Section V: Biomedical Sciences Science Hall E1049 Seyed H. Hosseini, Presiding.
8:15 EFFECTS OF RESVERATROL AND QUERCETIN ON B[a]P- AND 3MC-INDUCED CYP1A1 REPORTER ASSAY**, Caitlin N. Meads* and Jennifer C. Schroeder, Young Harris College, Young Harris, GA 30582. The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor involved in the regulation of multiple cellular pathways, including transcription of the cyp1a1 gene related to drug metabolism. Benzolaipyrene (BlalP) and 3-methylcholantherine (3MC) are both known agonists of the AHR. Bioflavonoids, such as quercetin, and the stilbenoid resveratrol have also been characterized as ligands for the AHR. Resveratrol and quercetin are naturally found in grapes and red wines; both are also available as nutritional supplements. Many plant-derived compounds, including these two, have been touted as chemo-preventive agents. In this study we explore the competitive inhibition of 3MC- and BlalP-induced AHR activation with resveratrol and quercetin by utilizing a cyp 1a1 promoter-driven luciferase assay. The degree of inhibition is determined in mouse hepatocytes exposed to varying concentrations of the compounds. Both 3MC- or BlaIP- activated luciferase activity are inhibited in a dose-dependent manner by quercetin. Cotreatment of resveratrol with 3MC or B[alP also modified expression levels. Funded by the Young Harris College Science Research Initiative.
8:30 EXTRACTS FROM GANODERMA LUCIDUM, TRAMETES VER-SICOLOR, GR1FOLA FRONDOSA, AND LENTINULA EDODES DECREASE XRE-MEDIATED TRANSCRIPTION INDUCED BY B[A]P**, Matthew D. Sudderth" and Jennifer C. Schroeder, Young Harris College, Young Harris, GA 30582. Pharmacological activity of mushrooms is widely unknown due to their numerous chemical constituents. Modern herbalists revere mushrooms for their medicinal activity and even claim they can be effective in treating cancer. Ganoderma lucidum (reishi) is one of the most widely aclvertizecl mushrooms for purported anti-carcinogenic effects. Tra metes versi-color (turkey tail). Lentinula edodes (shiitake), and Grifola frondosa (maitake) are also regularly claimed as effective treatments of various cancers. Benzolalpyrene (BialP) is a carcinogen found in cigarette smoke, which is also a strong agonist of the AHR. Binding of the AHR to XRE sequences regulates transcription of many genes, including cyplAl. In these studies mouse liver cells stably transfected with a cyplAl-XRE luciferase reporter construct are exposed to B[a]f), mushroom extracts, or combinations of BlalP and extracts. While none of the extracts alone activate the reporter, all four were able to inhibit B[a]P-induced activity. These results suggest that components of the mushroom extracts tested here may exhibit their chemoprotective properties via competitive inhibition of the AHR. Funded by the Young Harris College Science Research Initiative.
8:45 DOES FLUOXETINE-HCL PROMOTE NEUROGENESIS IN ADULT ZEBRAFISH SUBJECTED TO STRESS?**, Nina Bi Couch*, Ryan D. Shepard* and Linda G. Jones, Young Harris College, Young Harris, GA 30582. Stress is an everyday event. However, chronic or severe stress can be the cause of physical and mental disorders ranging from headaches, insomnia, depression and anxiety to more deleterious effects such as heart disease, hypertension, and even neurologic damage. Excitotoxicity and oxidative stress have been suggested to promote damage along the hypothalamic-pituitary-adrenal axis (HPA axis) resulting in degradation of areas such as the amygdala and the hippocampus. Studies have also suggested that the neurotransmitter serotonin has neurogenic properties. We are interested in determining whether treatment with fluoxetine-HCI (a selective serotonin reuptake inhibitor or SSRI) will increase neurogenesis in adult zebrafish (Danio rerio) subjected to stress relative to fish subjected to stress only or to non-stressed control fish. We established four tanks of adult zebrafish (two of which were subjected to stress). One tank from the stressed group and one from the non-stressed group received fluoxetine-HCI treatment. Evidence of neurogenesis will be determined by immunohistochemistry of paraffin-embedded and sectioned tissues using an anti-acetylated tubulin antibody as a marker of newly formed neurons and their axons. Density levels of new neurons will be quantified, and the data analyzed to determine whether differences exist among the experimental groups. Funding for this project was from the Young Harris College Undergraduate Research Initiative.
