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CBU Biology 1999 Senior Research Presentations at
Collegiate Division of Tenn. Academy of Sciences (Host: Rhodes College)
| '99 Titles |     | '99 Abstracts
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1999 Best Paper Awardees (with Dr. Kesler)
CBU students received 3 of the 5 Best Paper awards: 
Ash Rasoul
(2nd from left), Marlo Anderson (3rd from left), and Andy Briese (not pictured above).

 

 

Titles CBU Biology Student Research Papers ~ TAS 1999
Class of 1999
J. TN Acad. Sci. Vol 74 (3-4) pgs 103-109, 1999. 
Presented at Rhodes College
  • The Effects of Pueraria Lobata on the Diversity of Plant Species Elizabeth Graham and Scott Franklin
  • Effects of Herbicidal Transline Treatment of Pueraria Lobata Heather Carpenter and Jack Grugbaugh
  • Tick Cement Cones David Conner, Lou Boykins and Lewis Coons.
  • *A Study of Seasonal Variation in Overall Abundance of Macroinvertebrates Andrew G. Briese and Jack Grubaugh
  • Effect of IFN-y Antibody on the Latency Period of Gryptosporidiosis Infection in Scid Mice Michael Gipson, Jerold E. Rehg, Tony Capizzani and Mark McGaw
  • Cytosolic Phospholipase A is Phosphorylated and Activated by Calcium/Calmodulin Dependent Proteinkinase Matthew A. Dress, Mubarack M. Muthalif, Jason L. Harper and Kafait U. Malik.
  • Induction of Cancer Cell Apoptosis by Doxorubicin and N-benzyladriamycin-14-Valerate  Brinson Boyte and Len Lothstein
  • The Importance of Polyamines in Retinal Pigmented Epithelial Cells Jonathan R. Keith and Dianna Johnson.
  • Reduction of Nitrate to Nitrate: Does Varying Nadph Concentration Improve Nitrate Recovery? Greg Grokulsky, Y.B. Wang, E.S. Kang
  • Classification of Nonpyramidal Neurons Based on the Correlation of Morphology and Physiology Angel Barritt, Shaul Hestrin and Mario Galaretta.
  • *Effectiveness of Primer R105 as a Genetic Mark for Canine Hip Dysplasia. Marlo Anderson, Amy Miller and Keith Murphy
  • Photoreceptor Cell Development in Dissociated Retinal Cultures Tim E. O'Leary, C.M. Withrow, Dr. A. Johnson
  • Anthropological Analysis of Sharp Trauma in Cartilage and Bone. Gina A. Horton and S.A. Symes
  • Contributions of Arachdonic Acid Metabolites of Cytochrome P-450 and Ras Gtpase to Deoxycorticosterone-Salt Induced Hypertension in Rats. Suzana Malik, Mubarik Muthalif, K. U. Malik and Ibrahim Benter
  • Autogenic Biofeedback Training as a Potential Treatment for Patients with Chronic Gastrointestinal Pain. Vinh Q. Dang, Hani Rashed and Debra Reser
  • Cytokine Pesonse of Macrophages Exposed to Orthopaedic Implant Particles. Alfredo Narvaez, Steven J. Charlebois and Richard A. Smith
  • *Menstrual Cycle Affects Tilt-Table Tests in Women. Ash Rasoul, Judith Soberman, Pamela Jordan and Matthew Robinett
  • Effects of Prenatal Alcohol Exposure on the Sizes of the Corpus Callosum, Hippocampal Commissure and Anterior Commissure.   Uyen Vu, Dan Livy and Adrea Elberger

  • Correlates of Nursing Homes Residents’ Pain. Yin H. Chan and Veronica Engle.
 
Tennessee Academy of Sciences
Collegiate Division, Western Regional Meeting
This meeting is a forum for undergraduate college students to present their research.  Undergraduate student researchers in all areas of science will present talks describing their research. 
  • What is TAS, Collegiate Division and what happens at a TAS meeting?

