3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Poster No 2 Poster The protective effect of ethyl Presentations pyruvate on liver following experimental partial hepatectomy. Poster No 1 T. Artış1, A. Abdülrezzak1, F. Mutlu1, How the cortisol level and brain A.Seda Artış2, K. Deniz3, Z. Yılmaz1 antioxidant activity change in chronic REM-sleep deprivation. Departments of General Surgery1, Physiology2 and Pathology3, Erciyes 1 1 2 A. Seda Artış , L. Şahin , A. Çetin , N. University Medical Faculty, Kayseri 1 1 Dolu , C. Süer 38039, .

1 Departments of Physiology and 2 Biochemistry , Faculty of Medicine, Following liver resection improving the hepatic University of Erciyes, Kayseri 38039, blood flow and controlling a series of Turkey. inflammatory process are nowadays gaining more importance. Ethyl pyruvate is secreted from cells and found in the extracellular fluid as In this study, it was aimed to measure the its anionic conjugate, pyruvate. It is a stabile oxidative damage and any changes that might form of pyruvic acid which is the last product in occur in antioxidant defense system in the brain glycolysis, and at the same time an effective together with plasma cortisol levels following 21 free radical scavenger. In this study we aimed days of REM-sleep deprivation. to investigate its antioxidant, anti-inflammatory, Rats were randomly assigned to one of three and protective effects on tissue against the free groups (n=10 for each): cage control (Group1), oxygen radicals and cytokines. tank control (Group2) or sleep-deprived Rats were equally divided into two as the (Group3). Rats were subjected to sleep control and the experimental groups. Each deprivation by multiple platform method. group was then subdivided into two (n=8 for Group2 rats were kept in a similar environment each). The experimental rats were given 40 as Group3 rats, but they were able to sleep. At mg/kg ip ethyl pyruvate after liver resection of the end of the 21st day the rats were sacrificed. 70%. Physiologic saline was administered to Plasma cortisol changes were determined control groups as 40 mg/kg ip. Rats were together with levels of malondialdehyde (MDA), th th sacrificed at the 48 and 72 hours to measure superoxide dismutase (SOD) and glutathione AST, ALT, PT, IL–6, IL–10, and TNF-α from peroxidase (GSH-Px) from the whole brain plasma and MDA from liver samples. homogenates. th th After 48 and 72 hours AST, ALT, PT, IL–6, No significant difference between the groups TNF-α, and MDA levels were found to be was found for cortisol levels. There was a significantly lower, whereas IL-10 levels was significant group effect for MDA, SOD and significantly higher in the experimental groups. GSH-Px activities in the whole brain Our findings demonstrated that ethyl pyruvate, homogenates. In the Group3 MDA levels were which inhibits secretion of IL-6 and TNF-α elevated, and the activities of SOD and GSH-Px cytokines and has anti-inflammatory and were significantly lower in comparison to the antioxidative effects, shows a protective effect Group1 or 2. In the Group2, MDA levels were on liver following partial hepatectomy. also elevated and the activities of SOD and

GSH-Px were also decreased in comparison to Poster No 3 the group1.

We speculate that an adaptation in the HPA axis occurs in chronic REM-sleep deprivation. Influence of vanadium Chronic REM-sleep deprivation caused a supplementation on oxidative stress decrease in the antioxidant defenses in brain in factors in the muscle of STZ-diabetic the present study. So, we think it is also rats noteworthy to investigate to what extend these changes occur in different brain regions.

1 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Ö. Kurt1, T. Yılmaz1, N. Özsoy1, S. Tunalı2, 1Marmara Univ. School of Medicine, Ayşe Can1, R. Yanardağ2 Microbiology Dept. 2Marmara Univ. School of Medicine, 1Department of Biochemistry, Faculty of Biophysics Dept. Pharmacy, University, 34116, Istanbul, TURKEY. Extremely low frequency electromagnetic fields 2Department of Chemistry, Faculty of (ELF-EMF) have been shown to change Engineering, , 34320, physicochemical properties of cell membranes. Istanbul, TURKEY. Cell surface hydrophobicity and cell surface charge have been implicated to biofilm formation. Staphylococcus epidermidis, are the In recent years the role of free radical damage predominant cause of implanted medical-device consequent to oxidative stress is widely related infections. The formation of adherent discussed in diabetic complications. In this multilayered biofilms on implanted devices is aspect, the protection of cell integrity by trace believed to be essential for the pathogenesis of elements is a topic to be investigated. S. epidermidis infections. Staphylococcus Vanadium is a trace element believed to be epidermidis was subjected to 50Hz, 1mT ELF- important for normal cell function and EMF and the change in hydrophubicity, development. The aim of the present study was membrane charge and biofilm formation was to investigate the effect of vanadyl sulfate evaluated. Quantitative determination of biofilm supplementation on the antioxidant system in production was performed by using a microtiter the muscle tissue of diabetic rats. Diabetes was assay. induced by intraperitoneal injection of Bacterial cell-surface hydrophobicity was streptozotocin (STZ, 65 mg/kg body weight) to determined by measuring the bacterial male Swiss albino rats. The rats were randomly adhesion to xylene in a xylene-water system. divided into 4 groups: Group I, control; Group II, Change in surface charge was determined by vanadyl sulfate control; Group III, STZ-diabetic elektrokinetic measurements. This work untreated; Group IV, STZ-diabetic treated with determined the effect of ELF-EMF on biofilm vanadyl sulfate. Vanadyl sulfate (100 mg/kg) formation and its correlation to bacterial surface was given daily by gavage for 60 days. At the hydrophobicity and surface electrical properties. last day of the experiment, rats which were fasted overnight were sacrificed, muscle tissue o was taken, homogenized in cold saline to make Poster N 5 a 10% (w/v) homogenate. Antioxidant enzymes, catalase (CAT), glutathione peroxidase (GPx), Effects of desflurane and vitamin C and superoxide dismutase (SOD) as well as E combination on element and oxidative myeloperoxidase activities and protein carbonyl stress levels in blood of operative content were determined in muscle tissue. The patients results were evaluated using an unpaired t-test and ANOVA variance analysis with the NCSS Berit G. Ceylan1, M. Nazıroğlu2, 3, A. C. statistical computer package. It was shown that Uğuz2, C. Barak4, B. Erdem1, L. Yavuz1 vanadium supplementation significantly decrease antioxidant enzyme levels which were 1Department of Anesthesiology and elevated in the muscle tissue of untreated diabetic group, showing that this trace element Reanimation, Medical Faculty, Suleyman Demirel University, Isparta, Turkey. could be used as preventive for diabetic 2 complications. Department of Biophysics, Medical Faculty, Suleyman Demirel University, Poster No 4 Isparta, Turkey. 3Neuroscience Research Center, Suleyman Effect of extremely low frequency Demirel University, Isparta, Turkey. electromagnetic fields on biofilm 4Department of Chemistry, Science formation of S.epidermitis; Institute, Balıkesir University, Balıkesir, correlation to surface charge and Turkey. hydrophobicity Volatile agents are frequently used in general 1 2 anesthesia practice without complications. B. Aksu , M.E. Cüce , Günseli Ayşe İnhan Clinical trials of desflurane on oxidative stress Garip2

2 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels and antioxidant systems are still limited (1). In a oxidative stress. Cell Biochem Funct 28;211- recent study we observed that desflurane 216. produced lipid peroxidation (LP) by decreasing vitamin E levels (2). In this study we aimed to Poster No 6 investigate effect of the new volatile anesthetic’s oxidant property on element, LP The oxidative stress effect of thermal and antioxidative response (as measured by water on natural distribution GSH-Px, GSH, -carotene, vitamins A, C and asphoedolus aestivus Brot. plant in E) in blood of patients. Forty adult patients are scheduled for elective surgery, ASA I or II Kestanbol Tuzla (Çanakkale-Turkey) Physical Status. The patients were randomly geothermal field and the effect of divided into two groups. First group was used water on the regional public as control and single dose of intravenous VCE were preoperatively given to the patients Berrak Damla Yağan1, A. Gönüz1 consisting second group. Tidal volume, ventilation frequency and intraabdominal 1 pressure were kept constant during the Department of Biology, Çanakkale 18 operation. Baseline values in venous blood Mart University, Çanakkale, Turkey samples were preoperatively taken from control and treatment group before VCE administration. Kestanbol-Tuzla is one of the most important The blood samples from the control and VCE geothermal fields of our country within the groups were also taken post operatively at the st th th boundaries of the town of Canakkale Ayvacik. 1 , the 24 and 72 hours of desflurane Area is quite low considering the diversity of exposure. Erythrocytes and plasma lipid flora is observed. Asphoedolus aestivus Brot. peroxidation (LP) levels in the postoperative st th th species are distributed in an area close to the period at the 1 , 24 and 72 hours of control source. The observations show that; the plant group were higher in baseline values while st th has lost its ability to produce its flowers, so vitamin E levels at the 1 and 24 hours, vegetative reproduction occurs to depends on erythrocyte glutathione peroxidase (GSH-Px) st th th stress effect. The plant that has resistance activity at the 1 , 24 and 72 hours in against osmotic and salt stress, so that situation postoperative period of control were lower than suggested us to oxidative stress related cellular in preoperative period of control. Erythrocyte damage cause the inhibit flowering period. In GSH-Px activity, plasma vitamin A, C and E st th th this study; plants growing in the geothermal levels at the 1 , 24 and 72 hours were higher area were compared to a pilot plant that grows treatment groups than in control whereas healthy plants in the region was chosen as an erythrocyte and plasma LP levels were lower in example in terms of biochemical properties. treatment group than in control. Erythrocyte and Analysis carried out in this study, the plants plasma reduced glutathione, plasma -carotene showing the geothermal distribution accumulate and serum copper, zinc, selenium, aluminum, Mn and Fe relatively high amounts then pilot iron, magnesium and calcium levels did not plant, thus these elements cause oxidative different between pre- and postoperative stress and also that results supported by periods. In conclusion, VCE combination multiple literature. This plant has been used as prevents the desflurane-induced vitamin E and fodder crops and folk remedies in the region; glutathione peroxidase decrease to strengthen and that cause the transfer of stress agent from the antioxidant levels in the blood of operative plant thought the other organisms including patients. human by food chain. Although soil and water is transported to agricultural land for agricultural References activities; depends on geographical position 1. Dikmen B, Unal Y, Pampal HK, Nurlu N, and seasonal fluctuations the area is Kurtipek O, Canbolat O, Ozoğul C, Kavutcu M. contaminated. As a general opinion; the (2007). Effects of repeated desflurane and increasing cancer cases in this region has close sevoflurane anesthesia on enzymatic free relation with the current situation. radical scavanger system. Mol Cell Biochem 294:31-36. Poster No 7 2. Eroglu F, Yavuz L, Ceylan BG, Yilmaz F, Eroglu E, Delibas N, Naziroğlu M (2010). New Adaptation of rat gastric tissue against volatile anesthetic, desflurane, reduces vitamin indomethacin toxicity E level in blood of operative patients via Beyzagul Polat1, H. Suleyman1, H.H. Alp2.

3 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

1Ataturk University, Faculty of Medicine, on glutathione peroxidase activity in Jurkat cell Department of Pharmacology, 25240 line has been investigated. Jurkat cell line (E- Clone E6) is grown in RPMI1640 medium and Erzurum, Turkey. 6 the cell number is adjusted to 1x10 cells in 2Ataturk University, Faculty of Medicine, each well of the plate. The cells were incubated with different doses of melatonin: 1mM, 2mM, Department of Biochemistry, 25240 5mM and 10mM at 37C in 5%CO2 incubator Erzurum, Turkey. for 24 hours. Then the GSH-Px activity was measured by microplate reader This study investigated whether adaptation spectrophotometrically. GSH-Px activities were occurred after various dosages of repeated diminished in a dose-dependent manner (chronic) indomethacin in rats to the gastro compared with the control group, however the toxic effects of indomethacin. It also examined dose of 5mM were more effective than all the whether the adaptation was related to oxidative other groups to lower the activity. As a result, DNA damage in gastric tissue. To illuminate the 5mM melatonin is found to be the most effective adaptation mechanism in the gastric tissue of dose to decrease the oxidative damage in rats given various dosages of chronic Jurkat cell lines. And this study is a pre-trial to indomethacin, the levels of oxidative DNA 5 investigate further effects of melatonin on damage (8-OHd Gua/ 10 Gua) were oxidatively-stressed Jurkat cell lines. measured. Results were compared to the control group, which received a 25-mg/kg single Poster No 9 dose of indomethacin, and the role of oxidative DNA damage in the adaptation mechanism was Store-operated calcium entry evaluated. The average ulcer areas of gastric tissue of the 1-, 2-, 3-, 4-, and 5-mg/kg dosages increases in shTRPC1-expressing of chronic indomethacin given to rats were vector-transfected hepatocellular 19.5±3.7, 12.5±3.3, 10±5.2, 4.5±3.6, 8.6±2.4, carcinoma cell line and 9.5±2.1 mm2, respectively. This rate was 2 measured as 21.3±2.6 mm in the control Cigdem Selli1, Y. Erac 1, B. Kosova2, E. group. Consequently, after various dosages of Erdal3, M.Tosun1 repeated (chronic) indomethacin administration in rats, it was observed that a clear adaptation 1Departments of Pharmacology, Faculty of developed against gastric damage and that 2 gastric damage was reduced. The best Pharmacy and Medical Biology, Faculty of 3 adaptation was observed in the gastric tissue of Medicine, , Medical Biology, the 3-mg/kg chronic indomethacin group. In Faculty of Medicine, Dokuz Eylul parallel with the damage reduction oxidative University, 35100, Izmir. 5 DNA damage (8-OHd Gua/ 10 Gua) were reduced. This circumstance shows that We previously observed that TRPC1 and oxidation play a partial role in the adaptation TRPC6 expression levels were reciprocally mechanism. altered along with potentiated adrenergic 1 o receptor responses in aging rat thoracic aorta . Poster N 8 Furthermore, TRPC6 and store-operated Ca2+ were upregulated in TRPC1-silenced A7r5 Glutathione peroxidase activities in embryonic rat aortic smooth muscle cell line2. Jurkat cell line: The antioxidant Therefore, the purpose of this study was to effects of melatonin investigate any regulatory role of transient 1 2 receptor potential canonical 1 (TRPC1) in store- Canan Aslan , M.İnanır , G.Ş. 2+ 3 2 operated Ca entry in hepatocellular carcinoma Karatoprak , M.B. Yerer-Aycan cell (HCC) lines. For this purpose, HCCs were University of Erciyes, Faculty of cultured in RPMI-1640 and transiently Pharmacy, Department of transfected with shRNA-expressing vector Biochemistry1, Pharmacology2, (pSUPERIOR) targeting TRPC1 mRNA for in Pharmacognosy3 vitro post-transcriptional gene silencing. After 72 h incubation period, total RNA and protein As an endogenous indolamin melatonin is samples were isolated. To determine the known to be potent antioxidant in addition to its changes at transcriptional and protein level, regulatory roles in many functions via biological quantitative real-time PCR (LightCycler 2.0, rhythm. In this study, melatonin and its effects Roche Diagnostic) and immunoblot analyses

