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CBU Biology 1998 Senior Research Presentations at 
Collegiate Division of Tenn. Academy of Sciences (Host: Union Univ.)

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Titles CBU Biology Student Research Papers ~ TAS 1998
Class of 1998  
J TN Academy Sci., Volume 73, Number 3-4, pgs 115-120 July-October 1998. Presented at Union University, Jackson.

DOES NITRIC OVIDE PRODUCTION LOWER MEAN ARTERIAL BLOOD PRESSURE DURING PREGNANCY?  Swanette Anderson and Robert A. Ahoaks.

DEXAMETHASONE INHIBITION OF INDUCIBLE NITRIC OXIDE SYNTHASE PRODUCTION IN MACROPHAGES STIMULATED WITH GENITAL MYCOPLASMAS.  Karima T. Causey, Dennis T. Crouse, B. Keith English, Cindy Newman, Elizabeth Mals, and Lisa Livingston.

NITRIC OXIDE SYNTHASE ACTIVITY IN THE RED BLOOD CELL:  IS IT POSSIBLE?  Kevin Ford and Ellen S. Kang.

CARNITINE LEVELS IN RAT RED BLOOD CELLS, WHITE BLOOD CELLS, AND BLOOD SERUM.  Nichole Koenigsknecht, M. Volk, and M. Banschbach.

MECHANISM OF POTASSIUMCHOLORIDE-INDUCED RELEASE OF ARACHIDONIC ACID AND CONTRACTION OF BLOOD VESSELS:  SELECTIVE ACTIVATION OF OLIPOXYGENASE AND INHIBITION OF PROSTAGLANDIN SYNTHESIS.  Zachary Maxwell, Mubarack M. Muthalif, Jason Harper, Nour Karzoun, and Kafait U. Malik.

IMMUNOASSAYS OF GLYCOPHORIN A:  AN ERYTHROCYTE MEMBRANCE PROTEIN.  L. Gregory Meriwether, Dennis Carrigan, and Douglas P. Blackall.

THE ROLE OF SERUM EOSINOPHILIC CATIONIC PROTEIN IN EXERVISE-INDUCED ASTHMA.  Bobby Myers, K. V. Leeper, J. Soberman, T. T. Mastin, and P. Jordan.

*UP-REGULATION OF CADHERINS FOLLOWING CENTRAL NERVOUS SYSTEM INJURY IN RATS.  Felix Vazquex-Chona and Eldon E. Geisert, Jr.

*THE DISLOCATED KNEE.  Trent A. Gullett, Fred Azar, Schott Sharpt, and Marc Mihalko.

INFLUENCE OF THE NORTH HOLLYWOOD DUMP ON WOLF RIVER SEDIMENT METALS CONCENTRATIONS.  Thomas Briggs, C.J. Leppanen, and K. J. Maier.

LOW ANTIOXIDANT POTENTIAL AND PSYCHOLOGICAL DISTRESS AMONG THE MORBIDLY OBESE.  Michael E. Cole, Cynthia K. Buffington, Richie J. Geuers, Ronald W. Hard, and George S. M. Cowan.

THYROID HORMONE REGULATION OF UCP-2 AND THE OB GENE IN RAT ADIPOSE TISSUE.  Linda Nguyen and Suleiman Bahouth.

LOCALIZATION OF D2 DOPAMINE RECEPTORS IN AVIAN STRIATUM USING IMMUNOHISTOCHEMICAL LABELING.  Rodney A. Paullus.

DIFFERENTIAL DIAGNOSIS FOR PATIENTS WITH PRESYNCOPE SYMPTOMS.
Felicia A. Hayes and H. Rashed.

RELATIONSHIP OF ACOUSTICAL PROPERTIES OF THE VOICE AND HEALTH STATTUS IN OLDER ADULTS.  Yong Q. Lin, Veronica F. Engle, Barbara Fuller, Marshall J. Graney, and Douglas Connor.

THE EFFECTS OF MANUAL LYMPHATIC STIMULATION IN WOMEN SUFFERING FROM SECONDARY LYMPHEDEMA.  Lynsday Woodward, Cathy Sandord, Laurel Means, Kris Valentine, and Tammy Gibson.