9:15 EFFECT OF XYLITOL ON FUSOBACTERIUM NUCLEATUM AND STREPTOCOCCUS SP. GROWTH AND BIOFILM FORMATION**, L.N. Johnson* and A.L. Kwiatkowski, Young Harris College, Young Harris, GA 30582. Fusobacterium nucleatum is commonly found in human dental plaques that can potentially cause dental caries. It is a middle colonizer that can co-aggregate with early colonizing bacteria to form a biofilm. Xylitol is an artificial sweetener that has been shown in our lab to reduce the growth of F nucleatum monospecies cultures, yet increase the biofilm formation. In this study, experiments were performed to determine the effect of xylitol on mixed species biofilms containing F nucleatum and early colonizing Streptococcal species. One mixed culture consisted of F. nucleatum and Streptococcus salivarius while another one consisted of F nucleatum and Streptococcus sanguinis. Cultures were grown for 72-hours anaerobically in 96-well plates at 37[degrees]C in Schaedler broth containing xylitol concentrations of 0%, 0.50%, 1%, or 2%. A spectrophotometer was used to measure the optical densities of the growth at 600nm and biofilrn stained with safranin at 492nm. The Biology Department of Young Harris College funded this research.
9:30 THE INFLUENCE OF SECRETIONS FROM MACROPHAGE-LIKE CELLS PULSED WITH BACTERIAL GHOST ON PROSTATE CANCER CELLS**, 0. Martinez*. D. N. McKeithen', A. Stevens and G. A. Ananaba, Department of Biology, Clark Atlanta University, Atlanta, GA 30314. Prostate cancer (PCa) is one of the most common types of cancer in men and is most prevalent among African American men. Some types of PCa grow slowly and may need minimal or no treatment, others are aggressive and can spread quickly. Previously, our lab has shown increased levels of Tumor necrosis factor-alpha when THP-1 cells, a human monocytic Leukemia cell, are pulsed with bacterial ghost. This study focuses on the effects THP-1 cell secretions have on PCa cells and testing the hypothesis; THP-1 cells may secrete factors that can influence proliferation of PC3 prostate cancer cell line. Since bacterial ghosts have adjuvant effects and elicit secretion of THP-1 cytokines, we treated the PCa cells with secretions from bacterial ghost pulsed THP-1 cells. The collected condition media was then cultured with the PCa. The treated THP-1 cells' cytokines can activate T cells having anti cancer properties. Our results suggest factors from innate immune response are essential in stimulating adaptive long-term immunity against endogenous antigens and the importance of robust innate immunity in cancer immunotherapy. This research was funded by RISE Grant 115R25GM060414-11.; NST/CREST/CFNM Award #HRD-1137751: Title III Grant HBGI #22211K- Enhancing Graduate Students Academic and Professional Development.
9:45 BIOFLAVANOIDS AS FULL OR PARTIAL AGONISTS OF THE AHR AND THEIR EFFECTS ON LEVELS OF LACTIC ACID AS A MODEL OF DIABETES**, Whitney R. Marcus*, Alejandra Escamilla*, Madison L. Perdue* and Jennifer C. Schroeder, Young Harris College, Young Harris, GA 30582. Diabetes is a condition in which insulin cannot properly regulate glucose transport from the bloodstream into cells. In these studies we aim to validate the findings of Wang, et a!, who reported in 2011 that exposure of mouse hepatocytes to 11-naphthollavone (BNF) caused a decrease in glucose metabolism, while those levels could be restored by co-treating the cells with a PPAR-c4 inhibitor (GW6471). We further seek to determine if bioflavonoids such as apigenin, hesperetin and quercetin modify this metabolism and counteract the effects of BNF by acting as a competitive inhibitor of the AHR, for which BNF is a known ligand. Results shown here are unable to confirm the earlier findings, but did provide unique insight as to the interaction of BM.: and the bioflavonoids in this system. All three of the bioflavonoids examined here significantly diminish the levels of glucose metabolism. Current studies are looking to determine if this effect is related more to the regulation of AHR or PPAR-u. Funded by the Young Harris College Science Research Initiative.
