Health sciences.

Chair: Hamed Benghuzzi, University of Mississippi Medical Center 

Vicechair: D. Olga McDaniel, University of Mississippi Medical Center

THURSDAY MORNING

Lake View II

8:10 Introduction

8:15 EXPRESSION OF TRANSFORMING TGF-B III SPARC-NULL

Nikeya Carter (1) *, Gregory Copper (2), Lynn Opperman (2), and Joseph M. Wahome (1), (1) Mississippi Valley State University, Itta Bena, MS 38941, and (2) Baylor College of Dentistry, Dallas, TX

Osteonectin or secreted protein acidic and rich in cysteine (SPARC) is a secreted [Ca.sup.2+] ion binding glycoprotein found in a variety of cell types including osteoblast, endothelial cells, ligament fibroblasts, pariental yolk sac cells and basement membrane tumors. Osteonectin is important in maintenance of bone mass in vertebrates and in growth of bone in sutures hence affecting cranial facial development. Postnatal mouse craniums at 15 d and 30 d with either wild type or SPARC Osteonectin (-/-) knockout genotypes were fixed, sectioned and stained. Staining was scored from 1 to 3 and compared with the controls. The wild type interfrontal sutures had some TGF-B2 immuno-positive fibroblasts at 15 d, which disappeared at 30 d. The knockout mouse sutures were negative. Minimal TGF-B2 immuno-staining was noted within the interfrontal suture in the knockout compared to the wild type mice at 30 d. These data will be interpreted in light of cranial development.

8:30 GENOMIC MARKERS ASSOCIATED WITH ISCHEMIC CARDIOMYOPATHY AND THE OUTCOME OF CARDIAC TRANSPLANTATION

Vernetta Coleman (1) *, Sani Yamout (1), D. Perrin Roten (1), Charles K. Moore (1), Georgio Aru (1), Maria E. Oropeza (1), Joseph A. Cameron (2), and D. Olga McDaniel (1), (1) University of Mississippi Medical Center, Jackson, MS 39216, and (2) Jackson State University, Jackson, MS 39217

The effects of cytokine gene polymorphism were evaluated in African-American (AA) and Caucasian (CAU) patients that had undergone cardiac transplantation (CTx). It has been suggested that allograft rejection is mediated by cytokines. Our working hypothesis is that the clinical condition of recipients before CTx might affect the outcome of allograft function through pre-existing genetic factors that are unique to each individual. Genomic DNA samples from 19 AA and 46 CAU recipients were tested by either single or multiplex PCR. Frequency distributions of genotypes were analyzed in respect to pre-CTx clinical characterization including Coronary artery disease (CAD) and non-ischemic cardiomyopathy. The rejections were scored based on endocardial biopsy and histopathological assessment: rejection grades were IA, 2, 3A, or 3B and they were analyzed for cytokine genotype association. Overall, the IL-10 high producer genotype was present in all patients graded as 1A and the frequency was reduced significantly in gra de 2, 3A, and 3B patients (28.6%, p = 0.02; 23%, p = 0.001; and 20%, p = 0.04, respectively). IFN-[gamma] A/A and IL-10 low producer genotypes were significantly increased in AA patients, compared to CAU patients with grade 3A rejection (47% vs 16.1%, p = 0.04; 76.5% vs. 32.2%, p = 0.006 respectively). There was a direct relationship between IFN-[gamma] T/T high producer and ischemic CAD as compared with IFN-[gamma] A/A low producer and non-ischemic cardiomyopathy, supporting the influence of immunological factors in coronary vasculopathy. These data might provide a valuable information regarding the cellular and molecular mechanisms of graft response and might predict the outcome of allograft function.

8:45 GROWTH FACTOR PRODUCTION IN EARLY STAGES OF BENIGN PROSTATIC HYPER-PLASIA UPON EXPOSURE TO SUSTAINED DELIVERY OF ANDROGENS

Melanie C. Pollan *, Hamed Benghuzzi, and Michelle Tucci, University of Mississippi Medical Center, Jackson, MS 39216

Benign prostatic hyperplasia (BPH) is a condition that affects a significant population of older males, yet its pathogenesis is not clearly understood. This study was designed to give broader insight into the early development of BPH by looking at changes in growth factor production in the ventral prostate. To accomplish this, Sprague Dawley rats (n =16, 250-300 g) were randomly divided into four equal groups. Three treatment groups were each implanted with ceramic drug delivery devices that were designed to deliver continuous physiologic doses of testosterone (T), dihydrotestosterone (DHT), and androstenedione (AED) for ninety-day duration. Immunohistochemical analysis for epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) indicated whether these growth factors were involved in early processes of BPH induced by the delivery of androgens. Histopathological evaluation demonstrated increased positive reactivity for both EGF and bFGF in all steroid treated animals compared to controls. A sim ilar trend was observed in the vascular endothelium. This information could be helpful in developing new methods for early diagnosis of BPH, but more importantly this knowledge provides the literature with clues about the cellular responses encountered at the initiation of the disease process.

9:00 LDL OXIDATION BY THE EXPOSURE ANDROGENS USING MRC-5 CELL LINE AS A MODEL

Felicia Magee Tardy *, Hamed Benghuzzi, and Michelle Tucci, University of Mississippi Medical Center, Jackson, MS 39216

The relationship between LDL oxidation and the development of atheroma has not been fully investigated. The objective of this study was to investigate the effect of androgens (testosterone, TE) on the intactness of LDL upon incubation with MRC-5 cells. A total of 72 tubes plated with MRC-5 fibroblasts (50,000) were divided into four equal groups. The four groups were treated with one of the following: serum only (control), low-dose LDL, high-dose LDL, and LDL + TE. The experiment was divided into 24, 48, and 72-hour periods. At the end of the incubation periods, representative samples from each group were used for the determination of LDL oxidation (Dow Coming units/lipoprotein kits). This was accomplished through lipoprotein electrophoresis procedure. Results of the lipoprotein electrophoresis indicate that LDL was slightly modified during the incubation periods with low-dose LDL, and high-dose LDL treatments. Extensive electrophoretic mobility was observed in the combination of DL and TE treatment. Results from this study suggest that MRC-5 cells are capable of modifying the native form of LDL. However, further investigation is needed to determine the exact mechanism of LDL modification and the role of MRC-5 cells in its modification.

9:15 LOCALIZATION OF PROINFLAMMATORY CYTOKINES ON CARDIOVASCULAR TISSUES EXPOSED TO SUSTAINED DELIVERY OF ANDROGENIC STEROIDS IN RODENTS

Shontell Credit (1) *, Hamed Benghuzzi (2), Joseph A. Cameron (2), Michelle Tucci (2), and Audrey Taso (2), (1) Jackson State University, Jackson, MS 39217, and (2) University of Mississippi Medical Center, Jackson, MS 39216

Recent studies have shown that cytokines are over-expressed within heart failure cases. It is also well documented that various cytokines have an important role in biological processes such as inflammation, wound healing, and tumor progression. The objective of this study was to deliver continuous levels of testosterone (T), dihydrotesterone (DHT), and androstenedione (AED) in a sustained manner using ceramic drug delivery systems and determine their role as risk factors for cardiovascular diseases. A total of 16 rats were subdivided into four equal groups. Groups 1-3 were implanted with TCPL capsules loaded with T, DHT, and AED respectively. Group 4 animals were untreated and served as control group. At the end of 90 days post treatment, heart tissue were harvested and subjected to H&E and immunohistochemistry procedures following standard lab protocols. The data obtained from this study demonstrated that IL-6 was the most prominent cytokine expressed in the tissue in group 2 animals compared to control and other experimental groups. No significant difference in IL-I induction was observed among all groups. In the ventricular sleeve, T caused the most significant evidence of hypertrophy (p <0.05). In contrast, hypertrophy was prominent in the apical region in animals treated with AED compared to the control and T treated animals. The results of this investigation uncovered evidence that various regions of the heart have specific binding receptors for certain steroids to act upon.