  
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?
  • CBU students won two of the three Best Paper awards in 1998: 
    Trent Gullet, 1st place (center) 
    Felix Vazquez-Chona,  3rd place (left)

     

     

    Abstracts CBU Biology Student Research Papers ~ 1998
    J TN Academy Sci., Volume 73, Number 3-4, pgs 115-120 July-October 1998. Union University.

    1. 
    DOES NITRIC OXIDE PRODUCTION LOWER MEAN ARTERIAL BLOOD PRESSURE DURING PREGNANCY?  Swanette Anderson1 and Robert A. Ahokas2, PhD. 1Dept. of Biology, Christian Brothers University, Memphis, TN; 2Dept. of Obstetrics and Gynecology, and Physiology / Biophysics, University of Tennessee at Memphis.
     Pregnancy is normally associated with vasodilation, and in hypertensive animals, such as the spontaneously hypertensive rat (SHR), causes a profound decrease in blood pressure. To test the possibility that enhanced basal vascular nitric oxide (NO) activity has a role in the vasodilation of pregnancy, we measured the changes in mean arterial blood pressure (MAP) induced by NG-nitro-L-arginine methyl ester, a specific inhibitor of NO synthesis, in conscious postganglionic   c blocked non-pregnant and pregnant SHR's. In the pregnant rats, litter size was adjusted to 1-12 on day 7 of pregnancy. On days 14 and 21 of pregnancy, MAP was measured before and 30 minutes after Hexamethonium (20 mg/kg, IV). MAP of the day 21, but not 14 day pregnant rats was significantly lower than that of the non-pregnant rats. Hexamethonium decreased MAP 50-60 mm Hg in all rats. NG-nitro-L-arginine methyl ester (10 mg/kg, IV) increased MAP greater in the non-pregnant than in the 21 day pregnant rats. These results suggest that vascular NO activity is not increased in pregnancy, and is not responsible for the decrease in MAP. Rather, vasodilation is due to a late pregnant decrease in sympathetic neurogenic vascular tone which is modulated by some unknown mechanism by the fetoplacental unit. 
    Supported by: the Department of Obstetrics and Gynecology, UT Memphis
     

    2.  
    INFLUENCE OF THE NORTH HOLLYWOOD DUMP ON WOLF RIVER SEDIMENT METALS CONCENTRATIONS.  T. Briggs1, C. J. Leppanen2, and K. J.  Maier2  1Dept. of Biology, Christian Brothers Univ., Memphis, TN; 2Dept. of Biology, Univ. Memphis, Memphis TN.
     The North Hollywood Dump in Memphis, TN is a closed municipal/industrial landfill and federally listed superfund site.  We examined core Wolf River sediments for metal concentrations upstream, adjacent to, and downstream of the dump to determine the influence of the dump on Wolf River sediment toxicity. Metal concentrations were determined using graphite furnace atomic absorption for Pb, and flame atomic absorption for Al, Ba, Cu, Fe, Mn, and Zn.  Aluminum, iron, and zinc concentrations were highest along side of the dump.  Copper and lead concentrations peaked at the downstream site.  Manganese concentrations were greatest upstream and Barium was not present in any sediment. More frequent sampling, as well as taking a greater number of samples will aid in pinpointing the source(s) of contamination.
     

