Health Sciences.

Chair: D. Michelle Tucci, University of Mississippi Medical Center 

Vice-chair: Audrey Tsao, University of Mississippi Medical Center

THURSDAY MORNING

Auditorium

Clinical Diagnostics I

8:30 GENETIC MARKERS FOR EVALUATION OF SIC PATIENTS AT RISK OF SEPTIC COMPLICATIONS

James Hamilton*, Lee Y. Tee, Lynn J. Calcote, Marjolyn Brock, Christine Toves, Ginger Hogg, Roger Blake, Robert Schmieg, Gregory Timberlake, and D. Olga McDaniel, University of MS Medical Center, Jackson, MS 39216

Background: Severe trauma injury often leads to the development of sepsis and organ failure. A challenge for appropriate treatment of sepsis is identification of the patients who are at increased risk for sepsis. Clinical findings support the fact that despite comparable risk factors, post trauma sepsis and organ failure develops in some patients but not others. Hypothesis: In clinical settings, interindividual genetic differences associated with host immune response appear to be a major contributing factor to the development of trauma induced infection and subsequent organ failure in surgical patients. Methods: All blunt and penetrating trauma, with injury severity scores of > 15 were included in this study. Thirty eight patients (17 African American and 21 Caucasian patients) were studied. Peripheral blood mononuclear cells (PBMCs) were used for genotype analysis. Genotypes for cytokines including IL-6, TNF-[alpha], IL-10, IL-18, IFN-a and Toll-like receptor genes, such as TLR-2 and TLR-4 were determined. Results: A majority of patients have developed sepsis. In terms of cytokine genotypes, the high producer genotype of IL-6 was found in 90.5% of patients who developed sepsis (p < 0.05). No correlation was observed among Toll-like receptor mutations and the development of sepsis. In conclusion, a high number of patients that met the criteria for the study (ISS > 15) developed sepsis. While the IL-6 high producer genotype shows a statistically significant correlation to sepsis, other factors including Toll-like receptors are important in the development of sepsis. A larger sample size is needed to determine the significance of the Toll-like receptors in sepsis.

8:45 MHC CLASS II-RESTRICTED PROCESSING AND PRESENTATION OF A CYTOPLASMIC ANTIGEN

Jeremy Lott (1,2*), Joseph A. Cameron (2), and Janice S. Blum (1), (1) Indiana University School of Medicine, Indianapolis, IN 46202 and (2) Jackson State University, Jackson, MS 39217

The Major Histocompatability Complex (MHC) encodes genes necessary for host defense. These genes code for many molecules involved in protecting the body from foreign pathogens and antigens. MHC class II proteins typically present peptides derived from extracellular antigens, which have been internalized by antigen presenting cells (APC). Upon uptake into an APC, these antigens are processed within endosomes and lysosomes by acidic proteases called cathepsins. Studies by our lab and others have shown that cytoplasmic antigens normally presented by MHC class I, may also be presented by class II. In this study, the processing and presentation of SMA, a mutant form of the IG Kappa light chain variable domain was monitored. Studies have shown that endogenous SMA gives rise to peptides such as kappa II (residues 145-159), which are then presented by the class II proteins to T-cells. Here, investigations determined that cytoplasmic SMA is presented via an endogenous pathway for display in the context of class II. Thus, release of SMA of its antigenic peptides were not detected with SMA transfected APC. Current studies also suggest a role for specific proteases in processing SMA for class II presentation. Investigations are ongoing in terms of defining specific steps in cytoplasmic antigen presentation as this pathway has relevance to viral, tumor and auto-immunity. Supported in part by: NIH R25 GM067592-02

9:00 MODULATION OF PHASE 1 AND BRCA PROTEIN EXPRESSION IN RESPONSE TO TREATMENT WITH ALCOHOL AND VERNONIA AMYGDALINA EXTRACT

Alden Hopkins (1,2*), Jacqueline Samuel (1), Joseph A. Cameron (1), and Carolyn B. Howard (1), (1) Jackson State University, Jackson, MS 39762 and (2) Hinds Community College, Jackson, MS 39217

Useful anti-cancer agents affect Phase 2 metabolic enzymes without affecting Phase 1 enzymes. Phase 1 enzymes are relevant in cancer studies in that they are involved in chemical reactions, whereas phase 2 enzymes are involved in clearance. Previous studies revealed that treatment of MCF-7 breast cancer cells with low concentration of ethanol alone induced cell growth, whereas very high concentrations of ethanol were inhibitory. Studies have shown that Vernonia amygdalina (V.A.) extract induces Phase 2 enzymes, without affecting Phase 1 enzymes, providing evidence supporting the chemotherapeutic potential of V.A. We hypothesize that treatment with a combination of V.A. extract and ethanol will change responses which take place following treatment with ethanol alone. Twenty-four female mice were exposed to ethanol in the drinking water for sixteen weeks. Eight were continued for four additional weeks of ethanol exposure, eight others received V.A. plus ethanol for the four additional weeks, and the other eight, which served as the control group received drinking water alone. Liver and mammary tissues samples were removed from animals following treatment and immunohistochemistry was performed. Results indicated V.A. treatment affected ethanol-induced responses by leading to a decrease in apparent mammary tumors. Studies are being conducted to further characterize these responses. This work is supported in part by GM50117.

9:15 PROTECTION AGAINST PNEUMOCOCCUS BY PspA

Quincy Moore* and Larry S. McDaniel, University of Mississippi Medical Center, Jackson, MS 39216

Pneumococcal infections are prevalent in children under the age of 5 and the elderly (> 65 years of age). Prevention of diseases that result from pneumococcal infection is of great medical importance. Our work focuses on the use of the pneumococcal surface protein A (PspA) as a vaccine candidate. The DNA fragment encoding the alpha-helical domain of PspA/EF5668 was cloned into an eukaryotic expression vector designated pJB100EF. We examined the effect of priming with pJB100EF and boosting with purified recombinant protein. We also tested the hypothesis that the genetic background of the pneumococcus affects the ability of anti-PspA antibodies to protect in a systemic model. We used a genetically modified variant of WU2 that expresses PspA from EF5668, WU2, and EF5668 as challenge strains. Anti-PspA levels were enhanced when mice were primed with DNA and boosted with protein. Western blot analysis demonstrates that the immune serum was cross-reactive with PspA from several different pneumococcal isolates representing different PspA clades. Survival of immunized mice following pneumococcal challenge demonstrated the ability of prime-boost immunizations with the alpha-helical domain of PspA to elicit protective immunity.

9:30 Break

Environmental Influences on Health I

9:45 HARMFUL HEALTH EFFECTS OF HIGH DOSE IONIZING RADIATION

Amin Haque, Alcorn State University, Alcorn State, MS 39096

The most common forms of ionizing radiation are alpha and beta particles and photons of gamma and x-rays. Effect depends on the type, dose, and the dose rate of the incident radiation and the composition of the human tissue. The mechanism which causes health effect is the direct ionization of the atoms and molecules of the cells, while indirect effects are mainly produced by highly reactive free radicals [H.sub.2][O.sup.+], [H.sup.+], and H[O.sup.-] created by the radiation when it interacts with water molecules. This often leads to damage to the membrane, nucleus, chromosomes, or mitochondria of the cell that either inhibits cell division, results in cell death, or produces a malignant cell. A dose of only 300 J of x-ray or [gamma]-ray radiation is fatal for the average human, even though this radiation raises the temperature of the body by only 0.001 [degrees]C. Some of the people who died at Chernobyl received a high dose, 100-250 rem, in a short period. External sources of ionizing radiation include cosmic rays from the sun and other stars, [alpha]-particles or [gamma]-rays emitted from rocks and soil. Internal sources include nuclides that enter the body when we breathe ([.sup.14]C, [.sup.85]Kr, [.sup.220]Rn, and [.sup.222]Rn) and through the food chain ([.sup.3]H, [.sup.14]C, [.sup.90]Sr, [.sup.131]I, and [.sup.137]Cs). The cells of bone marrow, the reproductive organs, the epithelium of the intestine, and the skin suffer the most damage from radiation.

10:00 THE DEVELOPMENTAL EFFECTS OF BENZO(A)PYRENE ON FUNDULUS HETEROCLITUS EMBRYOS

Joseph Wahome (1*), Kimberley Jefferson (2), and Kristine L. Willett (2), (1) Mississippi Valley State University, Itta Bena, MS 38941 and (2) University of Mississippi, University, MS 38677

This experiment was done to study growth and developmental effects of the carcinogenic polycyclic aromatic hydrocarbon benzo(a)pyrene on Fundulus heteroclitus embryos. The Medaka Embryo Larval Assay was adapted for use of studying the chemical effects on the saltwater teleost, Fundulus heteroclitus. Hatching and sterilization processes were adjusted. The affects of 1 and 10 ppb concentrations of BaP in ethanol and DMSO carrier solvents were studied along with ethanol and DMSO controls. Ethanol had adverse affects from DMSO treatments and proved to be the worse solvent. Ethanol treatments were statistically different when comparing stage developments to the other treatments. A basic morphological scoring system was used to score the embryos with craniofacial, cardiovascular, skeletal, organ, body part, pigmentation, movement disorders, and stage delay. BMS scores for 1 and 10 ppb BaP in DMSO treatments were higher than the controlled groups meaning BaP dosed fish were affected by the chemical. Also percent survival of embryos in 1 and 10 ppb BaP in DMSO treatments were lower when compare to controls. When observing abnormal swimming behavior the 1 and 10 ppb treatments, the BaP dosed fish displayed a higher percentage of abnormal swimming behavior. Ethanol was an unacceptable carrier solvent for benzo(a)pyrene. Benzo(a)pyrene does cause effects on growth and development.