9:30 TETRAHYDROPAPAVEROLINE IN VIVO: A POSSIBLE BIOCHEMICAL LINK TO ALCOHOLISM IN MAMMALS

Jacob C. Strawbridge *, Kenneth D. McMurtrey, John G. McCoy, and Christopher P. Ward, University of Southern Mississippi, Hattiesburg, MS 39406

The role of tetrahydropapaveroline (THP), a condensation product of 3,4-dihydroxyphenyl-acetaldehyde with dopamine, in the regulation of alcohol consumption has been the subject of much debate among addiction researchers. In this paper, 31 adult male Sprague-Dawley rats received intraventricular injections of either racemic THP hydrobromide (0.65 or 1.3 mg/mL), R-(+)-THP hydrobromide (0.66 or 1.4 mg/mL), or an equal volume of vehicle. The lower doses of both ([+ or -]-THP and (+)-THP significantly increased volitional alcohol intake. For animals treated with the racemic compound, the increase was significant at 7-13% (v/v% ethanol solution) concentrations. The R-(+)-enantiomer increased consumption at 4-11%, and 15-20% concentrations of ethanol. The higher doses of both compounds did not significantly alter alcohol preference. Whether or not endogenously formed THP participates in the etiology of alcohol addiction remains unclear. Nonetheless, there are few known compounds that induce a preference for unsweet ened alcohol solutions over water in laboratory animals.

9:45 SPATIAL ANALYSIS OF PESTICIDE EXPOSURE AND RISK OF BREAST CANCER MORTALITY iN MISSISSIPPI

Mohamed H. Abdulla *, Mary Lou Gutierrez-Mohammed, and Ibrahim O. Farah, Jackson State University, Jackson, MS 39217

Breast Cancer is the most common form of cancer in women and its incidence increase with age. Established risk factors include advancing age, early menarche, late menopause, positive first relative, late age at first birth and socioeconomic status. Mississippi as state has a combination of risk factors making it a suitable candidate for elucidating the pathways of breast cancer etiology. The purpose of this study was to analyze pesticide exposure and the risk of breast cancer mortality in Mississippi. Data comprised county-level, state economic areas (SEA), breast cancer mortality data from NCI and pesticide use data from USDA (1970-1994). Methods included descriptive statistics, unit analysis at the county level, and geospatial analysis (GIS), Results showed significant correlation for three SEA's including Yazoo, Vicksburg and Cornith, Total acres planted vs. female cancer mortality showed Spearman values (P-value) of 0.674 (0.030) in Yazoo, 0.604 (0.0062) in Vicksburg for acreage/white females, and 0.667 ( 0.049) in Cornith for acreage/black females. We conclude that there are moderate statistically significant associations between number of acres planted crops in Mississippi and the mortality rate of breast cancer, the association varies by economic area, race, and crop planted, Spatial analysis using GIS will demonstrate a better estimate for this correlaion through visual display of topography, crops, and mortality rates.

10:00 Break

10:30 Divisional Poster Session

A NOVEL TREATMENT FOR EARLY STAGES OF AVASCULAR NECROSIS

Audrey Tsao (*), James H. Hughes, and Ruth Ann Buckhalter, University of Mississippi Medical Center, Jackson, MS 39216

This study is to investigate a novel surgical implant to treat early femoral head Avascular Necrosis (AVN). Methods: Patients between the ages of 18 to 75 with Stage I or II AVN as classified by Steinberg/UPenn system are entered into one of three groups (FDA approved IDE #G990234). Group I with unilateral disease are randomized with core decompressions. Groups II and III with bilateral disease receive the implant; Group II having both hips eligible for the procedure. Group III, the contralateral hip is excluded. The AVN Implant is made of a porous tantalum with elasticity similar to bone and evidence of osteoconductive, osteoinductive and vascular invasion properties. Clinical outcomes are measured by Harris Hip Scores (HHS), SF-12 and/or radiological studies completed at pre-op, three, six, twelve, and twenty-four months, and then annually. Results: University of Mississippi Medical Center has enrolled seventeen patients, with seventeen implants, and one core decompression. One patient dropped secondary to non-compliance. One year follow-up for five hips, six month follow-up for thirteen hips reveal an increase in the HHS, function, and a decrease in pain and no radiographic progression of disease, except in two cases. The core decompression has experienced a similar outcome. Discussion: Preliminary data appears promising, with no progression of the disease, a decrease in pain and an increase in function for the majority of cases. Patients will be monitored for five years.

ADMINISTRATION OF DIOSGENIN TO OVARIECTOMIZED RATS PROTECTS KIDNEY FUNCTIONS

Anita Winfield (1)*, Hamed Benghuzzi (2), Michelle Tucci (2), and Joseph A. Cameron (1), (1) Jackson State University, Jackson, MS 39217, and (2) University of Mississippi Medical Center, Jackson, MS 39216

The use of over the counter progesterone creams is on the rise and is thought to be a safe and effective means for controlling menopausal symptoms. The active ingredient in these preparations is the compound diosgenin. Diosgenin, which is synthetically derived from wild yams, is converted to human progesterone through a series of enzymatic steps. The efficacy and safety of these products has not been fully investigated in vivo. The objective of this study was to supplement ovariectomized rats with sustained release of diosgenin for 47 days and evaluate its effect on kidney structure. A total of 15 animals were randomly divided into three equal groups. Animals in group I served as intact control, animals in groups II were ovariectomized and animals in group III were ovariectomized and supplemented with a TCPL capsule containing 500 mg of diosgenin. Blood and body weights were recorded weekly and at the end of 47 days kidneys were collected and analyzed morphometrically. The results indicated significant increa ses (p <0.05) in the body weight of ovariectomized animals compared to control and diosgenin treated animals. Significant increases were observed in the kidney wet weights of ovariectomized animals. Further analysis of the kidneys showed remarkable changes in both the glomerular length and area in ovariectomized animals. The results suggest that diosgenin was able to protect the kidney from morphological changes associated with ovariectomy. The mechanism is most likely responsible for this protection is the conversion of diosgenin to progesterone.

APOPTOSIS: CINNAMON EXTRACTS AND BUCCAL EPITHELIAL CELLS

Ayesha Alam (1) *, Hamed Benghuzzi (2), and Michelle Tucci (2), (1) Jackson State University, Jackson, MS 39217, and (2) University of Mississippi Medical Center, Jackson, MS 39216

Buccal epithelial cells line the oral cavity, and are the first cells to encounter cinnamon compounds in food upon ingestion. Previous experiments have shown ETOH and [H.sub.2]O extracts of cinnamon to cause increase in cell death. However, mechanisms for the observed cell death have not been identified. Potential mechanisms for cell death have not been identified. Potential mechanisms for cell death include cellular lysis due to buffer interaction with cells (hypertonic/hypotonic) and/or apoptosis (programmed cell death). The nature of suspension buffer was dismissed as a possible means for cellular death in this series of experiments because vehicle treated cells did not undergo either cellular death or lysis. The object of the work was to identify whether extracted cinnamon compounds caused higher incidence of apoptosis in isolated human buccal epithelial cells challenged with ETOH, ETOH + Cinnamon, [H.sub.2]O and [H.sub.2]O + Cinnamon for 15 and 60 minute periods. Results indicated that heat soluble and E TOH soluble compounds of cinnamon induce greater degree of apoptosis in cells compared to cells treated with ETOH or PBS. Various fractions from cinnamon extracts are currently being isolated based on liquid chromatography. (Supported by grant from the Howard Hughes Medical Institute).

BIOLOGICAL IMPLICATIONS OF SYNTHESIS OF CALCIUM PHOSPHATE

Kenisha DeLoach (1) *, Jason Ritchie (2), and Joseph M. Wahome (1), (1) Mississippi Valley State University, Itta Bena, MS 38941, and (2) University of Mississippi, University, MS 38677

Calcium Phosphate is an important component of bone and teeth matrix. An artificial method of synthesizing bone and teeth component may serve as a source of replacement of natural parts in situations where biological synthesis is deficient. Calcium phosphate was synthesized by converting the hydroxyl group in monomethyl-polyethyleneglycol (MPEG) to an alkyl chlorine using thionyl chloride and pyridines. The resultant MPEG-Cl was converted to the alkyl phosphonate precursor MPEG-P+(OEt)3 hydrolyzed to MPEG-[PO.sub.3][H.sub.2] using an ion exchange chromatography. The resulting porous material can serve as a matrix for bone to grow around forming living bone tissue. The application of this research will be discussed.