    3.  
    DEXAMETHASONE INHIBITION OF INDUCIBLE NITRIC OXIDE SYNTHASE PRODUCTION IN MACROPHAGES STIMULATED WITH GENITAL MYCOPLASMAS. Karima T. Causey 1, Dennis T. Crouse2, 3, B. Keith English2, 3, Cindy Newman2, 3, Elizabeth Meals 2,3, Lisa Livingston 2,3. 1Christian Bro. Univ., Depart. of Biol, Memphis, TN. 2 UT, Memphis, TN., Dept. Pediatrics & OBGYN, & 3Crippled Children's Foundation Research Center, LeBonheur Children's Medical Center.
     In preterm infants, mycoplasmas are associated with the chronic lung disease, Bronchopulmonary Dysplasia. Dexamethasone is a corticosteroid often administered to these infants to suppress inflammation.  Nitric oxide (NO) formation, in macrophages and  increased intrinsically in humans during inflammation, is controlled by an enzyme called inducible nitric oxide synthase (iNOS).  Thus, studying iNOS production could be used as a marker for inflammation that occurs due to the stimulation of macrophages by mycoplasmas and the effect of dexamethasone on this process.  We hypothesized that dexamethasone would decrease iNOS production in mycoplasmal-stimulated macrophages. Macrophage cell cultures were incubated with sterile media alone, Lipopolysaccharide (LPS) (1ng or 100ng), Mycoplasma hominis (Mh) (104, 105 and 106 colony forming units (cfu)/ml, or Ureaplasma urealyticum (Uu) (104, 105, 106 cfu/ml) for 16 hours.  Immunoelectrophoresis was used to measure iNOS.  Incubation of the macrophages with 1U (unit) or 10U Interferon Gamma and either LPS, or Uu, or Mh produced iNOS in a dose-dependent manner.  Dexamethasone reduced the production of iNOS in all experiments.  Thus, genital mycoplasmas stimulate iNOS in a dose-dependent fashion, which can be inhibited by dexamethasone.  
     

    4.  
    LOW ANTIOXIDANT POTENTIAL AND PSYCHOLOGICAL DISTRESS AMONG THE MORBIDLY OBESE.  Michael E. Cole**, Cynthia K. Buffington, Ph.D.+, Richie J. Feuers, Ph.D.*, Ronald W. Hart, Ph.D.*, and George S.M. Cowan, M.D.+.  **The Dept. of Biology, Christian Brothers University, Memphis, TN, +The Dept. of Surgery, UT, Memphis, TN, and *The National Center for Toxicological Research, Jefferson, AR.
     Free radicals such as peroxides cause irreversible damage to membranes, DNA, mitochondria, and other cellular components.  Glutathione plays an important role in the destruction of organic peroxides and other free radicals through a reaction catalyzed by Glutathione peroxidase (GPx).  In our studies, we measured the activities of GPx and enzymes important in the reduction of glutathione, glucose-6-phosphate dehydrogenase (G6PDH) and glutathione reductase (GR), in isolated red blood cells of morbidly obese (MO) and lean patients (LC).  We found that neither G6PDH nor GR activities (using student t-tests) significantly differed (p>0.05) between the MO and LC groups, suggesting normal levels of NADPH and reduced glutathione in our MO population.  GPX activities of the MO, however, were found to be significantly (p<0.001) below LC values.  As GPx is important in catalyzing the removal of hydrogen peroxide, low activities of this enzyme would thus reduce antioxidant potential.  We therefore conclude that the ability to detoxify free radicals within our morbidly obese population is impaired, producing a potential for the accumulation of oxidative damage.  This may contribute to the development of degenerative diseases often associated with obesity such as arthritis, diabetes, and artherosclerosis.
     

    5.  
    NITRIC OXIDE SYNTHASE (NOS) ACTIVITY IN THE RED BLOOD CELL: IS IT POSSIBLE?  Kevin Ford1 and Ellen S. Kang2, 1Christian Brothers Univ. Memphis, TN; 2Dept. of Medicine, Univ. of Tennessee, Memphis, TN.  
     Nitric oxide (NO) is a powerful mediator of physiological processes in the vascular system.  It is tightly regulated by rapid inactivation to NO2 upon interaction with reducing substances such as the iron in hemoglobin and cysteine and tyrosine as amino acids or peptide residues. Each of these mechanisms for inactivation is present in the RBC.  We studied whether the RBC had NOS activity by a direct assay, unlike two reports of NOS activity in RBC by increase in NO2 which could result from nitrosothiol groups released from peptide residues.  Normal human RBCs recovered after washing with Tris buffer (pH 7.2) were sonicated at 4oC, spun at 15,000g for 30 min., aliquoted and kept at -70oC until analysis.  NOS activity was measured with mM NADPH by recovery of 3H L-citrulline from 3H L-arginine as substrate.  Time and protein (Lowry) were varied with incubated at 37oC, detected by beta counting of 3H L-citrulline.  NOS activity was detected in the supernatant fraction but not in the pellet fraction.  Optimum conditions for NOS were 50 ug protein using 0.5 mM 3H L-arginine incubated for 5 min. at 37oC.   These findings with a more direct assay support the eariler reports that the RBC does express NOS activity which is limited to the cytoplasmic fraction and is not expressed by the membrane.  We speculate that this capability gives the RBC a vital role in the regulation of processes controlled by NO.  Supported by:  a grant from LeBonheur Childern’s Hospital 
     