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    Abstracts CBU Biology Student Research Papers ~ 1999
    CLASSIFICATION OF NONPYRAMIDAL NEURONS BASED ON THE CORRELATION OF MORPHOLOGY AND PHYSIOLOGY. Angel Barritt2, Shaul Hestrin1, and Mario Galarreta1. Univ. of TN Memphis1, Dept Anat. & Neurobiol., Memphis, TN, & Christian Brothers University2, Biol. Dept., Memphis, TN.
    An accurate classification of neurons of the cerebral cortex is yet to be accomplished. The purpose of this research was to determine if a correlation could be found between the morphology and the physiology of nonpyramidal neurons to more accurately classify these cell types. Physiological studies were conducted on cortical brain sections of fourteen to twenty-one day old rats. Recorded neurons were identified using the intracellular label biocytin. Immunologically identified biocytin positive pyramidal and nonpyramidal neurons were classified on the basis of morphological and physiological characteristics. Nonpyramidal cells were classified as bitufted, bipolar, or multipolar. Physiologically, the nonpyramidal cells were classified as fast-spiking or regular-spiking. Bitufted and bipolar cells characteristically exhibited regular-spiking action potentials, with multipolar cells showing fast-spiking action potentials. Although it cannot be concluded that there is a direct causal relationship between the morphology and the physiology of these neurons, distinct characteristics of nonpyramidal cells observed in this study could be used to further explore the function of cortical neurocircuitry. Supporting grant EY-09120. 

    EFFECTIVENESS OF PRIMER R105 AS A GENETIC MARKER FOR CANINE HIP DYSPLASIA. Marlo Anderson1, Amy Miller 2, Keith Murphy 2. 1Dept. of Biol., Christian Brothers University, Memphis TN and 2Dept. of Biol., Univ. of Memphis, Memphis, TN
    Canine Hip Dysplasia (CHD) is a polygenic disease that causes abnormal development of the hips. It results partly from an increase in joint laxity. DNA from the Boykin Spaniel breed has been previously analyzed by randomly amplified polymorphic DNA technique using the primer r105. That analysis showed primer r105 gave a high percent of correctness in identifying increased joint laxity. This current study was conducted to expand on previous data by using the same primer r105, but modifying that variables, sample size and selection of subjects. Our results showed that primer r105 was inconsistent in the larger sample of non-pedigree dogs. In our samples, primer r105 produced a large margin of error in identifying dogs with increased joint laxity. We can conclude primer r105 is a poor genetic marker for CHD, but may prove useful for other genetic traits in a pure pedigree line.
     

    INDUCTION OF CANCER CELL APOPTOSIS BY DOXORUBICIN (DOX) AND N-BENZYLADRIAMYCIN-14-VALERATE (AD 198). Brinson Boyte1 and Len Lothstein2 1Univ. of TN., Memphis, Dept. of Pharmacology and 1Dept. of Biol., Christian Brothers Univ., Memphis TN.
    The regulation of cancer cell apoptosis by various anthracycline derivatives is an ongoing area of research. The purpose of this experiment is to help determine if there are different mechanisms of cytotoxicity in anthracyclines based on structural modifications. The two drugs used in this experiment were Doxorubicin (DOX) and N-benzyladriamycin-14-valerate (AD 198). Human leukemic cells of the CEM cell line were exposed to both drugs at various concentrations and time frames. The cells were then counted and examined for evidence of apoptosis using either Trypan Blue stain and a hemocytometer or bis-benzimide stain and fluorescence microscopy. Each cell count was used to determine an apoptotic index for that sample. The apoptotic index, or number of apoptotic cells per three hundred counted, was then used to compare the cytotoxicity of the two drugs. In repeated trials AD 198 showed a greater apoptotic index and therefore greater cytotoxicity no matter which concentration and time frame was used. These results help to confirm the previous belief that DOX and AD 198, although structurally similar, have different methods of apoptotic induction and mechanisms of cytotoxicity. This conclusion can help open the door for further research and improvement of anticancer drugs based on structural modifications. 
     

    EFFECTS OF HERBICIDAL TRANSLINE TREATMENT OF PUERARIA LOBATA. Heather Carpenter1, and Jack Grubaugh2, 1Dept. of Biol., Christian Brothers Univ., Memphis, TN and 2Dept. Of Biol., Univ. of Memphis, Memphis, TN.
    Pueraria lobata, commonly known as the kudzu vine, was introduced to the southern United States in 1876. Due to its progressive expansion, many persons have tried to control the spread and possibly eliminate kudzu. Benthic Invertebrates can be used as a bioindicator, where high diversity is indicative of a healthy environment. In this study, the distribution of benthos from two ponds was investigated before and after one pond area was treated with a legume specific herbicide, Transline. This was done in order to observe the pollutant effects, if any, of the treatment on the environment. The first pond was surrounded by the kudzu vine (Kudzu pond), and the other pond was not (Payne's). The area adjacent to Kudzu pond was treated with Transline. Pre and post-treatment samples from both ponds were collected, washed, stained, and preserved before taken to the laboratory where the benthos were visualized, extracted and identified. Data was collected for analysis and manipulated using percent dominance, number of taxa per sample collection, and Simpsonís index of diversity. Results show no decrease in diversity, evenness or richness between the benthic invertebrate samples taken pre and post-treatment. In conclusion, these results can be generalized to no apparent immediate effects, pollutant or other, on the immediate environment by the herbicidal treatment of kudzu with Transline. Long term effects are unknown at this time. 
     