10:00 Section Business Meeting
10:30 NLRP3 INFLAMMASOME'S ROLE IN IL-10K0 MICE FOLLOWING CHLAMYDIA INFECTION, DN McKeithen (1.2), Y Omosun (1), EC Kibakaya (1), F Eko (1), CM Black (3), JU Igietseme (1.3), GA Ananaba (2) and, Q He (1), (1) Department Microbiology, Biochemistry, and, Immunology, Morehouse School of Medicine, Atlanta, GA 30310, 'Department of Biology, Clark Atlanta University, Atlanta, GA 3031.4 and (3) Centers for Disease Control & Prevention (CDC), Atlanta, GA 30333. Chlamydia is a public health concern due to its prevalence and devastating reproductive consequences, including inflammation and infertility. In previous studies we have shown that Interleukin 10 knock out (IL-10K0) mice were less likely to develop Chlamydia induced inflammation and infertility. "lhe purpose of this study is to determine the immunoregulatory role of NLRP3 inflammasome within the bone marrow dendritic cells (BMDCs) of IL-10 KO mice fol- lowing Chlamydia infection. BMDCs were harvested from wild type (WT) and IL-10K0 mice and infected with C. muridarum. Our results show that Chlamydia infected IL-10 deficient DCs down-regulated NLRP3 inflammasome assembly, inhibited apoptosis, and, increased the lifespan of the DCs. Taken together, our results suggest that IL-10 deficiency yields enhanced immunity against Chlamydia infection. This research is supported by RISE Grant #5R25GM060414-11, Title Ill Grant 422210J and NIH Grant 111SC1A1103041-01A1.
10:45 PHYTOCHEMICAL CAPABLE OF INHIBITING PHIP INDUCED CYTOTOXICITY, Ashok Jain* and Abhilash Samykutty, Department of Natural Sciences, Albany State University, Albany, GA 31705. Breast cancer is the second leading cause of cancer-related deaths for women. Heterocyclic amines (HCAs) are formed when meat products are cooked at high temperatures and have been shown to be carcinogenic. 2-Amino-1-methyl-6-phenylimidazol 14, 5-b]-pyridine (PhIP) is the most abundant HCA found in well-done and grilled meats. Phytochemicals are wide variety of compounds that occur naturally in plants and known for their antioxidant properties. Phytochemicals are promoted for the prevention and treatment of many health conditions, including cancer. The objective of this study is to understand which antioxidant is more effective in inhibiting the PhIP toxicity. The culture of breast epithelial (MCF 10A) cells was initiated and treated with PhIP in the presence and absence of antioxidants namely ascorbic acid (AsA), Glutathione (GHS), N-acetyl-cysteine (NAC), alpha-tocopherol (Vitamin E), [61-gingerol, -gingerol, lycopene, Vitamin K3, Piperine and Curcumin for 24 hours, The interaction of PhIP and phytochemicals were evaluated by cell proliferation assay; and for DNA damage by comet assay. The phytochemical showed promising results was further analyzed for Reactive Oxygen Species (ROS), DNA adduct formation and cell cycle. The core signaling pathways evaluated by RT PCR and/or Western blotting which included Nrf2 (GSR, GPX, NQ01), FOX() (Catalase. GADD45, PRDX3) targets; DNA repair genes/ proteins BRCA1, H2AFX and PARP-1; and tumor suppressor P16 gene expression. Expression of antioxidants genes was induced by PhIP whereas curcumin significantly suppress the PhIP induced ROS activation, DNA strand breaks and DNA adduct formation. The presence of Curcumin with PhIP inhibits both DNA double strand breaks and DNA adducts formation. Curcumin also induces the tumor suppressor P16 gene. In conclusion, it appears that Curcumin anti-cancer actions are via multiple molecular targets.