BONE SPARING SURGICAL OPTIONS FOR TOTAL HIP REPLACEMENT

Audrey Tsao *, Tracy Pesut, Charity Peacock, and Ruth Ann Buckhalter, University of Mississippi Medical Center, Jackson, MS 39216

This study is to investigate a new Conservative Hip Prosthesis designed for younger patients with a primary diagnosis of Avascular Necrosis (AVN). This prosthesis is designed to spare bone and make revisions easier. Methods: Patients between the ages of 18 to 60 with the primary diagnosis of AVN are considered prime candidates for this prosthesis. However, young patients with other diagnosis will be considered for this prosthesis. The Mayo Conservative Hip Prosthesis is made of Tivanium[R] To-6AI-4V Alloy, with a dual-plane taper. Clinical outcomes are measured by pre-operative Charnley Scores, and Harris Hip Scores (HITS), SF36, and/or radiographic studies completed preoperatively and post-operatively at three, six, twelve months, and annually thereafter for a minimum of five years. Results: University of Mississippi Medical Center has presently enrolled thirty patients, with thirty-six implants, six being bilateral. Post-operative follow-ups include three patients at three years, four patients at two years, eighteen patients at one year, and eleven at various other stages ranging from six weeks to near one year. All follow-ups reveal an increase in HHS, increase in quality of life, and a decrease in pain. Discussion: Data shows the patients increase in function, decrease in pain, and an overall improved quality of life. No revisions have been required to date, but should the need arise we hope to prove that this hip prosthesis will allow for easier revisions and more bone stock to prolong the hip function as the patient matures.

CLONING OF pspA/EF3296 FOR USE AS A DNA VACCINE AGAINST STREPTOCOCCUS PNEUMONIAE

Urseline A. Hawkins (1) *, Quincy Moore (2), and Larry S. McDaniel (2), (1) Tougaloo College, Tougaloo, MS 39174, and (2) University of Mississippi Medical Center, Jackson, MS 39216

The prevention of pneumococcal infection is currently a topic of great medical interest because of the significant morbidity and mortality associated with Streptococcus pneumoniae (pneumococcus). The current polysaccharide vaccine is limited in its coverage and is ineffective in the elderly and children less than two years of age. One of the alternatives being considered, is the formulation of a genetic vaccine using pneumococcal surface protein A (PspA). PspA is an effective antigen because it is present on all pneumococci, and it is required for full pneumococcal virulence. Based on sequence analysis, PspA can be grouped into six clades. We have previously demonstrated that pspA from dade 2 or dade 4 can elicit protection when used in mouse pneumococcal sepsis model. It has been hypothesized that a mixture of PspA from clades 2,3, and 4 would protect against> 90% of all pneumococcal infections. Therefore the main goal of this study was to clone pspA/EF3296 which is a prototypic dade 3 PspA. We cloned pspA/E F3296 into pNGVL3, a eukaryotic expression vector, for use in mouse immunization studies.

COMPARISON OF HIGH FAT, CARBOHYDRATE, AND PROTEIN DIETS REGARDING RELEASE OF CYTO-KINES IN INTESTINAL TISSUE AND BLOOD SERUM OF RATS

Kim Parker * and A. Kurt Thaw, Millsaps College, Jackson, MS 39120

Blood serum and intestinal tissue from rats given one of four diets, consisting of 60% fat, 60% carbohydrate, 60% protein, and standard rat chow were examined to determine the release of cytokines produced by each type of diet. The samples were examined using ELISA kits designed to determine the concentration of cytokines interleukins (IL-1[beta],IL-6) and Tumor Necrosis Factor [alpha](TNF-[alpha]). There was a clear release of cytokines in response to ingested nutrients. IL-1[beta] and TNF-[alpha] were released in larger quantities, while the release of IL-6 was minimal, yet still present. The current research is applicable to future medical and nutritional studies because of the information known regarding the release of cytokines and control of appetite. Though the release of numerous factors have been reported in response to ingested nutrients, this is the first report on the effect of specific nutrients on cytokine release.

COMPARISON OF NPR EXPRESSION WITH NP1 AND NARP IN ADULT RAT BRAIN

Deidra Jones *, Richard Cho, Radhika Reddy, and Paul Worley, Johns Hopkins University, School of Medicine, Baltimore, MD 21287

Pentraxins are a functionally diverse group of secreted calcium dependent lectins originally named for their ability to form pentamers of five non-covalently bound subunits (Gewurz et al. 1995; Goodman et al. 1996). The two subfamilies of pentraxins are the classic short pentraxins and the relatively long pentraxins which Narp, NPl and NPR are members. Neuronal activity-regulated pentraxin (Narp) is a secreted immediate early gene regulated by synaptic activity in the brain (O'Brien et al. 1999); Neuronal pentraxin 1 (NP1) is also a secreted protein but is constituitively expressed in some regions of the brain (Hopf et al. 2002). Neuronal Pentraxin Receptor (NPR), a 494 amino acid protein, has properties that are unique to Narp and NP1 that may confer special function. Evidence suggests that these two proteins may be synaptogenic factors. The goal of my project was to broadly characterize NPR expression in rat brain and compare it to Narp/NP1 expression. In addition, NPR expression regulation during developme nt was also examined. Sprague-Dawley rats were sacrificed at various postnatal time points during development. Cortices and hippocampi were collected for a western blot analysis of pentraxin expression in vivo. The co-expression of long form pentraxins during development and in regions of adult rat brain hippocampi support our hypothesis that pentraxins co-function in their ability to modulate synapses formation by clustering AMPA receptors.

CYTOKINE ANTIBODIES (ANTI-IL-1[beta], ANTI-IL-6, AND ANTI-TNF-[alpha]) INCREASE APPETITE IN FREE-FEEDING RATS

Kim Parker * and A. Kurt Thaw, Millsaps College, Jackson, MS 39120

Recently, investigators have become interested in the immune systems' role as a regulator of physiological processes such as sleep and appetite. Data revealing a specific lengthening of the interval between meals related to the administration of excess cytokines has already been established. When the cytokines interleukin- 1 beta (IL-1[beta]), tumor necrosis factor alpha (TNF[alpha]), or interleukin-6 (IL-6) are given to a rat immediately following a meal the subject will wait an average of 50% longer before eating the next meal (compared to control injections). Thus, the se 3 cytokines appear to be playing a role in regulating the feelings of fullness or satiety between meals. The current research administered three concentrations of antibodies for each cytokine to rats during their first nocturnal meal. The amount of time elapsed between that first meal and the initiation of a second meal and the size of each meal measured in time spent eating were recorded. The effect of the antibodies on the interval betw een meals as well as the relationship between the size of the first meal and the interval between meals (satiety ratio) was able to be calculated. Results indicate cytokine antibodies increase total food intake as well as decreasing the amount of time between meals.

DELAYED APOPTOSIS UPON THE TREATMENT OF HEP-2 CELLS WITH BS

Charla Corder *, Hamed Benghuzzi, Michelle Tucci, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

Nigella sativa (Black seed, BS) has been used to promote health and fight disease for centuries. The objectives of this investigation were: (1) to study whether agents such as cortisol and LPS alone or in combination induce cellular (HEp-2, laryngeal carcinoma) damage with time in culture (24, 48, and 72 hours) using apoptosis as a marker, (2) to determine if an immune stimulant such as BS, can protect HEp-2 cells from insult and ultimately thwart the programmed cell death mechanism. A total of 54 HEp-2 cell/tubes (50,000 cells per tube) were divided into six equal groups. Group one served as untreated control, while groups 2-6 were treated with either cortisol (10 ng/ml), LPS (10 [micro]g/ml), BS (25 [micro]g/ml), or a combination of LPS and cortisol and cortisol plus LPS plus BS, respectively. At the end of each phase the cells were harvested, heat fixed and stained with H&E to evaluate morphological changes. Immuno-histochemistry, using antibodies against caspace-3 to evaluate cells undergoing apoptosis wa s conducted in all groups. The results of this study showed evidence of cells undergoing apoptosis at different magnitudes in all groups. However, the most dramatic change was seen in groups containing cortisol and LPS alone or in combination. This was supported by the fact that there were several adaptive responses observed in all phases. In addition, the exposure of BS to cells pretreated with cortisol and LPS showed evidence of protection against the progressive apoptosis.