4 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels were performed, respectively. Changes in of cells with 1 mM trolox, an α-tocopherol intracellular Ca2+ levels were monitored via front analogue which works as an antioxidant, for 30 surface spectrofluorimetry (PTI QM8/2005) in minutes, reduced significantly both caspase fura-2-loaded cells grown on coverslips2. In activation. Similar results were obtained when shTRPC1-coding vector-transfected HCCs, the cells were pretreated with 1 mM N-acetyl- TRPC1 protein expression significantly cysteine for 30 min, a derivative of cysteine decreased (p<0.05) without any change in which is well-known for playing a role as an mRNA levels. Furthermore, Ca2+ released from antioxidant. endoplasmic reticulum (p<0.05) and entered via Our results suggest that TNFα-induced store-operated Ca2+ both induced by apoptosis in human myeloid cell line HL-60 is cyclopiazonic acid, a selective sarco- dependent of intracellular ROS production. endoplasmic Ca2+ ATPase blocker, were significantly increased (p<0.01). These results Supported by Ministery of Science and suggest that TRPC1 may play a regulatory role Innovation grant BFU 2007-60091. in store-operated Ca2+ entry in HCCs. Despite the decreased TRPC1 protein levels, no Poster No 11 apparent change in mRNA levels may be due to the presence of alternative mRNA spliced Nutritional and functional isoforms of TRPC1 in cancer cells. characterization of Crimson globe This study was supported in part by The Scientific and Technological Research Council Japanese plums: Antioxidant effect of Turkey (TUBITAK, SBAG-3953 to MT; of their consumption. BIDEB-2211 to CS) and Ege University (BAP 08ECZ009, MT). D. González1, B. Velardo2, M. Garrido1, D. González-Gómez2, M. Lozano2, M.C. Poster No 10 Ayuso3, C. Barriga1, S.D. Paredes1, 1 TNF-α-induced apoptosis in human A.B. Rodríguez myeloid HL-60 cells is dependent of 1 ROS generation Department of Physiology (Neuroimmunophysiology and D. González-Flores, C. Barriga, A. B. Chrononutrition Research Group), Rodríguez, J. A. Pariente Faculty of Science, University of Extremadura, 06006, Badajoz, Spain. 2 Department of Physiology, Technological Institute of Food and Neuroimmunophysiology and Agriculture of Extremadura, 06071, Badajoz, Spain. Crononutrition Research Group, 3 University of Extremadura, Badajoz, Agricultural Engineering School, 06006, Spain. University of Extremadura, 06007, Badajoz, Spain In the present work the authors investigated whether oxygen reactive species (ROS) play a Plums are considered a source of role in the activation of extrinsic pathway phytochemical compounds with beneficial apoptosis induced by TNFα in human leukemia health effects. The study was aimed at cell line HL-60. characterizing nutritionally and functionally We have analysed ROS generation and Japanese plums (Prunus salicina Lindl. cv. caspase -3, and -8 activity by fluorimetric Crimson globe) and evaluating the antioxidant methods using the fluorescent dye effect of a diet enriched in these fruits in young dihydrorhodamine 123 and specific fluorigenic (20±10 yr-old), middle-aged (45±10 yr-old) and substrate, respectively. We also determined cell elderly (75±10 yr-old) individuals. Participants survival using the MTT-method. consumed 200 g of plums twice a day (as the Our results show that the treatment of HL-60 lunch and dinner desserts) for 5 days. First-void morning urines were collected before treatment cells with TNFα (100 ƞg/ml) produces a time- (basal values), the immediate day after the last dependent increase of intracellular ROS ingestion of plums (assay), and one day production followed as well as caspase -3 and - afterwards (post-assay). Urinary 6- 8 activation and finally cell death. Pretreatment sulfatoxymelatonin and total antioxidant

5 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels capacity were measured by means of with oxidative stress. In this study, it was aimed commercial ELISA and colorimetric assay kits, to determine the effect of water-pipe respectively. Nutritionally, Crimson globe plums consumption on the activities of lipid were shown to contain lower carbohydrate peroxidation and antioxidant enzyme activities. values than the reported levels for other 24 water pipe users (experimental group) and cultivars. From a functional point of view, 25 nonsmokers (control group) were included to detectable levels of serotonin, phenolic the study (nT = 49). All subjects were selected compounds and anthocyanins were found. among the people without a habit of smoking or Melatonin was detected but not quantifiable. any other use of tobacco. Catalase and The consumption of plums induced a significant superoxide dismutase (SOD) activities were increase in the participants’ urinary 6- investigated to determine the antioxidant statue sulfatoxymelatonin and total antioxidant and MDA level for lipid peroxidation. All blood capacity levels in relation to their corresponding samples were collected from both groups prior basal and post-assay values. Japanese plums to taking any food in the morning and of the cultivar Crimson globe may be seen as processed to separate serum. Catalase activity source of antioxidants with potential properties was determined utilizing the method of Aebi. for counteracting oxidation. SOD activity was found by the method of Sun et al. Finally, MDA levels were investigated using Funded by INTERREG 0318-RITECA-4E; Ohkawa’s method based on thiobarbituric acid Programa POCTEC, 053/09. S.D. Paredes was reaction. The data obtained for two groups were beneficiary of a grant by Junta de Extremadura- compared statistically using SPSS v16.0 and Fondo Social Europeo (REI09009). *Both Independent T-Test. In conclusion, catalase authors contributed equally to this work. and SOD activities were found to be lower and MDA level was determined to be higher in Poster No 12 water-pipe smokers than the control group (p<0.05). Therefore, it can be suggested that Effects of water pipe smoking on water pipe consumption reduces antioxidant oxidative stress enzyme activities and increases lipid peroxidation. In turn, it induces oxidative stress 1 2 and triggers many diseases as with cigarette Ebru Derici Eker , M. Berköz , N.Ö. smoking. 1 2 Şahin , E. Jaroch

Mersin University, Faculty of Pharmacy, References 1Department of Pharmaceutical 1. Burlakova EB, Zhizhina GP, Gurevich SM, 2 Fatkullina LD, Kozachenko AI, Nagler LG, Technology, Department of Zavarykina TM, Kashcheev VV. Biomarkers of Pharmaceutical Biotechnology, oxidative stress and smoking in cancer patients. Yenisehir Campus, Mersin 33169, J Cancer Res Ther. 2010; 6: 47-53. Turkey. 2. Nakkash R, Khalil J. 2010. Health warning labelling practices on narghile (shisha, hookah) waterpipe tobacco products and related Water-pipe smoking (WPS) has been practiced accessories. Tob Control. 2010; 19:235-239. extensively for almost five hundred years. It is a common habit in Turkey, Middle Eastern countries and Asia. In recent years, there has Poster No 13 been a revival of WPS, notably among adolescents. The aerosol of water pipe smoke Calcium impression on proliferation and has been reported to have a composition of carbon monoxide, nicotine, tar, and heavy phenotype changes of differentiated metals. Therefore, it can not be stated that cardiomyocytes in vitro water pipe smoking sets a safe alternative to 1 1 1 cigarette smoking. Cigarette smoking induces Esin Akbay , G. Ayaz , H. Sevim , E. M. lipid peroxidation resulting in suppression of Albayrak1, Ö. A. Gürpınar1, M. A. Onur1 antioxidant system. Although there is intensive research conducted on the effect of cigarette 1Hacettepe University, Faculty of Science, smoking causing oxidative damage, no Department of Biology, 06800 Beytepe, significant study was found in literature Ankara, Turkey investigating the relation of water pipe smoking

6 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Intracellular calcium (Ca2+) signaling plays a ADR were injected intraperitoneally for 8 days role in cardiac physiology and modulates (ADR and Se+ADR groups, 4 mg/kg, every cardiac gene expression. Changes in alternate day); Se were injected ip for 21 days intracellular Ca2+ levels play an important role in (Se and Se+ADR groups, 0.15 mg/kg, every the cardiac myocyte physiology. Although the day) and Saline were injected ip for 21 days activities of intracellular Ca+2 on the physiology (Control groups, everyday). Mitochondrial and metabolic functions of heart cells have membrane potential (MMP), ATP levels were been studied extensively, the extracellular Ca+2 determined with Cambrex and Cayman kits, activities has been analyzed scarcely. The aim respectively. Cytosolic, mitochondrial total of this study was to investigate the effects of antioxidant (TAS) and oxidant status (TOS) various Ca+2 concentrations on the proliferation were measured with Rel assay kit. The and phenotype regulation of cultured oxidative stress index (OSI) was defined as cardiomyocytes. Differentiated cardiomyocytes TOS to TAS ratio for both cytosol and were cultured in media with different mitochondria. concentrations of Ca2+ supplements in 24-well. The cultures were maintained at 37˚C in a ADR administration caused oxidative stress, humidified atmosphere of 5% CO2 and 95% O2. showing that increased total oxidants and The cell proliferative activity was evaluated after decreased total antioxidants in cytosol and 24, 48, and 72 h, counted with hemositometer mitochondria. Selenium normalized oxidative by using trypan blue dye exclusion method. stress induced by ADR treatment. ADR Cellular morphology of live cultures was decreased the ATP level in myocyte observed by inverted microscope. mitochondria. Coadministration Se with ADR Results of analysis indicated that, at a calcium improved ATP levels. ADR decreased the concentration of 1, 8 mmol L-1 enhance cell mitochondrial membrane potential and, proliferation rate as from second day. But the coadministration Se with ADR improved the other doses of calcium showed toxic effects on mitochondrial membrane potential. cell morphology and proliferation. Our The results provide evidence for the role of consequence means that the differentiated selenium in renoprotection against ADR cardiomyocytes proliferative activity may nephropathy in rats possibly by changing regulate by intracellular calcium, whose oxidant/antioxidant balance and modulating concentration depends on the inflow of ATP production and apoptosis. extracellular calcium through various ionic channels without involvement of intracellular 1.Durrin OD and Kuloglu T.2009. Effects of calcium stores. Vitamin D3 (Cholecalciferol) on Adriamycin- Induced Nephrotoxicity. Renal Failure, 31:400– Poster No 14 405. 2. Oteki T, Nagase S, Shimohata H, Hirayama Protective effect of selenium on A, Ueda A, Yokoyama H, and Yoshimurat adriamycin-induced mitochondrial T.2008. Nitric oxide protection against adriamycin-induced tubulo interstitial injury. damage of rat renal tissue Free Radical Research, 42(2): 154-161.

1 1 N. Dursun , Eylem Taşkın , M. B. Yerer Poster No 15 Aycan2 Oxidative stress in the blood of farm Department of Physiology1 and workers following intensive pesticide Pharmacology2, Faculty of Medicine exposure and Pharmacy, University of Erciyes, 1 2 3 Serdal Ö. , Fatih Gültekin , A.N. Kişioğlu , Kayseri-TURKEY. E. Küçüköner 4 1Süleyman Demirel University, Faculty of It is well known that oxidative stress is related Medicine, Blood Bank, 32260, Turkey to the pathogenesis of adriamycin (ADR) 2Süleyman Demirel University, Faculty of nephropathy1, 2. The aim of the study was to show the protective effect of selenium (Se) on Medicine, Department of Biochemistry, 32260, Turkey ADR-induced mitochondrial–mediated 3 apoptosis in nephropathic rats. Süleyman Demirel University, Faculty of Medicine, Department of Public Health, 32260, Turkey