11:00 VIBRIO CHOLERAE GHOSTS AS AN ADJUVANT TO ENHANCE IMMUNE RESPONSE DURING TREATMENT OF CHLAMYDIA INFECTION", A Stevens (1), GA Ananaba (1), JU Igietseme (2.3) and FO Eko (2), (1) Clark Atlanta University, Atlanta, GA 30314, (2) Morehouse School of Medicine, Atlanta, GA 30310 and (3) Centers for Disease Control and Prevention. Atlanta, GA 30329. Chlamydia trachomatis is one of the leading sexually transmitted infections (STI) worldwide. These are serious infections that lead to infertility in women of childbearing age. Current treatments for the infection are limited to antibiotics and a vaccine, neither of which provide long-term immunity. and thus increasing the rates of infertility in women. Previous studies have shown that clearance of the infection depends on T helper 1 (Th 1) response along with complementary antibody response in the primary infection area. This study seeks to continue the investigation in finding a safe potent adjuvant to boost and sustain chlamyclial immune responses for long-lasting protective immunity. We hypothesize that stimulation of monocytes and macrophages with Vibrio cholera ghosts (VCG) induces the secretion of multiple immune responders. THP-1 monocytes or macrophages were pulsed with VCG (10 ug/ml) for 24 h. After stimulation, cellular supernatant was collected and analyzed for Thi and Th2 cytokine secretion. THP-1 cells tend to produce increased Th1 cytokine secretion levels vs. Th2 cytokine secretion during immune response. We conclude that stimulation of THP-1 cells with VCG leads to enhanced secretion of Thl cytokines. This work is supported by grant RO1 A141231 from National Institutes of Health (NIH) and Grant #1 C06 RR18386 from National Centers for Research Resources, NIH; Title Ill Grant FIBGI 422211k - Enhancing Graduate Students Academic and Professional Development.
11 : 15 MEASUREMENT OF THE INTERACTIONS OF LOW ENERGY GAMMA RAYS WITH DENSE METALS FOR APPLICATIONS IN NUCLEAR CARDIOLOGY IMAGING**, G.C. Passmore, T.F. Lynam* and J.L. Spradlin*, Georgia Regents University, Augusta, GA 30912. Gamma camera imaging of myocardium perfusion with either TI-201 or Tc-99m is widely used for the detection of coronary artery disease (CAD). Myocardium perfusion imaging studies allow the clinician to differentiate between healthy and damaged myocardium based on the amount of accumulated radionuclide represented in the images. Both radioisotopes, Tc-99m and TI-201. can indicate the perfusion characteristics of the myocardium. However. Tl-201 has the additional capability to indicate cardiac tissue viability. Simultaneous imaging of Tl-201 stress and Tc-99m rest images would have the benefits of optimal diagnostic perfusion imaging and tissue viability signaling. However, there is a difficulty in trying to image the lower TI-201 energy photons in the presence of the higher energy Tc-99m photons when using a standard lead collimator because interactions between the Tc-99m photons and lead create down-scatter in the TI signal region. The objective of this project is to measure the transmission and interactions of Tc-99m photons and Tl-201 photons separately and simultaneously using high density metal discs (different from lead) as attenuators for application in gamma camera collimators. Correction of the down-scatter problem would allow the clinician to take advantage of a simultaneous dual-isotope approach, increasing the detectability of reversible myocardium defects. Methods include using NIST attenuation data to identify multiple high density metals that meet the criteria for reduced Tc-99m down-scatter with K-shell x-rays signals that are different from the Tl-201 energy signal. Spectra for lead (Pb) have been collected. Continued analysis will be based on comparisons of spectra using multiple metallic attenuators.
11:30 PHAGE THERAPY: A POSSIBLE PARADIGM SHIFT FROM ANTIBIOTICS?, V. L. Chivukula* and D. O'Bryant, Atlanta Metropolitan State College, Atlanta, GA 30310. With the increase in antibiotic resistance among various bacterial species, scientists are investigating alternatives to fight bacterial infections. Using viruses that infect bacteria (bacteriophages) to treat bacterial infections could be more effective than antibiotics. Phage therapy dates back to the 1890s with Ernest Hankin's observations on cholera followed by Twort's studies suggesting viruses as the agents killing the cholera bacteria. D'Herelle discovered the first phage and used it to cure patients of dysentery. Though a number of studies followed that first discovery in the treatment of bacterial infections using phages, phage therapy never gained the importance that was received by antibiotics due to batch inconsistencies and manufacturing issues at that time. Phage therapy has recently regained importance with companies such as Intralytix, Inc. producing and developing products that inhibit pathogens in various food products. Advantages of using phage therapy include exponential growth of phages at the site of application depending on bacterial numbers, natural existence of phages for mutated bacteria, ease of using phage mixtures to inhibit different strains of a specific bacterial disease, and causing no harm to the normal flora. Currently, our lab is developing techniques for isolating bacteriophages from wastewater influent samples against Klebsiella pneumoniae and Streptococcus pyogenes and identifying the best phage titer values for a known bacterial concentration.