    CORRELATES OF NURSING HOMES RESIDENTS’ PAIN
    Y. H. Chan 2 and Veronica Engle1 PhD, RN, 1The University of Tennessee, Memphis, College of Nursing and 2Department of Biology, Christian Brothers University, Memphis, Tennessee. 
    Previous research has documented inadequate pain treatment of nursing home residents, especially Black residents. This study described the relationship of resident pain frequency and intensity with mental status, activities of daily living, depression, communication, and demographic characteristics. Data were collected from 104 residents as admitted sequentially to two large nursing homes serving indigent Black and White older adults. There were 74% Black and 52% male residents, with an average age of 68.4 (SD = 15.9) years. The federally-mandated resident assessment instrument, the Minimum Data Set, was used to measure the study variables. Residents with better long-term memory (r = -0.26), or greater ability to feed themselves (r = -0.23), or who made negative statements about themselves (r = 0.21), or who had less education (r = -0.21) reported significantly (p > .05) more intense pain, and thus may be under-treated for pain. No variables were significantly correlated with pain frequency.
    Research supported by the NIH National Institute of Nursing Research (1 R01 NR04299)
     

    TICK CEMENT CONES. David Conner 1, Lou Boykins2, and Lewis Coons2 Dept. of Biol., Christian Brothers University1, Memphis, TN, and Integrative Microscopy Center, The University of Memphis2, Memphis, TN.
    Ticks are of medical as well as economical importance, because they are the number one vector for vector-borne transmittable diseases. The purpose of these studies was to investigate the relationship between the tick and its attachment to the host. Part of this attachment is a cement cone that secures the tick. Partially fed female Dermancentor variablis (5-7 days) were processed and viewed with the scanning electron microscope. Samples of individual cement cones were also dissected from these ticks. Micrographs reveal that the tick mouthparts are anchored in the cement cone forming a sealed food canal for blood uptake and shielding the tick from the host. Micrographs also reveal that the cement cone has an amorphous surface. The cement cone has several functions and plays a vital role in the relationship between the tick and the host. 
     

    CYTOSOLIC PHOSPHOLIPASE A2 (cPLA2) IS PHOSPHORYLATED AND ACTIVATED BY CALCIUM/CALMODULIN DEPENDENT PROTEIN KINASE (CaMK II).
    Matthew A. Dress 1, Mubarack M. Muthalif 2, Jason L. Harper 2, and Kafait U. Malik 2. 1Biology Dept., Christian Brothers Univ., Memphis, TN and 2Dept. of Pharmacology, College of Medicine, The Univ. of TN, Memphis, TN
    It has been reported that norepinephrine stimulates calcium/calmodulin dependent protein kinase II (CaMK II), which activates cPLA2 and mitogen activated protein kinase (MAPK) and releases arachidonic acid (Muthalif et al, 1996). Arachidonic acid metabolites have been implicated in numerous physiological processes. The products of arachidonic acid generated via cytochrome P-450 and lipoxygenase activate MAPK which amplifies cPLA2 activity by a positive feedback mechanism (Muthalif et al, 1998). The purpose of this study was to determine if CaMK II directly phosphorylates and activates cPLA2. CaMK II obtained from rat brain caused phosphorylation of purified recombinant cPLA2 (3 µg) within 2 minutes.  Phosphorylation continued in a time-dependent manner, with a maximal phosphorylation occurring at 60 minutes.   CaMK II  failed to phosphorylate cPLA2 in the absence of calcium ions (Ca2+) and calmodulin. cPLA2 immunoprecipitated from rabbit vascular smooth muscle cells was also phosphorylated in vitro by purified CaMK II. Addition of MAPK further enhanced phosphorylation of recombinant cPLA2. Phosphoamino acid analysis showed that cPLA2 is phosphorylated by CaMK II on serine residues. These data suggest that CaMK II directly phosphorylates and activates cPLA2, which is further activated upon phosphorylation by MAPK. 
    Supported by the American Society for Pharmacology and Experimental Therapeutics
     