7 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

4Süleyman Demirel University, Faculty of “paclitaxel” to rats. Also we know that β-lactam Engineering and Architecture, Department antibiotics increase EAAT’s expression via of Food Engineering, 32260, Turkey gene activation. This prompted us to try to heal paclitaxel induced allodynia by increasing The aim of this study was to evaluate oxidative EAAT’s expression via β-lactam antibiotics. stress in workers using organophosphate, Two different groups of rats were injected with synthetic pyrethroid and carbamate pesticides paclitaxel to induce allodynia and another one in their farm. In this survey, 94 farm workers was injected with distilled water (exposed to pesticides at least 5-year during intraperitoneally. The pilot studies we did show that pain thresholds markedly reduce and the apple and cherry production) and 45 control th subjects (living in the same region with no allodynia becomes most severe on 30 day after the first paclitaxel injection we started exposure to pesticides) were included to the th study. Lipid peroxidation level, catalase, Penicillin G treatment to first group on 30 day superoxide dismutase and glutathione and continued for 10 days. 30 days after the peroxidase activities in erythrocytes were first Penicillin G injection we observed that analysed as biomarkers of oxidative stress. In Penicillin G increase the decreased pain addition, the acetylcholinesterase activity was thresholds (from 75g to16, 5g±5,8to 35g±14, 4) measured as a biomarker of toxicity. As a result significantly by comparison paclitaxel injected of the analysis, lipid peroxidation level and group treated with distilled water. This study activities of antioxidant enzymes, catalase and shows that glutamate transporter or receptors superoxide dismutase significantly increased may be potential targets to treat neurological (p<0.05). Acetylcholinesterase activity did not disorders like allodynia, hepatic show any significant changes between the two encephalopathy. groups (p>0.05). Consequently, chronic o exposure to pesticides may negatively affect Poster N 17 antioxidant system. Baseline and salt-stimulated Poster No 16 paraoxonase, arylesterase and platelet activating factor The antiallodynic effect of Penicillin G in acetylhydrolase activities and their paclitaxel induced mechanoallodynia relationship with oxidized LDL in

polycystic ovary syndrome A. Hacimuftuoglu1, Fatih Saruhan2, D. 1, 1 Cetin H. Suleyman 1 1 1 S. Uysal , Ferah Armutcu , A. Department of Pharmacology, Faculty of 2 2 Medicine, Ataturk University, Erzurum, Çarlıoğlu , F. Karakurt 1 Turkey Department of Biochemistry and 2Department of Pharmacology, Faculty of 2Department of Endocrinology, Fatih Pharmacy, Ataturk University, Erzurum, University Medical Faculty, TURKEY Turkey Polycystic ovary syndrome (PCOS) is Insufficient selectivity of anticancer agents for characterized by hyperinsulinemia and tumor cells makes them toxic for both tumor hyperandrogenemia. These metabolic and healty body cells. If nerves are affected by abnormalities in women with PCOS cause the anticancer agents neuropathy may occur and increase in risk of some diseases with high patients may feel pain for even a light touch mortality and morbidity, including stimuli which are perceived as innocuous cardiovascular disease and obesity. We studied normally. This condition is named as “allodynia” a total of 31 patients with polycystic ovary and often resistant to standard analgesics. We syndrome and 30 healthy comparison subjects. know that expression of glutamate transporters Baseline and salt-stimulated Paraoxonase 1 (EAAT’s), responsible for transporting (PON1), aryl esterase ARE, and platelet excitotoxic major neurotransmitter glutamate activating factor, acetylhydrolase (PAF-AH) thought to be responsible for a variety of activities were determined in control and neurological disorders from synaptic cleft to patiens. PON1192Q/R phenotypes also were astroglias to avoid increasing its concentrations determined by using results of ARE activity. to excitotoxic levels in synaptic cleft, down PAF-AH activity and oxidized LDL levels were regulate after administration of anticancer agent significantly higher in patients with PCOS than

8 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels in control (p<0.001, and p<0.003 respectively) effect of the calcium on the growth of MSCs, although PON1 activity was significantly 10⁴ cells/ml were seeded into the 96-well (p<0.002) lower than in the control group. ARE culture plates. Different concentration of activity was also significantly higher (p<0.007) calcium (1.8 mM, 3.6 mM and 7.2 mM) were in patients with PCOS, whereas there was no used in the experiments. After 24, 48 and 72 significant difference in salt-stimulated PON1 hours, the cell proliferative activity and viability activities. In PCOS group the phenotypic of MSCs were determined by MTT assay. Our distributions of paraoxonase compared with the study showed that the 1.8 mM calcium controls were QQ 45.2% - 46.7%, QR 48.4% - concentration enhanced cell proliferation in 53.3%, and RR 6.4% - 0%, respectively. These MSCs and it was time dependent. However, phenotypes (QQ, QR, RR) and plasma PON1 higher concentration of calcium (7.2 mM) would levels were statistically (p<0.05) significant decrease the viability of MSCs. lower in patients with PCOS than in the control group. Oxidized LDL levels and ARE activities Poster No 19 were elevated; on the contrary baseline and stimulated PON1 were reduced in patients with Sport climbing causes oxidative polycystic ovary syndrome. These results stress in sedentary individuals. suggest that these parameters are closely related, and could be suitable biomarkers for 1 2 3 the evaluation of the risk of atherosclerosis in Gökhan Cesur , E. Atay , S. Öğüt , M. patients having polycystic ovary syndrome. Polat3, H. Oruç2.

o Poster N 18 1Department of Physiology, 2 3 The effect of calcium on the Department of Sport Sciences, Blood proliferation and viability of rat Bank, University of Süleyman Demirel, mesenchymal stem cells Isparta, Türkiye.

Sport climbing is a worldwide sport with Gamze Ayaz , E. Akbay, H. Sevim, particular physiological and physical demands. M.E. Albayrak, Ö.A. Gürpınar, M.A. The topic of exercise-induced oxidative stress Onur has received considerable attention in recent years. This study was aimed to investigate the , Faculty of effects of sport climbing on oxidative stress, antioxidant levels and hematologic parameters. Science, Department of Biology, 06800 Seven voluntary male students whose average Beytepe, Ankara age is 20,57 ± 1,27 years old, participated in this study. None of the subjects had performed Several in vitro studies have shown that regular exercise. These subjects were climbed calcium is involved with a range of cell functions 28±3.13 times during eight weeks. Blood including attachment, tissue morphology, signal samples were collected at rest, 24h before, and transduction and proliferation. Different 24 h after climbing protocols to analyze total concentrations of calcium in the culture medium oxidant status (TOS), total antioxidant status may change the permeability of plasma (TAS) and hematologic parameters membrane and intracellular signaling systems (Hemoglobin (Hb), hematocrit (Hct), platelet significantly (Osipenko et al., 2007). The aim of (Plt) and leukocyte (Wbc)). Resting heart rates this study was to determine the effect of were measured at the same time. According to increasing extracellular calcium ion our findings, climbing increased TOS (1,49 ± concentrations on growth and viability of 0,04, 1,52 ± 0,03), decreased TAS (1,54 ± 0,06, mesenchymal stem cells (MSC) in vitro. MSCs 1,51 ± 0,06) and significantly increased were isolated from 5-6 week old rat’s tibias and oxidative stress index (TOS/TAS) (0,97 ± 0,01, femurs. The cells were primarily cultured in 0,99 ± 0,02) (p<0.05). Only Hct levels (45,01 ± DMEM/Ham’s F-12 media supplemented with 1,42, 44,80 ± 1,39) of hematologic parameters 15% fetal bovine serum (FBS) and 0.5% increased after climbing protocols (p<0.05). penicillin /streptomycin in a humidified There has been no statistically relation among atmosphere of 5% CO₂ , 95% O₂ at 37 C. The the other parameters. These data demonstrate media was changed every 3 or 4 days and the that sport climbing leads to increased plasma cells were subcultured two times before oxidative stress in sedentary individuals and employing them in the study. To evaluate the negatively affect the athletes’ performance.

9 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

References stress for 3 days. Rats were injected with either saline or oxytocin antagonist (atosiban, 1mg/kg) intraperitoneally for 3 days following stress. On 1. Araneda OF, García C, Lagos N, Quiroga G, rd Cajigal J, Salazar MP, Behn C. 2005. Lung the 3 day, following decapitation colonic and oxidative stress as related to exercise and gastric tissues were examined to measure altitude, Lipid peroxidation evidence in exhaled myeloperoxidase (MPO) activity and superoxide breath condensate:a possible predictor of acute dismutase (SOD), catalase (CAT), glutathione mountain sickness. Eur J Appl Physiol. 95(5- (GSH) and malondialdeyde (MDA) levels. The 6):383-390. data were analyzed using ANOVA. Stress- 2. Erel O. 2004. A novel automated method to induced increased colonic MPO activity, gastric measure total antioxidant response against and colonic MDA levels, as well as decreased potent free radicals reactions. Clin Biochem; colonic GSH levels were reversed by exercise 37:112–9, (p<0.05), while reduced gastric SOD level in the 3. Erel O. 2005. A novel automated colorimetric ovariectomized rats was increased by exercise. method for measuring total oxidant status. Clin On the other hand, stress-induced gastric and Biochem; 38:1103–1111. colonic injury, which was aggravated by ovariectomy, was partially alleviated by 4. Merrells KJ, Friel JK, Knaus M, Suh M. 2008. atosiban (p<0.01). The results demonstrate Following 2 diet-restricted male outdoor rock that stress-induced gastric and colonic injury climbers: impact on oxidative stress and appears to be ameliorated by exercise in the improvements in markers of cardiovascular risk. presence of ovarian hormones, which may be Appl Physiol Nutr Metab. 33(6):1250-1256. acting though increased antioxidant enzymes.

Moreover, the findings show that oxytocin is an

important mediator in the regulation of HPA Poster No 20 axis.

The effect of chronic swimming exercise on several target tissues of the stressed References 1. Taylor SE, Gonzaga GC, Klein LC, Hu P, female rats with or without functioning Greendale GA, Seeman TE: Relation of ovarian hormones: the role of the HPA oxytocin to psychological stress responses and axis hypothalamic-pituitary-adrenocortical axis activity in older women. Psychosom Med. 2006; Gülsün Memi1, S. Özgür2, A. Yarat2, B.Ç. 68(2):238-45. Yeğen1 2. Park E, Chan O, Li Q, Kiraly M, Matthews SG, Vranic M, Riddell MC: Changes in basal 1 School of hypothalamo-pituitary-adrenal activity during exercise training are centrally mediated. : Am J Medicine, Department of Physiology, Physiol Regul Integr Comp Physiol. 2005; Haydarpaşa, Istanbul 289(5):R1360-71. 2Marmara University School of 3. Marques CM, Nascimento FA, Mandarim-de- Dentistry, Department of Biochemistry, Lacerda CA, Aguila MB: Exercise training Nişantaşı, Istanbul attenuates cardiovascular adverse remodeling in adult ovariectomized spontaneously hypertensive rats. Menopause 2006 13(1):87- When a threat to homeostasis occurs, 95. hypothalamus-pituitary-adrenal (HPA) axis is activated as a regulatory mechanism of the stress response, where oxytocin plays an o important role. The present study was aimed to Poster N 21 investigate the role of oxytocin receptors in stress-induced inflammation in intact or Zinc deficiency and 8-hydroxy-2- overiectomized rats that were sedentary or deoxyguanosine levels as a marker exercised prior to chronic stress exposure. of DNA oxidation in children Female Sprague-Dawley rats underwent ovariectomy (n=30) or sham operation (n= 30). Halil Ibrahim Atasoy1, G. Bugdayci2, Then, subgroups were divided as sedentary or 1 swimming groups (9 weeks). Each group was A.P. Uyan . further divided as “non-stress” group or “stress” group exposed to psychological electric shock

10 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Abant Izzet Baysal University, Izzet 1 Department of Medical Biology, Baysal School of Medicine, Department Medical Faculty, Yuzuncu Yil of Paediatrics1, Department of University, Van, Turkey Biochemistry2, Bolu, Turkey. 2 Department of Biology, Science & Art Faculty, , Sanliurfa, Zinc deficiency is known to result in retardation Turkey of growth, immune and cognitive deficits in children. Zinc has antioxidant properties. Coal dust causes lung diseases in occupational Deficiency may cause imbalance in the exposure. Reactive oxygen species have been prooxidant-antioxidant status. We aimed to implicated in the pathogenesis of its toxicity. In demonstrate DNA oxidant damage in zinc this study serum enzymes, lipid profile and deficient children by calculating 8-hydroxy-2- other biochemical values with deoxyguanosine as a marker of genetic oxidant/antioxidant status in whole blood and damage. serum of central heating system workers Serum zinc levels were calculated by (CHSW) (the persons who are responsible for colorimetric method in 190 male and 274 heating the apartments with coal) were female children ages of which were between 5 determined to reflect the cell injury. Blood to 15 years. Out of 466 children sex and age samples were obtained from CHSW (n= 25) matched 35 children with zinc deficiency (serum and healthy individuals (n= 25). All values were zinc levels less than 70 mcg/L) and control measured in whole blood and serum. ANOVA plasma 8OH2dG levels were calculated by was used for the estimation of statistical data. enzyme immunoassay. In the group of CHSW, creatinine, ferritin, alanin Work-up of serum zinc levels 35 healthy age aminotransferase, aspartate aminotransferase, and sex matched zinc deficient children and creatine phosphokinase, gamma glutamil control children showed statistically significant transferase, lactate dehydrogenase and increase of 8-OH-2-dG levels in zinc deficient glutathione reductase activities as well as group (p < 0.05). 8-OH-2-dG levels in zinc triglyceride, very low density lipoprotein, protein deficient and non-deficient children were 1,94 ± carbonyl and malondialdehide were significantly 0,47 and 1,53 ± 0,45 ng/mL (mean ± SD) higher, while transferrin, high density lipoprotein respectively. and catalase activities were lower than the Oxidative stress affects cellular lipids, proteins group of healthy individuals. This result is and nucleic acids. 8OH2dG is valuable marker consistent with hypothesis that respirable coal frequently used for revealing damage to nucleic dust generates lipid and protein oxidation and acids. When nucleic acids are oxidized point induces leakage of serum enzymes by cell mutations and chromosomal breaks could form damage. It also leads to imbalance in which might result in carcinogenesis. We used antioxidant defense system, lipid profile and 8-OH-2-dG levels as markers of DNA damage other biochemical parameters. in zinc deficient children and showed a significant increase compared to control children which might pose them to worse Poster No 23 clinical consequences. Zinc deficiency in children should be screened and corrected if consequences are being Gastroprotective effect of small avoided. centaury (Centaurium erythraea) on aspirin induced gastric damage in Poster No 22 rats

Increased occupational coal dust Y. Tuluce 1, Halil Ozkol1, I. Koyuncu2, H. toxicity in blood of central heating Ine2 system workers 1 Department of Medical Biology, Y. Tuluce 1, Halil Ozkol1, I. Koyuncu2, H. Faculty of Medicine, Yuzuncu Yil Ine2 University, Van, Turkey