POSTERS SCREENING OF SOME NATURAL PRODUCTS AND NANOPARTICLES FOR THEIR ANTI-BREAST CANCER ACTIVITIES, C. T. Ahweyevu*, L. Wrensford and M. A. Taha, Albany State University, Albany, GA 31705. Cancer refers to a broad group of diseases that involve unregulated cell growth that form malignant tumors, which ultimately lead to death. Breast cancer is the most common type of cancer in women living in Western nations. It is estimated that approximately 1 in 8 North American females will develop breast cancer throughout their lifetime. The best way to battle cancer or any disease is prevention. One of the most researched aspects of chemoprevention is the use of natural products due to its cost effectiveness and little toxic side effects as opposed to other synthetic methods of chemo-treatment. Herein, we employed both biochemical and microscopic techniques to investigate the anti-breast cancer activities of seven different preparations namely; Milk thistle extract (MTE), Ginkgo biloba extract (GBE), Selenium (Se). Se-MTE, Se-GBE, chitosan, and Se-chitosan nanoparticles. These natural products have previously shown to have various forms of anti-cancer activities. Our results showed that only selenium and chitosan have promising anti-breast cancer activities according to the MTT and microscopy results. Further studies are required to confirm this result. This research is supported by the MBRS-RISE program to Department of Natural Sciences, Albany State University.
EXAMINATION OF SODIUM FLUORIDE'S EFFECT ON YEAST CELL GROWTH AND GENOMIC STABILITY**, A.R. Arvidsson* and A. L. Abclulovic-Cui, Georgia Regents University, Augusta, GA 30912. Sodium fluoride is a toxic, highly electronegative compound that floods public drinking water, lurks in toothpaste, produce, and pharmaceuticals. The compound sodium fluoride has an electronegativity of three due to the combination of fluorine's electronegativity of four forming an ionic bond with sodium's electronegativity of one. As a result of fluorine's tendency to gain an eighth electron to become stable sodium fluoride acts as a free radical within cells and will take electrons from molecules with only one electron in their valence shell. We believe this tendency may cause sodium fluoride to take electrons from sodium molecules within cells and change the electric charge within cells. It is very important that certain cells, like neurons. maintain a certain internal charge in relation to their surrounding .environment to function properly. The charge altering effect in neurons may cause or contribute to dementia and specifically Alzheimer's. To better understand the toxicity of sodium fluoride our aims to explore the effect sodium fluoride has on Saccharomyces cerevisiae. the budding yeast. We are exposing the yeast cells to varying concentrations of sodium fluoride solutions and examining results. Specifically, we will be looking for a decrease in cell growth over varying sodium fluoride concentrations and if DNA mutations accumulate in genes within the yeast cells caused by the exposure to sodium fluoride.
EFFECT OF A RNR1 MUTATION ON DNA MICROSATELITE STABILITY IN YEAST**, A.I. Nam*, N.A. 1-lashmi and A.L. Abdulovic-Cui, Georgia Regents University, Augusta, GA 30912. Microsatellites are repetitive DNA sequences within eukaryotic genomes. The repeated sequences can consist of two, three, or four base pairs and can be repeated up to a hundred times (e.g. CACACACACACACACACACA) Microsatellite sequences are known to be hotspots for DNA mutations, especially frameshift mutations. Importantly, mutations within microsatellite sequences can be very detrimental to human health and are known causes of colorectal cancer, Huntington's disease, and Fragile X Syndrome. In this study, we will be investigating a genetic pathway in the budding yeast, Saccharomyces cerevisae, which we hypothesize will increase the rate of mutations within microsatellite sequences. We have integrated a mutation into the RNR1 gene. RNR1 encodes the large subunit of the enzyme ribonucleotide reductase whose primary function is to create the precursors of DNA, the dNTPs, in a balanced manner. The mutation we used is rnrlY285A which causes an imbalance in the dNTP pools. Previous studies have shown that an imbalance in the dNTP pools causes increased mutations on non-repetitive DNA and we predict that the increase in mutations caused by imbalanced dNTP pools would be amplified on microsatellite DNA. We integrated mr1285A. into the yeast genome in addition to a 10 repeat unit GT microsatellite and a 10 repeat unit CA microsatellite sequences. Using an in vivo mutation assay and molecular DNA techniques we will measure the mutation rate of our new mutant strain and investigate the different mutations that were created.