    AUTOGENIC BIOFEEDBACK TRAINING AS A POTENTIAL TREATMENT FOR PATIENTS WITH CHRONIC GASTROINTESTINAL PAIN. Vinh Q, Dang 1, Hani Rashed 2 and Debra Reser 2. 1Dept. of Biol., Christian Brothers University, and 2Univ. of TN, UT Bowld Hospt. Autonomic Function Lab, Memphis,TN.
    Gastrointestinal (GI) symptoms such as nausea, vomiting, abdominal paon, and dizziness or fainting are common in patients with motion sickness. In this study we evaluated a method called Autogenic Feedback Training (AFTE) developed by NASA to reduce GO symptoms experienced by astronauts in space travel. AFTE provides individuals with the ability of modulating their physiological response levels from their own resting baseline. We studied 10 patients (age 37+ 3.8) before and after 4-6 sessions of AFTE. All patients had no known cause for their symptoms. In AFTE sessions conducted over an 8-10 week period, hear rate was utilized as a guide in measuring stress. Each session consisted of 30 min. of training preceded and followed by 6 min. of baseline. Autonomic function test and electrogastrogram were performed non-invasively as previously reported (GE 196;945-950, 1994). Nausea, b;oating, abdominal pain anorexia, and early satiety were quantified by the patient as 0=no symptoms, 1=mild, 2=moderate, 3=severe. Heart rate and blood pressure were measured as previously reported (Clin. Auton. Res. 7;93-96, 1997) and were analyzed by t-tests and expressed as mean + SEM. Data collected revealed that 1) 60% of all patients had improved as determined by total GI scores. 2) All patients had improved in at least three of the measured parameters and were close to normal levels seen in healthy subjects. Our preliminary results suggest that AFTE could be a potential method in alleviating common GI symptoms in patients suffering from chronic motion sickness problems.
     

    EFFECT OF IFN-g ANTIBODY ON THE LATENCY PERIOD OF CRYPTOSPORIDIOSIS INFECTION IN SCID MICE. Michael Gipson1 Jerold E. Rehg2, Tony Capizzani2,and Mark McGaw2; Christian Brothers University1, Memphis, Tennessee and St. Jude Children’s Research Hospital2, Memphis, Tennessee. 
    Cryptosporidiosis is an opportunistic disease that causes severe dehydration and ultimately death in many immunocompromised patients. The effect of IFN-g antibody on the latency period of cryptosporidial infections in SCID mice was evaluated. Different titered amounts of IFN-g antibody secreting hybridoma cells were injected subcutaneously into SCID mice to determine the optimal level of cells that would evoke a reduction in the latency period. Mice were randomly selected from either 14 or 28 days post-oocyst inoculation for assessment of infection. The ileum from each mouse was examined for the presence of infection. At both time periods no significant sign of infection was seen in any of the hybridoma cell-injected mice. Since the dexamethosone control groups did show infection as expected, it is proposed that the hybridoma cells lost their ability to secrete the IFN-g antibody due to continuous cell culturing. By blocking IFN-g , the infected mouse would show a peak level of infection, possibly as early as two weeks post-inoculation, thus decreasing the latency period of infection. This allows the researcher to run more experiments and test for the effectiveness of more chemotherapeutic agents than would be possible with current constraints. This experiment was funded by the NIH and ALSAC. 
     