    6.  
    CARNITINE LEVELS IN RAT RED BLOOD CELLS, WHITE BLOOD CELLS, AND BLOOD SERUM.  N. Koenigsknecht1, M. Volk2, M.S., and  M. Banschbach3, Ph.D) 1Dept. Of Biology, Christian Brothers Univ. Memphis, TN; 2,3 Dept of Microbiology and Biochemistry, Oklahoma State Univ. College of Osteopathic Medicine, Tulsa, OK.
     Carnitine is an essential carrier molecule of fatty acids that plays a key role in the human body by serving as an accessory for burning fat energy.  When the levels of carnitine are low, the body exclusively depends on glucose as the energy source.  Severe carnitine deficiency can result in muscular dysfunction with serious clinical implications.  The purpose of this study was to determine if carnitine could be measured in either red blood cells or white blood cells using a new coupled enzyme spectrophotometric assay that we developed.  Serum and washed red or white blood cell fractions were prepared from rat blood.  The spectrophotometric assay using alpha-ketoglutarate dehydrogenase, carnitine acetyl transferase, and diaphorase was used to determine free carnitine (in untreated samples) and total carnitine (in base hydrolyzed samples).  While carnitine was easily detected in serum, no carnitine was detected in red or white blood cell extracts.  If future experiments to improve extraction of the carnitine from the red and white blood cells do not yield satisfactory results, one of the more costly but more sensitive carnitine assays will need to be used to assay these samples.
     

    7.
    THE DISLOCATED KNEE.  Trent A. Gullett, Dept. of Biology, Christian Brothers Univ., Memphis, TN; Fred Azar, M. D., Campbell’s Clinic, Memphis, TN; Scott Sharp, M.D., Campbell’s Clinic, Memphis, TN; Marc Mihalko, Univ. Tennessee, Memphis, TN.
    Dislocation of the knee is a traumatic injury that involves ligament damage.  In this retrospective study, the charts of 38 knee dislocation patients over the last twelve years (the Med, Memphis TN) were reviewed.  The purpose of this research was to determine if one form of treatment (surgery and/or physical therapy) was more beneficial to the healing of the knee than another type of treatment (physical therapy without surgery).  Background, associated injuries, and treatment were recorded for each patient.  This research revealed two interesting aspects.  The first was the comparison between associated injuries (arterial and/or nerve damage) and the velocity of the knee dislocation (high velocity or low velocity).  There was a higher incidence of associated injuries with low velocity dislocations as compared high velocity injuries.  Also, research revealed that the range of motion of patientís knees that received surgery was less than those who did not receive surgery.  This study revealed that low velocity knee dislocations can be very traumatic, and that a follow-up of at least two years is needed for determining the health and the progress of a re-cooperating dislocated knee.
     