2 Department of Biology, Faculty of Science & Art, Harran University, Sanliurfa, Turkey

11 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

hypoxic conditions on nerve conduction Small centaury (Centaurium erythraea) (family: velocity. Gentianaceae), a medicinal plant is used owing to its digestive, stomachic, tonic, depurative, This study was approved by the Pamukkale sedative and antipyretic properties in folk University Ethics Committee of Animal Care medicine. In this study, the gastroprotective and Usage. A total of 35 Sprague Dawley male effect of small centaury (SC) extract was rats, were divided into 5 groups: Control (K), investigated in the acetylsalicylic acid (ASA)- hypoxia (H), hypoxia + DHA (HD), hypoxia + induced gastric ulcers in rats at a dose of 200 exercise (HE), hypoxia + exercise + DHA mg/kg body weight. Twenty one Sprague- (HED). All of the rats, with the exception of K Dawley albino rats were divided into three group, exposed to hypoxia for 28 days. A groups of 7 rats each as follows: (1) the control treadmill exercise was performed as 1.8 km/h, group; (2) the acute ASA treated group and (3) 0% incline, 20 min/day in exercise groups. DHA ASA plus SC group. At the end of the 4 h drug were applied by gavage (36 mg /kg/day) to HD administration, ulcer index, oxidant and and HED groups for every day. At the end of antioxidant levels were measured and the experimental period, it was removed sciatic compared between the groups. The percentage nerves of anesthetized rats. Nerve conduction of lesion area to total gastric surface area (ulcer velocity was measured in sciatic nerve. One index) was significantly reduced (77 %) in ASA Way ANOVA and Post Hoc Tukey tests were plus SC group as compared with the acute ASA used for the differences between groups, p treated group. The oral administration of ASA values <0.05 accepted as statistically decreased catalase (CAT), reduced glutathione significant. (GSH), and increased lipid peroxidation (LPO) levels. Although myeloperoxidase (MPO) Nerve conduction velocity of H group was activity was increased by ASA, it was found to decreased compared to group K. Nerve be lower in the ASA plus SC group. GSH and conduction velocity of HE, HD and HED groups Vitamin A levels were determined higher in the were increased compared to K and H groups. ASA plus SC group compared with ASA group. These results suggest that SC extract protects This study it was shown that exercise and against ASA-induced damage due to its anti- dietary intake of DHA can be used to reduce oxidizing activity. deleterious effects on nerve conduction velocity of hypoxic conditions. Poster No 24 This study was supported by Pamukkale The effect of exercise and University Research Fund (2009-SBE-001) docosahexaenoic acid on the nerve Poster No 25 conduction velocity in normobaric hypoxia Antimicrobial and Antioxidant Activities of Cynanchum acutum, Cionura erecta, H. Ali Erken1, O. Genç1, G. Erken1, R. Trachomitum venetum subsp. Çolak2 sarmatiense Grown Wild in Turkey

1 1 1 1 1Pamukkale University, Medical Hülya Demir , E. Ergin , L. Açık , M. Vural

Faculty, Department of Physiology. 1 2Pamukkale University, School of Sport Atatürk Vocation Training School, , 06500-Ankara, Turkey Sciences and Technology

Wild plants have been reported to have antioxidant properties for centuries. Indigenous Deleterious effects of hypoxia exposure on plants have been used in herbal medicine for many nervous system functions are known. curing various diseases (Sokmen et al). There are studies indicating the beneficial Oxidative damage to crucial cellular molecules effects exercise and docosahexaenoic acid induced by reactive oxygen species has been (DHA) on various neurophysiological functions. implicated as a possible factor in the etiology of The aim of this study is to investigate the several human diseases, including cancer, effects of exercise and DHA supplementation in cardiovascular disease, and aging (Halliwell and Gutteridge, 1989).

12 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

DPPH Radical Scavenging Activity:1,1- Department of Physiology, Diphenyl-2-picrylhydrazyl radical scavenging Neuroimmunophysiology Research capacity. DPPH radical scavenging capacity was determined according to the Blois method. Group, University of Extremadura, Reducing Power: The reducing powers of 06006-Badajoz, Spain. extracts were quantified by the method of Oyaizu. Ferric thiocyanate method-total Melatonin has many effects on a wide range of antioxidant activity: The total antioxidant physiological functions and is involved in a capacities of the extracts were determined number of pathological events including according to the ferric thiocyanate method in oncostatic and neoplastic processes. The tissue linoleic acid emulsion. protective actions of melatonin are attributed to Determination of the Amount of Total Phenolic its well-known antioxidant activity though Compounds: The phenolic compound content melatonin might also exert pro-oxidant effects, was determined as pyrocatechol equivalents particularly in tumor cells. Here, we evaluated using the following linear equation based on the the pro-oxidant effects of melatonin and their calibration curve role on caspases-3 and -9 activation in a http://pubs.acs.org/doi/full/10.1021/jf010357 human promyelocytic leukaemia cell line HL-60 2?cookieSet=1 - jf0103572e00001. cells. Intracellular reactive oxygen species (ROS) cell analysis was quantified by Antimicrobial activities: The antimicrobial effect of ethanol extract of C.acutum, C. erecta, T. rhodamine-123 fluorescence, MTT assay was venetum subsp. sarmatiense were tested performed in order to evaluate the cell viability against bacteria strains Staphylococcus aureus under melatonin treatments, in addition, DNA ATCC 25923, Pseudomonas aeruginosa ATCC cellular was PI stained to discriminate died cells 27853, Escherichia coli ATCC 35218, from alive unstained cells. To determine Escherichia coli ATCC 25922, Bacillus subtilis caspase-3 and -9 activities the cleavage of ATCC 29213, Bacillus cereus NRLL B-3008. fluorogenic specific substrates, DEVD-AMC and Enterobacter fecalis ATCC 292112 and two AC-LEHD-AMC respectively, were measured. yeast strain Candida albicans ATCC 10231 and Melatonin treatment is able to stimulate Candida tropicalis ATCC 13803.The compound production of intracellular reactive ROS, as revealed by the increase in rhodamine-123 effect was carried out by the agar diffusion fluorescence, which was associated with method significant cytotoxicity and activation of caspase The results indicate that the antioxidant activity -1 activities. Furthermore, pretreatment of cells of ascorbic acid (8.2 µg mL ) was higher than the ethanol extract of C. acutum (196.09 µg mL- with well-known antioxidants, such as N-acetyl- 1), C.erecta (240.14 µg mL-1), T.venetum subsp. L-cysteine (NAC), trolox, PEG-catalase and sarmatiense (570.71 µg mL-1 ). Total antioxidant reduced glutathione (GSH), reversed the effects activities of 54 µg mL-1 of C. acutum, C. erecta, of melatonin on both intracellular ROS T. venetum subsp. sarmatiense extracts, production as on the cytotoxicity and caspase activation. This pro-oxidant action may be determined according to ferric thiocyanate significant in light of the findings that melatonin method. The antimicrobial results obtained has pro-apoptotic actions in tumour cells and it using the agar well diffusion method. The could increase the utility of melatonin as an highest inhibitory activity was seen against S.aureus using the ethanol extract of C.erecta anticancer agent. concentration of 100 mg/mL. This work was supported by MEC-DGI and Poster No 26 Junta de Extremadura grants BFU2007-60091 and PRI07-A024, respectively. I. Bejarano is supported by Junta de Extremadura grant Pro-oxidant effects of melatonin in (PRE06070). human myeloid leukaemia HL-60 cells. Poster No 27

Bejarano I, Espino J., Barriga C., Protective effect of antioxidants on Pariente J.A., Rodríguez A.B. indomethacin-Induced gastric mucosal injury in rats

Ismet Burcu Turkyılmaz, R. Yanardag

13 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Istanbul University, Faculty of 3Manager Scientific Department of Engineering, Department of Chemistry, I+D+i. Grupo Ordesa. Barcelona, Istanbul, Turkey. Spain.

Non-steroidal anti-inflammatory drugs (NSAIDs) It is known all the advantages of the human are some of the most widely used drugs in the milk for the development of the breast-fed baby, world. It is well-known that gastric damage is between the nutritional components they find the major NSAID-induced side effect. Reactive the vitamins A, E and C, characterized by the oxygen species (ROS) also play a vital role in defense antioxidant. In the newborn children mucosal lesions associated with NSAIDs. the stress oxidative is major due to the Antioxidant defense systems and their immaturity of the mechanisms of antioxidant components, including antioxidant enzymes, defense and of the digestive system. It is known foods and drugs are important in preventing the that it humanizes the nutritional components of toxic and disease causing effects of oxygen- the milk, they are changeable throughout the derived free radicals. In this study, the effects of day, being intimately tied to the ingestion of the antioxidants on indomethacin-induced gastric mother. damage were evaluated in rats. Male Sprague- The aim of the authors was to analyze the Dawley rats were randomly distributed into 4 antioxidant capacity of the mother milk along groups. Group I; intact animals. Group II; the period of 24 hours, the antioxidant changes control animals receiving vit C (100 mg/kg), between the day and the night. vit.E (100 mg/kg), β-carotene (15 mg/kg) and Samples of milk colostrum of healthy mothers sodium selenate (0.2 mg/kg) for 3 days, orally, (n=7), quiet (-80 °C) along a period of 24 h, in daily. Group III; rats receiving 25 mg/kg the Service of Neonatalogy (S.E.S.), Badajoz. indomethacin. Group IV; animals receiving The antioxidant capacity decided for the vit.C, vit.E, β-carotene and sodium selenate for improved spectroscopic method TEAC (Trolox 3 days (in the same dose and time), 2 h prior to equivalent antioxidant capacity). By this method the administration of indomethacin. 6 h after there is calculated the percentage of inhibition the indomethacin administration all the animals of the radical cation ABTS - + by means of the were sacrificed. Stomach glutathione Trolox, the analogous soluble one in water of peroxidase, catalase acitivities and protein the alfa-tocopherol, which is the antioxidant carbonyl and sialic acid levels were increased standard. For the statistical analysis it was in indomethacin groups, also, stomach used; descriptive statistics (X±DS) and glutathione-S-transferase activity and inferential test not parametric Kruskal-Wallis. A glutathione and mucus levels were decreased. value of p <0.00 was considered to be Administration of antioxidants reversed these significant. effects. In conclusion, the result of present We found variations in the antioxidant activity study indicates that antioxidant administration between the night and diurnal samples of milk. protects gastric tissue against indomethacin- Being the levels of Trolox Equivalent higher , induced damage. statistically significant (p <0,00), in the samples collected at 24:00 h opposite to the samples of Poster No 28 milk collected at 09:00 h and 21:00 h. We observed the increase in the antioxidant Analysis of the antioxidant activity in capacity in the samples of the night period human milk, day vs night. opposite to the rest of samples of the diurnal period, probably as consequence of the immunological mother activity and the ingestion J. Cubero1, C.L. Sánchez1, R. Bravo1, 2 1 of vitamins and proteins realized during the day. J. Sánchez , A.B. Rodriguez , M. Rivero3, C. Barriga1 Poster No 29

1Laboratory of Chrononutricion Faculty Melatonin blocks intracellular of Sciences, Uex. Badajoz, Spain. calcium overload- and ROS- 2Laboratory of Metalopathies Hospital dependent caspase activation in Perpetuo Socorro. S.E.S. Badajoz, human leucocytes. Spain. Javier Espino, I. Bejarano, C. Barriga, J.A. Pariente, A.B. Rodríguez.

14 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Poster No 30 Department of Physiology, Neuroimmunophysiology and In vitro exposure to melatonin Chrononutrition Research Group, improves sperm motion parameters.

Faculty of Science, University of 1 2 1 Extremadura, 06006 Badajoz, Spain Javier Espino , A. Ortiz , I. Bejarano , F. Monllor2, G.M. Lozano2, J.F. García2, 1 1 Emerging evidence suggests that melatonin J.A. Pariente , A.B. Rodríguez . may be involved in the protection of different cell types against damage-induced apoptosis. 1Department of Physiology, We aimed to evaluate the effect of melatonin on Neuroimmunophysiology and caspase activation evoked by increases in both intracellular calcium and reactive oxygen Chrononutrition Research Group, species (ROS) levels in human leucocytes. Faculty of Science, University of Human leucocytes were separated from whole Extremadura, 06006 Badajoz, Spain, blood using Ficoll-Hypaque. ROS generation and 2 Extremadura Centre for Human was quantified with the non-fluorescent, cell- Assisted Reproduction, 06010 Badajoz, permeable probe dihydrorhodamine-123 (DHR- 123). Caspase-3 and -9 activities were Spain determined from the cleavage of their respective specific fluorogenic substrate. Although human seminal fluid contains Our results show that the treatment of human melatonin and spermatozoa reportedly possess neutrophils with the specific inhibitor of calcium membrane melatonin receptors, there are no reuptake thapsigargin (TG, 1 µM) for 60 experimental studies that ascertain the minutes induced a rise in intracellular ROS relationship between melatonin and sperm levels, which was forestalled by melatonin (1 motility. To this end, we aimed to evaluate the mM, 60 minutes) pre-incubation. Likewise, TG- in vitro effects of melatonin on human induced ROS production was also abolished by spermatozoa function by analysing several pre-treatments with the well-known antioxidant, sperm motion parameters. N-acetyl-L-cysteine (NAC, 15 mM, 60 minutes), Human semen was obtained from fifteen men the intracellular calcium chelator, dimethyl attending infertility counselling and collected by BAPTA (10 µM, 30 minutes), and the specific masturbation after 4-5 days of sexual blocker of calcium uptake into mitochondria, abstinence. Motility kinematic parameters were Ru360 (10 µM, 30 minutes). Moreover, our assessed by a computer-aided semen analysis results indicated that TG was able to raise the (CASA) system in both non-treated and caspase-3 and -9 activities. TG-induced melatonin-treated samples. caspase-3 and -9 activation was again inhibited Melatonin treatment (1 mM, 30 min) improves by melatonin pre-incubation, but also by NAC, motile and progressive motile cells percentage, BAPTA and Ru360 pre-treatments. Similar and decreases static cells count, mainly results were obtained in human lymphocytes. promoting the proportion of rapid cells, because In sum, our results suggest that melatonin melatonin enhances sperm velocity, in general, prevents caspase-3 and -9 activation induced and straightness, in particular. by both increases in intracellular calcium and Briefly, short-term exposure to melatonin ROS levels in human leucocytes. ameliorates sperm motion parameters, which is Funded by MEC-DGI (BFU2007-60091) and extremely important because melatonin Junta de Extremadura (PRI07-A024). I. supplementation may be potentially used to Bejarano is supported by Junta de Extremadura obtain a successful assisted reproductive (PRE06070). techniques (ART) outcome. Funded by Merck, S.L. J. Espino and I. Bejarano are supported by Merck, S.L. and Junta de Extremadura (PRE06070), respectively.