DOWN REGULATION OF CD14 EXPRESSION THROUGH PRETREATMENT WITH GLUCOCORTICOIDS 

Randee K. Ivy *, Michelle Tucci, Hamed Benghuzzi, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

Glucocorticoids such as cortisol are potent immunosuppressive agents that act on many cells of the body, including macrophages. Macrophages express CD14 in response to lipopolysaccharides (LPS) extracted from bacterial coats. The objectives of this study were: (1) to determine if pretreatment of macrophages with cortisol for 30 minutes prior to challenging the cells with endotoxin results in increased cell loss, cell damage (MDA), and suppression of CD14 receptors; and (2) to determine if CD14 receptor expression is able to recover with time. An experimental design incorporating RAW264.7 cells (RAW) was used in order to evaluate our objectives. The cells were plated on 24 well plates and subsequently divided into four groups. The first group was untreated and served as the control. Group two was treated with LPS, group 3 with 10 [micro]1 of cortisol, and a combination of LPS and cortisol was used in the treatment of the fourth group. The cells were recovered at 24,48, and 72 hours. Results showed that there w as a significant decrease in the proliferation rate in RAW cells exposed to cortisol and LPS either alone or in combination when compared to the untreated cells. Cell damage was also increased in the treated cells. LPS caused increased receptor expression at all time points. CD 14 expression was down regulated at 48 hours in cells pretreated with cortisol, however, this suppression was no longer evident at 72 hours.

EFFECTS OF RIBOFLAVIN AND/OR ASCORBIC ACID SUPPLEMENTATION ON PLASMA, LEUCOCYTE AND URINARY ASCORBIC ACID IN UNIVERSITY STUDENTS

Felix Adah *, and O.A. Ajayi, University of Mississippi Medical Center, Jackson, MS 39216, and University of Ibadan, Nigeria

A study was carried out to observe the effects of riboflavin and/or ascorbic acid supplementation on plasma, leukocyte and urinary ascorbic acid in healthy young volunteers. A total of 36 students were assigned for this study (age ranges between 23-33 years) and further divided into three equal groups (random selection). A base line for ascorbic acid (AA) levels in the plasma, leukocytes, and urine were established. Each volunteer in groups I, II, and Ill was given orally riboflavin (5 mg/day), AA (100 mg/day), and a combination of AA (100 mg/day) plus riboflavin (5 mg/day), respectively. At the end of experimental phase (10 weeks post supplementation), blood and urine were collected and processed for AA levels in plasma, leukocytes and urine following standard procedures. Data obtained demonstrated that (I) riboflavin supplementation resulted in no significant change in AA in the urine and plasma (p> 0.05). In contrast, AA levels in leukocytes were increased (p <0.01), (II) the supplementation of AA alone o r in combination with riboflavin resulted in a remarkable increase in AA at the plasma, urine and leukocytes levels. In conclusion, results of this study revealed that the supplementation of riboflavin provided stimulatory pathway of AA production and this could be helpful in promoting the immune system and preventing the development of symptoms of AA deficiency.

FACTORS INFLUENCING BREAST AND CERVICAL CANCER SCREENING AMONG AFRICAN AMERICAN WOMEN

Ina Chapman (1), Thelma Hurd (1), Courtni Newsome (2) *, and Sharita Womack (3), (1) Roswell Park Cancer Institute, Buffalo, NY 14263; (2) Tougaloo College, Tougaloo, MS 39174; and (3) State University of New York at Buffalo, Buffalo, NY 14260

Breast and cervical cancer are major public health issues that are gaining significant importance in the field of research. Studies have shown that minority women are less likely to undergo screening than Caucasian women and are frequently diagnosed with more advanced disease. This could be due to differences in the attitudes of women of various ethnic groups and other factors. These differences and factors have caused minority women to have a higher mortality rate than Caucasian women, although the number of Caucasian women who get cancer of the breast and/or cervix is higher than minority women. The goal of this project was to evaluate the relationship between socioeconomic factors, the perceived attitudes of health care providers, and mammography and cervical cancer screening utilization by minority women. A 62 item telephone survey containing questions about socioeconomic factors, attitudes, and breast and cervical cancer screening utilization was administered to 59 African American women participating in the Witness Project of Buffalo. Participants' responses were entered into a database and analyzed using SPSS and chi square analysis. The data indicate d that the level of education completed and regular church attendance positively correlated with both breast and cervical cancer screening. Cervical screening, but not mammography, was positively associated with peer group support.

IN VIVO ANTIMUSCARINIC ACTIONS OF THE ANTIHISTAMINERGIC AGENT, DESLORATADINE

LaShondra Powell (1) *, Trey Howell (1), Jamekia Holloway (2), Colibri Jenkins (2), Brandon Lineberry, (1) David Yokum, (1) and Rob Rockhold, (1) (1) University of Mississippi Medical Center, Jackson, MS 39216, and (2) Tougaloo College, Tougaloo, MS 39174

Desloratadine (DES), a metabolite of the antihistamine, loratadine (Claritin [C]), is marketed as Clarinex [R], for management of seasonal rhinitis. In vitro tests indicate that DES can block muscarinic receptors and possibly contribute to adverse effects, such as dry mouth or tachycardia. Two murine and one rat model were used to evaluate antimuscarinic actions of DES in vivo. In the pithed rat, DES (1 mg/kg, i.v.) inhibited negative inotropic (left ventricular dP/dt) and chronotropic effects caused by oxotremorine, a nonselective muscarinic agonist (p < 0.05). Urethane-anesthetized (1.5 g/kg, i.p.) male ICR mice (25-35 g) were pretreated i.p. with vehicle, DES (1, 5 mg/kg) or a muscarinic antagonist, either atropine (AT; 0.5 mg/kg), atropine methyl nitrate (AMN; 0.5 mg/kg), diphenhydramine (1 mg/kg), methoctramine (0.5 mg/kg), or 4-DAMP (1 mg/kg). Oxotremorine (0.5 mg/kg, i.p.) was given 15 min. later and salivation evaluated for 30 min. Only AT and 4-DAMP inhibited salivation. In conscious mice, oxotremori ne (0.5 mg/kg, s.c.) was given 15 min. following pretreatments and tremor, salivation and lacrimation evaluated. Only AMN and DES (at doses below 5 mg/kg) were ineffective at moderating tremor, while only AT, AMN and 4-DAMP inhibited salivation and lacrimation. DES exerts significant antimuscarinic actions in the heart, but not on salivation, lacrimation or tremor. (Supported by A ventis Pharmaceuticals).

MORPHOLOGICAL EVALUATION OF MRC-5 FIBROBLASTS AFTER STIMULATION WITH STATIC MAGNETIC FIELD AND PULSATING ELECTROMAGNETIC FIELD

Kimberly Pate *, Hamed Benghuzzi, Michelle Tucci, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

The quality of tissue repair and the speed with which that repair can be accomplished are the two major variables in the healing of any injury. Today, magnetic field exposure to traumatized areas has shown to be a promising tool in the healing process. The exact mode of action by which radiating and unchanging magnetic fields still has to be elucidated. The objective of this study was to evaluate the morphology of MRC-5 fibroblasts after stimulation with static and pulsating magnetic fields. Under sterile environment, a total of 24 wells were loaded with 50,000 MRC-5 cells each and further divided into three groups. Groups land 2 were exposed to magnetic fields, static and pulsating respectively. Group 3 wells were unexposed and served as the control group. The cells were monitored at 0,24,48, and 72 hours and representative views were captured using digital analysis techniques. The live cells were screened for cellular mobility, cell distribution, and cellular morphology (size, shape, lysis, and background). After 72 hours, the supernatants and cells of all three groups were collected and MDA analysis was performed to determine possible cellular damage. Group I cells continued to grow at a reasonable rate, but there was substantial cell membrane damage (high MDA levels, p < 0.05). Group 2 cells, appeared to be very stressed under these conditions especially at the initial phase (24 hours). In conclusion, the use of pulsating magnetic stimulation can be beneficial in the healing process of soft tissues.