10:15 HOUSEHOLD ELECTROMAGNETIC FIELDS EFFECTS ON HUMAN HEALTH

Amin Haque, Alcorn State University, Alcorn State, MS 39096

Exposure levels of electric and magnetic fields (EMFs) around the home are in the range of 0.01-0.25 [micro]T, near powerlines 0.5-1.0 [micro]T, and under the power lines, 6.0-10.0 [micro]T. The frequency and energy of radiation produced by time-varying electric and magnetic fields are very low (< 300 Hz) to damage a cell or DNA, and therefore in principle are not capable of initiating cancers. The only known interaction between the EMFs and the human body is the induction of an electric current which is proportional to the magnetic field. Laboratory studies on animals and cell cultures have shown that weak magnetic fields can have effects on several biological processes. For example, they may alter hormone and enzyme levels and the rate of movement of some chemicals through living tissue. By themselves, these changes do not appear to constitute a health hazard. Most studies have produced inconclusive results or no increased cancer incidence in laboratory animals following exposures to EMFs. The radio waves and microwaves used for communications, such as radio, television, cell phones, radar, telephone, etc, are of much higher frequency and energy, but the intensity used in these applications are low and therefore safe for the general public. The evidence for a link between EMFs and adult central nervous system cancers, male breast cancer, and leukemia is also very weak. More research to produce more reliable information is needed before any conclusions can be drawn.

10:30 ASSESSMENT OF MICROBIOLOGICAL WATER QUALITY OF PEARL RIVER AND THE GBNERR IN MISSISSIPPI

Zakiya Coleman (1,3*), Stephen S. Kishinhi (1), Ibrahim O. Farah (1), Paulinus Chigbu (1), Paul B. Tchounwou (1), and Mark Woodrey (2), (1) Jackson State University, Jackson, MS 392171; (2) Grand Bay National Estuarine Research Reserve, Moss Point, MS 39562; and (3) Hinds Community College, Raymond, MS 39211

The Grand Bay NERR and the Pearl River Basin are important water bodies in the state of Mississippi for recreation as well as being sources for providing aquatic food for the population. Health considerations with regards to these two water bodies and the diversity of their ecological systems are attracting research in the area of water quality. The purpose of this study was to evaluate levels and temporal variations of heterotrophic and indicator microorganisms in these bodies. We have established 15 sampling stations on the GBNERR and ten stations for the Pearl River. Samples were analyzed by membrane filtration and incubation on selective media, within 8 hours of collection, for total coliforms (ENDO), fecal streptococci (ENTERO), fecal coliform (FC/E. coli) and total heterotrophic (HPC) counts. Summary of the Pearl River data showed averages of 1578 for ENDO, 127 for ENTERO, 150 for FC/E. coli, and 67,335 for HPC. Corresponding results for the GBNERR were 381, 74, 118, and 27,297 respectively. Data presented here showed variability in ranges and averages within sampling stations and within sampling duration. It also indicates the presence of significant numbers of coliforms, fecal coliforms, and fecal streptococci at both water bodies. The Pearl River data also showed higher numbers for all groups as compared to the GBNERR. This is an alarming call for further research and intervention. Supported in part by NIGMS R25 GM50117.

10:45 Break

Outcomes Research

11:00 EFFECTS OF STANDARD POPULATION IN COMPUTING DISPARITY IN HEALTH STATUS OF RACIAL MINORITY IN MISSISSIPPI

Abu Khan* and Fazlay S. Faruque, Jackson State University, Jackson, MS 39217 and University of Mississippi Medical Center, Jackson, MS 39216

This study uses total mortality as an indicator of health status, and compares it between whites and non-whites of 82 counties of Mississippi. When the rates are age-adjusted, the minority non-white population experiences significantly higher mortality rate than the white population, suggesting that a disparity in minority health status exists in Mississippi. The need and rationale of converting mortality observed in a population to that of a standard population (age-adjustment), is well established. The 1940 standard population was in use for quite some time. The distribution of this standard was such that the proportion in lower age groups was more than that in higher age groups. It is recently suggested that with the increase in life expectancy, the average population has increased in age (i.e., proportion of people in higher age groups has increased), and thus, a new standard that reflects this change is required. As a result, the 2000 standard population came in use. When the age distribution of non-white and white population of each of the 82 counties of Mississippi were compared to the standards of 1940 and 2000, it was found that the distribution of non-white population resembles more to the 1940 standard suggesting that the non-white population has not substantially changed in its age distribution. If the 2000 standard is used, the computed mortality rates of non-whites are underestimated. The reflected disparity in health status markedly differs as well. This suggests that the 1940 standard population may be more reflective of the non-white population and the 2000 standard population may fit better to the white population.

11:15 FACTORS INFLUENCING EXERCISE AND BMI IN UNDERGRADUATES

Rhiannon Neff (1*), Carol Jones (2), and Reid Jones (2), (1) Southern Arkansas University, Magnolia, AR 71753 and (2) Delta State University, Cleveland, MS 38733

Recent reports from the Centers for Disease Control have cited an alarming increase in obesity among young Americans. Lack of exercise was considered a key determinant influencing obesity, as defined by elevations in body mass index (BMI). The present research examined the role of exercise patterns and personality characteristics on BMI among undergraduates. Eighty-five students at a regional state university in the Southeast volunteered to complete surveys involving demographics, exercise patterns, and Wallston's Multidimensional Health Locus of Control (HLOC). An 'internal' HLOC indicated that the person expected that their health was most likely influenced by their own personal behavior. An 'external' HLOC indicated that the person expected that their health was most likely influenced by external factors, beyond their control. Results showed that students with an internal HLOC were significantly (p < .05) more likely to exercise, and that this effect was far stronger (p < .01) among female students. Further, upperclassmen were significantly (p < .05) less likely to believe that external factors influenced their health than were Freshmen.

11:30 HEALTH INSURANCE CHARACTERISTICS OF COLLEGE STUDENTS

Jessica C. Olive, University of Southern Mississippi, Hattiesburg, MS 39406

Lack of health insurance is a persistent problem in the United States. Due to escalating costs, many people can afford health insurance only if it is offered as a benefit of employment. Even then, employers are cutting back on benefits to employees and their dependants and are requiring more cost-sharing for those benefits. The consequences of having no insurance include reduced use of preventive health services, less access to a regular source of care, inappropriate use of emergency services, and a higher probability of not receiving needed medications and treatment. The purpose of this study was to characterize the insurance status and needs of full-time undergraduate and graduate students at the University of Southern Mississippi. An online survey was developed and pre-tested using a focus group. Incentives were provided to encourage participation. A total of 79 people responded to the invitation to participate in the survey. The data was analyzed using SPSS. The majority of students that participated in the survey was found to have health insurance and had a regular place to go for medical care.

11:45 THE USE OF ORAL HISTORY METHODOLOGY FOR EDUCATIONAL AND PROFESSIONAL DEVELOPMENT

Cynthia K. Scott* and Ruth M. Burgess, University of Mississippi Medical Center, Jackson, MS 39216

This study explores the educational and professional benefits of student participation in oral history research. Subjects include five Master of Physical Therapy students who participated in a project to document the development of physical therapy in Mississippi. Using an oral history format, accounts from Mississippi pioneers in physical therapy were documented. Contributions from two deceased pioneers were documented via proxy. Four months after completion, the subjects were asked to reflect on the value of this project to their educational and professional development. This was done via an open ended questionnaire of nine questions developed by the researchers and validated by faculty members with research and educational backgrounds. Responses were anonymous and participation was not grade related. Results: Qualitative analysis revealed 3 themes resonating with subjects: (1) importance of professionalism; (2) effective teamwork; and (3) importance of understanding professional history. Subjects reported a new appreciation of those who "paved a road" allowing us to practice as we do today. This was an excellent method to actively teach professionalism, teamwork, and respect for the profession of physical therapy. Gaining insight into these issues may strengthen long term professional development. Professionalism, teamwork and an appreciation of professional contributions are essential qualities which allow physical therapists to function as part of a team. Participation in this project strengthened these qualities for these students.

THURSDAY AFTERNOON

Auditorium

Mini-Symposium Drug Delivery

1:30 OPENING INTRODUCTION TO DRUG DELIVERY DEVICES

Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Keynote: Historical perspective of the discovery of ceramic delivery systems and major applications in the biomedical field.

1:45 INVITED GUEST SPEAKER, OSMOTIC PUMP DRUG DELIVERY

Ateegh Al-Arabi, JCCC, Overland Park, KS

Novel Applications for Osmotic Pump Delivery Systems.

2:00 EVALUATION OF THE MALE REPRODUCTIVE ORGANS AFTER TREATMENT WITH CONTINUOUS SUSTAINED DELIVERY OF STATIN FOR FRACTURE HEALING

Felix I. Adah (1*), Hamed Benghuzzi (1), Michelle Tucci (1), George Russell (1), Audrey Tsao (1), and Barry England (2), (1) University of Mississippi Medical Center, Jackson, MS 39216 and (2) University of Michigan Medical School, Ann Arbor, MI 48104

The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) are widely used for the treatment of hyperlipidemia, and recent in vitro and animal data suggest that statins promote bone formation and increase bone strength. We examined the relationship between sustained continuous delivery of statin and fracture healing rates in adult male animals with either femoral defect or segmental fractures. Because statin affects the production of cholesterol we also evaluated the influence statin, on adrenal and testicular steroidogenesis and the morphology of the reproductive tract tissues in animals receiving statin for periods of 4, 8 and 12 weeks post surgery. Simvastatin significantly increased fracture healing and reduced total cholesterol without influence on the plasma HDL-cholesterol levels. Basal plasma LH, FSH and testosterone levels were not affected by active treatment with simvastatin. Reproductive tissue morphology was unchanged by local sustained release of statin. In conclusion, long-term simvastatin treatment, at doses effective in improving lipid profile, did not influence testicular reproductive and endocrine function, but was able to effectively heal simple and complicated fracture types. * Graduate student in Clinical Health Sciences

2:15 ANDROGENIC MODULATION OF ANGIOGENESIS IN THE TISSUE IMPLANT RESPONSE

Kenneth R. Butler and Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Several previous studies on the effect of steroid hormones on the tissue-implant response have recently appeared in the scientific literature. The objective of this project was to demonstrate that androgens affect angiogenesis in the fibrous tissue surrounding TCP implants. It is hypothesized that the number and size of blood vessels formed within the fibrous tissue surrounding TCP implants loaded with androstenedione (A), dihydrotestosterone (D), and testosterone (T) is greatly affected by the androgens. Sixteen animals in four experimental groups (n = 4/group) were implanted with one TCP implant each. Group I animals were implanted with the sham TCP ceramic (Control). Group II animals received the T-TCP ceramic. Group III animals were implanted with the D-TCP bioceramic. Group IV animals received the A-TCP bioceramic. At 90 days post-implantation, the animals were euthanized and the fibrous tissue surrounding the implants were evaluated microscopically following staining with routine hematoxylin and eosin (H & E), modified Papanicolaou, and PAS. Using digital analysis software, data were collected and evaluated comparing hormonal effects on number and size of blood vessels within the fibrous tissue collected from all four groups. The presence of androgens greatly affected the angiogenic response within the fibrous tissue. Both testosterone and androstenedione appear to suppress or limit angiogenesis. The results of this study demonstrate that some steroid hormones have a strongly varying affect on angiogenesis within the fibrous tissue surrounding TCP implants.