15 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

effects / symptoms of schizophrenia through Poster No 31 improvement in the reduction of purine metabolism. Effects of paliperidone on purine o catabolizing enzymes and level of Poster N 32 nitric oxide in rat brain Effects of paliperidone on

antioxidant system in rat brain Kadir Demirci1, R. Özcankaya1, H. R. 2 2 1 Yılmaz , A. Yiğit , K. Karakuş 1 2 Kadir Demirci , H. R. Yılmaz , R. 1 2 1 1 Özcankaya , A. Yiğit , K. Karakuş Department of Psychiatry, Medical Faculty, Süleyman Demirel University, 1 Department of Psychiatry, Medical Isparta, Turkey. Faculty, Suleyman Demirel University, 2Department of Medical Biology, Isparta, Turkey. Medical Faculty, Süleyman Demirel 2 Department of Medical Biology, University, Isparta, Turkey. Medical Faculty, Suleyman Demirel

Schizophrenia is a serious mental illness that University, Isparta, Turkey. affects how a person thinks, feels, and behaves. In treatment, antipsychotic drugs are Paliperidone, is one of the most commonly generally used. Paliperidone is a new drug used atypical antipsychotic drugs, and it is the which includes basically the same active active metabolite of risperidone. metabolite of risperidone which generally used The aim of this study is to investigate the as an antiphysotic drug to treat schizophrenia. effects of paliperidone on the enzyme activities The main objective of the current study is to of superoxide dismutase (SOD), catalase investigate the effects of paliperidone on (CAT), and glutathione peroxidase (GSH-Px) enzyme activities of adenosine deaminase and malondialdehyde (MDA) levels. (ADA) and xanthine oxidase (XO), and nitric Twenty Sprague Dawley male rats were used in oxide (NO) levels, which has important roles on this study. Rats were randomly divided into two purine catabolism in rat brain. Twenty Sprague equal groups as control (n=10) and Dawley rats were used in this study. Rats were paliperidone group (n=10). Physiological saline randomly divided into two equal groups as solution was given to the control group as control (n=10) and paliperidone group (n=10). intraperitoneally (i.p.) once a day during 14 Physiological saline solution was given to the days. control group as intraperitoneally (i.p.) once a 1 mg/kg per day paliperidone was given to day during 14 days. 1 mg/kg per day paliperidone group once a time in a day during paliperidone was given to paliperidone group 14 days. 24 hours later after last enjection, rats once a time in a day during 14 days. 24 hours were sacrificed and brain tissues were taken. later after last injection, rats were sacrificed and The enzyme activities of SOD, CAT and, GSH- brain tissues were taken. The enzyme activities Px and MDA levels were measured from brain of ADA and XO, and NO levels were measured tissues as spectrofotometrically. from brain tissues homogenates as When we compared paliperidone group with spectrofotometrically. control group; CAT activity was significantly When we compared paliperidone group with decreased, SOD activity and MDA levels were control group, ADA and XO enzyme activities increased but not statistically significant. GSH- were significantly decreased. NO levels were Px levels were also decreased but also not increased but it was not statistically significant. statistically significant. In conclusion we observed that ADA and XO As a result, paliperidone could change enzyme activities had significantly decreased in antioxidant status. According to this result, paliperidone group compared to the control paliperidone may be a risk factor for oxidative group. This may show us the reduction of the damage. purine metabolism. This reduction in the purine metabolism may diminish the activity of dopamine by increasing the amount of adenosine. Paliperidone treatment is thought to be useful to taking under control of the positive

16 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Poster No 33 Funded by PDT0A008-Junta de Extremadura. S.D. Paredes was beneficiary of a grant by Comparison of Jerte Valley cherry- Junta de Extremadura-Fondo Social Europeo based nutraceutical product vs. (REI09009). placebo-controlled trials in terms of Poster No 34 6-sufatoxymelatonin and total antioxidant capacity in urine Urinary 6-sulfatoxymelatonin and total antioxidant capacity increase 1 2 M.Garrido , D. González-Gómez , M. after the intake of a grape juice 2 2 1 Lozano , T. Hernández , C. Barriga , obtained with high hydrostatic 1 1 S.D. Paredes , A.B. Rodríguez pressure techniques.

1 Department of Physiology D. González1, E. Gamero2, María (Neuroimmunophysiology and Garrido1, D. Moreno2, E. Valdés2, C. Chrononutrition Research Group), Barriga1, S.D. Paredes1, A.B. Faculty of Science, University of Rodríguez1 Extremadura, 06006, Badajoz, Spain. 2 Technological Institute of Food and 1Department of Physiology Agriculture of Extremadura, 06071, (Neuroimmunophysiology and Badajoz, Spain. Chrononutrition Research Group), Faculty of Science, University of The consumption of fruits rich in the antioxidant Extremadura, 06006, Badajoz, Spain. melatonin increases the circulating and urinary 2 levels of this indole. Jerte Valley cherries Technological Institute of Food and (Prunus avium) contain melatonin as well as its Agriculture of Extremadura, 06071, precursors, the amino acid tryptophan and the Badajoz, Spain. neurotransmitter serotonin, which also act as antioxidant bioactive compounds. This study Red grapes contain elevated concentrations of was aimed at comparing the effect of a Jerte antioxidant compounds (polyphenols) that may Valley cherry-based nutraceutical product potentially prevent cell aging, cardiovascular (patent nº P200803761) and a placebo product disease and oxidation-related disorders. Since on the urinary 6-sulfatoxymelatonin (aMT6s) functional drinks are presently one of the most and total antioxidant capacity (TAC) in young dynamic sectors of the market, the aim of the (20-30 yr-old), middle-aged (45-55 yr-old) and present work was to evaluate the possible elderly (65-75 yr-old) participants. The antioxidant effect of an experimental grape nutraceutical product consisted of equal parts of (Vitis vinifera cv. Tempranillo) juice in terms of the pulp of 4 cultivars of Jerte Valley cherries 6-sulfatoxymelatonin and total antioxidant from Cáceres, Extremadura, Spain (94%), capacity in the urine of young (20±10 yr-old), maltodextrin (5%) and ascorbic acid (1%). The middle-aged (45±10 yr-old) and elderly (75±10 placebo product was a commercial cherry- yr-old) individuals. The grape juice was flavored soft drink. Volunteers ingested either obtained by means of high hydrostatic pressure the placebo or the nutraceutical product twice a (HHP) techniques. Participants consumed 200 day for 5 days. aMT6s was analyzed using a ml of grape juice twice a day (as the lunch and commercial ELISA kit in first-void morning dinner desserts) for 5 days. First-void morning urines. TAC was quantified by means of urines were collected before treatment (basal colorimetry in urines collected at 21.00 h. The values), the immediate day after the last consumption of the nutraceutical cherry-based ingestion of juice (assay), and one day product augmented the participants’ 6- afterwards (post-assay). For the quantification sulfatoxymelatonin levels and total antioxidant of 6-sulfatoxymelatonin, a commercial ELISA kit capacity as compared to the values obtained was used. Total antioxidant capacity was before and after the treatment. The placebo did evaluated using a colorimetric assay kit. The not modify their urinary antioxidant levels. The intake of grape juice cv. Tempranillo obtained Jerte Valley cherry-based nutraceutical product with HHP induced a significant increase of may be used as a source of antioxidants. urinary 6-sulfatoxymelatonin and total antioxidant capacity in the three groups of age

17 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels analyzed as compared to their corresponding Echt Violet and H+E histostaining for basal and post-assay values. These investigating the tissue morphology. In the functional/nutraceutical properties may be of trauma group was observed common interest for a prospective commercialization of hemorrhage, necrosis, severe degeneration of the grape juice. motor neurons and leukocyte infiltration in the Funded by INTERREG 0318-RITECA-4E; gray matter. In the trauma + Huperzine A group Programa POCTEC, 053/09. S.D. Paredes was was decreased seen cavitations area, beneficiary of a grant by Junta de Extremadura- hemorrhagic, and edema in the gary matter Fondo Social Europeo (REI09009). compared to trauma groups. Apoptotic shows were mostly observed in the glial cells and motor neurons in the trauma group. In the Poster No 35 control group decreased number of apoptotic cells per unıt area is observed compared to The effects of Huperzine A, on trauma group (p<0.05). In the Huperzine A traumatic spinal cord injury in rats. groups (3 and 7 days) were determined significant decrease compared to trauma 7th day

(respectively p<0.01 p<0.01). Groups were also Ş. Yüceli1, Mediha Eser2, G. 2 3 evaluated semi quantitatively in terms of TGF-β Tanrıverdi , O. Akdemir , M. and BDNF immunopositiveness. While it was 4 5 2 Tunçdemir , M. Kaya , F. Alkan discovered that TGF-β positivity was higher in trauma group than in the group applied 1Taksim Emercency Education and Huperzine A, it was determined that the number of BDNF positive cells was higher in Huperzine Research Hospital, Neurosurgery A applied group than in trauma group. In Department, conclusion, in rats local application of İstanbul, Turkey, Huperzine A after SCI is effective on decreased 2İstanbul University, Cerrahpaşa School secondary tissue damage and therefore on of M edicine, Department of Histology protection of the motor function. This effect can and Embryology, İstanbul, Turkey, be explained by the inhibition of apoptotic death 3 of total cell types in spinal cord. Taksim Emercency Education and Research Hospital, Neurosurgery References Department,İstanbul, Turkey, Agrawal SK, Fehlings M G: Role of NMDA and 4İstanbul University, Cerrahpaşa School non NMDA ionotropic glutamate receptors in traumatic spinal cord axonal injury. J Neurosci of Medicine, Department of Medical 17: 1055-1063, 1997. Biology, İstanbul, Turkey, Agraval SK, Nashmi R, Fehlings M G. Role of L 5 Kars , Neurosurgery and N type calcium channels in the Department, Kars, Turkey, pathophysiology of traumatic spinal cord white matter injury.Neuroscience 99:179-188, 2000. The aim of this study is to investigate the Aguayo AJ: Axonal elongation in peripheral and neuroprotective and antiapoptotic effects of central nervous system transplants. Advancesin Huperzine A on the traumatic rat spinal cord cellular neurobiology 3, New York: Academic tissue. Thirty six Wistar type albino rats were Press: 215-234 1982. studied in 3 groups of 12 animals: sham- operated control animals (group 1); trauma-only o control animals (group 2); and rats subjected to Poster N 36 trauma + Huperzine A treatment (group 3). Funalia Trogii Application on Spinal cord injury was produced at the thoracic Preventing Oxidative Damage in level using the clip compression technique. Brain Produced by Deltamethrin Responses to injury and the efficacy of Exposure Huperzine A were assessed by light microscopy, apoptosis and cellular damage 1 2 were assessed by staining with TUNEL and Mehmet Berköz , S. Yalın , Ü. immunostaining for BDNF and Tgf-beta. The Çömelekoğlu3, B. Mazmancı4, M. Ali functional recovery was assessed by field Mazmancı5, P. Eroğlu2, A. Ünyayar5 locomotor test at the 3 and 7 days after surgery. Also, the samples were stained with Cresyl

18 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Mersin University, Faculty of Pharmacy, mutagenic effects of Coriolus versicolor and 1Department of Pharmaceutical Funalia trogii extracts on mammalian cells. Technology, 2Department of Drug Chem Toxicol. 29:69-83.

Biochemistry, 3Faculty of Medicine, 4 Department of Biophysics, Faculty of Art and Science, Department of Poster No 37 Biology, 5Faculty of Engineering, Department of Environmental The Antioxidant Effect of Ocimum Engineering, 33169, Mersin Basicilum on E.coli Exposed to Different Doses of Radiation Deltamethrin is a dibromo pyrethroid pesticide M. Cihan Yavaş1, M.S. Çelik1, V. Akpolat1, that has widespread using fields in agriculture 2 3 industry. For reducing the harmful effects of N. Akpolat , S.B. Zincircioğlu , M. H. 3 insecticides on living organisms, using medical Doğan plants could be a helpful treatment approach. In this study, we investigated the protective effects 1Dicle University, Medical Faculty, Dept. of of Funalia trogii that is a medicinal mushroom Biophysics, 2Dept of Microbiology, 3Dept. of species, on oxidative damage in rat brain Radiation Oncology, Diyarbakır / Turkey produced by deltamethrin exposure. Totally 21 Wistar albino rats were used for our study and In this study, the E.coli in agar medium and we separated the animals to 3 groups that agar + ocimum basicilum extracts medium were contains 7 animals in each groups. Any subjected to different doses of gamma application wasn’t done to the control group radiation. The antioxidative effects of ethanolic during the experiment. Deltamethrin group was extracts of Ocimum basicilum on the E coli taken 1.28 mg/kg deltamethrin during 30 days exposing to irradiation was investigated. and deltamethrin+fungus group was taken 1.28 mg/kg deltamethrin and 0.5 mL Funalia trogii Ocimum basilicum plant used in the study was extract during 30 days. At the end of the study, dried and powdered by a mechanical chopper all the animals were sacrificed and superoxide of 5 gr, which was later incubated by a dismutase (SOD) and catalase activities for mechanical shaker within ethylalcohol for 48 detecting the antioxidant state and hours at 25 ˚C. After filtration, the ethanol was malondialdehyde (MDA) levels for definition of removed at evaporator at 50 ˚C at 200 rpm. the lipid peroxidation state were measured. It After preparing the Blood Agar Base by both was found that MDA levels of deltamethrin and checking the plant extract and agar’s pH, to deltamethrin+fungus groups were higher than each plate was added 83 ± 2 μl plant extract. control group. MDA level of 0,5 cc bacteria culture was added to the deltamethrin+fungus was lower than medium which has 4, 5 cc pure Mueller Hinton deltamethrin group but it wasn’t statistically Broth. After it was diluted at the rate of 106, 0,5 significant (p>0.05). SOD activity of cc broth culture was planted to agar medium. deltamethrin group was higher but catalase 100, 200, 500, 1000, 3000, 6000 cGy doses activity was lower than control group. SOD was performed with Co 60 teletherapy machine activity of deltamethrin+fungus group was lower (exception of control group). After incubated 18 but catalase activity was higher than hours at 37 ˚C, colony counts were performed. deltamethrin group, statistically (p<0.05). The colony numbers of E.coli were decreased Finally, it is said that treatment with Funalia as the dosage of irradation increased. On the trogii extract helps preventing the living agar medium contaning extracts of Ocimum organisms from the oxidative stress on basilicum, irradiated at different doses, the deltamethrin exposure. bacterial colonies were grown enormously on the surface of petri plates by spreading all over. References: The reason for the increase of the colonies in the presence of the extracts of Ocimum Abdollahi M, Ranjbar A, Shadnia S, Nikfar S, basilicum at the irradiated medium is likely due Rezaie A. 2004. Pesticides and oxidative to antioxidant effects of the plant extracts. stress: a review. Med Sci Monit.10:141-147. However, the studies on the subject are being Unyayar A, Demirbilek M, Turkoglu M, Celik A, carried out. Mazmanci MA, Erkurt EA, Unyayar S, Cekic O, Atacag H. 2006. Evaluation of cytotoxic and