MORPHOMETRIC ANALYSIS Of MRC-5 FIBROBLAST LIKE CELLS EXPOSED TO INTERMITTENT UV RADIATION

Pamala Jones *, Hamed Benghuzzi, and Michelle Tucci, University of Mississippi Medical Center, Jackson, MS 39216

The goal of this study was to develop a tissue culture model with and without cellular protection (antioxidants) in an attempt to prevent or treat squamous damage as a result of free radical generation. The specific aim was to evaluate the morphological responses of MRC-5 fibroblast cells exposed to UV-radiation and vitamin E supplementation. The criteria for the morphological evaluation was as follows; cell and nucleus shapes, size, N/C ratio, nucleus pleomorphism, hydropic swelling, as well as other features or characteristics of membrane and cytoplasmic alterations. The experimental design was divided into two phases. In the first phase the cells were exposed to radiation for 30, 45, and 60-minute intervals, the morphological evaluation was assessed using Image Pro Digital Analysis technique. In the second phase, vitamin E was administered to the cells before and after exposure to 45 min of radiation. Data obtained demonstrated that, 45 minutes of radiation exposure caused traumatic stress to the cells, sw elling, cellular debris and fragmentation. However, treatment with vitamin E pre and post exposure provided protection for the cells. Vitamin E treatment resulted in less cellular aggregation, more abundant eosinophilic cytoplasm and an overall healthy appearance. The data suggest that the use of vitamin E may minimize skin damage by protecting the integrity of the fibroblast cells for up to 72 hours in culture.

MORPHOMETRIC EVALUATION OF OVARIAN TISSUE EXPOSED TO PCB CONVENTIONALLY AND IN A SUSTAINED MANNER

Alisa Nixon *, Hamed Benghuzzi, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

Most natural estrogens are short-lived, do not accumulate in tissue and are easily broken down in the liver. In contrast to natural estrogens, estrogenic drugs such as ethynylestradiol, diethylstilbestrol, synthetic environmental estrogens such as beta-hexachlorocyclohexane, polychlorinated biphenyls (PCBs), and phytoestrogens, are more stable and remain in the body longer than natural estrogens. Because most of these compounds are lipophilic, they tend to accumulate within the fat and tissue of animals and humans. Thus, depending on the natural estrogen levels, environmental estrogens may have different influences (mimicking, or blocking estrogen's effects) on estrogen activities. A total of 14 adult female rats were divided randomly into three groups. Animals in group I (n = 4) served as control. Animals in group II (n = 5) were injected with PCBs, and animals in group III (n = 5) were implanted with TCPL capsules containing PCBs. Pap smears were obtained daily, and at the end of the experimental phase the animals were sacrificed and vital organs as well as reproductive organs were obtained and wet weights were recorded. Significant reductions in ovarian wet weights were found in all animals treated with either sustained release of PCBs or injection of PCBs. Ovarian tissue was further analyzed histologically to determine the effects of PCBs. Histomorphometric data revealed significant reduction (p <0.05) in the total ovarian area of animals treated with PCBs. Measurements of cross-sectional lengths confirmed the reduction seen in the area. Overall, the data suggest that PCB pollutants have produced detrimental effects on endocrine function as well as fertility regardless of the route of administration.

POOR PHYSICAL ACTIVITY IN RENAL TRANSPLANT RECIPIENTS: IS THERE A LINK TO CARDIOVASCULAR DISEASE?

Tori S. Hill * and Sylvia Rosas, Alcorn State University, Alcorn State, MS 39096, and University of Pennsylvania, Philadelphia, PA 19104

In the end stage renal disease (ESRD) population, cardiovascular disease (CVD) tends to occur at an increased rate compared to the general population. Physical inactivity is a common cause of muscle atrophy and may be related to muscle abnormalities and reduced functional state of the dialysis population. Information about physical activity levels could be important for identifying patients in need of physical therapy or other interventions designed to increase mobility. This multi-center prospective cohort study is concerned with the level of physical activity in RTR prior to, six months, and one year after transplantation. Physical activity levels are assessed using Physical Activity Survey for the Elderly (PASE), a brief, valid, self-report of physical activity during a one-week recall period. The control group for this study consists of "healthy" (free from renal disease and other chronic illnesses) individuals whose level of physical activity is measured against the activity of RTR. We hypothesize that t he physical activity in renal transplant recipients is lower than that of their healthy counterparts. The control group did indeed have increased physical activity compared to RTR. Physical activity promotes good health and helps to prevent or fight against many chronic illnesses such as CVD. Physical activity pro grams should be encouraged in every population, and also as a part of post transplant care.

SINGLE CASE REPORT OF THE EFFECTS OF REFLEXOLOGY ON BLOOD PRESSURE, HEART RATE, CORTISOL, AND IMMUNOGLOBIN

Margaret Drake *, Min Huang, Hamed Benghuzzi, Neva Greenwald, and Renee Rosenfeld, University of Mississippi Medical Center, Jackson, MS 39216

Eastern and Western philosophies of medicine and science have caused a controversy over effectiveness and appropriateness of using complementary and alternative therapies with Western treatments. More research has been done on reflexology in Asian countries. This case-based study of reflexology on a normal healthy adult involved a series of measures of the subject's blood pressure, heart rate, and collection of two saliva samples each workday afternoon at 3:30 for 20 days. Biochemical markers such as total protein, IgG, and cortisol levels were measured following standard lab protocols. On days two, four, seven, nine, twelve, fourteen, seventeen, and nineteen, a trained reflexologist administered approximately 40 minutes of foot massage before the measurements were taken. The results of this study demonstrated that blood pressure, heart rate, stress level proteins, and immunology are affected by reflexology. There was no significant difference in the cortisol levels during and after the treatment period.

THE CONSTRUCTION OF A CHANNEL CATFISH INSULIN-LIKE GROWTH FACTOR HORMONE, IGF-1

Clorissa Davis (1) *, Abigail S. Newsome (1), Joseph M. Wahome (1), and Geoffrey Waldbieser (2), (1) MississippiValley State University, Itta Bena, MS 38941, and (2) United States Department of Agricultural, Agricultural Research Service, Stoneville, MS 38776

Channel catfish production, which has excellent growth compared to other catfish currently being used by producers and is recommended for food production, has become an emerging commercial industry in the Mississippi Delta. Commercial producer are seeking various ways to increase growth production and yield. This is done primarily through studies of growth performance, carcass composition, and fillet yield. Of particular interest are serum hormone levels such as the insulin-like growth hormone IGF-1. Isolation and/or the construction of a DNA probe of the gene can provide an important tool which can be used to improve techniques for the selection of superior growing lines of channel catfish for the aquaculture industry. Using the polymerase chain reaction and manipulations of the process, we created a small construct of an insulin-like growth factor hormone of channel catfish, IGF-1. This construct was designed using cDNA made from mRNA extracted from channel catfish's liver and a bacterial artificial chromos ome library of channel catfish. The construct was prepared using primers which correlated to the insulin-like growth factor hormone sequence which had previously been reported in literature and several created in house. The created construct consisted of the mature protein cloned into a TOPO TA vector. Implications of these findings will be discussed.

THE DESIGN AND EVALUATION OF A PROGRAM TO ASSESS THE ACCURACY OF CYTOTECHNOLOGISTS IN TRAINING IN THE DIAGNOSIS OF CERVICAL/VAGINAL SMEARS

Zelma Cason * and T.G. Nick, University of Mississippi Medical Center, Jackson, MS 39216

The purpose of this study was to design a program to improve the competency of cytotechnology students in assessing cervicovaginal smears using kappa statistic to evaluate inter-rater agreement among cytotechnology students. Kappa statistic used as a measurement of agreement, training programs can employ a more accurate measure of assessing student performance. Study Design: Cervicovaginal smears (n = 3,075) were randomized for evaluation by eight cytotechnology students (Fall, 2000). The students were instructed to evaluate and apply standard cytologic criteria. Following evaluation, students were instructed to assign one of the five-targeted diagnoses according to the Bethesda System classification. In addition, students determined smear adequacy and the presence of pathologic organisms. The final results were analyzed using SPSS and Stata statistical software. Results: The kappa, false positive (FP), and false negative (FN) rates for the students' evaluations were generated, as well as smear adequacy and the detection of pathologic organisms. The kappa for the first grouped session was 0.07 [+ or -] 0.08 to 0.49 [+ or -] 0.31 (class average 0.23 [+ or -] 0.08) and for the last grouped session was 0.43 [+ or -] 0.22 to 0.69 [+ or -] 0.18 (class average 0.57 [+ or -] 0.07. The average weighted kappa for the grouped sessions were 0.25 [+ or -] 0.07 to 0.56 [+ or -] 0.05. As expected, the values are lower than the unweighted kappa due consideration of the closeness to the targeted diagnosis. Conclusions: The results can assist in selecting appropriate corrective action when student's competency falls below acceptable levels. Consequently, these observations can improve a cytotechnology student's competency, leading to improved health outcomes for patients and improved cost effectiveness in healthcare organizations.