    8.  
    DIFFERENTIAL DIAGNOSIS FOR PATIENTS WITH PRE-SYNCOPE SYMPTOMS ((F. A. Hayes, H. Rashed)) Dept. of Biology, Christian Brothers Univ. Memphis, TN; Dept. of Autonomic Function, Univ. Tennessee, Memphis, TN.  Syncope or fainting is a common clinical disorder whose main etiology is deficient cerebral blood flow resulting from peripheral circulatory failure.  Using tilt table testing, a non-invasive method, subjects with histories of frequent fainting and dizziness were studied.  Two other groups were also studied; those with pre-syncope and age-matched control groups were compared.  Because syncope is mainly a peripheral autonomic dysfunction, monitoring brachial blood pressure alone during tilt table testing was often insufficient in diagnosing syncope.  Ankle blood pressure, when expressed as the ankle/brachial ratio has been shown helpful in the assessment of the peripheral circulatory dysfunction during tilt table testing.  When this ratio was used to investigate patients during tilt table testing, significant differences were found in the ankle/brachial ratios of control subjects, syncope patients, and pre-syncope patients.  Systolic blood pressure ratios of patients with syncope and control patients were compared; they differed in both supine and tilted positions.  When the controls were compared to the pre-syncope patients a significant difference was found in the systolic ratio only in the tilted position.  The observed results suggest that using ankle blood pressure, in a ratio with brachial blood pressure, to monitor the autonomically-controlled peripheral cardiovascular responses to posture is a valuable method to diagnose, evaluate and predict peripheral autonomic dysfunction in patients with syncope and pre-syncope.
     

    9.  
    RELATIONSHIP OF ACOUSTICAL PROPERTIES OF THE VOICE AND HEALTH STATUS IN OLDER ADULTS.  Y.Q. Lin**, Veronica F. Engle+, Barbara Fuller*, Marshall J. Graney+, Douglas Connor*, **Dept. of Biology, Christian Brothers University, Memphis TN, + College of Nursing, UT Memphis, and * University of Colorado.  
          The purpose of this study was to identify the relationship between acoustical properties of the voice and health status (HS) of older adults. Acoustical properties of the voice (jitter, shimmer, tenseness) are non-invasive, physiological measure of stress that are influenced by HS, but have not been studied in older adults. Older adult volunteers from the community (N = 28, 50% men) with a mean age of 73.46 years (SD= + 6.62years) and average educatiion of 13.61 years (SD = + 3.44 years) were recruited from a Senior Citizen Center. HS and demographic data were collected using a questionnaire. Participants produced three sets of four phonations of the vowel ‘a’ and ‘e’. A uniform five second segment of each vowel phonation was digitized and processed using computerized speech laboratory (CSL) and Horii computer programs. The mean of 12 samples for each subject were calculated and used for statistical analysis. In the vowel ‘a’, jitter, shimmer, and tenseness were significantly correlated with male gender, while only jitter and shimmer were significantly correlated with age; and no significant correlations were observed between HS and the voice acoustics. For the vowel ‘e’ only shimmer was significantly correlated with male gender, and only jitter was significantly correlated with smoking and surgery. Results indicate that there is a differential effect of HS and demographic characteristics of the participant on acoustical properties of the voice for vowels ‘a’ and ‘e’ which should be considered when conducting future research.  
    Supported by a grant from the University of Tennessee, Memphis of College of Nursing.
     

    10.
    Mechanism of Potassium Chloride induced release of arachidonic acid and contraction of bloodvessels: Selective activation of lipoxygenase and inhibition of prostaglandin synthesis
    Zachary Maxwell, Mubarack M. Muthalif, Jason Harper, Nour Karzoun and Kafait U. Malik  Dept. Biology, Christian Brothers University, Memphis, TN 38104 and the Department of Pharmacology, College of Medicine, The University of Tennessee , Memphis 38163.
     This study was conducted to elucidate the mechanism by which potassium chloride (KCl), which causes depolarization and increases extracellular calcium (Ca2+) influx, promotes arachidonic acid release for prostaglandin synthesis and their contribution to vascular smooth muscle contraction.  In cultured rabbit vascular smooth muscle cells (VSMC), KCl (60 mM) increased the activity of Ca2+/calmodulin dependent protein kinase II which was followed by an increase in mitogen activated protein kinase activity, cytosolic phospholipase A2 activity, and arachidonic acid release.  However, KCl failed to increase prostaglandin synthesis in VSMC or intact aortic rings.  In both VSMC and aortic rings, exogenous arachidonic acid was converted to prostaglandins, but their production was inhibited during exposure to KCl.  Interestingly, in the presence of the lipoxygenase inhibitor, baicalein, KCl increased prostaglandin production in both VSMC and aortic rings.  KCl-induced contraction of aortic rings was reduced by the prostaglandin endoperoxide receptor antagonist, ifetroban.  These data suggest that KCl promotes arachidonic acid release by increasing cytosolic phospholipase A2  activity via activation of Ca2+/calmodulin dependent protein kinase II and mitogen activated protein kinase.  Moreover, the arachidonic acid released by KCl is converted by lipoxygenase, and the products of lipoxygenase inhibit production of prostaglandins and cause accumulation of prostaglandin endoperoxides, which contribute to KCl induced contraction of rabbit aorta.  
    Supported by grants from the NIH (USPHS-19134) and the American Heart Association.