19 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Acknowledgements: This study was supported nerves were taken from animals. Then, by Research Foundation of sections were taken for stereological and (DUBAP-08-TF-08). histological analysis. There were no any significant differences (p Poster No 38  0.05) between PG, SG and DG in respect of myelin thickness, axon cross section area, The Effect of Prenatally Exposure of axon numerical density, total section area of a Nonsteroidal Anti-inflammatory optic nerve and axon number. Altogether, the Drug on the Optic Nerve of Female study results showed that DS have no a toxic effect on the structure and myelinization of Rats in the Late Postnatal Life: optic nerve of female rats if it is administered A Stereological, Histological and during the gestational period. Electron Microscopic Study Both histological and stereological results from present study show that DS as well as S. Kaplan1, M. Eşrefoğlu2, A. Aktaş3, M. saline do not cause to undesirable effect on 2 1 female rat optic nerve development and Gül , Mehmet Emin Önger , E. myelinization in respect of morphology. 1 3 Altunkaymak , M. B. Ulkay , M. Ç. 4 Rağbetli Poster No 39

1Department of Histology and Effect of estrogens and Embryology, Medical School of progesterone on preadipocyte Ondokuz Mayis University, differentiation 2Department of Histology and 1 1 1 Embryology, Medical School of Inonu Mehtap Pektaş , A. H. Kurt , İ. Ün , K. University, Malatya Büyükafşar1 3Department of Histology and Embryology, Veterinary School of Department of Pharmacology, Medical Istanbul University, Istanbul Faculty, Mersin University, Mersin 4Department of Histology and Embryology, Medical School of Sex hormones are the major regulators of adipocyte development and functions (1). It has Yuzuncu Yil University, Van been suggested that estrogens has differential effects on adipocyte differentiation (2, 3). Nonsteroidal anti-inflammatory drug However the effect of progesterone has not (NSAID)s can have adverse effects in both completely been understood (4). In the present mother and fetus following administration study we investigated the effects of estrogen during the prenatal period. Diclofenac sodium and progesterone on the differentiation of (DS), a NSAID, is given during the mouse embryonic fibroblasts, 3T3-L1, to white pregnancy, may also affect development of adipocyte cells. 3T3-L1 cells were cultured in the central nervous system (CNS) or related %10 CS/DMEM. After confluency (regarded as structures. In this study, the effects of DS on the day 0), the cells were incubated with a the developing optic nerve following differentiation medium (0.5 mM IBMX, 0.25 µM administration of the prenatal period were dexamethasone and 1μM insulin with 10 % investigated using stereological methods. FBS/DMEM) at the day 0-2, with only insulin in Pregnant rats were separated into the pure 10 % FBS/DMEM at the day 2-4 and with 10 % control group (PG), saline group (SG) and FBS/DMEM at the day 4-8. 17β-estradiol (10-9- diclofenac group (DG). A dose of 1mg/kg of 10-4 M), estradiol-BSA (10-9-10-4 M) or DS and 1 ml/kg saline daily was injected progesterone was applied to the confluent cells intraperitoneally to the DG and SG for 8 days. Adipocyte differentiation and respectively at beginning from the 5th day for intracellular lipid deposition were evaluated with a 15 day of period. PG received no any Oil Red-O staining at the day 8. For statistical treatment. After spontaneously delivery, analysis ANOVA and Dunnet post hoc test were female offspring were obtained from PG, DG used. A P value of less than 0.05 regarded as and SG. After at the end of 20th week of significant. Estrogen treatment for 8 days postnatal life, the animals (n=6 for each suppressed adipocyte differentiation at the low group) were perfused and the right optic (10-9 and 10-8 M) and high (10-5 and 10-4 M)

20 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels concentrations. However intermediate cells by using an inverse metabolic engineering concentrations (10-7 and 10-6 M) had no effects strategy; evolutionary engineering. The yeast on the differentiation of 3T3-L1 cells to strain CEN.PK113.7D was mutagenized first by adipocytes. Estradiol-BSA, which is a cell- ethyl methane sulfonate (EMS) to increase impermeable conjugate of estrogen almost genetic variation. The mutant yeast cell completely, suppressed adipocyte population was then exposed to increasing differentiation, similar to lipophilic estradiol. hydrogen peroxide pulse stress for successive -8 -6 However, progesterone (10 -10 M) did not batch cultivations. The sub-lethal levels of H2O2 change adipocyte differentiation. The was determined and H2O2 resistant generations suppression of adipocyte differentiation by the were obtained by applying increasing levels of long-term exposure of estrogenic hormones pulse stress to each generation for 90 min until could be mediated via membranal and/or the survival rate of the population was below cytosolic estrogenic receptors. 0.15. Initial experiments showed that EMS This study was supported by TÜBİTAK (SBAG- mutagenized S.cerevisiae population was 109S126). tolerant up to 5 mM H2O2, thus initial stress level was chosen as 5 mM H2O2. After that References step, the individual mutants were selected 1. Cooke SP, Naaz A, 2004. Role of Estrogens randomly from the final populations and tested in Adipocyte Development and Function. for their H2O2 resistance by using 5-tube MPN Experimental Biology and Medicine 229: 1127- method. The survival of the final population 1135. increased up to about 11,000 and 2x106-fold of 2. Ryo O, Daisuke I, Minako S, Mieko S, the wild type under continuous 0.1 and 0.2 mM Shinsuke K, Hikari O, Hirofumi T, Yoshikazu S, H2O2 stress, respectively. The survival of the Toshio M, 2002. Estrogen Promotes Early final population increased up to about 7800-fold Osteoblast Differentiation and Inhibits of the wild type at pulse 0.1 M H2O2 stress Adipocyte Differentiation in Mouse Bone level. The results showed that oxidative stress Marrow Stromal Cell Lines that Express resistant S.cerevisiae mutants were Estrogen Receptor (ER) α or β. Endocrinology successfully obtained by using evolutionary 143: 2349–2356. engineering. 3. Liu H, Aylin C, Ioana AP, 2006. Modulations of 17-β Estradiol on Osteogenic and Adipogenic Differentiations of Human Mesenchymal Stem Poster No 41 Cells. Tissue Engineering. Volume 12, Number 10. Do Non-Thermal Electromagnetic Fields 4. Mayes JS, Watson GH, 2004. Direct effects cause Nitrosative Stress in male and of sex steroid hormones on adipose tissues and female rats? obesity. Obesity reviews 5, 197–216. 1 1 2 o M. Arda Eşmekaya , B. Sırav , Ç. Özer , N. Poster N 40 Seyhan1

How to obtain hydrogen peroxide 1Department of Biophysics, Gazi University, resistant Saccharomyces cerevisiae Ankara, TURKEY, 2 Department of by evolutionary engineering? Physiology Gazi University, Ankara, TURKEY. Meliha Merve Hız1, 2, Z.P. Çakar1 Nitrosative stress occurs when the generations 1Department of Molecular Biology, of highly Reactive Nitrogen Species (RNS) such as nitric oxide (NO) exceed the ability of the Genetics and Biotechnology, İstanbul human body to neutralize and eliminate them. It Technical University, Istanbul, 34469, may lead to nitrosylation reactions that can alter Turkey protein structure by inhibiting normal body 2Present address: Department of functions. In this study, we aimed to investigate Biology, Çanakkale 18 Mart University, whether RF Fields at non – thermal level can affect total NO (NOx) and antioxidant levels in Çanakkale, Turkey plasma thus cause nitrosative stress. Twenty- four male and female Wistar rats were divided In this study, the aim was to obtain hydrogen into four groups: Group I and Group II: male peroxide resistant Saccharomyces cerevisiae (n=6) and female (n=6) control groups, Group

21 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

III and Group IV: male (n=6) and female (n=6) induced by pentylenetetrazole (PTZ) injection. RF radiation exposed groups. The exposed Eight weeks old mice were used in this study. groups of rats were exposed to non - thermal Mice were divided randomly into three groups RF radiation 20 min/day for a month. NOx as follows; Group I: Control group treated with (nitrite/nitrate) levels were measured by Griess PTZ, Group II: 15 minute mobile phone assay using Elisa reader. The total sulfhydryl radiation + PTZ treatment + 30 minute mobile (RSH) levels were determined phone radiation, Group III: 30 minute mobile spectrophotometrically by Kurtel method. The phone radiation + PTZ treatment + 30 minute exposure to RF Radiation for a month caused mobile phone radiation. The RF radiation was significant increase in NOx plasma levels in produced by a mobile test phone. Lipid male and female rats (p < 0.05). Conversely peroxidation, which is the indicator of oxidative RSH levels were significantly lower in exposed stress was quantified by measuring the rats compared to control rats (p < 0.05). No formation of thiobarbituric acid reactive statistically significant difference was observed substances (TBARS). The Glutathione (GSH) in the rectal temperature between control and levels were determined by Ellman method. The RF - irradiated rats (p > 0.05). In the present absorbances of the samples were measured study, we showed that plasma NOx level may using a spectrophotometer. The accepted level be increased in both male and female rats due of significance was set at p < 0.05. Lipid to RF radiation exposure. The results of our peroxidation levels of brain tissue increased study are evident that RF Fields at non – significantly in group II and III compared to thermal level can induce nitrosative stress by group I. On the contrary, GSH levels were increasing NOx and decreasing antioxidant significantly lower in group II and III than group levels. I. However, no statistically significant alterations in any of the endpoints were noted between Poster No 42 group II and Group III. These results indicated that exposure to RF fields emitted from cell Effects of Cell Phone Radiation on phones may lead to oxidative stress in mouse Oxidant and Antioxidant status in brain during epileptic activity by enhancing lipid peroxidation and reducing the antioxidant epileptic mouse model (GSH) level. Oxidative injury may play a key role in the initiation and progression of epilepsy. Meric Arda Esmekaya1, A. G. Our results showed that mobile phone radiation Canseven1, E. Yücel2, Z. Aktuna3, S. may increase the oxidative damage during 4 1 epileptic activity in mouse brain. Keskil , N. Seyhan Poster No 43 1 Department of Biophysics, Gazi University, Ankara, TURKEY, Quercetin partially improves renal 2Department of Neurosurgery, Baskent dsyfunction in diabetic nephropathy University, Alanya Training and Research Hospital, Antalya, TURKEY, Mustafa Edremitlioğlu1, M. F. Andiç2, D. 3Department of Medical Pharmacology, B. Sayin3, O. Korkut4 and Ü. Kisa5 Kırıkkale University, Kırıkkale, TURKEY, 4Department of 1Department of Physiology, Çanakkale Neurosurgery, Bayındır Hospital, Onsekiz Mart University Medical Ankara, TURKEY. Faculty, Turkey; 2 Department of Physiology, Kirikkale Scientists have been conducting experimental University Medical Faculty, Turkey studies on whether Radiofrequency Radiation 3 Department of Medical Genetics, (RFR) emitted from cellular mobile phones induces oxidative damage on biological tissues Kirikkale University Medical Faculty, or cells. According to the current assumption Turkey RFR may initiate oxygen free radical 4Pharmacology, Kirikkale University intermediates in cells or tissues. In this study, Medical Faculty, Turkey and we investigated the effects of whole body RFR 5Biochemistry, Kirikkale University exposure on oxidant and antioxidant levels in epileptic mouse brain. The epileptic seizure was Medical Faculty, Turkey

22 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Effects of Vitamins C and E Diabetes mellitus (DM) can cause very serious Combination on Element complications such as nephropathy. One of the Levels in Blood of Smoker and factors of diabetic nephropathy pathogenesis is increase of oxidant stress. Another antioxidant Nonsmoker Radiology agent which has been shown to have beneficial X-Ray Technicians effects in cases of DM is quercetin. The aim of this study is to examine the effect of quercetin Mustafa Kayan¹, M. Nazıroğlu², C. on diabetic nephropathy rats and the change in Barak³ balance of oxidant-antioxidant capacity in this process. Five groups were used: Control, DM 8 wks (DM8), DM 16 wks (DM16), DM 8 wks + ¹Department of Radiology, Medical quercetin (QUER8), DM 16 wks + quercetin Faculty, Süleyman Demirel University, (QUER16) DM was induced by intraperitoneal Isparta, Turkey (IP) injection of streptozotocin. Rats in QUER8 ²Department of Biophysics, Medical and QUER16 were treated with IP quercetin (15 mg/kg/day) until the end of the experiment. Faculty, Süleyman Demirel University, MDA increased in DM8 and DM16 groups. On Isparta, Turkey contrary, it decreased in groups which were ³Department of Chemistry, Science administered quercetin. Accordingly, increase in Institute, Balıkesir University, Balıkesir, NADPH oxidase activity, and decrease in SOD Turkey and CAT activities in diabetic rats were determined. In QUER8 and QUER16 groups, NADPH oxidase activity decreased while Cigarette paper and cigarette smoke contain antioxidant enzyme activities increased. It is toxic elements, whereas vitamins C and E observed that functional kidney parameters (VCE) may have regulator effects on the were considerably corrupted in diabetic rats. elements. We investigated effects of VCE Especially the decrease in reabsorption of administration on X-ray-induced element sodium and water is remarkable. It is possible changes in blood of smoker and nonsmoker X- to say that kidney dysfunction depends on the ray technicians. increase of oxidant damage. Administration of quercetin totally prevented the increase of Twenty technicians and 30 healthy age- oxidant damage. In these groups, partial matched control subjects were used in the improvement in kidney functions was observed. study. 10 of the X-ray technicians and 15 of the The present experiment showed that renal control were smokers. Blood serum samples dysfunction in diabetes was attenuated, were taken from the control. Oral vitamins C although not completely prevented, by the use (500 mg) and E (150 mg) were supplemented of quercetin. daily to the smoker and nonsmoker X-ray technicians for 5 weeks. Serum samples were taken from the X-ray technicians before and Functional kidney parameters after 5 weeks. GFR Plasma Copper, zinc, selenium, aluminum, iron, GROUPS FENa (%) FEwater (%) creatinin magnesium, and calcium levels were ( L/min) (mg/dL) investigated in control and X-ray technicians, CONTROL 1525 137 0.81 0.16 1.36 0.14 0.33 0.02 both smokers and nonsmokers. Copper, zinc, DM8 274 55 3.86 0.63 6.05 1.37 0.87 0.26 and selenium levels were lower in the total X-