* Graduate Student in CHS 

THE EFFECT OF EXERCISE ON THE RELAXATION RESPONSE

William C. Mahone * and Takesia Turner, Mississippi Valley State University, Itta Bena, MS 38941

It is now widely accepted that exercise is a valid way to alleviate many of the effects of environmental and emotional stress. Various studies have shown dissipation of stress chemicals after exercise. We also know that exercise stimulates growth and repair processes by stimulating the release of certain chemicals. It is also known that exercise causes the release of brain chemicals called endorphins that tend to make one relax. In this research effort we attempt to document a linear connection between simple exercises and relaxation markers like the galvanic skin response GSR. The results of our research in this area will be presented. We have preliminary data indicating that people who exercise regularly can relax more easily than someone who leads a purely sedentary life style. In other these tests we tested subjects prior to exercise then had them do mild exercise then re-tested them immediately following the exercise. These tests indicate that mild to moderate exercise can have an immediate effect on the relaxation response.

THE EFFECT OF VARIOUS CORTISOL CONCENTRATIONS ON THE MITOTIC ABILITY OF MRC-5 CELL LINE

Nourelhoda I. Farah (1) *, Hamed Benghuzzi (2), Michelle Tucci (2), and Ibrahim 0. Farah (1), (1) Jackson State University, Jackson, MS 39217, and (2) University of Mississippi Medical Center, Jackson, MS 39216

Fibroblast cells are an essential component of the acute inflammatory process and ultimately wound healing. Fibroblasts in normal metabolic conditions constitute a simple cellular model for the study of anti-inflammatory drugs in culture, and for this reason this model was selected to investigate the effects of cortisol dosage on an established fibrolast cell line. MRC-5 cells were obtained in culture tubes plated with 50,000 cells per tube. Tubes were divided into four groups and each group was further divided into 5 tubes per time phase (24, 48, and 72 hours). Group 1 cells were treated with 10 [mu]L of vehicle, cells in group II-IV were treated with 1, 10, and 50 [mu]g cortisol; respectively. At the end of 24, 48, and 72 hours, live cellular images were captured before the cells were harvested for cell number and cell metabolic profile determinations (protein determinations and markers of cellular damage). An immediate depression of the rate of cell proliferation occurred upon addition of glucocorticoster oids to cultures of fibroblasts in the early growth stages. High dose administration caused the most significant level of cellular suppression. Surviving cells at 72 hours began to show signs of cellular proliferation and recovery. In conclusion, cortisol suppresses the cellular proliferation of MRC-5 fibroblast cells after 24 hours and the cellular recovery seen at 72 hours is most likely caused by the metabolism of the cortisol into an inactive metabolite.

THE EFFICACY OF TCPL AS E PLUS P REPLACEMENT THERAPY MODEL USING OVARIECTOMIZED RATS AS A MODEL

Zelma Cason (1) *, Hamed Benghuzzi (1), Michelle Tucci (1), and Barry England (2), (1) University of Mississippi Medical Center, Jackson, MS 39216, and (2) University of Michigan Medical Center, Ann Arbor, MI 48109

Tricalcium phosphate lysine (TCPL) delivery system was used to deliver various organic compounds at sustained levels in many different models. The specific aim of this investigation was to utilize TCPL delivery system as a model for estrogen (E) plus progesterone (P) replacement therapy in post-ovariectomized adult rats mimicking a postmenopausal condition. A total of 13 adult female rats were used in this study. The animals were randomly divided into four different groups: groups 1,2, and 3 were overiectomized (OVX), OVX plus E (20 mg loaded TCPL), and OVX plus E (TCPL, 20 mg) plus P (TCPL, 60 mg) treatment. Group 4 animals (n = 4) served as intact control group. Blood samples were collected biweekly for 33 days. Vaginal smears were taken and screened daily during the entire investigation. The total serum levels of E, P, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were measured by means of radioimmunoassay procedure. Data obtained from this investigation suggest the following: (I) OVX re sulted in an increase in total serum levels of LH and FSH within 2 days post-ovariectomy, (II) TCPL were capable of releasing sustained levels of E (10-40 pg/ml) and P (2.30-3.75 ng/ml) at the end of second day and continued until the 33rd day, (III) the sustained levels of E plus P were able to suppress the post ovariectomy rise of LH and ESH to almost undetectable levels, (IV) sustained delivery of E resulted in maturation of vaginal epithelium and the smears exhibited the estrus phase throughout the investigational period (V) E + P treatment induced no estrus and the epithelial changes resembled the OVX group. * Graduate Student in CHS

THE PATHOPHYSIOLOGY OF DIOSGENIN REPLACEMENT THERAPY

Rebecca Eckie *, Michelle Tucci, Hamed Benghuzzi, and James H. Hughes, University of Mississippi Medical Center, Jackson, MS 39216

Diosgenin is asteroidal saponin, which is extracted from the root of wild yam (Dioscorea villosa). It has been reported to have tremendous medical applications and as an herbal extract appears to be free of any major adverse effects. Recently, wild yam has been used to alleviate postmenopausal symptoms and is sold unregulated over the counter at a variety of pharmacy and health food stores. The specific objectives of this study were to continuously deliver diosgenin to adult ovariectomized female rats for 45 days and follow changes in body weight, organ weight and histopathological changes in the adrenal gland. Adult female Sprague Dawley rats were divided into three equal groups. Rats in group I served as the control group, animals in group II were ovariectomized and animals in group III were ovariectomized and supplemented with tricalcium phosphate (TCP) drug delivery system loaded with 500 mg diosgenin. Serum analysis of animals supplemented with diosgenin showed levels ranging between 3.98-5.31 nglml dios genin. The results indicated that ovariectomized animals had a significant increase in body weight and spleen weights. Slight increases in wet adrenal weights were observed in the ovariectomized group compared to the control animals. Histopathological evaluation of the adrenal gland revealed an increase in the cortical and medullary adrenal areas of the ovariectomized group and a significant decrease in area in the diosgenin treated animals (p < 0.001). The information is important because reduction in adrenal mass poses a potential risk for major endocrinological complications.

THE ROLE OF BLACK SEED IN THE PROLIFERATION AND BIOCHEMICAL MARKER LEVELS OF HEP-2 CELLS

Jeffrey T. Hansen *, Hamed Benghuzzi, Michelle Tucci, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

For centuries, people in the Middle East and Southeast Asia have used Nigella sativa, also known as black seed (BS), for its homeopathic effects. The objective of this study was to investigate the role BS might have on the metabolic biomarkers of the Hep-2 cell line. The experimental design entailed six groups of five wells each (50,000 cells). Groups II through VI were treated with BS, lipopolysaccharide (LPS), cortisol, LPS + cortisol, and BS + LPS + cortisol, respectively. Group I was the untreated control group. At the end of 24, 48, and 72 hours, the total cell count, protein and MDA levels were measured by following standard lab protocols. Data collected from this study revealed that Hep-2 cells exposed to LPS and cortisol (group V) resulted in a decrease in cell proliferation compared to the control. BS treatment induced a higher proliferation rate than group V. Similar trends were observed in the metabolic behavior of Hep-2 cells as evidenced by the total protein and MDA levels. The exposure of BS sho wed a shift in the metabolic pathways. In conclusion, this study showed that exposure to LPS resulted in an alteration in the metabolic function and this phenomenon was further escalated under stressful conditions (increased cortisol exposure). In addition, the use of BS reversed the traumatic condition.