    THE EFFECTS OF PUERARIA LOBATA ON THE DIVERSITY OF PLANT SPECIES. Elizabeth Graham 1 , and Scott Franklin 2 Dept. of Biol., Christian Brothers University 1, and Dept. of Biol., University of Memphis 2, Memphis, TN.
    The kudzu vine, Pueraria Lobata, has the ability to destroy valuable forests by reaching up to heights of 40 feet or more depending on the substrate the kudzu is growing on, thereby preventing underlying trees and plants to receive sunlight. This physical blockage prevents photosynthesis from occurring and causes trees to die. The purpose of this study was to investigate the effects kudzu had on the diversity of plant species under the kudzu vine. The reestablishment of plants after the eradication of the kudzu by using a herbicide was also investigated. Two areas adjacent to ponds were used in this study at Meeman Biological Station in Shelby Forest, Payne’s Pond, which served as a control having no kudzu surrounding it, and Kudzu Pond which was completely surrounded by kudzu. A third area, Kudzu slope, was also a control which was covered in kudzu but no herbicide was applied to this area. Forty plots of soil adjacent to each pond were studied for diversity of plant species and percent coverage of the foliage of each plant. The herbicide, Transline, was then applied to the area around Kudzu Pond and the kudzu was eradicated, while no herbicide was sprayed at Payneís Pond. The plots at Kudzu Pond were then monitored for regrowth of plants. The results showed that the initial diversity of plant species was much greater at Payneís Pond as expected, showing that kudzu is harmful to other plant species living in that area. There were no plants found growing under the kudzu before its eradication. Plants were observed growing after herbicide treatment and seeds must have originated from dispersed by wind, water, or animals to that area. In conclusion, the kudzu vine has devastating effects on plant life where the vine grows, and if it is eradicated to save valuable forests it is destroying, new seedlings will most likely have to be introduced for reforestation to occur.
     

    REDUCTION OF NITRATE TO NITRITE: DOES VARYING NADPH CONCENTRATIONS IMPROVE NITRITE RECOVERY? G. Grokulsky 2, Y.-B. Wang 1, E.S. Kang 1. Depts. Pharmacol. & Peds. 1, Univ. of TN, Memphis, TN and Dept of Biol, Christian Brothers Univ. 2, Memphis, TN. 
    Nitric oxide (NO) is essential for the regulation of many physiological and pathological processes. Quantification of NO itself is difficult because NO is readily oxidized by molecular oxygen to nitrite (NO2) and nitrate (NO3). The Greiss reaction can be used to quantify NO levels as NO2, but not as NO3. Prior to assay, Reduction of NO3 to NO2 using nitrate reductase in the presence of nicotinamide adenine dinucleotide phosphate (NADPH) is possible before the Greiss reaction. A second enzyme/substrate system, glutathione reductase and oxidized gultathione, can oxidize excess NADPH to NADP. NADP would not interfere with the Greiss assay. Determining the optimal NADPH concentration for recovery of NO2+NO3 by this 2-step reaction was the objective. Mixed standards of NO2 and NO3 from 10-100 micromolar (uM)were analyzed by the 2-step method with 0.25, 0.5, 0.75 and 1 millimolar (mM) NADPH. Recovery as NO2 of NO2+NO3 greater than 20 uM diminished as NADPH was decreased. Recovery of NO2+NO3 less than 20 uM was not diminished across the NADPH concentrations studied. Recovery of NO2 at all concentrations of NO2+NO3 was close to 100% at 1mM NADPH. These findings indicate that optimal recovery of total NO2 by the Greiss reaction after reduction of NO3 to NO2 is achieved using 1.0 mM NADPH.
     

    THE CONTRIBUTION OF p70s6k IN ?-HEXOSAMINIDASE A (Hex A) STIMULATED PROLIFERATION OF BOVINE AIRWAY SMOOTH MUSCLE CELLS. J R Jackson 1, Y Zhao 2 and D B Lew 2. 1Dept of Biology, Christian Brothers University, Memphis,TN, 2Dept. of Pediatrics, University of Tennessee, Memphis, TN.
    Ribosomal p70s6k (p70s6k) is a mitogen activated serine/threonine kinase that is required for cell growth and G1 cell cycle progression. We investigated the role of p70s6k in Hex A stimulated bovine airway smooth muscle cell (ASMC) proliferation. Cellular proliferation of airway muscle is a key factor in airway remodeling, a well-recognized pathological change in asthma. Rapamycin (1.0 nM, 30 min pretreatment), a known inhibitor of p70s6k, inhibited DNA synthesis (100%) and Hex A (50nM) induced cellular proliferation (53%) as measured by 3H-thymidine incorporation and tetrazolium salt reduction (MTT) assays. Rapamycin also inhibited p70s6k activation in Hex A stimulated ASMC as measured by immune complex in vitro kinase assay. These data suggest that p70s6k is important to cellular proliferation in Hex A stimulated ASMC which may assist us in developing a drug that affects airway remodeling in asthma. 
    Research supported by NIH, HL-56812.
     