2:30 COMPARISON OF CONVENTIONAL AND SUSTAINED DELIVERY OF L-DOPA ON SH-SY5Y NEUROBLASTOMA CELLS

Tina Martin*, Hamed Benghuzzi, and Michelle Tucci, University of Mississippi Medical Center, Jackson, MS 39216

L-dihydroxyphenylalanine-(L-DOPA) has been used for the treatment of Parkinson's disease for decades. Recently it has been shown that L-DOPA, at concentrations of 0.25 X [10.sup.(-4)] M or larger, can be toxic for human neuroblastoma cells. Toxicity has been associated with the productions of high levels of quinones. The reactive oxygen or nitrogen species generated in the enzymatic oxidation or auto-oxidation of an excess amount of dopamine or L-DOPA induce neuronal damage and/or apoptotic or non-apoptotic cell death. The objective of this study was to investigate if the delivery method could reduce the toxicity associated with higher levels of L-DOPA. SH-SY5Y neuroblastoma cells were challenged with a bolus administration of 5, 50, 100, or 500 [micro]M of L-DOPA or a sustained delivery of L-DOPA to release the desired concentrations by the end of one incubation period. The cells were incubated for periods of 24, 48, or 72 hours, and at the end of each phase cell number, cell morphology, and cellular glutathione levels were determined. Conventional administration of L-DOPA showed reductions in cell number at 72 hours in cells treated with 50 and 500 [micro]M L-DOPA. A similar trend was observed using drug delivery administration of L-DOPA. Glutathione levels were reduced in the treatment groups after 24 hours and showed recovery by 48 hours. Morphologic evaluation revealed that the lower dose groups appeared similar to control in both conventional and drug delivery whereas cellular distortion was observed at higher doses regardless of delivery method. Conclusion: Lower dose L-DOPA was demonstrated to be less cytotoxic in SH-SY5Y neuroblastoma cells. * Graduate Student in Clinical Health Sciences

2:45 THE EFFECTS OF CORTISOL CONCENTRATION ON THE RC/4B PITUITARY CELL LINE

Lisa Haynie* and Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Pituitary adenomas are benign tumors that arise exclusively within the anterior pituitary. Treatment for any pituitary hormone-secreting adenoma has three main goals: (1) remove/shrink tumor mass, (2) remove/inactivate any hypersecreting pituitary tissue, and (3) provide hormone replacement. Surgery generally accomplishes the first two goals. The replacement hormone therapy regime is the most cumbersome and often leaves the patient at serious risk for peaks and valleys in hormone levels. Without the hypothalamic-pituitary-adrenal axis (HPA) the patient's ability to produce an anti-stress effect may be altered. Glucocorticoids (GCs) are regulated by HPA and mediate diverse physiological effects. GC level is elevated after infectious/inflammatory stress and exerts a suppressive effect on the immune function. The objective of the study was to evaluate RC/4B/C pituitary cells after 24, 72, and 96-hour incubation periods with low, medium and high doses of cortisol. Cell viability, number, morphology, and metabolism were evaluated. The results of the study showed a significant decrease in cell number as early as 24 hours in the treated groups. Increases in cellular damage were evident at 72 and 96 hours in all treated groups. Cellular protein levels were unremarkable for the duration. Reduced cell numbers in the treated groups along with evidence of cellular membrane oxidation suggests the possibility that GC may reduce or suppress neuroendocine regulation by causing cellular apoptosis. The results show a need for further studies on the cell line to establish the mechanism by which GC suppress the immune system in times of infectious/inflammatory stress.

4:00 Divisional Poster Session

THE EFFECTS OF CORTISOL ON THE VIABILITY AND PROLIFERATION OF CERVICAL TUMOR CELL LINES, SW 756 AND HELA, AND NORMAL CERVICAL CELLS, ECT 1/E6E7, IN CULTURE

Melissa Daniel* and Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Cervical cancer remains a health issue worldwide, and the role of steroid hormones on cervical cancer cells is not clearly defined. This study investigated the effects of the physiological and supraphysiological levels of cortisol, on the viability and proliferation of two cervical tumor cell lines, SW 756 and HeLa, and cervical cells, Ect 1/E6E7 cells, that were HPV transformed. In this study, all cell lines were treated cortisol and examined biochemically and morphologically at 24, 48, and 72 hours. Results from this study indicated the following: the MTS Assay for cell counts revealed supraphysiological doses of cortisol (0.05 [micro]M) produced a statistically significant decrease in cell proliferation in all cell lines for the duration of the study (p < 0.05). Assays for cellular damage revealed the following: MDA and LDH levels were increased at all time periods following the addition of supraphysiological levels of cortisol for SW 756, HeLA and Ect1/E6E7 cells. Protein levels were unremarkable. Morphological changes were seen in all cell lines given supraphysiological doses of cortisol. Cytoplasmic changes included slight increases in vacuolization, fragmentation, and intracytoplasmic debris. Nuclear changes included a slight increase in bizarre and aberrant nuclear shapes. This investigation provides significant information regarding the interrelationship between the stress hormone, cortisol and the viability and proliferation on cervical tumor cells and normal cells transformed with HPV.

ELECTRICAL STIMULATION AT THE ACUPUNCTURE POINTS FAILS TO IMPROVE DELAYED ONSET MUSCLE SORENESS 

Min Huang*, Mark D. Weber, William R. Woodall, Cynthia K. Scott, Felix I. Adah, and Neva F. Greenwald, University of Mississippi Medical Center, Jackson, MS 39216

The purpose of the present study was to determine whether electrical stimulation at acupuncture points can decrease muscle pain and increase muscle strength at different time periods after delayed onset muscle soreness (DOMS) induced by exercise. Subjects consisted of 19 untrained female volunteers that had no known medical conditions. A single blind experimental design was used during this experiment. Each subject was randomly placed into either a treatment or control group. DOMS was produced in subjects by having them perform eccentric flexion of the elbow using free weights. Elbow range of motion, muscle strength of biceps and pain were measured daily for 3 days. All subjects were connected to electrical stimulation machines by electrodes placed at eight pre-determined places based on acupuncture points and pathways. The experimental group received stimulation over the sensory threshold. The control group received no stimulation. The main effect within subjects over time was found to be significant for passive extension (p < 0.01), strength (p < 0.001), and pain (p < 0.001). There was no significant difference between the treatment group and control group for passive extension (p > 0.05), strength (p > 0.05), and pain (p > 0.05). The results of this study showed an increase in subjective pain and a decrease in strength of biceps and range of motion of the elbow over the time, but failed to show any beneficial treatment effect of electrical stimulation at the acupuncture points on DOMS.

INCORPORATION OF LIPID PANELS, WEIGHT MANAGEMENT PROGRAMS, AND DIETARY CHANGES IN THE TREATMENT OF OBESE PEDIATRIC PATIENTS

Felisa Wilson* and Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Critical measures are being explored to combine laboratory data, clinical guidelines, weight management programs, and nutritional counseling to establish more effective treatment plans for obese pediatric patients. Clinical practice alone is not enough to remedy the rapidly increasing number of obese children and adolescents. The efficacy of incorporation of laboratory data, weight management program, and dietary changes was implemented in approximately 25 patients 6 to 13 years of age who were obese with average weight 20% greater than standard guidelines. Treatment included use of lipid panel, nutrition counseling and therapeutic exercises guided by licensed physical therapist for 8 weeks increments. Prior to program admissions, lipid panels were drawn to establish risk for coronary heart disease. These levels were monitored to ensure the patient was at minimal risk for heart attack prior to initiating any type of exercise regime. After a median follow-up of 2 months, changes in weight and laboratory profiles were seen. Combining laboratory expertise, exercise, and diet the pediatric population can initiate resolution strategies in the fight against obesity in the pediatric population. Thus, from a medical viewpoint, clinical evidence indicates that a significant reduction and modification of risk factors can be obtained by incorporation of lipid panel monitoring, exercise, and healthy dietary changes in daily lifestyle. Furthermore, healthier children result in healthier adults and decreased healthcare cost.

ACUPRESSURE FAILS TO IMPROVE DELAYED ONSET MUSCLE SORENESS

Min Huang*, Mark D. Weber, William R. Woodall, Cynthia K. Scott, Felix I. Adah, and Neva F. Greenwald, University of Mississippi Medical Center, Jackson, MS 39216

The purpose of this study was to determine if acupressure treatment at the acupuncture points can decrease muscle pain and increase muscle strength faster after DOMS was induced by exercise. Twenty subjects with no complicating medical history volunteered for this study. The control and treatment group consisted of 3 males and 7 females. Delayed onset muscle soreness was induced through eccentric exercise of the subject's non-dominant biceps. Range of motion, pain, and strength were measured each day. Subjects were randomly placed into either a treatment or control group. Subjects in the treatment group received acupressure treatment and subjects in the control group received no treatment. A repeated ANOVA test showed a significant difference for strength between days 0 and 1 (p < 0.01), 1 and 2 (p < 0.01) and 3 and 4 (p < 0.05). Pain was significant between days 0 and 1 (p < 0.05), 1 and 2 (p < 0.01), and 3 and 4 (p < 0.05). There was no significant difference between the treatment group and control group for passive extension (p > 0.05), strength (p > 0.05), and pain (p > 0.05). Conclusion: The protocol used to induce DOMS was effective, but acupressure treatment did not significantly improve DOMS as compared to the placebo treatment. A larger patient population is needed to determine if acupressure is truly effective in treating DOMS.