USE OF THE PAP STAIN VERSUS THE DQ STAIN ON THE DISTINCTION OF CELLULAR PATTERNS UPON EXPOSURE TO ESTROGEN

Leernesto R. Vassar *, Hamed Benghuzzi, Michelle Tucci, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

The literature is lacking on the exact distribution of cornified cells during the estrus cycle upon exposure to E in adult fertile female rats. The specific objectives of this study were to investigate the effects of E on the cyclic activity of adult female rats, and to compare the use of the Papanicolaou (PAP) stain versus the Diff Quik (DQ) stain on the distinction of cellular behavior upon exposure to E for three days. Six different laboratory rats (three control and three experimental) were labeled from l-6 (R1-R6). Cyclic activity (2, 4, 8, 12, 24, 36, 48, 72 hours) was assessed by using 0.5 ml Hank's solution unloaded in the vaginal canal for few seconds followed by gentile pipet suction. This mixture was then smeared onto microscopic slides and stained using PAP and DQ stains. Once the slides were screened, it was found that the PAP stain proved to be a better staining technique than the DQ stain in both nuclear and cytoplasmic details. In the second stage of this experiment, the rats labeled R1-R3 wer e used as controls, whereas lab rats R4-R6 had estrogen administered (2 mg/ml) to them for three days. The results showed a significant increase in the proliferation of degenerative cells in the E treated rats compared to control animals.

VASCULARIZED FIBULAR GRAFT

Audrey Tsao *, William Lineaweaver, Sheila Lindley, Vipul Sud, Kye Higdon, Javad Tavassoli, Ruth Ann Buckhalter, and Charity Peacock, University of Mississippi Medical Center, Jackson, MS 39216

This study is to investigate the success of Vascular Fibular Grafts (VFG), a standard care option for the treatment of Avascular Necrosis (AVN). Methods: Patients with a minimum age of 18, with AVN in Stage II or above based on Steinberg/UPenn system are considered for this procedure. Patients are offered a variety of surgical options depending upon the progression of their disease and what best will suit their lifestyle. The VFG utilizes a piece of fibular bone, including the attached blood vessels, inserted into a core decompression of the femoral head. The harvested vessels are then anastomosed to the hip area vessels, hence improving blood flow to the femoral head. Clinical outcomes are measured by Harris Hip Scores (HHS), SF36, and/or radiographic evaluations pre- and post-operatively at six weeks, three, six, and twelve months and annually thereafter. Results: The University of Mississippi Medical Center has presently enrolled eleven patients with twelve procedures. At present, there has been no progres sion of disease, increased HHS, increased quality of life, and a decrease in pain. Discussion: The improvement in HHS correlates well with radiographic vascularization of the femoral head and patients returning to normal activities of daily living (ADL). We hope to prove this procedure will decrease the need for future total hip replacements.

DELTA OPIOID AGONIST IMPROVES SURVIVAL FOLLOWING HEMORRHAGE IN RATS

R. Sumrall *, J. Penton, A. Smitherman, and D. Hildebrandt, University of Mississippi Medical Center, Jackson, MS 39216

Mortality from hemorrhage is due to ischemic end-organ damage. Therapy is aimed at increasing systemic oxygen delivery; few studies have addressed reducing demand. This study was designed to determine if the deltaopioid agonist DADLE (D-ALA2-LEU5-enkephalin), which reduces metabolism and increases survival of ischemic tissues, increases survival in a rat shock model. Experiments were done in conscious rats (300-350 g) with catheters in a femoral artery for blood removal and measuring arterial pressure (AP) and heart rate (HR), and in a femoral vein for drug infusion. Following control measurements, 3.25 ml/100 g body weight of blood was removed over 20 minutes, followed by i.v. DADLE (1 mg/kg), saline or norepinephrine (7 [micro]g/kg; NOR) in a 200 [micro]l bolus. None of the saline rats (n=3), but 4 of 5 NOR and 6 of 7 DADLE rats lived 180 minutes post-hemorrhage. Mean AP before (124[+ or -]4 mmHg NOR; 121[+ or -]3 DADLE) and following hemorrhage (94[+ or -]4 mmHg NOR; 94[+ or -]5 DADLE) were similar in the surviving groups, as were HR before (277[+ or -]9 bpm NOR; 261[+ or -]4 DADLE) and after (260[+ or -]19 bpm NOR; 247[+ or -]10 DADLE). Plasma lactate, the most sensitive measure of tissue ischemia, increased in NOR (1.1[+ or -]0.3 to 4.5[+ or -]2.7 mmol/L) but not in DADLE (0.8[+ or -]0.2 to 1.0[+ or -]0.3). Thus, DADLE increased survival similar to NOR, a common treatment, but without the metabolic cost of increased lactate, which should improve long-term survival. Therefore, DADLE could be a clinical treatment for hemorrhagic shock.

INCREASED SKIN TRANSPLANT SURVIVAL WITH DELTA OPIOLD AGONIST

J. Penton *, A. Smitherman, R. Sumrall, D. Hildebrandt, University of Mississippi Medical Center, Jackson, MS 39216

A major cause of failure of transplanted or transposed tissue is ischemia from inadequate blood supply. Most treatments focus on increasing blood supply, little has been done on decreasing demand. This study was designed to determine if the delta-opioid agonist DADLE (D-ALA2-LEU5-enkephalin), which reduces metabolism of tissues, increases survival of muscle-skin flaps. Pedicled rectus abdominus muscle-skin (TRAM) flaps were created in rats. All rats received 1 ml total of saline, with or without DADLE (1 mg/kg flap weight), injected into the underside of the flap. Rats were examined daily and the extent of necrosis estimated to the nearest quartile for each of 4 zones (Z). None of the DADLE rats (n = 9), but 4 of the control (CON; n = 9) rats developed Z4 necrosis (Z4N) on day 1, one of these also developed Z3N. By day 3 all CON had necrosis, greater than 25% in Z4 in 6 animals; 4 also with Z3N. Only 7 DADLE rats had necrosis, 1 greater than 25% Z4N, only 2 of these also Z3N. By day 6 all rats in each group h ad Z4N, 7 greater than 50% Z4N in CON, and these 7 also Z3N. Only 5 DADLE had Z3N, and only 3 greater than 50% Z4N. Even a single treatment with DADLE improved TRAM flap survival compared with saline, noticeable even out to 6 days post-surgery. Thus, DADLE may prove to be a valuable treatment option for increasing survival of pedicled flaps for reconstructive surgery.

THURSDAY AFTERNOON

Lake View II

2:00 RENAL FUNCTION CURVE IN NORMOTENSIVE AND DOCA-SALT HYPERTENSIVE RATS DURING CHRONIC MINOXIDIL TREATMENT

Min Huang (*) and Robert D. Hester, University of Mississippi Medical Center, Jackson, MS 39216

This study tested the hypothesis that the chronic-salt loading pressure-natriuresis curve determines the level of arterial blood pressure in both normotensive and DOCA-salt hypertensive rats during chronic minoxidil treatment. The rats were divided into 8 groups: control, minoxidil, salt, salt-minoxidil, DOCA , DOCA-minoxidil, DOCA-salt and DOCA-salt minoxidil. DOCA-pellets (75 mg) were implanted subcutaneously in the DOCA animals. Minoxidil, 3 mg/day, was given orally to the rats in the minoxidil groups. The rats in the salt groups drank saline while the rats in the non-salt groups drank tap water. Sodium intake was measured every 24 hours. Mean arterial blood pressure was measured at the end of 6 weeks. There were no significant differences in salt intake among any of the non-salt groups. Minoxidil treatment did not significant change salt intake in any of the tap water or saline animals. Arterial pressures were 119[+ or -]4 mmHg (control), 117[+ or -]4 mmHg (minoxidil), 111[+ or -]3 mmHg (salt), 111[+ or - ]3 mmHg (salt minoxidil), 139[+ or -]8 (DOCA), 133[+ or -]4 (DOCA minoxidil), 160[+ or -]5 (DOCA-salt), 146[+ or -]9 (DOCA-salt minoxidil). There was a significant effect of DOCA and an interaction between DOCA and salt treatment to result in a significant increase in MAP. However, neither saline nor minoxidil treatment alone had a significant effect on MAP. Chronic minoxidil treatment did not shift the chronic salt-loading pressure-natriuresis curve in either normotensive or hypertensive rats. These results indicate that chronic salt-loading pressure natriuresis curve plays a central role in long term control of arterial blood pressure and the development of DOCA-salt hypertension.