    11.  
    IMMUNOASSAYS OF GLYCOPHORIN A: AN ERYTHROCYTE MEMBRANE PROTEIN.  L. Gregory Meriwether1, Dennis Carrigan2, and Douglas P. Blackall2, 1Christian Brothers Univ., Memphis, TN, & Dept. Pathology, 2UT Memphis, Memphis, TN.  
     Glycophorin A is an erythrocyte integral membrane protein which carries the human MN blood group antigens.  This protein serves as a receptor for several pathogens, including reovirus and Plasmodium falciparum, the causative agent of malaria.  Recombinant glycophorin A was expressed in a variety of eukaryotic cell lines through transfection techniques.  Chinese hamster ovary (CHO) cells were mainly utilized for this purpose, and stable CHO cell lines were established expressing either the M or the N allele of glycophorin A.  Eight glycophorin A-specific monoclonal antibodies were assessed for their ability to bind with either red blood cell or recombinant glycophorin A through immunoblotting, immunofluorescence, and immunomagnetic bead agglutination.  These monoclonal antibodies included three specific for the M allele of glycophorin A (6A7, M2A1, AO9), four specific for the N allele of glycophorin A (N61, NN3, N92, AH7), and one MN nondiscriminatory antibody (10F7).  Immunoblotting revealed that six of the eight antibodies were capable of binding normal red blood cell glycophorin A, while only two antibodies bound recombinant glycophorin A.  Similarly, by immunofluorescence, only three antibodies recognized recombinant glycophorin A in stable CHO cells, but six antibodies recognized the glycoprotein when expressed transiently.  Three of the antibodies conjugated to immunomagnetic beads agglutinated CHO cells expressing recombinant glycophorin A, while the other five failed.  These results suggest a fundamental difference between red blood cell and recombinant glycophorin A.  Future studies will be directed to expressing a recombinant glycophorin A molecule more similar to the red blood cell membrane version.  
    Supported in part by: the National Blood Foundation.
     

    12. THE ROLE OF SERUM EOSINOPHILIC CATIONIC PROTEIN (ECP IN EXERCISE-INDUCED ASTHMA (EIA).  B. Myers, K.V. Leeper, J. Soberman, T.T. Mastin, & P. Jordan.   Dept. of  Biology, Christian Brothers Univ. Memphis, TN; Dept. of  Pulmonary and Critical Care Medicine, UT, Memphis, TN; Dept. of Cardiology, Heart Station, UT, Memphis, TN.
         Asthma is known to be a disease characterized by chronic inflammation.  One of the many key cells involved in the inflammatory process in the lung is the eosinophil.  Eosinophils and their mediators, specifically eosinophilic cationic protein (ECP), are potential markers of the severity of the inflammatory response.  The aim of this work was to ascertain whether patients with exercise-induced asthma (EIA) demonstrate not only an immediate increase in ECP, but also one that is sustained up to 30 minutes after exercise is ceased.  We studied five asthma patients and seven healthy nonasthmatics (controls) which were age, race, and sex matched.  Three different blood samples from each subject were obtained, one prior to exercise (baseline), the next immediately after exercising at 1/2 predicted VO2 max for five minutes (5 minutes), and a third after exercise was stopped and the participant had rested for 30 minutes (35 minutes).  A total of 36 samples were analyzed using a RIA kit.  Comparison of the EIA group and the non-EIA group were done with the Kruskall-Wallis test.  In comparison with studies by Venge et.al., their results showed elevated ECP levels in asthmatics.  Therefore, it might be possible to determine the severity of a disease based on ECP levels.  Supported by: Dept. of Pulmonary & Critical and Dept. of Cardiology, Univ. of TN, Memphis, TN. 
     