DEG/ENaC PROTEINS ARE REQUIRED FOR MY OGENIC CONSTRICTION IN MOUSE INTERLOBAR ARTERIES

Nikki Jernigan* and Heather Drummond, University of Mississippi Medical Center, Jackson, MS 39216

Autoregulation is the local adjustment of vascular resistance to maintain blood flow and is important in protecting the kidney against injury. A component of autoregulation, the myogenic response, is the inherent ability of vascular smooth muscle (VSM) to contract in response to increases in intraluminal pressure. Although mechanosensitive ion channels are thought to initiate VSM stretch-induced contraction, the molecular identity of these channels is unknown. Recent reports suggest Degenerin/Epithelial [Na.sup.+] Channels (DEG/ENaC) are required for normal mechanosensation in sensory neurons, however, the role of DEG/ENaC proteins in myogenic constriction has not been established. We hypothesized that DEG/ENaC proteins are mechanosensors in VSM and required for myogenic vasoconstriction. To determine if ENaC transcripts and proteins are expressed in cultured mouse renal VSM cells, we used reverse transcriptase PCR, immunoblotting, and immunolocalization. We detected expression of [alpha], [beta], and [gamma]ENaC transcripts and proteins. Further, we examined myogenic constriction and agonist-induced constriction and dilation in isolated mouse renal interlobar arteries in the presence or absence of the specific DEG/ENaC inhibitors, amiloride and benzamil. The DEG/ENaC inhibitors blocked myogenic constriction in a concentration dependent manner, without interfering with agonist-induced reactivity. Our findings suggest DEG/ENaC subunits are 1) expressed in renal vessel VMC and 2) required for myogenic vasoconstriction. DEG/ENaC proteins may be components of mechanosensitive ion channels required for myogenic vasoconstriction. This work was supported by the NIH and NHLBI.

PHARMACOLOGICAL BLOCKADE OF CORTICOTROPIN RELEASING FACTOR DOES NOT ALTER DEPENDENCE ON BUTORPHANOL

George Howell III, Melissa Parker*, and Robin Rockhold, University of Mississippi Medical Center, Jackson, MS 39216

We hypothesized that corticotropin releasing factor (CRF) facilitates both behavioral signs and neuronal activation which occur upon withdrawal from physical dependence on butorphanol or morphine. Dependence was induced in male Sprague-Dawley rats by administration, via osmotic minipump, of either morphine or butorphanol (28 nmol/[micro]l/hr; i.c.v.) for 72 hours. At the end of 72 hours, animals were treated with saline (5 [micro]l i.c.v.) or the CRF antagonist, alpha-helical CRF ([alpha]hCRF; 10 [micro]g/5 [micro]l i.c.v.), 15-20 minutes prior to naloxone (48 nmol/5 [micro]l i.c.v.) precipitated withdrawal. Behavioral signs of withdrawal were counted for 30 minutes and scored. Pretreatment with [alpha]hCRF significantly decreased the occurrence of teeth-chattering in butorphanol-dependent animals, however, it did not significantly decrease the overall withdrawal scores in butorphanol- or morphine-dependent animals. Immunohisto-chemical analysis revealed a significant increase of Fos immunopositive cells, an indication of neuronal activation, in the paraventricular nucleus of the hypothalamus (PVN), central nucleus of the amygdala (CeA), periaqueductal grey (PAG), and supraoptic nucleus (SON) of saline-pretreated, morphine-dependent animals compared to saline-pretreated, butorphanol-dependent animals. Our data indicate chronic administration of butorphanol, as opposed to morphine, elicits a physical dependence in which one or more behavioral signs of withdrawal are facilitated by CRF. Paradoxically, morphine withdrawal elicits a greater degree of neuronal activation in most of the measured brain nuclei than butorphanol withdrawal.

EFFECT OF ADJOINING LINKER SEGMENTS UPON THE STABILITY OF DOMAIN 2 OF EPITHELIAL CADHERIN

Alka Prasad*, Corey Nichols, Tex Song, Gregg Anazia, Nicole Housley, and Susan Pedigo, University of Mississippi, University, MS 38677

Epithelial cadherin is a cell adhesion protein that is essential in calcium-dependent cell-cell recognition and adhesion. It is contains 5 to 7 modular domains in its extracellular region that have a 7-strand [beta]-sheet immunoglobulin-type fold. These domains, particularly Domain 1, are responsible for the protein-protein interactions that lead to adhesion between cells in tissues. Recently stability studies of Domain 2 were undertaken with the motivation that it is a representative modular domain for this family of proteins. Because the definition of a domain boundary is somewhat arbitrary, we have undertaken studies to examine the effect of the adjoining linker regions that connect Domain 2 to the adjacent domains in the physiological context of the molecule. These short 7 amino acid segments are highly conserved and offer 3 to 4 oxygens that are known to chelate the calcium ions that bind at the interface between the domains (Nagar et al. (1995) Nature 380; 360 4). We performed temperature induced denaturation studies monitored by circular dichroism. Each of these linker regions destabilize Domain 2 by 4 to 5[degrees]C in the apo state such that the construct with both linkers attached is destabilized by 8[degrees]C. This represents a decrease in stability of 1.7 kcal/mol for the construct with both linkers over just Domain 2 alone. In addition, all constructs with linkers are stabilized to approximately 6 kcal/mol by the addition of 5 mM calcium to solution. Calcium stabilization of constructs with Linker 1 was confirmed through limited proteolysis. Contrary to similar studies on other modular proteins these studies show that the adjoining sequences actually destabilize the modular domain.

EVALUATION OF MRC-5 AND MG-63 CELL GROWTH RATES USING THE ALAMAR BLUE ASSAY

Joyce Belcher (1), Alexis Hand (1), Laura Franklin (2), Michelle Tucci (2), Hamed Benghuzzi (2), and Joseph A. Cameron (1), (1) Jackson State University, Jackson, MS 39217 and (2) University of Mississippi Medical Center, Jackson, MS 39216

The process of cell counting is a valuable tool that enables researchers to study cellular proliferation and optimize cell culture conditions. Traditionally, the hemocytometer has been a widely used to manually count cells; however, there is an increased chance of human error in performing this technique. An alternative method of counting cells is the Alamar Blue Assay (ABA). The specific aim of this experiment was to examine the growth rates of normal lung fibroblast (MRC-5), and an osteosarcoma cell line (MG63), utilizing the ABA. Use of the ABA presents numerous advantages over established proliferation measurements, but we also observed several drawbacks to the routine use of the ABA. First, we showed a decrease in variability in the spectrophotometric data with minimal differences within the group, but the ABA was not sensitive enough to detect differences between the groups. Our data showed the reduction rate of Alamar Blue varies according to the cell line. Cells with higher metabolic rates (MG63 cancer cells) reduce the dye much quicker than primary non-transformed cell lines leading to a over estimation of cell number. We also found that the cells growing within the 96 well plates interfere with absorbance values reducing the accuracy of the total cell numbers. Varying the method of cellular treatment and analysis with the plate reader may help to improve the sensitivity of the ABA. Supported in part by NIH GM50117. *Graduate Student

EVALUATION OF INSULIN-LIKE GROWTH FACTOR-1 AND PARATHYROID HORMONE ON GROWTH CHARACTERISTICS OF MG-63 CELLS

Laura Franklin*, Michelle Tucci, Hamed Benghuzzi, George Russell, Ashraf Ragab, and Audrey Tsao, University of Mississippi Medical Center, Jackson, MS 39216

Insulin-like Growth Factor-1 and Parathyroid Hormone are hormones secreted in the body that have roles in bone formation. The purpose of this experiment was to examine MG-63 cells after treatment with PTH and IGF-1 in low (1 [micro]g), medium (5 [micro]g), and high (50 [micro]g) dosage levels and to make comparisons between the two hormones. MG-63 cells were plated onto a 24 well tissue culture plate at a density of 1 X [10.sup.5] cells per well. The experiment was designed to evaluate cell counts, MDA, protein levels, calcium levels, alkaline phosphatase levels, and cellular morphology after 24, 48, and 72 hours post incubation with IGF-1 and PTH. Both hormones stimulated cellular division as evidenced by morphology and cell numbers. There was an inverse relationship between dose and cell number with the lower dose of IGF-1 and PTH causing the most increase. In both hormones, the highest dose level showed the largest MDA level increase. However, in the protein levels, few changes in protein levels were found with IGF-1, but PTH showed an increase of protein levels over the time periods. Morphological evaluation showed prominent nucleoli and cellular division throughout both treatments, however, the cells with IGF-1 became extremely elongated and the cells with PTH became rather plump. The information gathered suggests that IGF-1 and PTH have an anabolic effect on MG-63 and the effect is dose dependent with both treatments with the lower dose being more effective.

CALCIUM BINDING TO E-CADHERIN TWO DOMAIN CONSTRUCT

Susan Pedigo* and Huaying Zhao, University of Mississippi, University, MS 38677

Cadherins are a family of calcium binding cell-adhesion molecules. Cadherin is a transmembrane protein that has 3-7 independently folded beta-barrel extracellular domains. Cadherins mediate calcium-dependent cell-cell adhesion through the most N-terminal extracellular domains. Calcium ions bind at the interface of the extracellular domains changing the relative disposition of the domains. Our work focuses on the first two-extracellular domains of epithelial cadherin (ECAD12). The crystal structure of ECAD12 (Nagar et al. (1996) Science 380: 360-4) showed 3 calciums binding at the interface between the domains and requiring residues from both domains as well as the short linker peptide that joins them. We use tryptophan-fluorescence and circular dichroism to monitor the local and global conformational change as a function of calcium concentration. Time-dependent proteolytic susceptibility studies of ECAD12 were performed at various levels of calcium using chymotrypsin. Titration endpoints were not well-defined in the fluorescence data, but the midpoint was estimated and compared to results from CD experiments. In our CD work, the spectrum of ECAD12 was monitored from 210 nm to 240 nm at 20 calcium concentrations. Titration curves were analyzed in two groups 212 nm to 222 nm and 230 nm to 236 nm and individually according to two models (Equal/independent, and Unequal/cooperative). There was no difference between the two groups indicating that tryptophan signal at 235 nm monitors 2[degrees] structure. This is consistent with fluorescence experiments. Proteolytic susceptibility studies revealed that in the absence of calcium the protein (ECAD12) was digested gradually through multiple cleavage events, however, one of the fragments was very stable even after 2 hours of digestion. In the presence of calcium, ECAD12 was totally protected by calcium.