    THE IMPORTANCE OF POLYAMINES IN RETINAL PIGMENTED EPITHELIAL CELLS. Jonathan R. Keith 2 and Dianna Johnson 1, Dept. of Biol., Christian Brothers University 2, Memphis, TN & Dept.Ophthalmol., Univ. of TN 1 Memphis, TN. 
    The Retinal Pigmented Epithelial Cell layer serves and protects the integrity of the neural retina to carry out proper function. Many diseases are considered to be solely a result of retinal dysfunction, but may actually be the result of RPE dysfunction. One possibility for dysfunction are compounds known as polyamines which aid in regulation of cellular processes such speed and accuracy of gene transcription and protein phosphroylation. Therefore, we examined the importance of polyamines in RPE development. RPE cells were isolated from fetal rabbits. Once cultures had been established, the cells were grown in serum free culture over a three day period in DFMO to block polyamine synthesis. The cells were fixed in 4% Paraformaldehyde overnight and then stained using the fluorescent tag Rhodamine conjugated phalloidan, which binds f-actin filaments and examined them under a fluorescent microscope to observe cell morphology. When compared to RPE grown in normal cell culture, the DFMO treated cells appeared deformed. These results suggest that polyamines may be essential to maintain normal function in RPE cells and that polyamine depletion could be a trigger for retinal dysfunction. 
    Supported by NEI grant 1655 and by Research to Prevent Blindness.
     

    PHOTORECEPTOR CELL DEVELOPMENT IN DISSOCIATED RETINAL CULTURES. (T.E. O’Leary 1, C.M. Withrow2, and D.A. Johnson2) Dept. of Biol., Christian Brothers Univ1., Memphis, TN and Dept. of Ophthalmol., Univ. of TN2 Memphis, TN 
    Purpose. We have examined cell cultures of developing rabbit retina in order to investigate the cellular aggregation. We have also examined the preferential separation of inner and outer retina layers in culture. Methods. Dissociated retinal cultures were grown in control medium. Some cultures were co-cultured with retinal pigment epithelium (RPE) to identify any relation between retinal cells and RPE. The samples were stained with toluidine blue stain, a general cytoplasmic stain, that allows for visualization of the retinal cell bodies. Results. The dissociated cell cultures formed aggregates of two types. One containing cells from the inner retina and the other with cells from the outer retina. The outer retinal aggregates, termed rosettes, consisted of a horizontal cell surrounded by photoreceptor cells. When RPE was added to the cultures, the RPE cells preferentially appeared with rosettes.The two types of clusters were attached to different regions of the cover slip. The aggregates formed within one hour in culture. Given longer time intervals, the aggregates became more compact. Conclusions. The rapid separation of the cell aggregates shows preferential cell to cell adhesion is taking place. Formation of two separate cell cluster types could result from a specific binding site. 
    Supported by NEI grant 1655 and by Research to Prevent Blindness. 
     

    CONTRIBUTION OF ARACHIDONIC ACID METABOLITES OF CYTOCHROME P-450 AND RAS GTPASE TO DEOXYCORTICOSTERONE-SALT INDUCED HYPERTENSION IN RATS. Suzana Malik 1, Mubarik Muthalif 2, K.U. Malik2 , and Ibrahim Benter 2, 2Dept. of Pharmacology, Univ. of TN, & 1Dept. of Biol., Christian Brothers Univ., Memphis, TN.
    Administration of deoxycorticosterone acetate (DOCA) to rats drinking 1% sodium chloride (NaCl0 solution causes hypertension. The vasoactive agents produced are known to stimulate release of arachidonic acid (AA) from tissue phospholipids. The AA metabolites generated via cytochrome P-450 pathway mainly, 20-hydroxycorcortico-steronic acid (20-HETE) increase vascular tone, and stimulate Ras GTPase and mitogen activated protein kinase (Ras/MAPK). The purpose of this study was to determine if specific AA metabolites are involved in DOCA/NaCl induced hypertension in Sprague Dawley rats. Administration of DOCA to the rats (all animals in the study had an uninephrectomy) drinking 1% NaCl solution for five weeks increased mean arterial blood pressure (MABP) which was higher than rats treated with vehicle of DOCA. Administration of CYP-450 inhibitor aminobenzotriazole (ABT) reduced MABP, and infusion of farnesyl proteint transferase inhibitor, FPT III, also decreased MABP. Data suggests one or more metabolites of AA formed through CYP-450, most likely 20-HETE, contributes to development of DOCA/NaCl hypertension probably by activating Ras via MAPK pathway. This study was supported by the NIH Grant 19134-24.
     