2:15 COMPARISON OF TEN TUMOR ANTIGENS FOR THE SERODIAGNOSIS OF PANCREATIC CANCER

Margot Hall (1) * Sabrina Bryant (1), Margaret Jackson (1), James T.Johnson (1), Harold Schultze (1), Shawn Clinton (1), Kevin Beason (1), Cynthia Wilson (2), Debbie Fortenberry (1), Cynthia Bright (1), Helen Hua (1), Jiarong Ying (1), Paul Sykes (1), Kay Hollifield (3), and Chariton Vincent (3), (1) University of Southern Mississippi, Hattiesburg, MS 39406; (2) University Medical Center, Jackson, MS 39216; and (3) Laurel (3) Clinic for Women, Laurel, MS 39442

Pancreatic cancer (CA) is a serious health problem. Due to its non-specific early symptoms, pancreatic cancer is frequently not diagnosed until late stage disease when the prognosis is poor. A noninvasive early detection method is sought. In this study, sera from 554 patients (16 pancreatic CA, 128 other GI CA, 216 other CA, and 195 non-CA) were assayed for the presence of tumor antigens and the results correlated with diagnoses established pathologically. Immunoassay test kits from Hybritech (CEA, CA195), Centocor/Fujirebio Diagnostics (CA125, CA19-9, CA72-4, CA15-3, CA27.29, Cyfra2l-1), CIS Bio International (CA50), and Abbott (AFP) were used to test for the concentration of these antigens. Using the manufacturer's decision values the following diagnostic sensitivities were obtained for pancreatic cancer: CA195 100.0%, CA19-9 66.7%, CA5066.7%, CA 12540.0%, CA27.2940.0%, CEA 37.5%, CA72-4 31.3%, Cyfra2l-1 26.7%, CA15-3 26.7%, and AFP 18.2%. Diagnostic specificities were above 75%. Nine/sixteen pancreatic CA patients had a CA195 concentration> 50x the upper limit of normal (ULN). Two patients had CA 195 concentrations> 1000x ULN prior to their diagnosis by conventional methods (imaging and biopsy). We conclude that CA195, CA19-9, and CA50 were especially useful in the diagnosis of pancreatic CA.

2:30 THE EFFECTS OF METABOLESS AND OXIDATIVE STRESS ON THE SURVIVAL OF MCF-7 BREAST CANCER CELLS

Ibrahim O. Farah *, Rowshan Begum, and Joseph A. Cameron, Jackson State University, Jackson, MS 39217

Metaboless is available as over the counter nutritional supplement. It contains many antioxidants and is used for weight control. There have been no studies done with regards to its protective/adverse effects on breast cancer. Our objective therefore was to expose MCR-7 breast cancer cells to its extracts (alcohol and water) and monitor their survival under various concentrations and also in combination with 11202 as an oxidative stressor. Using standard cell culture techniques, exposure protocols in 96 well plates and an Accent fluoroscan, percent survival was computed from the data comprising three replicates of three different experiments. Contrary to our expectations, the results showed that the alcohol extract alone or in combination with 11202 caused adverse effects to these cells. The effective concentrations ranged from 1.95 to 1000 ppm. At the range of 500-1000 ppm, alcohol extract and its combination with [H.sub.2][O.sub.2] were able to completely inactivate these cells. Water extract of this produc t was not found to be as effective in inactivating MCF-7 cells even at the highest concentration of 1000 ppm. The combination of both extracts and 11202 was found to be as effective as [H.sub.2][O.sub.2] alone in inactivating these cells defying the protective theory of antioxidants with regards to breast cancer (no antagonism or synergism). In conclusion, Metaboless as a diet pill showed adverse effects on the survival of MCF-7 cells. Metaboless extracts applied alone or in combination with oxidative stress showed a potential preventive role in breast cancer, a finding that warrants further studies.

2:45 USING GEOGRAPHIC INFORMATION SYSTEMS (GIS) TO CORRELATE LOCATION OF MAINSTREAM MEDICAL PRACTITIONERS WITH COMPLEMENTARY AND ALTERNATIVE THERAPY PROVIDERS

Margaret Drake * and Fazlay S. Faruque, University of Mississippi Medical Center, Jackson, MS 39216

As complementary and alternative medicine (CAM) providers increase in number and acceptability to the mainstream American healthcare users, it is important to be able to compare the practice locations of CAM practitioners and traditional physicians. This is valuable for healthcare planning and economics, as well as for those attempting to pinpoint areas of service deficit. A nationwide survey reports a 40% increase of CAM customers between 1991 and 1997 without any increased acceptability of CAMs among the physician group. In an attempt to fill the void of studies conducted on CAM practitioners in Mississippi, this pilot was taken from an insurance company database. This company insures over 100,000 workers in Mississippi. The study mapped both alternative providers and mainstream medical practitioners. Identified were areas with a high concentration of CAM and traditional practitioners. The demography of those areas was analyzed for correlation with practitioner locations. Most patients do not inform their p hysicians about using CAMs; in addition, most physicians do not ask patients about CAM use. This study will help the physicians to be aware of CAM practitioners in the vicinity of their practice locations and the probability of their patients being CAM users.

3:00 SPATIAL ANALYSIS OF TOXIC EXPOSURE SURVEILLANCE SYSTEM DATA OF PCC, UMMC, MS

Fazlay S. Faruque *, Harpal Singh, Michael P. Hughes, and Frederick B. Carlton, Jr., University of Mississippi Medical Center, Jackson, MS 39216

This study analyzes the spatial distribution of the callers to the state's Poison Control Center (PCC) using Geographic Information Systems (GIS). Inappropriate use of chemical substances can have ill effects, which may range from mild to the most severe outcomes, even leading to death depending on the amount and type. Since 1978, the PCC at the University of Medical Center has been rendering prompt advice to the public and health care providers on matters of exposure to toxic substances, thereby helping in reducing not only morbidity and mortality but also in saving considerable time and money, which otherwise would have incurred in providing ambulance services and emergency room visits. Based on the geographic distribution and pattern of calls from different locations in the state, a community-based comprehensive plan could be undertaken for proper resource allocation to reduce potential dangers. This study analyzes one full year of data to identify geographic areas that are statistically significant in mak ing calls related to toxic exposures and also generates hypotheses to explain the significant differences in number of calls.

3:15 STUDENTS' PERCEPTIONS AND USAGE PATTERNS OF DIETARY SUPPLEMENTS

Emily S. Dix *, James K. Glisson, Jay Pitcock, Rob Rockhold, and Warren May, University of Mississippi Medical Center, Jackson, MS 39216

Medical, nursing, and pharmacy schools across the country have begun to integrate courses or lectures on alternative therapies including herbals and dietary supplements. However, a survey of student perceptions regarding these supplements is yet to be published. This study examined baseline usage patterns, attitudes, and knowledge levels of dietary supplements and herbal products among first year medical and pharmacy students at the University of Mississippi; this same cohort will be studied longitudinally through graduation (August 2003, May 2003, April 2005) to monitor changes in usage patterns and/or knowledge levels throughout their course of study. Baseline assessments were conducted in September of 2002 using self-administered questionnaires in classroom settings. Ninety-six medical (58.5%) and 68 pharmacy (41.5%) students completed the survey. Fifty-four percent of medical students reported the use of dietary supplements either daily or at least once a week, compared to 47% of pharmacy students. When a sked if "Dietary supplements can be unsafe during pregnancy," nearly 20% all students responded that they "don't know." Approximately 80% of medical students agreed that "It is possible for dietary supplements to contain plants/chemicals that are not intended to be in them," compared to 69% of pharmacy students. These preliminary findings suggest that pharmacy and medical students should receive detailed lectures on appropriate uses of dietary supplements as well as their regulation and approval processes.

3:45 Divisional Business Meeting