EXPRESSION ANALYSIS OF TUMOR PANELS: DIFFERENT ARRAYS AND METHODS

Margot Kaelbling*, Christoph Klett, Warren May, and Charles Streckfus, University of Mississippi Medical Center, Jackson, MS 39216

Our aims were to (1) assess if microarray analysis of tumors detects altered gene expression and (2) evaluate different microarrays, RNA isolation kits, and labeling methods. We used tumors containing mutations in PTEN/MMAC1/TEP1 which leads to down-regulation of many proteins and to cell cycle arrest and/or apoptosis. We analyzed three primary cancers and one uterine tumor along with uninvolved tissue from the same patients; all but the uterine tumor contained mutations in PTEN. Total RNA was isolated from tumor and uninvolved tissue using one of three kits, reverse-transcribed into cDNA, and labeled with fluorescent dyes (either Cyanine 3 and Cyanine 5 or Alexa 594 and Alexa 488). The labeled tissues of each panel were hybridized to a microarray. Panels labeled with Cyanine dyes were hybridized to Human Oncogene and Tumor Suppressor Gene Microarrays that contained at least 281 known human cancer-related genes of average 2,200-bp cDNA length spotted twice including PTEN. Panels labeled with Alexa dyes were hybridized to Human SS-H19k7 microarrays containing 19,008 human ESTs (Expressed Sequence Tags) of 100-150-bp, supposedly spotted once but actually spotted two-four times; no PTEN sequences were found. Expression ratios were calculated for each slide and statistically analyzed using SPSS software. Most genes with altered expression were underexpressed. RNA isolation methods were equally satisfactory. Expression from duplicate gene sequences provided confidence in uniform hybridization across a slide. The longer fluorescence of Alexa versus Cyanine dyes was of no advantage, but our labeling method for Alexa dyes required more RNA than labeling with Cyanine. For PTEN expression analysis, EST miroarrays are not useful. Research and specimen collection were supported by MFGN and CHTN.

A NOVEL TECHNIQUE FOR THE QUANTITATIVE IMMUNO-STAINING

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

The use of histochemical immuno-staining has been widely used to identify the presence of certain proteins in animal tissue. The results gained from this type of examination are purely qualitative, and are based on the ability to visualize a stain tagged to a second antibody under a microscope. Problems that arise when using this method include: expense of reagent, visualization of minute quantities of protein, reaction time, and difficulty reporting results. We describe a new method that employs both the specificity of the traditional double antibody immuno-stain along with the sensitivity of a Western blot. In this procedure the tissue slide is flooded with the primary antibody synthesized in rabbit. After an overnight incubation, anti-rabbit IgG with conjugated peroxidase (second antibody) is added to the slide. To this point the procedure is identical to a traditional immuno-stain, except that the concentration of antibody used is reduced due to sensitivity of the detection agent, a chemiluminescent substrate. The chemiluminescent properties of the substrate allow the tagged proteins to develop on X-ray film in a relatively short period of time (about 30 minutes). The exposed x-ray film can also be analyzed using a densitometer to extrapolate the protein concentration using a standard curve. This technique offers several advantages over the traditional staining methods including the use of less reagent, easily visualized results without a microscope, decreased turn-around time, and the ability to quantitate the results using a densitometer and standard curve. *Graduate Student in Allied Health

RESEARCHING AMIFOSTINE (ETHYOL) AND ITS USES IN COMBINATION WITH RADIATION THERAPY

Brian Comer (1,2*) and Raymond Wynn (3), (1) Mississippi Gulf Coast Community College, Gautier, MS 39533; (2) National Aeronautics and Space Administration, Stennis Space Center, MS 39522; and (3) Singing River Hospital Oncology Center, Pascagoula, MS 39563

The purpose of this project was to conduct a literature review of Amifostine. Amifostine is a cytoprotective agent manufactured by MedImmune, Inc. It is used to reduce levels of renal toxicity in patients receiving cisplatin. Amifostine is also used to reduce the occurrence of moderate to severe xerostomia in patients that are receiving radiation therapy. This presentation will provide information on the pharmacology and clinical uses of Amifostine.

FRIDAY MORNING

Auditorium

Clinical Diagnotics II

8:00 TYLESYNERGY[R]: BENEFITS IN CANCER DIAGNOSIS

Stacey Overstreet (1,2*) and Raymond Wynn (3), (1) Mississippi Gulf Coast Community College, Gautier, MS 39533, (2) National Aeronautics and Space Administration, Stennis Space Center, MS 39522; and (3) Singing River Hospital Oncology Center, Pascagoula, Mississippi 39563

Telesynergy[R] is a new technology from which both doctors and patients can benefit. A patient can be diagnosed faster when the doctor needs a second opinion with the use of this system. Telesynergy[R] has the ability to place conference calls, send x-rays, and send microscope views to doctors many miles away. The focus of this project was to discover the benefits of the use of Telesynergy[R] in the patient diagnosis.

8:15 PURIFYING POSsmLE PRIMAQUINE-BINDING PROTEINS FROM PNEUMOCYSnS CARINII USING AFFINITY CHROMATOGRAPHY

Pamela L. Ruffin (1,2*), William I. Sullivan, Jr. (2), and Sherry F. Queener (2), (1) Jackson State University, Jackson, MS 39213 and (2) Indiana University School of Medicine, Indianapolis, IN 46202

Primaquine, an approved treatment for malaria, is effective against the opportunistic fungal pathogen Pneumocystis carinii. P. carinii is one of the major opportunistic infections of AIDS patients. The goal of the study is to detemtine the mechanism of action of primaquine against P. carinii. Our hypothesis is that P. carinii contains primaquine binding proteins. To begin we isolated proteins from P. carinii-infected rat lungs that were retained by primaquine-linked Sepharose. A column of primaquine-linked Sepharose was loaded with extract containing the soluble proteins of P. carinii; bound proteins were eluted with primaquine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the eluted fractions shows a single protein band in a late eluting fraction. This band has been submitted for protein sequencing by mass spectrometry. Our results indicate that the primaquine-linked resin retained proteins from the P. carinii infected rat lung extract. Identification of the retained protein is being performed to determine if it is from P. carinii. Future studies include using extract from uninfected rat lung as control to compare with P. carinii infected lungs to ensure eluted proteins are from the pathogen. We also evaluated P. carinii antisera using a dot blot as a potential tool for use in future affinity chromatography experiments. Supported in part by NIH R25 GM067592.

8:30 INHIBITION OF IFN-[gamma] SIGNALING IN MACRO-PHAGES BY POXVIRUS PHOSPHATASE VHI 

Andre W. Hite, Jr. (1,2*), Alicia A. Cecil (1), Joseph A. Cameron (2), and Michael J. Klemsz (1), (1) Indiana University School of Medicine, Indianapolis, IN 46202 and (2) Jackson State University, Jackson, MS 39217

Poxviruses including Variola (small pox), and Vaccinia, which is used to vaccinate against small pox have developed numerous methods for evading an immune response. One way is the ability of the viral VH1 phosphatase to alter signaling cascades in infected cells. Our studies focused on showing if either the Vaccinia or Variola VH1 protein dephosphorylates STAT1 during activation of macrophages with Interferon-gamma. In order to answer this question we transfected the STAT1 responsive Tap-1 luciferase reporter plasmid (pLTP) into the murine macrophage line P388D1. We optimized DNA concentrations by titrating pLTP and determined that maximal light units were obtained using 1.0 [micro]g of pLTP. Our results show that a ratio of 3:1 of Fugene transfection reagent to pLTP yielded maximal light units. To determine if the Vaccinia VH1 protein affected Tap-1 promoter activity, pLTP was co-transfected with the VH1-PJ6 or PJ6 alone followed by stimulated with IFN-[gamma]. These results show that Vaccinia VH1 protein reduced pLTP expression compared to the empty vector following the addition of IFN-[gamma] for 4 hours. We also determined that the transfection reagent Fugene reagent had an effect on IFN-[gamma] induction of the pLTP plasmid. Our results suggest that VH1 may be blocking the IFN-[gamma] response, and the Fugene may alter how IFN-[gamma] activates cells. Supported in part by: NIH R25 GM067592-02.

8:45 Break

Environmental Influences on Health II

9:00 BENEFICIAL HEALTH EFFECTS OF LOW-LEVEL IONIZING RADIATION

Amin Haque, Alcorn State University, Alcorn State, MS 39096

We receive approximately 0.20-0.30 rem per year from natural radiation sources, depending on where we live. Several places are known in Iran, India (Kerala), Brazil (Araxa, Tapira), Sudan, China (Han) and Europe, where natural background radiation gives an annual dose of more than 5 rem and up to 79 rem/yr (at Ramsar in Iran). There is no evidence of any increase in cancer among people living in areas where natural, background radiation is several times higher than average. Also, the mortality rate due to cancer was significantly less than similar areas with a low background radiation level. The survivors in Nagasaki from the atomic bombs in Japan, who received 1 to 19 rem of radiation, had lower cases of cancer and are living longer than the non-irradiated population. It has been reported that the mortality caused by cancer at nuclear power plants in Canada and U.K. were lower than the national average. UNSCEAR Report on Chernobyl concludes that no increases in cancer incidence or mortality have been observed that could be attributed to ionizing radiation. Below an individual dose of 5 rem in one year or a lifetime dose of 10 rem risks of health effects are either too small to be observed or are non-existent. Low to moderate doses of radiation are not harmful and evidence has proven that radiation at these levels can actually be beneficial to one's health.

