have
a increase in Par-4 meaning there is a connection of the protein to ALS.
The transfected mutant cell line did not show an increase in Par-4 since
the level of the protein was high in all cells, not just those treated.
This was thought to be caused by the cells high rate of proliferation.
HIV causes multiple skin disorders by decreasing EGF-r Density.
PENG LEI, duPont Manual High School, 120 West Lee Street, Louisville, KY
40208.
Transgenic mice infected with the human immunodeficiency
virus type-1 (HIV-1) and human patients with Acquired Immunodeficiency
Syndrome (AIDS) both exhibit similar skin disorders. While the mechanism
by which HIV causes such disorders is yet unclear, it is known that the
skin problems and growth failures observed in the HIV- I transgenic homozygous
mice are similar to those of mice with low epidermal growth factor receptor
(EGF-r) expression. Moreover, the hyperproliferative skin disorder
in HIV- 1 transgenic heterozygous mice is comparable to the hyperactive
growth of tumors in mice with abnormally high EGF-r expression. Thus,
it is hypothesized that HIV affects the expression of EGF-r in mice skin.
The EGF-r densities in skin of nontransgenic and HIV- I transgenic mice,
both homozygous and heterozygous, are determined using an avidin-biotin
immunoperoxidase staining technique. The data show that HIV-1 transgenic
homozygous mice have EGF-r densities significantly less than those of non-transgenic
mice.
Resveratrol As An Estrogen Agonist or Antagonist in Era
and Erb. ROSEMARY SIMS, duPont Manual High School,
120 W Lee Street, Louisville, KY 40208.
Resveratrol is a phyto estrogen found in the
skins of red grapes. It is believed by many scientists to be an explanation
for the French-Paradox; the low occurence of heart disease despite a high
fat diet. This experiment was conducted to see if resveratrol would
act as an estrogen agonist, or antagonist in the presence of Estrogen receptor
alpha, and Estrogen receptor beta. Prior to experimenting, the belief
was held that estrogenic activity would be blocked by resveratrol in both
receptors, while showing greater reactivity with ERb.
CHO-Kl cells (containing tandem copies of EREc38 DNA) were transiently
transfected with the estrogen receptors, ethanol, 4-hydoxytamoxifen, estradiol,
and resveratrol. After treatment, the cells were harvested, and their
estrogenic activity was counted in a liminometer, which expressed the values
in light units of luciferase. These values were divided by the beta-galactosidase
activities, leaving the fold induction of luciferase. The results
showed that ERa was more active than ERb
in the presence of estradiol. Synergistic activity was noticed in
three and four EREc38, a trend not seen in one and two EREc38. Suprisingly,
resveratrol has similar activity in both ERa
and ERb. It is also apparent that resveratrol
has stronger agonist activity with ERb than estradiol. Due to the
ability of 4-OHT to inhibit its estrogenic activity in all cases, resveratrol
is proved to induce estrogen activities therefore making it an estrogen
agonist in the presence of both estrogen receptors.
Optimization of sickle cell anemia detection using PCR amplification
and Dde I digestion. EFFIE WANG, MST Department, duPont Manual High
School, 120 West Lee Street, Louisville, KY 40208.
Sickle cell anemia (SCA) is caused by a point
mutation in the gene of the beta chain of
hemoglobin. This mutation happens to occur at a DdeI endonuclease
digestion site. Therefore, while DdeI will digest the DNA from normal
individuals at this site, it will not digest the DNA from SCA patients
due to the mutation. Based upon this information, a scheme can be
developed for a quick clinical diagnosis by amplifying the DNA harboring
the SCA mutation, followed by DdeI digestion. The current study was
aimed at identifying the best primer pair for PCR amplification.
Human genomic DNA (normal, heterozygous, homozygous) and oligonucleotide
primers were obtained from UCSF. DNA concentrations were determined
by spectrophotometry. The DNA fragment harboring the mutation was
amplified using Taq DNA polymerase on a PCR cycler with the following program:
denaturing at 94oC for 45 seconds; annealing at 55oC
for 30 seconds, and extension at 72oC for 30 seconds.