THE EFFECTS OF NICOTINE ON CRANIOFACIAL DEVELOPMENT IN EMBRYONIC ZEBRAFISH**, Philip R. Uys* and Linda G. Jones, Young Harris College, Young Harris, GA 30582. Prenatal exposure to nicotine has been associated with craniofacial malformations, the most common of which is cleft palate. In this study we used embryonic zebrafish (Dcmio rerio) to determine whether exposure to nicotine promoted developmental abnormalities particularly with respect to cartilage formation in the pharyngeal arches. Eggs were collected and exposed to various concentrations of nicotine in order to determine the optimal sublethal close to use in comparison with control embryos. We also varied the time of treatment, both the total time of exposure and exposure at different phases of development, to determine if these variables affected the outcome. At specified time points, control and treated juvenile zebrafish were anethestized. fixed overnight in 4% paraformaldehyde before staining with Alcian blue to highlight formed cartilage. Preliminary evidence suggests that exposure to nicotine results in a delay in pharyngeal arch formation. Furthermore, a shortening and hypercontraction of the tail was noted in some instances following nicotine exposure. Further data collection will be required before firm conclusions can be made. Funding for this project was from the Young Harris College Undergraduate Research Initiative.
MEASUREMENT OF THE INTERACTIONS OF LOW ENERGY GAMMA RAYS WITH DENSE METALS FOR APPLICATIONS IN NUCLEAR CARDIOLOGY IMAGING**, J.L. Spradlin*, T.F. Lynam* and G.G. Passmore. Georgia Regents University, Augusta, GA 30912. Gamma camera imaging of myocardium perfusion with either TI-201 or Tc-99m is widely used for the detection of coronary artery disease (CAD). Myocardium perfusion imaging studies allow the clinician to differentiate between healthy and damaged myocardium based on the amount of accumulated radionuclide represented in the images. Both radioisotopes, Tc-99m and TI-201. can indicate the perfusion characteristics of the myocardium. However. TI-201 has the additional capability to indicate cardiac tissue viability. Simultaneous imaging of TI-201 stress and Tc-99m rest images would have the benefits of optimal diagnostic perfusion imaging and tissue viability signaling. However, there is a difficulty in trying to image the lower Tl-20 1 energy photons in the presence of the higher energy Tc-99m photons when using a standard lead collimator because interactions between the Tc-99m photons and lead create down-scatter in the TI signal region. The objective of this project is to measure the transmission and interactions of Tc-99m photons and TI-201 photons separately and simultaneously using high density metal discs (different from lead) as attenuators for application in gamma camera collimators. Correction of the down-scatter problem would allow the clinician to take advantage of a simultaneous dual-isotope approach. increasing the detectability of reversible myocardium defects. Methods include using NIST attenuation data to identify multiple high density metals that meet the criteria for reduced Tc-99m down-scatter with K-shell x-rays signals that are different from the T1-201 energy signal. Spectra for lead (Pb) have been collected. Continued analysis will be based on comparisons of spectra using multiple metallic attenuators.