9:15 NITRIC OXIDE INHIBITORS POTENTIATE DOMOIC ACID INDUCED HIPPOCAMPAL DAMAGE IN VIVO

Manju Pande (1*), A. Harps (1), M. Lopez (2), D. D'Souza (2), J. Wei (2), T. Jones (2), and M. Sundaram (2), (1) Mississippi Valley State University, Itta Bena, MS 38941 and (2) University of Mississippi Medical Center, Jackson, MS 39216

Domoic Acid (DA), a potent excitotoxin produces hippocampal damage and functional lesions as seen in temporal lobe epilepsy. Previous studies have suggested that nitric oxide (NO) may possess anticonvulsant properties and have a protective role against excitotoxic injury in epileptogenesis. The present investigation was done to further understand the role of NO in DA induced hippocampal degeneration, and if a single subconvulsive exposure to DA causes progressive damage to the hippocampus. Based on the data obtained from our earlier studies, the ICR mice were treated with subconvulsive dose of DA in the presence and absence of NOS inhibitors L-NAME and 7NI. After treatment, the animals were observed for 30-45 min for the stereotypic neurological effects in the form of scratching. Animals in various groups were sacrificed after 30 and 60 days of treatment. The brains were removed and processed for histological and immunohistochemical analysis. Some brains were stored at -80 degree C for biochemical analysis. The initial observation of animals showed that nitric oxide inhibitors potentiated DA induced stereotypic seizures indicating that NO may possess anticonvulsant properties. Tissues from the 60-day treatment groups were processed first. The CA1/2 region of hippocampus immunostained for glial fibrillary acidic protein (GFAP) showed reactive glial cells with large cell bodies and long processes in DA treated animals as compared to the controls. These changes were more pronounced in groups treated with NO inhibitors. The double immunostaining showed loss of calbindin D28K positive neurons in CA1/2 regions in L-NAME and 7NI treated animals. The data from 30-day treatment groups is currently being analyzed.

9:30 NONIONIZING ELECTOMAGNETIC RADIATION AND HEALTH EFFECTS

Amin Haque, Alcorn State University, Alcorn State, MS 39096

Nonionizing electromagnetic radiation photons have smaller frequency, and therefore, smaller energy than the photons of ionizing radiations (< 12.4 electron volts), which is insufficient to ionize atoms and molecules in human tissue. But they can excite the atoms and the molecules and break chemical bonds in the tissues, and can be hazardous to health if their intensity is large. These include with increasing frequency: radio waves, microwaves, infrared (heat), and visible light. The intensity as well as the frequency of the radiation, and exposure time are crucial to the health effects. Biological systems are sensitive to heat, which may come from a microwave oven, cooking range, sun, radio waves, or intense visible monochromatic light (laser beam). An increase of about 6 [degrees]C in body temperature as a result of radiation absorption would be fatal. Any part of the body that cannot dissipate heat efficiently or is heat sensitive may be damaged by microwave radiation. A 70-kg human requires 1.5X[10.sup.6] J of nonionizing radiation to kill. For a visible light of frequency 5X[10.sup.14] Hz, absorption of about seven moles of photons will be needed to kill an average human. The radiowaves used by analog and digital cellular phones have much lower frequency and the intensity level is very low to produce harmful biological effects even with long term exposure. National and international scientific opinion is that there is no substantiated evidence that using a mobile phone causes harmful health effects.

9:45 SYNTHESIS AND CHARACTERIZATION OF COMPLEXES OF VANADIUM ACETYLACETONATE WITH DI-2-PYRIDYLKETONE THIOSEMICARBAZONE

Miriam Thaggard (1,2*), Ramaiyer Venkatraman (1), Terrell Bradford (1,2), and Joseph A. Cameron (1), (1) Jackson State University, Jackson, MS 39217 and (2) Hinds Community College, Raymond, MS 39211

Vanadium complexes have been considered for use as insulin-mimetics. It has been shown that vanadium(V) and vanadium(IV) can stimulate glucose uptake/oxidation and glycogen synthesis. Vanadium complexes with organic ligands have proved to be less toxic with improved solubility and lipophilicity. As an organic ligand, thiosemicarbazones were found to exhibit antilukemic properties. Extensive research has been carried out on aliphatic, aromatic, heterocyclic and other types of thiosemicarbazones and their metal complexes due to their therapeutic value. In view of the extensive biomedical significance of these compounds, we have synthesized and characterized three new vanadium complexes of di-2-pyridylketone thiosemicarbazone and vanadyl acetylacetonate. These complexes were characterized using thermal, spectral, and molar conductivity methods. The molar conductivity studies indicate that the complexes exist in the nonionic form. Solubility studies indicate the nonpolar nature of the complexes. Further structural characterizations were carried out using infrared, UV-Visible and proton NMR techniques. These studies, combined with information available from previous X-ray studies in our laboratory, provided the structural characterization of these complexes. The results indicate that the trifunctional thione form of the ligand is chelated to the vanadium center through the pyridine nitrogen, imine nitrogen and thione sulfur atoms. The bidentate acetylacetonate ligand occupies the fifth and sixth metal coordination. Supported in part by NIH-NIGMS Bridges Grant GM50117.

10:00 Break

10:15 Divisional Poster Session

CYP 1A2 ENZYME ACTIVITY IN THE LIVERS OF RATS EXPOSED TO LEAD

Christopher Bennett*, Roger Holloway, Shang-Zhi Xu, and Bettaiya Rajanna, Alcorn State University, Alcorn State, MS 39096

The liver detoxifies xenobiotics. Liver enzymes such as, P450s play a significant role in detoxification. CYP 1A2 is abundant (15%) in the liver compared to other CYPs. Toxins are found in higher concentrations in the liver. This leads to liver damage, including decreased activities of essential enzymes. Lead (Pb) is an environmental contaminant and upon exposure is found in huge quantities in the liver. The objective of this study is to determine the effect of Pb on the CYP 1A2 enzyme in the rat liver. Wistar, male rats were treated with 0.1 M lead acetate in water for 45 days. Control groups received distilled water. All animals were provided with rat chow food ad libitum. Animals were sacrificed at intervals of 1, 5, 10, 15, 21, and 45 days. The livers were perfused with ice-cold saline through the gate vein until they became yellowish. The livers were then removed, and microsomes were fractionated as per procedures established in our laboratory. Enzyme activity was determined by measuring the fluorescent intensity of the livers. 7-ethoxy3H-phenoxazin-3-one was the substrate used. NADPH started the reaction and it was stopped by adding acetonitrile. The tissue Pb levels were determined. There was a gradual decrease in fluorescent intensity of exposed compared to normal rats and the response was time dependent. The tissue Pb levels were higher in Pb exposed rats confirming a higher Pb accumulation. The results also suggest that Pb decreases CYP 1A2 enzyme activity in liver over time. (Supported by NIH/NIGMS/MBRS-SCORE#GM 55356).

CARDIOVASCULAR HEALTH PROMOTION IN SCHOOLS: MODIFIABLE RISK FACTORS IN RURAL MISSISSIPPI FIFTH GRADE STUDENTS

Raven Worthy (1*), Deborah S. King (2), and T. Kristopher Harrell (2), (1) Murrah High School, Jackson, MS 39202 and (2) University of Mississippi Medical Center, Jackson, MS 39216

Hypertension and obesity are more prevalent in the Southeast than in any other region in the United States. These health disparities exist even among children, with rural Mississippi children at increased risk. This study evaluated body weight, height, and blood pressures among children from Forest and Morton, Mississippi. The body mass index (BMI) and height percentiles were calculated for each student and blood pressure was classified as acceptable, borderline, or high. Of the 205 fifth grade students, 57% were male and 43% female. Average systolic blood pressure (SBP) for Morton was 108 mmHg compared to 102 mmHg for Forest. Average diastolic blood pressure (DBP) for Morton was 65 mmHg while that for Forest was 75 mmHg. Both SBP and DBP were > to the 85th percentile in approximately 15% of both groups. Forest's average height percentile was 62% while Morton's was 58%. Forest's BMI average was 70% and Morton's 75%, with over 50% of children in both groups either at risk for overweight or overweight (> 85th percentile). Morton children had both higher SBP and BMI percentile averages. Forest had the higher DBP and height percentile averages. Rates of cardiovascular risk factors in rural Mississippi school children are well above reported national averages, mirroring the increased rates in Mississippi adults. Early intervention in this age group is crucial to reversing these risk factors and preventing progression to cardiovascular disease. Future studies are needed regarding successful interventions in this age group, as well as family and societal approaches for prevention.

THE INHIBITORY EFFECTS OF D-CARNITINE AND QUERCETIN ON TRYPANOSOMA LEWISI INFECTION IN RATS

Catherine Karlak*, Rebecca Rose, and Frank Lowell, Belhaven College, Jackson, MS 39202

Trypanosomes are a leading cause of death in third world countries, and most current drug therapies are impractical due to their toxicity and cost. Nutritional therapies are often more practical and recent research has explored such methods of controlling trypanosomal infections. Manganaro et al. (2003) suggested that D-carnitine, an amino acid derivative, is successful in controlling Trypanosoma lewisi infections in rats. Based on this study, the inhibitory effects of D-carnitine and Quercetin, a flavonoid, were investigated. A combined therapy of both drugs was considered for a possible synergistic effect. Sprague-Dawley rats were given I.P. injections of the drugs in dosages based on weight, beginning on day 3 of infection and ending on day 18. The level of trypanosomes in the rats' blood was measured through counts performed regularly according to standard procedures. The means of these counts demonstrate that rats given D-carnitine had significantly lower levels of parasitemia, in agreement with previous research. Quercetin and the combined therapy did not appear to significantly inhibit the trypanosomes. Further study is needed to develop the potential of such combined drug therapies as treatment for Trypanosoma infections.

THE EFFECTS OF ESTROGEN AND TESTOSTERONE ON MG-63 CELLULAR METABOLISM.