DM16 174 27 8.78 0.87 17.54 3.08 1.23 0.03 ray group and smoker X-ray group than in control and nonsmoker X-ray group, although QUER8 208 44 6.02 1.7 9.93 2.10 0.97 0.19 iron, magnesium, and calcium levels were QUER16 434 76 3.43 0.74 4.44 0.69 0.75 0.13 higher in X-ray group than in control. The copper, zinc, selenium, and aluminum levels were higher in the VCE treatment group than those in X-ray group, although magnesium and calcium levels were decreased by the

treatment. The serum zinc, copper, selenium,

o and magnesium levels were lower in smoker Poster N 44 control group when compared to nonsmoker control group. The serum zinc levels were lower in smoker X-ray group than nonsmoker X-ray

23 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels group, although iron level was higher in smoker were lower in the X-ray group than in control X-ray group than in nonsmoker X-ray group. although their concentrations were increased by VCE prevents the smoke and X-ray-induced the treatment. The erythrocyte GSH level and selenium, zinc, magnesium, and copper GSH-Px activity were found to be higher in the decrease to strengthen the antioxidant trace treatment group than in the X-ray group. element levels in the serum of the technicians. Plasma GSH level was not found to be different in all groups. Poster No 45 Free radicals are highly unstable substances produced in body through the metabolism of Vitamin C and E combination oxygen. They also result from exposure certain modulates oxidative stress induced environmental factors. GSH, GSH-Px, and antioxidant vitamins protect cells from oxidative by X-ray in blood of smoker and stress.9, 10 study clearly shows that smoking nonsmoker radiology technicians induced oxidative stress increase effect in smoke exposure to X-ray technicians. VCE Mustafa Kayan¹, M. Nazıroğlu², Ö. combination may play an antioxidant role Çelik², K. Yalman², H. Köylü³ against X-ray-induced oxidative injury. X-ray and cigarette smoke induces oxidative 1 stress by augmenting LP and diminishing the Department of Radiology, Medical antioxidant vitamin levels. VCE combinations Faculty, Suleyman Demirel University, recover LP damage probably through its free Isparta, Turkey radical scavenging in blood. 2Department of Biophysics, Medical Faculty, Suleyman Demirel University, Poster No 46 Isparta, Turkey 3 Department of Physiology, Medical Testicular apoptosis and Faculty, Suleyman Demirel University, histopathological changes induced Isparta, Turkey by a 2.45 GHz electromagnetic field

Mustafa Saygin1, S.Caliskan1, X-ray radiation is detrimental to human cells. 2 1 1 Cigarette smoke contains chemicals that N.Karahan , A. Koyu , N. Gumral , A.C. include oxidant compounds whereas vitamin C Uguz3 and E (VCE) have scavenger effects on the compounds. We investigated effects of VCE 1Department of Physiology, administration on X-ray-induced oxidative 2 3 toxicity in blood of smoker and nonsmoker X- Department of Pathology, Department ray technicians. of Biophysics, Faculty of Medicine Suleyman Demirel University, Isparta, Twenty technicians and 30 healthy age- Turkey. matched subjects control were used in the study. Ten of the X-ray technicians and 15 of There is a growing public concern about the the control were smokers. Oral vitamin C (500 potential human health hazard caused by mg) and E (150 mg) were daily supplemented exposure to electromagnetic radiation (EMR). for 5 weeks. Blood samples were taken from The objective of this study is to investigate the the X-ray technicians after and before 5 weeks. effects of 2450 mhz electromagnetic field on Plasma and erythrocytes lipid peroxidation (LP), apoptosis and histopathological changes on rat reduced glutathione (GSH) levels, erythrocytes testis tissue. Twelve weeks-old male Wistar glutathione peroxidase (GSH-Px), and plasma Albino rats were used in this study. Eighteen antioxidant vitamin concentrations were rats equally divided into three different groups investigated. which were named Group I, II and III. Cage control (Group I), sham control (Group II) and Plasma and erythrocyte LP levels were higher 2.45 GHz EMR (Group III) groups are formed. in the total X-ray group and smoker X-ray group Group III were exposed to 2.45 GHz EMR, at than in control and nonsmoker X-ray group, 3.21 W/kg Specific Absorption Rate (SAR) for respectively although the LP level was 60 minutes/day for 28 days. There was no decreased by the VCE treatment. The plasma difference among the groups for the diameter of vitamin C, A, E, and ß-carotene concentrations

24 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels the seminiferous tubules, pyknotic, and fixation of pancreas tissue by using 10% neutral karyolectic and karyotic cells. However, the formaldehyde solution the pancreas tissue number of Leydig cells of testis tissue of the rats samples were embedded in paraffin, cross- in group III was significantly reduced comparing sections were cut in 4–5 μm thickness and with the group I (p<0,05). Estimation of stained with hematoxylin-eosin. Preparations spermatogenesis using the Johnsen testicular were examined with light microscope. The data biopsy score revealed that the difference were analyzed by using a commercially between groups is statistically significant. available statistics software package (SPSS for The level of TNF-α, Caspase-3 and Bcl-2 were Windows). compared, and no significant difference was Basing on blood serum amylase, lipase, ALP found between the groups. When Bax and GGT enzyme activities significantly apoptosis genes and Caspase-8 apoptosis increased in EMR group as compared with enzyme were compared, there were significant cage and shame control groups (p<0.05). differences between the groups (p<0,05). Basing on blood serum amylase, lipase, ALP Electromagnetic field affects spermatogenesis and GGT enzyme activities significantly and causes to apoptosis due to the heat and decreased in EMR+Selenium and EMR+L- other stress related events in testis tissue. carnitine groups as compared with EMR group (p<0.05). In respect to the structural Poster No 47 appearances of pancreas tissues there was no difference between control and the other Protective effects of L-carnitine and groups. Selenium on 2450 MHz It is observed that the radiation generated by 2450 MHz causes some biochemical changes Electromagnetic radiation exposed on pancreatic enzyme activities. These Pancreas Tissue changes were not at structural levels. Basing on amylase, lipase, ALP and GGT enzyme activities, a significant difference was A.Koyu1, N. Gumral1, A. Gokcimen2, determined between EMR exposed group and Mustafa Saygin1, H. Vural3, M. treatment groups. It can be an evidence of 2 Selenium and L-carnitine have protective role Ozgocmen on EMR exposed pancreas tissue.

1Department of Physiology, 2Department of Histology and Poster No 48 Embriyology, 3Department of Biochemistry, Faculty of Medicine The Role of Free Radicals in Retinitis Suleyman Demirel University, Isparta, Pigmentosa Turkey. Mustafa Tümay1, F. R. Hatungil1, M. 2 3 Berköz , S. Yalın 1 In this study, we aimed to research protective Mersin University, Faculty of Medicine, effects of L-carnitine and selenium on rat’s Department of Physiology, Faculty of pancreatic biochemical and structural changes Pharmacy, 2Department of induced by 2.45 GHz electromagnetic radiation. Pharmaceutical Technology, In this study thirty male Wistar Albino rats were 3 used. Rats divided five groups randomly and Department of Biochemistry, 33169, groups were named as cage control, shame Mersin control, electromagnetic radiation (EMR), EMR+Selenium [(1.5 mg/kg/ every other day- Mutations in humans are associated with intraperitoneal (ip)], EMR+L-carnitine (100 several forms of inherited retinal dystrophies, mg/kg/day-ip). Except for cage and shame such as Retinitis Pigmentosa which lead to control groups all of the other groups were retinal cell death and irreversible loss of vision. exposed to 2450 MHz EMR 60 min per day In retinitis pigmentosa cases, photoreceptors during 28 days. At the end of the study blood can get lost as a result of apoptosis in retina. samples were obtained. Then amylase, lipase, Reactive oxygen products can have a role on alkaline phosphatese (ALP) and gamma retina cells apoptosis and trigger the retinitis glutamil transferase (GGT) enzyme activities pigmentosa. For this aim, venous blood were measured from the blood serums. After

25 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels samples of patients and controls were collected antioxidant levels in brain cortex of diabetic and analyzed for plasma malondialdehyde ovariectomized rats were investigated. Forty- (MDA) level as a marker of lipid peroxidation two rats were equally divided into seven and catalase, superoxide dismutase (SOD) and groups: (1) Control; (2) ovariectomized (OVX); glutathione peroxidase (GSH-Px) activities for (3) OVX+E2; (4) OVX+E2+vitamin E; (5) determining the antioxidant statue in a group of OVX+diabetic; (6) OVX+diabetic+E2 and (7) -1 30 patients with retinitis pigmentosa and OVX+diabetic+E2+vitamin E. E2 (40μg kg /day) compared with 45 age and sex-matched and vitamin E (100 mg kg-1/day) were healthy control subjects without retinitis administered. The activities of glutathione pigmentosa. Statistical evaluation of data was peroxidase (GSH-Px) and catalase (CAT), performed SPSS v10.0 programme and superoxide dismutase (SOD) and levels of independent t-test for the comparison of the glutathione (GSH), vitamin A and beta-carotene groups. The levels of MDA showed a significant were significantly reduced in brain (p<0.05) increase (p<0.05) in the patients when compared to control in OVX but level of MDA in compared to control. SOD, catalase and GSH- brain and level of glucose in plasma were Px activities were lower in retinitis pigmentosa significantly increased (p<0.05). The activities than in control group (p<0.001). As a result of of antioxidant in brain (p<0.05) increased this study, it can be suggested that lipid although MDA (p<0.05) in brain and glucose peroxidation increases and antioxidant enzyme (p<0.05, p<0.01) in plasma decreased in OVX activities decrease in retinitis pigmentosa after E2 and E2+vitamin E supplementation. The cases. In conclusion, it is assumed that being activities of GSH-Px (p<0.001, p<0.01), CAT exposed to free radicals or insufficient level of (p<005, p<0.001) and SOD (p<0.001) and the antioxidant enzymes can be the considerable levels of GSH (p<0.001), vitamin A and beta- reason of retinitis pigmentosa. carotene (p<0.001, p<0.05) were lower in the brain of OVX diabetic rats, while MDA in the brain and glucose in the plasma were higher References (p<0.001). The antioxidant enzymes (p<0.05), GSH, vitamin A and beta-carotene (p<0.01, 1.Wagreich H, Lasky MA, Elkan B. 1961. Some p<0.001) in brain increased while MDA in brain biochemical studies in retinits pigmentosa and glucose in plasma decreased (p<0.01, clinical chemistry. Clinical Chemistry. 7:143– p<0.001) in diabetic OVX after treatments. In 148. conclusion, the E2 and E2+vitamin E supplementation to diabetic OVX and OVX rats 2.Furukawa T, Morrow EM, Cepko CL. 1997. may strengthen the antioxidant system by Crx, a novel otxlike homeobox gene, shows reducing lipid peroxidation. photoreceptor-specific expression and regulates photoreceptor differentiation. Cell. Poster No 50 91:531–541. 17β-estradiol and vitamin E o Poster N 49 supplementation restore blood trace element and antioxidant 17β-estradiol and vitamin E decrease enzyme to control levels in oxidative stress in brain cortex of ovariectomized rats diabetic ovarictomized rats 1 2 1 2 Mustafa Ulas and M. Cay Mustafa Ulas and M. Cay

1 1 Department of Physiology, Faculty of Department of Physiology, Faculty of Medicine, Firat University, Elazig, Medicine, Firat University, Elazig, Turkey Turkey 2 2 Department of Physiology, Faculty of Department of Physiology, Faculty of Veterinary Medicine, Firat University, Veterinary Medicine, Firat University, Elazig, Turkey Elazig, Turkey In the present study, effect of 17β-estradiol (E2) In the current study, the effects of 17β-estradiol alone and separately vitamin E treatment on (E2) and vitamin E on lipid peroxidation and trace element status of rats following an