A total of 35 cycles of amplification were performed. Twelve different
primer pairs were tested in the experiment. The quality of the PCR
amplification were assessed using agarose gel electrophoresis. The
PCR amplified DNA fragments from normal individuals and patients were then
divided into two aliquots, one of which was used for digestion by 10 U
of DdeI and the other was undigested and used as a control. Both
samples were then separated on a 2.5% agarose gel. Based on the results,
primer pair 87N/43M2 gave the best result in terms of clarity and accuracy.
The effects of tumor necrosis factor-a and prostaglandin E2
on the expression of intercellular adhesion molecule-1 in eye microvascular
endothelial cells. SUDIP K. SAHA, duPont Manual High School, 120
West Lee Street, Louisville, KY 40242.
It has been established that during inflammation,
Tumor Necrosis Factor-a (TNF-a),
a proinflammatory cytokine, increases the expression of Intercellular Adhesion
Molecule- I (ICAM-1) in the eye's Microvascular Endothelial Cells (MVEC),
but the effects of Prostaglandin E2
(PGE2) on this process are unknown. ICAM-1 is a member
of the immunoglobulin gene superfamily with five extracellular Ig-like
domains. The purpose of this study was to examine the effects of
TNF-a and PGE2 on the expression
of ICAM-1.
Two six-well chambers were cultured with MVEC.
The groups that were being tested are as follows: (a) Tumor Necrosis Factor
(TNF-a); (b) TNF-a
and PGE2; (c) TNF-a, AH23848 (PGE2
antagonist), and PGE2 and (d) Mouse Serum, which contains various Immunoglobulin
G (IgG). After being stained with the primary antibody (ICAM- 1),
the cells were examined and photographed by a Nikon high-speed fluorescence
photomicroscope.
The groups treated with (a) TNF-a,
(b) PGE2 and TNF-a, and (c) TNF-a,
AH23848, and PGE2 showed fluorescence while the group stimulated
with (d) mouse serum showed none. There was no visible difference
in the expression of ICAM-1 in the groups stimulated with (a) TNF-a
and the group treated with (c) TNF-a, AH23848,
and PGE2. However, the group stimulated with TNF-a
and PGE2 expressed more fluorescence than both of those groups
did.
These findings suggest that the expression
of ICAM-1 by TNF-a is mediated predominantly
by PGE2 and that the EP4 receptor (a PGE2 receptor
subtype) exists in eye MVEC.
The synaptology of GABAB receptors in the developing gustatory
system. TONY STEFATER, duPont Manual High School, 120 West Lee Street,
Louisville, KY 40208.
There is substantial evidence that GABAergic
inhibition plays an important role in the processing of gustatory information
in the rostral nucleus of the solitary tract (rNST). Previous light
microscopic studies in the laboratory have demonstrated that GABAB
receptor labeling in the rostral nucleus of the solitary tract (rNST) is
present in neurons and processes. This study used electron microscopy
and immunohistochemistry to determine the synaptic location of GABAB
receptors in the adult and postnatal--day 10 (PND 10) rNST. Then,
the sizes of the post-synaptic profiles were measured. GABAB
receptors are located at pre-synaptic terminals and/or post-synaptic profiles.
Labeled terminals are associated with dendrites, dendritic spines, cell
somata. Two types of GABAergic terminals in the rNST have been defined
by their vesicular density: GABA-LD (low density) and GABA-HD (high density).
They have a differential distribution such that GABA-LD terminals contact
larger, more proximal dendrites and cell somata, while GABA-HD terminals
contact smaller, more distal dendrites. I determined that when GABAB
receptors are both pre- and post-synaptic label, the typical apposing dendrite
has the cross-sectional dimensions of proximal dendrites and thus is likely
GABA-LD. This is not true when GABAB receptor labeling
is only pre- or post-synaptic. These data suggests that the pattern
of labeling for GABAB receptors may be different for the two
previously defined GABAergic terminal types. It lends support to
the hypothesis that GABAergic inputs to the proximal and distal dendrites
may be different and reflect two GABAergic systems within the rNST capable
of influencing individual neurons.
Fine Motor Control Deficit in Patients with Parkinson’s Disease.
JARRETT S. STULL. Nicholas County High School, 103 School Drive, Carlisle,
KY 40311-9124.
More than half a million Americans suffer
from the neurodengerative disease known as Parkinson's disease (PD).
The cause of PD is related to the loss of cells in the substantia nigra
area of the brain; which results in a decrease in dopamine production.