Ameze Adah (1*), Hamed Benghuzzi (2), Michelle Tucci (2), and Felix I. Adah (2), (1) University of Mississippi, University, MS 38677 and (2) University of Mississippi Medical Center, Jackson, MS 39216

Estrogen (E) and testosterone (T) have complex effects on the skeleton, including regulation of modeling and maintenance of bone mass. Osteoblasts are key regulators of skeletal matrix synthesis and degradation. However, whether osteoblasts mediate E and T effects remain unclear. MG63 osteoblast-like cells were cultured in 24-well culture plates and treated with high, medium and low E or T at 24, 48, and 72 hours. At the end of each time point, cellular numbers, protein, damage (MDA), and glutathione levels were evaluated. The cell counts of the MG63 E-treated were increased at 48 and 72 hours in the medium dose and elevated at 72 hours in the high dose. The cell counts for T-treated cells showed a significant increase (p < 0.05) in the medium and high levels at 24 and 48 hours. Cellular damage was not evident in either treatment group as ascertained using MDA. The protein levels of the E and T-treated cells were similar to control for the duration of the study. The glutathione levels of the E-was reduced at 48 hours at the median concentration (p < 0.05). No significant difference in the glutathione levels for the cells treated with T were observed for the duration of the study. The results showed no significant difference between the two hormones (E and T), but both hormones undoubtedly increased cellular proliferation. The results revealed that E increased cell proliferation in a dose and time-dependent manner and testosterone in a dose-dependent manner. Both of their proliferative abilities suggest that estrogen and testosterone may contribute to bone loss prevention.* Undergraduate Student

THE PROTECTIVE EFFECTS OF UV BLOCKING AGENTS ON CELLULAR MORPHOLOGY AND CELLULAR FUNCTION OF FIBROBLAST CELLS EXPOSED TO UV LIGHT.

Montica Wilson (1*), Lolitta Crawley (1), Laura Franklin (2), 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 goal of this study was to determine the degree of cellular protection offered by sunscreens of different SPF ratings in an attempt to prevent squamous damage as a result of UV free radical induced generation. The specific aim was to evaluate the morphological responses of MRC-5 fibroblast cells exposed to UV-radiation in the presence of absence of sunscreen. 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 as follows: Twenty-four tubes per time period (24, 48 and 72 hours) were divided into four groups. Group 1 consisted of control unexposed cells, group 2 consisted of cells exposed to UV light, group 3 cell culture tubes were pre-treated with SPF 8 sunscreen prior to UV exposure, and cell tubes in group 4 were pre-treated with SPF30 sunscreen prior to UV exposure. Cell numbers were determined and the morphological evaluation was assessed using Image Pro Digital Analysis technique. Data obtained demonstrated that, forty-five minutes of radiation exposure caused traumatic stress to the cells, swelling, cellular debris and fragmentation. However, treatment with both SPF 8 and SPF 30 pre-exposure provided protection for the cells. The SPF 30 treatment resulted in less cellular aggregation, more abundant eosinophilic cytoplasm and an overall healthy appearance. Supported in part by NIH GM50117. *Undergraduate Student

THE EFFECTS OF [H.sub.2][O.sub.2] ON FIBROBLAST GROWTH PROPERTIES FOLLOWING PRE-TREATMENT WITH ANTIOXIDANTS

Mindy M. Bertucci*, Angela M. Phillips, Hamed Benghuzzi, Michelle Tucci, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

Cellular proliferation is a critical component of wound healing, and oxidative stress can lead to significant reduction in cellular proliferation. The goal of this study was to investigate the effects of antioxidants on MRC-5 fibroblast cells alone or in the presence of a potent oxidant, [H.sub.2][O.sub.2] for 24, 48 and 72 hours. Cells were evaluated using cytological criteria, protein concentrations, and cellular damage markers. The experimental design was divided into two phases. First, doses of vitamins E and C were evaluated to determine effective nontoxic dosages. Results from phase I showed moderate toxicity of vitamin C, which was due to the acidic nature of the vitamin and buffering capacity of the media. In the second phase, vitamin E low/high doses were added to the MRC-5 cells for thirty minutes prior to the addition of 180 mM [H.sub.2][O.sub.2] and the cells were evaluated at 24, 48 and 72 hours for changes in the aforementioned criteria. The results showed low and high doses of vitamin E were ineffective in protecting the cells from [H.sub.2][O.sub.2] damage. Decreases in cellular number, cellular protein concentrations and increases in cellular damage were evident at all time points in cells treated with [H.sub.2][O.sub.2]. Changes in cellular morphology included fewer cells, multiple nucleoli, bare nuclei, and web like cytoplasm. Overall, the results suggest oxidants play a strong role in reducing cellular proliferation which would result in decreased wound healing. *Undergraduate student, Allied Health

GROWTH AND CELL VIABILITY OF ESTRADIOL AND IP-6 TREATED HEP-2 LARYNGEAL CARCINOMA CELLS

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

Inositol 6-phosphate (IP6) has demonstrated novel anti-cancer activity using several different tumor models. IP6, a phytoestrogen, has estrogen receptor (ER) binding capabilities that are not known to cause cellular proliferation in hormone sensitive cells. It is hypothesized that IP6 can induce competitive inhibition with estrogen for estrogenic binding sites on cancer cells resulting in decreased proliferation. In this experiment, Hep-2 cells were treated with estradiol and IP6 in a dose dependant manner for 24, 48, and 72 hours. They were analyzed for changes in number, protein concentrations, damage, and morphology. There was an increase in cell proliferation in Estradiol treated Hep-2 cells. Cells treated with IP6 showed no change in cell proliferation in the 24 or 48 hour groups, but there was a decrease in number with the 72 hour group, particularly the ImM dose. Both the estradiol and IP6 treatments caused no membrane oxidation and the level of protein synthesis stayed consistent. The morphology showed small round to cuboidal, single cells with scant, dense, basophilic cytoplasm and hyperchromatic nuclei with smooth borders. Some cells showed anucleation and cellular degeneration. Although IP6 is a phytoestrogen, the results show that affinity for estrogen binding sites on Hep-2 cells is greatly decreased. However, over time given increased concentrations, IP6 can cause a decrease in the cellular proliferation of Hep-2 cells without initiating cellular apoptosis.

EFFECTS OF REPEATED ETHANOL ADMINISTRATION ON EXTRACELLULAR SEROTONIN CONCENTRATIONS IN THE HIPPOCAMPUS OF P AND WISTAR RATS

Samuel Haile (1*), K. Wratten (2), A.D. Moran (3), R.C. Marunde (3), L. Lumeng (4), W.J. McBride (3), and R.J. Thielen (3), (1) Jackson State University, Jackson, MS 39217; (2) Indiana University, Bloomington, IN 47405; (3) Indiana University School of Medicine, Indianapolis, IN 46202; and (4) Veterans Administration Medical Center, Indianapolis, IN 46202

Previous research has demonstrated that extracellular levels of serotonin (5-HT) in the hippocampus (hipp) increases in alcohol preferring (P) rats when exposed to ethanol (EtOH). Serotonergic innervation of the hipp appears to play a key role in the rate of tolerance development to the high dose effects of ethanol (hypothermia and motor incoordination). Wistar and P rats will be assigned to two groups. The ethanol group will receive 1 g/kg ethanol for 5 consecutive days while the saline control group will receive an equivalent volume of saline. On the sixth day, in vivo quantitative microdialysis using the no net flux procedure will be run. The no net flux procedure involves perfusing different concentrations of 5-HT levels into the hipp. We hypothesized that P rats will have lower 5-HT levels than Wistar rats in the hipp and that repeated ethanol will result in a larger increase in basal extracellular 5-HT levels in P rats compared to Wistar rats. In doing the experiment, we found that repeated ethanol administration resulted in a trend toward an increase in basal 5-HT levels in the vHipp. of Wistar rats; and significantly higher basal 5-HT levels in ethanol treated Wistar rats compared to ethanol treated P rats. Repeated ethanol administration resulted in a significant increase in 5-HT clearance within the vHipp. of P rats. The purpose of this study is to determine the effect of repeated ethanol administration on basal 5-HT levels in the hipp. Support Contributed By: AA 07611, HL 07802 and MH 16926.

EVALUATION OF THE CYTOTOXIC EFFECTS OF TANTALUM AND [K.sub.2][Cr.sub.2][O.sub.7] USING A FIBROBLAST CELL LINE

Angela M. Phillips*, Mindy M. Bertucci, Hamed Benghuzzi, Michelle Tucci, and Zelma Cason, University of Mississippi Medical Center, Jackson, MS 39216

Particulate wear debris is associated with aseptic loosening of the implant leading to implant failure. Inorganic metal compounds such as chromium and tantalum can be associated with implant debris triggering changes in cellular viability at the implant interface. The goal of this study was to determine the cytotoxic effects of chromium and tantalum (Ta) when co-incubated with fibroblast cells. A soluble hexavalent chromium salt ([K.sub.2][Cr.sub.2][O.sub.7]) was used at concentrations of 0.0001 M, 0.001 M, and 0.01 M, and insoluble Ta at concentrations of 0.005, 0.01, and 0.02 g/mL were incubated in the presence of MRC-5 fibroblast cells. At the end of 24, 48 and 72 hours cells were evaluated for number, protein, damage, and morphology and compared to non-treated cells. The results showed cell number was influenced positively in cells treated with the higher doses of metal salts. Levels of cellular protein and damage were unaffected by the metal salts. Cellular morphological evaluations of [K.sub.2][Cr.sub.2][O.sub.7] treated cells showed increased N/C ratios, multiple nucleoli, irregularity of nuclear membranes, mitotic figures, and wispy cytoplasm. Morphological evaluations of tantalum treated cells showed wispy and web-like cytoplasm compared to the control. Overall, the results indicate that cellularity was moderately elevated without alterations in cellular protein or damage. These results suggest that the concentration of metals tested were not cytotoxic to the fibroblast cells, one of two cell types at the tissue-implant interface. *Undergraduate student, Allied Health

INFLUENCE OF GLUCOSE, FRUCTOSE, VANADIUM AND THEIR COMBINATIONS ON METABOLIC TOXICITY AND SURVIVAL OF JURKAT CELLS

Jarret Smith (1,2*), Clement Yedjou (1), Ibrahim O. Farah (1), Ramaiyer Venkatraman (1), and Joseph A. Cameron (1), (1) Jackson State University, Jackson, MS 392171 and (2) Hinds Community College, Raymond, MS 39211

Studies showed that tumor growth and abnormal cell survival was associated with a number of metabolic abnormalities. Glucose metabolism is known to be deranged (an impaired oral glucose tolerance test), lipoprotein-lipase activity is depressed (hypertrigliceridemia after an exogenous lipid load) and protein metabolism showed abnormality as revealed by changes in plasma amino acid profile and evidenced by an increased plasma free tryptophan levels in patients with breast, lung, colon, stomach, and other cancers from various origins. The role of carbohydrate metabolism and Vanadium salts (as mimickers of insulin in glucose transport) were not clearly established in the literature, therefore, the purpose of this study was to evaluate metabolic toxicity to Jurkat cells exposed (48h) to glucose, fructose, vanadium salt (Vanadium (IV)-di-2-pyridyl-ethyl thiosemicarbazone) and their combinations, and to measure toxicity indices and survival trends using the MTT assay. Results showed that, both sugars and vanadium alone or in combination affected cell survival negatively and that LC50's ranged from 2133.96-3867.56 ug/ml. Death of these cells could be attributed to either an increase in pro-oxidant activity leading to the production of reactive oxygen species (ROS) or due to increased internal pH as a result of glycolytic metabolism. Future studies will focus on the mechanisms of death in response to manipulation of cellular metabolism in cancer aiming at prevention/control. Supported in part by NIGMS R25 GM50117.