IS LUTEINIZING HORMONE RECEPTOR (LHR) REDUCED IN MOUSE LEY-DIG TUMOR CELLS TREATED WITH PF0A?**, S.Y. Tadros* and J.D. Cannon, Georgia Regents University, Augusta, GA 30904. Perfluorooctanoic acid (PFOA) is a synthetic long-chain perfluorinated chemical used in numerous industrial and consumer products ranging from non-stick cookware to fire- and weather- resistant clothing. There is growing evidence suggesting that PFOA acts as an endocrine disruptor. Previous studies demonstrated that 24 h treatment of mouse leydig tumor (mLTC-1) cells with 100pM PFOA reduced human chorionic gonadotropin (hCG) - stimulated progesterone (P4) synthesis by greater than 16-fold compared to hCG-stimulatecl control, with no significant decrease in cell viability. It was hypothesized that PFOA-induced reduction in P4 synthesis may be clue to a decrease in expression of the luteinizing hormone receptor (LHR). To test this hypothesis. mLTC-1 cells were cultured with or without 100pM PFOA. cells collected, and RNA isolated and reverse transcribed. Primers for LHR and glyceraldehyde -3- phosphate dehydrogenase (GAPDH) were ordered and are currently being optimized against MgCl2 concentration, primer concentration, and cycle number. Once the primers have been optimized, mRNA levels will be determined using quantitative polymerase chain reaction (PCR). It is anticipated that LHR expression will be reduced in cells treated with PFOA.
REGENERATION OF RETINAL GANGLION CELLS IN EMBRYONIC ZEBRAF-ISH FOLLOWING GLUTAMATE EXCITOTOXICITY", Tiffany C. Goebel and Linda G. Jones, Young Harris College, Young Harris, GA 30582. The retina of the eye is composed of multiple layers of cells that work together to relay visual information to the brain. In most organisms, severe retinal damage can lead to permanent neuronal loss. Zebrafish (Danio rerio) have been shown to possess the ability to regenerate and replace retinal cells, specifically retinal ganglion cell (RGC) axons and photoreceptors from a population of progenitor cells, the Muller glia cells which are found in the retina of both zebrafish and mammals. In this study, we would like to determine whether RCG will regenerate following glutamate excitotoxicity and whether this response is limited to certain dose or time constraints of treatment or recovery. Immunohistochemistry of control and glutamate-treated embryonic zebrafish using an anti-acetylated tubulin antibody to newly formed neurons and their axons will be used to examine regeneration of the RGC and their axons. The Hu C/D antibody which labels RGC (in addition to a number of other cell types) will also be used to confirm regeneration of RGC (regardless of axonal regeneration). Understanding the regenerative processes exhibited by zebrafish could have important implications in potential treatments for ocular diseases such as glaucoma (found in mammals), in which RGC are destroyed. Funding for this project was from the Young Harris College Undergraduate Research Initiative.
ALPHA ENOLASE(EN01) EXPRESSION MIGHT BE A VITAL COMPONENT OF THE IMMUNE RESPONSE TO CHLAMYDIA TRACHOMATIS INFECTION**, Khamia Ryans, Camilla C. illsYusuf Omosun and Qing He, Clark-Atlanta University, Morehouse School of Medicine, Atlanta, GA 30314. Chlamydia trachomatis (C. trachomatis) is one of the most prevalent sexually transmitted diseases, in the United States with over one million Chlamydia infections reported in 2011. Chlamydia infection can cause pelvic inflammatory disease (PID), ectopic pregnancy, chronic pelvic pain, and infertility in women. A major challenge in Chlamydia vaccine effort is that Chlamydia infections do not only induce protective immunity but also immune mediated pathology. In this study we attempt to find the mechanism(s) underlying this immunopathology induced by Chlamydia. Preliminary studies in our lab have confirmed that IL-10 deficiency results in the rapid clearance of genital Chlamydia infection. Proteomic results indicated Alpha-Enolase (EN01) may be upregulated in C. trachomatis infected IL-1C) deficient cells. EN01 is a glycolytic enzyme associated with generating ATP and is present on the surfaces of neutrophils, monocytes and bacteria. To determine the mechanisms involved in the protection elicited in IL-10K0 mice we determined the levels of EN01 expression in BMDCs, from WT and IL-10K0 mice. infected for 1 and 2 hours with Chlamydia. Our study showed a significant increase in EN01 expression in IL-10K0 DCs compared to WT DCs. This result suggests that in IL-10K0 mice EN01 might be important in the early activation of the dendritic cells in the immune response against Chlamydia.
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|Publication:||Georgia Journal of Science|
|Article Type:||Conference notes|
|Date:||Mar 22, 2014|
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