    13.  
    THYROID HORMONE REGULATION OF UCP-2 AND THE ob GENE IN RAT ADIPOSE TISSUE.  Linda Nguyen1 and Suleiman Bahouth2. 1Dept. of Biology, Christian Brothers University, Memphis, TN; 2Dept. of Pharmacology, UT, Memphis, TN.
     UCP-2 is an uncoupling protein that has recently been discovered in white adipose as well as in extra adipose tissue.  UCP-2 belongs to the family of uncoupling proteins which function to burn excess fat and to generate heat from ATP.  Another gene product produced in white adipocytes is the ob gene product leptin which serves as a satiety signal.  Therefore, we sought to determine if UCP-2 and leptin expression could be regulated by thyroid hormone since thyroid hormone controls much of the bodyís metabolic functions.  Twenty-five (g samples of RNA were obtained from epididymal adipose tissue of euthyroid, hypothyroid, and hyperthyroid rats.  These samples along with markers and blanks, were then subjected to electrophoresis, transferred onto nylon membranes, and probed by Northern blotting with 32P-labeled probes for UCP-2 and leptin.  The blots were counted to determine the influence of altered thyroid status on UCP-2 and leptin mRNA levels.  The Packard InstaimagerTM revealed that the radioactive count in UCP-2 from hyperthyroid rats was 7, 386 cpm, the euthyroid cpm was 4,235, and the hypothyroid count was 4,003 cpm.  On the other hand, the cpm in leptin mRNA were highest in hypothyroid RNA and lowest in hyperthyroid conditions.  These radioactive counts were representative of the levels of UCP-2 and lepting mRNAs seen in altered thyroid states.  Consequently, the hyperthyroid condition resulted in much more UCP-2 than either of the other two conditions tested while the reverse was true for leptin.  Therefore, in hyperthyroidism there is a decrease in satiety signaling (increased food consumption) and an increase in fat utilization for heat production.  Supported by NIH grant HL-48169.

    14.  
    LOCALIZATION OF D2 DOPAMINE RECEPTORS IN AVIAN STRIATUM USING IMMUNOHISTOCHEMICAL LABELING.  Rodney A. Paullus, Dept. of Biology, Christian Brothers University, Memphis, TN.
     The purpose of this research was to provide evidence as to the neuronal location of the D2 subfamily of dopamine receptors in avian (specifically pigeon) striatal projection neurons in comparison with their mammalian counterparts (specifically rat).  The basal ganglia regulate motor output via dopamine connections from the substantia nigra to the striatum.  Neurodegeneration of part of the basal ganglia occurs in Parkinson’s and Huntington’s Disease and impairs the ability of the basal ganglia to originate movement within the striatum.  An Avidin Biotin Complex method of immunohistochemical labeling with antigen retrieval in a sodium citrate buffer was used to localize the D2 receptors.  Examination of the immunohistochemical labeling showed that the LPO and PA striatal regions possessed the greatest amount of labeling for D2 receptors.  The labeling appeared characteristic in labeling of cholinergic neurons.  These results are consistent with previous localization studies for D1 receptors in avain striatum using the phosphoprotein DARPP-32 in showing that the striatium is enriched in D1 and D2 dopamine receptors.  This study also suggests a possible antigenic difference between rat D2 and pigeon D2 receptors.
     