Dopamine, a neurotransmitter is important for normal movement.
This study was designed to determine the fine
motor capabilities of PD subjects
compared with normal aged subjects. As PD is associated with
people usually past the age of 55, these older subjects were compared to
normal aged subjects not having PD. PD subjects were tested on a
fine motor control testing apparatus called the Movement Assessment
Panel (MAP). It was hypothesized that the PD subjects would have
a diminished response in the fine motor functions as compared to normal
aged subjects.
The MAP tested four tasks; removal of a nut from four shaped
platforms in increasing complexity from a Platform Task to Corkscrew
Task. Times for completion of the Platform Task to Corkscrew
Task were measured by computer sensors. Dominant hand was measured
as well as non-dominate hand. Findings revealed that the PD subjects
did demonstrate the need for more time in removal of the nut in the more
complex tasks, but not in the simpler tasks.
These findings will hopefully lead to testing
measures that will result in early detection of patients with PD.
What ocular problems does diabetes present? SOWMYA SRINIVASAN,
duPont Manual High School, 120 W. Lee Street, Louisville, KY 40208.
Twenty records of diabetic patients were analyzed
to obtain relationship between diabetics vs. normal people getting afflicted
with each of the following diseases: background diabetic retinopathy, proliferative
diabetic retinopathy, macular edema, cataracts, and glaucoma. The
impact of age was also studied. Data was separated by gender and
then age groups. Statistics show that after having diabetes for ten
years, 50% of diabetics will get some degree of retinopathy. After
15 to 20 years, 90% of diabetics will have retinopathy. Among this
group 25% will even have proliferative diabetic retinopathy. Also,
in people 40 or younger, diabetics are 25 times more likely to develop
cataracts than non-diabetics. In people 60 or older, diabetics are
two to four times as likely to develop cataracts, and at this age people
with healthy eyes already have a 50% chance of developing a cataract.
Data obtained supported findings in most cases but there were exceptions
due to inadequate data for each age group. Findings show that it
is inevitable for diabetics to develop some degree of ocular problems in
their life time.
The Effects of Penny Composition on the pH of Stomach Acid.
MARGARET FENTON, duPont Manual High School, 120 West Lee Street, Louisville,
KY 40208.
This study explored the relationship between
the composition of pennies and the pH levels generated when they were bathed
in a hydrochloric acid solution for various periods of time (1-96 hours).
Pennies minted between 1962 and 1982 are a copper-zinc alloy; pennies minted
after 1982 are a copper-coated zinc core. Physicians saw an increase
in stomach ulcers caused by ingested pennies following the reformulation
of these coins. The cause of this increased occurrence may be linked
to a failure to maintain a pH balance between the gastric acids and the
surface of the epithelium of the stomach. The null hypothesis states
that the pH level of the simulated gastric acid will not be differently
affected by the reaction rate of an "old" penny versus a "new" penny when
bathed in a hydrochloric acid solution. Based on the data, the null
hypothesis was not rejected. Thus, a general change in pH is not
suspected as a cause for the formation of these gastric ulcers. However,
observations of rapid reactions of acid with the zinc core of new pennies
which produced hydrogen gas, suggests that local formation of hydrogen
gas bubbles may be connected to the development of ulcers around pennies.
Which Oils are the Healthiest? SUPRAJA PARTHASARATHY, duPont
Manual High School, 120 West Lee Street, Louisville, KY 40208.
Four different oils-olive oil, peanut oil,
soybean oil, and coconut oil- were analyzed using the Thin Layer Chromatography
method. Four different standards-- triglycerides (bad/good, depending
on the fatty acids), phospholipids (good), cholesterol (bad), cholesteryl
ester (good)- were expected inside the oils. These were the contents
of the oils and told whether the oil was healthy or unhealthy. It
was concluded that the peanut oil and coconut oil was unhealthy, because
it contained large amounts of cholesteryl ester and cholesterol, making
those oils unhealthy. The soybean oil was healthy because it contained
less amounts of cholesterol and triglycerides. The healthiness of
the olive oil could not be concluded because there was some cholesterol
and cholesteryl ester and lots of triglycerides. There are two different
types of fatty acids in triglycerides, one good and one bad and it was
not possible to determine which fatty acids were in that triglyceride.