THE EFFECTS OF ISOLATED ANTIOXIDANTS FROM BLACK SEED ON THE CELLULAR METABOLISM OF A549 CELLS.

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

Reactive oxygen species cause cytotoxic effects in biological systems. The body requires the uptake of exogenous compounds with antioxidant potential. Black seed is a plant that contains at least one active lipid soluble antioxidant, thymoquinone. Fractionation of the seed components yielded antioxidant compounds in both the water soluble and lipid soluble fractions. The OBJECTIVE of the study was to determine the safety of the fractionated compounds and compare their potency with pure thymoquinone and vitamin E on A549 cells in culture for 24, 48, and 72 hours. RESULTS: Black seed extracts and pure thymoquinone showed markedly reduced levels of MDA for the duration of the study. The vitamin E dosage used led to greater toxicity and cellular damage rather than cell protection. Cellular proteins levels after 24 hours showed cells in BS + ETOH extract group had the highest metabolic activity. However, at 72 hours, the activity was shifted and showed the least amount of protein synthesis. As for vitamin E, the results were consistent throughout all three phases showing slow metabolic activity. Cell number was decreased after 24 hours in thymoquinone treated cells, and remained reduced for the duration of the study. The BS+[H.sub.2]O fraction showed a similar trend to thymoquinone, where as the BS + ETOH fraction showed a negative shift in cell number at 48 hours when compared with the control. CONCLUSION: The study concludes that of the two BS extracts, the BS+[H.sub.2]O extract had the greatest effect on the cell viability and function.

THE EFFECTS OF PALMITIC ACID CONCENTRATIONS ON A549 CELLULAR INCORPORATION AN D CELLULAR FUNCTION

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

Neonates exhibit a high risk of developing acute and/or chronic lung disorder, often associated with surfactant deficiency. Alveolar type II cells (A549) synthesize dipalmitoyl phosphatidylcholine as the main surfactant phospholipid. The objective of our study was to administer various concentrations of palmitic acid (PA) to A549 cells and determine the level of cellular viability, cellular damage, cellular morphology, and degree of surfactant production at 24, 48 and 72 hours of incubation. The results of our study show that incorporation of [.sup.3]H-palmitic was approximately 96% after 24 hours. Our results also demonstrated concentrations of 0.01 M and 0.1 M PA caused marked reductions in cell number as early as 24 hours. The reduction in cellular numbers appeared to be dose dependent. Markers of membrane cellular damage also were elevated in all treatment groups after 24 hours and remained elevated for the duration of the study. Cellular morphology evaluations revealed cells treated with PA initially appeared in clusters, with hyperchromatic nuclei compared with control. No differences in nuclear/cytoplasmic ratios between control and treated cells were observed. After 72 hours the PA treated cells showed evidence of scant cytoplasm and hyperchromasia when compared to control. Overall, the results indicate that incorporation of palmitic acid ranging in concentrations from 0.001 M to 0.1 M caused increases in cellular damage and cellular death. This is most likely due to changing the distribution of the phosphatidyl moieties on the cellular membrane. *Allied Health Student

THE EFFECTS OF GENISTEIN CONCENTRATION ON Hep-2 CELLULAR FUNCTION

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

Genistein (GEN) is a phytoestrogen that has shown potential as a chemotherapeutic agent, which acts by inhibiting protein-tyrosine kinase and topoisomerase II enzymes. These particular enzymes are crucial for cellular proliferation. The goal of this study was to administer 0.5, 0.05 or 0.005 mg/mL to Hep-2 cells and evaluate cellular number, protein, damage and morphology at 24, 48, and 72 hours. Cell numbers were significantly reduced in low and medium concentrations after 24 hrs, and cell number appeared to rebound at 48 and 72 hrs in an inverse fashion. This data suggests GEN was metabolized by the cells, which left cells virtually without treatment after 24 hrs in both low and medium concentrations. Data also suggests continuous administration of the drug at therapeutic levels would serve as a better chemotherapeutic agent. Cellular damage was not evidenced in the experiment suggesting that the drug did not target the cellular membrane. Morphological changes such as anucleation were seen at 24 hrs in all dose treatments suggesting that GEN targets the cell nucleus. Interestingly, cellular function was able to recover in the lower doses of GEN treatment suggesting cellular metabolism of the drug. Also, this information suggests GEN targeted enzymatic activity as opposed to causing alterations within the cellular membrane leading to leakage of cellular contents and ultimately cellular death since the membrane did not show evidence of lipid peroxidation. *Undergraduate, Allied Health

FRIDAY AFTERNOON

Auditorium

Mini-Symposium Advances in Clinical Health Sciences

1:00 ADVANCES IN CLINICAL CHEMISTRY

La'Toya Ross Richards* and Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Over the years, the healthcare industry has made both rapid and numerous advances. These advances have resulted in better approaches to aid in the diagnostic work-up of patients as well as patient response to treatment. Clinical Chemistry serves as the major branch between basic and applied research, between the research scientist and the patient. The division of Clinical Chemistry provides an extensive array of highly sophisticated tests, which utilize current and advanced technology with unending intellectual changes. These changes have led to a better understanding of disease pathogenesis and more decisive roles in the diagnosis and interpretation of laboratory tests. Some examples of assays involved in Clinical Chemistry testing are assays for therapeutic drug monitoring, assays detecting various hormone levels such as thyroid and parathyroid hormone levels, triple-panel testing for neural tube defects, tests for cystic fibrosis diagnosis, and a vast majority of others. These tests are performed on instruments employing various technologies. Examples of some of these technologies include enzyme linked immunosorbent assays (ELISA), fluorescence polarization immunoassays (FPIA), microparticle enzyme immunoassays (MEIA), and chemiluminescence immunoassays. Regardless of the technological principles involved, all methods tend to serve as beneficial agents in the molecular pathogenesis of disease and patient management and improvement. Overall, advances in Clinical Chemistry have ultimately resulted in startling improvements in the field of laboratory medicine.

1:15 ADVANCES IN CLINICAL HEMATOLOGY

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

The role of the clinical laboratory in patient care has become increasingly more important over the last few decades. The increase in the number of laboratory tests ordered, in addition to the variety of laboratory tests being offered, has resulted in a demand for new and improved technology. Advances in hematology include the development of "all-inclusive" hematology analyzers that are capable of performing complete blood counts (CBCs) and reticulocyte counts, making peripheral blood smears, and staining the smears all within the same system. Other analyzers can perform body fluid analyses in a matter of seconds compared to manual methods, which take significantly longer. Advances in technology allow the clinical laboratory to provide accurate tests results more quickly, thus shortening test turnaround times. Technological improvements also increase employee productivity and reduce the number of medical errors.

1:30 ADVANCES IN BLOOD BANKING

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

The healthcare industry is growing vastly. The diversification of the field is extraordinary, the field of Laboratory Medicine is just one of the many outlets of the health care industry. Laboratory Medicine includes an array of departments, including blood bank. Human blood is required to save lives of people suffering from accidents and disease. Many years ago blood donation was limited to a few laboratory tests and a few preliminary screenings. Today the technology has expanded allowing for more adequate testing and donor screening. Some of the test that are performed today allow safe blood products to be transfused to patients. A few of these test include Hep B which was introduced in 1971, followed by Hep C, HTLV I and II which were introduced 20 years later. The first blood test for HIV was implemented in 1985, only a year after it was identified as a cause of AIDS. HIV-1 and-2 antibodies screening was implemented in 1992, and West Nile Virus in 2002. As time progress the new technology and available test have created a safer blood industry. Technology and research has extended the storage time of blood, in 1979 it was 35 days and 5 years later the shelf life of blood was extended to 42 days. Transfusion Medicine aspects of the industry has advanced tremendously, the time it takes to process blood has decreased significantly due to advanced testing and improved technology. Many years ago Blood Banking was mostly done by manual procedures, today it has become automated. Thanks to these advances, blood donation has become a safer practice.

1:45 MEDICAL LABORATORY ERRORS: HOW MANY HAVE YOU MISSED?

Stacy Vance*, Michelle Tucci, and Hamed Benghuzzi, University of Mississippi Medical Center, Jackson, MS 39216

Objective: To examine the type and frequency of laboratory errors and how errors can be prevented in the clinical laboratory. Design: Bibliographic review of the current literature through electronic search in Medline data-base from 1940 to 2004, with selection of the most relevant articles. A Medline search using terms medical laboratory errors (739), laboratory errors (3008), laboratory types (118), type of laboratory errors (215), delta check systems and laboratory (11), and proficiency testing (662) were performed. Results: The search revealed large differences in study designs and quality on this topic, as well as a lack of available data and the lack of a shared definition of "laboratory error" (also referred to as "blunder", "mistake", "problem", or "defect"). Despite these limitations, there was considerable concordance on the distribution of errors throughout the laboratory working process: most occurred in the pre- or postanalytical phases, whereas a minority occurred in the analytical portion. The reported frequency of errors was related to how they were identified: when a careful process analysis was performed, substantially more errors were discovered than when studies relied on complaints or report of near accidents. Conclusions: The large heterogeneity of literature on laboratory errors together with the prevalence of evidence that most errors occur in the preanalytical phase suggest the implementation of a more rigorous methodology for error detection and classification and the adoption of proper technologies for error reduction. Clinical audits should be used as a tool to detect errors caused by organizational problems outside the laboratory.

2:10 Divisional Business Meeting and Awards