     15.  
    PHOTORECEPTOR DAMAGE IN CULTURED ADULT RABBIT RETINAL EXPLANTS  M. Svadlenka, D.A. Johnson, M.E.C. Fitzgerald, M.M. Jablonski, S. Ashraf, T.O. Woods, Center for Vision Research, U.T. Memphis. 
     The purpose was development of an explant culture system for utilization in establishing an in vitro model of photoreceptor damage. To this end, sections of cornea were co-cultured with retinal explants and/or retinal pigmented epithelial cells (RPE). Morphology of processed tissue was assessed by light microscopy.  Explant cultures, in the absence of RPE, were also subjected to complement-mediated lysis of photoreceptors with B6-30N antiopsin.  Retinas were examined both light microscopically coupled with double labeling using indirect immunolabeling with WGA(wheat germ agglutinin)/PNA (peanut lectin agglutinin) followed by examination on the confocal microscope.  WGA stains rods green and PNA dyes cones red by binding to glycoproteins on the cell membranes.  The tissue co-cultures using cornea as a substrate did not remain in contact with each other.  The intrinsic qualities of the RPE were demonstrated as it repeatedly formed a tight ball of cells away from the cornea substrate.  In retinas exposed to anti-opsin/complement, the structure of the photoreceptor outer and inner segment complex was altered.  Outer segments were completely absent and the inner segments appeared to seep from the outer nuclear layer.  The lectin binding pattern was also modified and some photoreceptor cell loss was noted.  The cornea was determined a poor substrate for RPE and retinal explant cultures. A model of photoreceptor damage was successfully generated in a full thickness adult retinal explant.  This model will serve as a template in which potential photoreceptor rescue factors can be tested.  Funded by NIH grants EY01655, EY10853, Research to Prevent Blindness.
     

    16. 
    Up-regulation of Cadherins following Central Nervous System injury in Rats
    Félix Vázquez-Chona,1 and Eldon E. Geisert Jr.2  1Dept of Biology, Christian Brothers University, Memphis TN, and 2Dept of Anatomy and Neurobiol., UT, Memphis, TN.
     Following a lesion of the CNS, astrocytes become reactive and in many cases form a scar.  Defining the molecular mechanisms governing glial scar formation is critical to understanding how the CNS responds to injury.  In the present study, the role of cadherin at the glial scar was studied using immunohistochemistry and western blotting.  The expression of cadherin in astrocytes was confirmed in cultures of rat astrocytes where it was expressed at regions of cell-cell contact.  Cadherin was consistently upregulated at the site of gliosis following a stab wound in the CNS of the adult rat.  A quantitative analysis from injured and non-injured cortex revealed a threefold increase of a cadherin, at 135 kDa band, in the injured tissue.  The location of this band is consistent with the electrophoretic mobility of N-cadherin.  Due to its crucial role in the intercellular adhesion of most cell types, the high levels of cadherin at the site of CNS injury indicate that cadherin may play a role in the response of astrocytes to injury and in the formation and maintenance of the glial scar. 
    Supported by:  University of Tennessee Health Science Center.
     

    17. 
    THE EFFECTS OF MANUAL LYMPHATIC STIMULATION IN WOMEN SUFFERING FROM SECONDARY LYMPHEDEMA. 
    Lyndsay Woodward, Christian Brothers University; Cathy Sanford, PT, Laurel Means, PT, Kris Valentine, PT, Tammy Gibson, OT, Huntsville Hospital East, Huntsville, AL
     Secondary lymphedema is a disease that occurs due to some kind of external conditions, usually a result of surgery that occurred in surrounding areas to lymphatic tissue.  Patients suffering from breast cancer are at high risk for lymphedema.  These studies were conducted on a cohort of twenty patients, female, suffering from secondary lymphedema subsequent to breast cancer surgery.  Manual lymph drainage (MLD) is the main technique used for the treatment of this disease.  The treatment consisted of outpatients undergoing a series of exercises and massages, in order to release some of the excess fluid and proteins from the affected extremity.  Measurements are taken on both the affected and unaffected extremities at 5-cm increments.  The first 4-6 weeks, therapy was received three times a week.  Once positive results are seen, follow-up therapy is performed at 1 month, 3 months, 6 months and 12 months.  The 20 patients were subdivided into 3 groups: 0-23 months, 24-59 months and 60-120 months, based on the onset of lymphedema.  Based on our results the percent edema was reduced the most (89%) in patients who obtained treatment within two years of the onset or in individuals that have been afflicted with the disease for at least six years (79%) prior to the initiation of therapy.  The least percent of improvement (34%) was obtained from the group in which therapy was initiated 3-5 years from the onset of the disease. 
    [end]

      
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