    CYTOKINE RESPONSE OF MACROPHAGES EXPOSED TO ORTHOPAEDIC IMPLANT PARTICLES. Alfredo Narvaez 1, Steven J. Charlebois 2, and Richard A. Smith 2, 1Dept. of Biol, Christian Brothers Univ., Memphis, TN and 2The Univ. of Tennessee-Campbell Clinic Department of Orthopaedic Surgery, Memphis, TN
    The purpose of this study was to measure cytokine levels from macrophages exposed to particles of two orthopaedic implant materials. The cytokines measured (IL-1b, IL-6 and TNFa) are known to be involved in osteolysis, a major contributor to orthopaedic implant failure. Macrophages are currently thought of as the major cellular mediator of osteolysis. A transformed mouse macrophage cell line (ATCC, IC-21) was cultured on glass cover slips coated with fibronectin in six well tissue culture plates. Macrophages were exposed to cobalt-chrome or commercially pure titanium particles in a 1:1 cell/particle ratio. Zymosan and lipopolysaccharide (LPS) particles (10µg/ml) were used as positive controls. After 24 hours supernatants were assayed for cytokines. Cytokines from cells exposed to implant particles were 10 to 100 times lower than for the positive controls (LPS and Zymosan). Image analysis showed that the macrophages were highly associated with the particles. Confocal microscopy was used to confirm that the cells were engulfing the particles. Discrepancies between this work and previous studies which have shown high levels of these cytokines in response to cobalt-chrome and titanium particles may be due to endotoxins such as LPS adsorbed to their surface. A current hypothesis is that the particles, though sterile, may not be clean. The surface of these particles may contain adherent endotoxin that could trigger the high cytokine responses that have been observed in other in-vitro studies. 
     

    MENSTRUAL CYCLE AFFECTS TILT-TABLE TESTS IN WOMEN
    Ash Rasoul 1, Judith Soberman MD 2,Pamela Jordan MSN 2, & Matthew Robinett, BS 2 1Dept. of Biol., Christian Brothers University , and 2Dept. of Cardiology, University of TN, Memphis, TN.
    Background: A previous study in UT Bowld's Cardiology Lab, illustrated that there maybe differences in tilt-table testing results due to cyclical hormonal variation in the women. This study investigated this hypothesis with tilt table studies on women with normal menstrual cycles. Methods: Women, ages 18-35, with regular menstrual cycles and no previous history of syncope were randomly assigned to both a control (80 degrees) and either a premenstrual or preovulatory (70 degrees) trial. The test ended with either syncope or completion of protocol. Results: Approximately 1/3 of the subjects tested in the control experienced syncope. In the preovulatory trials, 1/2 of the subjects fainted. In the premenstrual trial, no fainting occurred. Conclusions: Cyclical variation in hormone levels due to the specific phase of the menstrual cycle can significantly affect the results of tilt table testing in women. 
     

    EFFECTS OF PRENATAL ALCOHOL EXPOSURE ON THE SIZES OF THE CORPUS CALLOSUM, HIPPOCAMPAL COMMISSURE, AND ANTERIOR COMMISSURE. Uyen Vu 1, Dan Livy 2, and Andrea Elberger 2, 1Christian Brothers University, Biology Department, Memphis, TN, and 2The University of Tennessee at Memphis, Department of Anatomy and Neurobiology.
    Fetal alcohol syndrome (FAS) is a disorder associated with a pattern of behavioral, facial, and central nervous system abnormalities caused by alcohol consumption in the womb. Previous autopsy studies have indicated that one particular region in the brain, the corpus callosum, is especially susceptible to the toxic effects of alcohol. Recent studies using the noninvasive resonance imaging technique have found a decrease in corpus callosum area in humans. In this study, the sizes of the corpus callosum (CC), hippocampal commissure (HC), and the anterior commissure (AC) were measured in Sprague-Dawley rats from prenatal and control treatment groups. Using a Kodak DCS 460 digital camera, images were taken of the mid-sagittal section. From these images, measurements were taken with the analysis program Lasersharp. There was no significant difference observed in sizes between the alcohol and control treatment groups. Thus, this data suggests that other factors in addition to alcohol exposure may have contributed to the reduction in the CC seen in previous studies.
    Supported by: NIH Grant AA-11325

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