26 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels ovariectomic operation were investigated. Forty In this study, we investigated the apoptotic and rats were equally divided into four groups: (1) antiproliferative effects of α-tocopherol on control; (2) ovariectomized (OVX) rats, which ECV304 human endothelial cells, combined were ovariectomized under general anesthesia. antiproliferative agents frequently used in (3) OVX+E2 rats, which received subcutan E2 at glaucoma surgery. Side effects of this current a dose of 40µg kg-1 per day. (4) antifibrotics on endothelium, limits their use in OVX+E2+vitamin E rats, which received glaucoma surgery. Vitamin E is an important intraperitoneal vitamin E at a dose of 100 mg natural antioxidant and its most common and -1 kg as well as subcutan E2 at a dose of 40µg biologically active form α-tocopherol, prevents kg-1 per day. At the end of the 30-day tenon’s fibroblast proliferation and it also effects treatment, the rats were sacrificed and their similar pathways with other antifibrotics as well. blood was collected for biochemical Antiproliferative and apoptotic effects of α- measurements. The levels of zinc, copper, iron, tocopherol were comperatively evaluated, both phosphorus, selenium, calcium, chromium and solely and in combination with antifibrotic manganese and activities of copper-zinc agents for long and short time applications. superoxide dismutase (SOD), manganese- Mitomycin-C, 5FU and paclitaxel were used as superoxide dismutase (Mn-SOD), glutathione antifibrotic agents in vitro. Apoptotic indexes peroxidase (Se-GSH-Px) and catalase (CAT) found with DAPI staining in fluorecence were lower in the OVX when compared to microscope. Apoptotic cells were evaluated in control group, but magnesium level was molecular level with RNA isolation, bcl2 and unaffected. However, zinc, copper, iron, bax gene expression were evaluated with PCR phosphorus, selenium, calcium, chromium and and also determined with caspase3 manganese levels and SOD, Mn-SOD, Se- immunohistochemical staining. Mitotic indexes GSH-Px and CAT activities were higher in E2– were also determined. Mitomycin-C and treated and E2+vitamin E–treated groups. There paclitaxel groups’ demostrated a significant AI was no significant difference between the increase which were augmented by α- treated-OVX groups the OVX group in the level tocopherol addition. Bcl2/Bax ratio decrease or of magnesium. In conclusion, E2 treatment has caspase activity was found only in mitomycin an ameliorating effect on the trace element group and further augmention was achieved status in OVX, and this effect may be enhanced with α-tocopherol addition. Mitotic cells were with the addition of vitamin E. observed in α-tocopherol groups. We observed noticable decreased in mitosis for short and long term applications of mitomycin-C and 5FU Poster No 51 groups. As a result, combination with α- tocopherol did not reduce the apoptotic effects of antifibrotics on human endothelial cells. 5FU Apoptotic and antiproliferative and α-tocopherol have been found to be safe in effects of single and combined use glaucoma surgery for therapy because of little of alpha tocopherol and antifibrotic apoptotic and antimitotic effects. agents, on human endothelium cells: a comparative evaluation References; 1.Brigelius-Flohe R, Traber MG. Vitamin E:

1 1 Function and metabolism. FASEB J. 1999; M. Beyza Çetin , S. Karadenizli , U. 13:1145-1155. Serbes2, K. Akgün- Dar2, G. Ö. Arıcan2, 2. Nagata M, Kawazu K, Midori Y, et al. K.N. Engin3, S. E. Kuruca1, Intracameral and lenticular penetration of locally applied stable isotope-labeled vitamin E. Jpn J 1 Ophthalmol. 2001; 45:125-127. Istanbul University, Istanbul Faculty of 3. Takahashi O. Haemorrhagic toxicity of a Medicine, Department of Physiology, large dose of - α-, -β, -γ ve α-tokoferols, Istanbul, Turkey ubiquinone, β carotene, retinol acetate ve L- 2Istanbul University, Faculty of Science, ascorbic acid in the rat. Food Chem. Toxicol. Department of Biology, Istanbul, Turkey 1995; 33:121-128. 3 Bağcılar Education and Research Poster No 52 Hospital, Department of Ophthalmology, Istanbul, Turkey The effect of smoke and smokeless tobacco use as Maraş powder on

27 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels saliva adenosine deaminase, 1University of Kahramanmaraş Sütçü xanthine oxidase, and İmam University, Faculty of Science malondialdehyde levels Department of Chemistry, Division of Biochemistry, Kahramanmaraş, Turkey Naciye Kurtul1, E. Gökpınar 1 2University of , Faculty of Medicine, Department of 1University of Kahramanmaraş Sütçü Neurology, Gaziantep, Turkey İmam, Faculty of Science Department of Chemistry, Division of Biochemistry, Migraine a common and disabling disorder. It has suggested that the release of nitric oxide Kahramanmaraş (NO) is an important trigger mechanism in migraine. In this study, NO level and lipid The use of smokeless tobacco (ST) is common peroxidation (LPO) as malondialdehyde (MDA) worldwide. Also, in Kahramanmaraş, a city were measured in the serum samples obtained Southern Turkey, ST used as Maraş powder from the 22 healthy controls without migraine (MP) has been consumed widely instead of (mean age: 30.40±6.28 years) and 26 migraine cigarette smoking for a long time. The habit is patients (mean age: 31.21±6.55 years) in a becoming increasingly popular, especially due crisis-free period. Also, migraineurs were to banning smoking in our country. MP is grouped as with and without aura. Mean MDA prepared from a tobacco of species Nicotiana level was found to be significantly higher in rustica L. In this study, xanthine oxidase (XO), migraine group than that of control subjects adenosine deaminase (ADA), and lipid (p<0.001). Even though migraine patients group peroxidation levels as malondialdehyde (MDA) had NO values higher than that of control were measured in saliva samples obtained from groups, this difference wasn’t statistically smokers (Group II), MP users (MPU; Group III), important (p>0.05). At these parameters there and healthy control subjects (Group I) who were wasn’t a statistically difference between nonsmokers and nonusers of ST and the migraine groups with aura and without aura. As results were compared. The salivary XO, ADA, a result it was observed that migraine had effect and MDA levels were found to be significantly on the levels of the measured parameters in higher in smokers and MPU than that of control serum. While NO normally functions as a subjects and also in MPU than that of smokers physiological neuronal mediator, excess (p<0.001). In Group III, statistically important production of NO mediates brain injury. As a correlation was found between MDA levels and free radical, NO mediates cellular toxicity and ADA, and XO activities. We have also observed reacts with superoxide to form peroxynitrite that as the number of cigarettes and MP which an even more potent oxidant. Thus the amount increases, the salivary XO, ADA, and findings of the present study suggest that NO MDA levels increase. We have concluded that and LPO are possibly important factors in the salivary XO, ADA, and MDA levels are migraine pathogenesis. increased by smoking and MP use. Results obtained from this study can help evaluate harmful effects of various use of tobacco. It is Poster No 54 important to point out that bigger change in the measured parameters observed for MP use. This finding may be an indication of harmful Attenuation of Ischemia-Reperfusion effects of ST use as MP and draw attention to a Injury by Iloprost on Apoptosis and significant potential public health hazard. Antioxidant Capacity in Kidney as a Distant Organ Poster No 53

Necmiye Canacankatan1, N. Sucu2, S. Evoluation the nitric oxide level and 3 1 4 lipid peroxidation in the serum of Şahan-Fırat , A. Görür , Ş.E. Antmen , B. Korkmaz3, B. Tunçtan3, OE. Gül1, L. migraine patients 5 5 2 6 Tamer , L. Ayaz , Ö. Vezir , A. Kanık 1 1 2 Naciye Kurtul , A. Seyhan , A. Akçalı 1 Mersin University, Faculty of Pharmacy, Department of Biochemistry, Mersin, Turkey.

28 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

2Mersin University, School of Medicine, Influence of grape seed oil on Department of Cardiovascular Surgery, paraoxonase/arilesterase activities Mersin, Turkey. of the erytroleukemia cell line treated 3Mersin University, Faculty of with methotrexate Pharmacy, Department of Pharmacology, Mersin, Turkey Nefise Özlen Şahin1, E.D. Eker1, M. 4Mersin University, Medical Services Berköz2, S. Yalın3, B. Pomierny1,4, K. Vocational School, Mersin, Turkey Przejczowska1,4 5Mersin University Medical Faculty, Department of Biochemistry, Mersin, Mersin University, Faculty of Pharmacy, Turkey 1Department of Pharmaceutical 6Mersin University, Medicine Faculty, Biotechnology, 2Department of Department of Biostatistics, Mersin, Pharmaceutical Technology, Turkey 3Biochemistry, Yenisehir Campus, Mersin 33169, Turkey. 4Jagellonian Activation of apoptotic cascade is crucially University, Faculty of Pharmacy, involved in post-ischemic inflammation which Krakow, Poland. causes tissue damage and organ deficiency. In this context, we investigated the cytoprotective and antioxidant effects of iloprost, a There is a balance between a free radical prostacyclin analogue in kidney as a distant formation and endogenous antioxidant defense organ after ischemia-reperfusion (I/R) injury. mechanisms. Disturbance of this balance For this purpose; apoptosis was evaluated by results in oxidative stress. This, in turn, causes measurement of caspase -3, -8 and -9 injury to all the important cellular components enzymes, antioxidant capacity was evaluated like proteins, DNA and membrane lipids which by measurement superoxide dismutase (SOD) can cause cell death. Recently, significant enzyme activity, and nitric oxide (NO) evidence in experimental and clinical data was production was evaluated by measurement of provided to indicate the involvement of nitrite content in kidney tissues. Wistar albino oxidative stress in carcinogenesis. On the other rats were randomized into five groups as sham, hand, research conducted on toxicity of ischemia, I/R, iloprost (10 g/kg) and I/R with anticancer drugs indicates compelling evidence on oxidative stress as well. The most significant iloprost (10 g/kg). 12 rats were included in drug with cytotoxicity is methotrexate (MTX), each group. A 4 h reperfusion procedure was which has been widely used in treatment of carried out after 4 h of ischemia. 10 g/kg various types of cancer. Additive effect of iloprost was administered to the rats in 1ml of oxidative damage caused by MTX to oxidative saline from the tail vein in iloprost (10 g/kg) stress induced by cancer makes the situation group. Rats in I/R with iloprost (10 g/kg) group dramatically bad. Oxidative stress has been received 10g/kg iloprost in the same way 10 reported responsible for the adverse reaction min before reperfusion. There were statistically mechanism of MTX in liver, kidney, intestine, significant increase in caspase -3, -8 and -9 CNS, and hematopoietic cells. In order to enzymes in both ischemic and I/R groups to the reduce the damage, several medical sham group. Iloprost prevented these increase approaches have been suggested. The use of in caspase activities. SOD enzyme activity and products with antioxidant activity (e.g. the nitrite levels also increased in ischemic rats melatonin, vitamin E, methionine) can reduce compared to the sham group; but only SOD the influence of MTX-induced tissue damage. activity decreased by iloprost. As a conclusion Grape seed is one of the most significant iloprost may be considered as a cytoprotective profilactic agents due to its antioxidant and agent to apoptosis. However, further studies bioflavonoids composition. However, there is no are needed with different doses of iloprost to study regarding the protective effect of grape emphasize these results. seed oil on the MTX-induced oxidative stress. Therefore, the aim of this study was to

o investigate the protective effect of grape seed Poster N 55 oil against MTX-induced oxidative stress in K- 562 human erythroleukemia cell lines. Paraoxanase and aryl esterase enzyme

29 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels activities indicating antioxidant properties in cell Reactive oxygen substances (ROS) and lipid lines were investigated by the method of peroxides cause injury by reacting with Eckerson et al. Protein assay was conducted by biomolecules as well as by depleting enzymatic Lowry method at the 24th and the 48th hour and/or nonenzymatic antioxidants, plasma following the treatment. The difference between membrane dependent microsomal Ca2+- groups was statistically evaluated using SPSS ATPase in the brain. Caloric restriction rescues v10.0 program and ANOVA test. It was neuronal cells from mitochondrial redox determined that MTX treatment reduced both impairment and ROS because ROS produce in PON and ARE activities at both time points. mitochondria by metabolism of proteins, lipids Involvement of grape seed oil administration did and carbohydrates. The mode of action on not change PON and ARE activities for 24 neuron protection by caloric restriction is hours. However, a significant increase was unclear. We investigated effects of caloric observed at 48th hour. In conclusion, grape restriction on Ca2+-ATPase and antioxidant seed oil administration for 48th hours following redox system in rat brain. Thirhty eight rats MTX treatment gives rise to paraoxanase and (four month) were divided into four groups. The aryl esterase activities in erythroleukemia cells. first, second, third and fourth groups were used Thus, it may be suggested that use of grape for the control, caloric restriction, obese and seed oil along with MTX in cancer patients may obese+caloric restriction groups, respectively. provide a protective effect against oxidative Initial body weight of control and obese groups stress and xenobiotics that are the substrates of were 250±20 g and 400±20 g, respectively. these enzymes. Second and fourth groups were exposed to 60% caloric restriction for 10 weeks. After 12 References hours fasting, brain samples were taken from the four groups and brain microsomal samples 1. Rubio M, Alvarez-Ortí M, Alvarruiz A, were obtained by ultracentrifugation. Lipid Fernández E, Pardo JE. Characterization of oil peroxidation levels of brain cortex were obtained from grape seeds collected during increased in second group although lipid berry development. J Agric Food Chem. 2009; peroxidation levels were decreased in fourth 57: 2812-2815. groups by caloric restriction. Brain vitamin A 2. Sahin F, Avcı CB, Avcu F, Ural AU, Sarper and C concentrations, and brain microsomal M, Hışıl Y, Omay SB, Saydam G. Red grape Ca2+-ATPase activities were decreased in seed extract and its compound resveratrol exert second groups although Brain vitamin A, C and cytotoxic effect to various human cancer lines. E concentrations, and brain microsomal Ca2+- Turk J Hematol. 2004; 24: 102-109. ATPase activities were increased in fourth groups by caloric restriction. Brain glutathione Poster No 56 peroxidase and reduced glutathione values did not change in the four groups. Effects of caloric restriction on brain In conclusion, caloric restriction in obese rats antioxidant redox system, microsomal seems to have protective effects on the Ca2+-ATPase activity and nitric oxide oxidative stress-induced brain toxicity by levels in rats inhibiting free radicals and supporting the antioxidant redox system.

M. Nazıroğlu1, A. C. Uğuz1, Ö. Çelik1, Nigar 2 3 4 Acknowledgement: Dr. Nigar Yılmaz was Yılmaz , M. Yılmaz , H. Vural supported by TUBITAK for the presentation.

1 Department of Biophysics, Medical Poster No 57 Faculty, Süleyman Demirel University, Isparta, Turkey. Investigation of the Effects of 2 Department of Biochemistry, Medical Electromagnetic Field at 2.45 GHz on Faculty, Mustafa Kemal University, Rat Skeletal Muscle Tissue with Hatay, Turkey. 3 Protective Effects of Selenium and L- Department of Neurology, Antakya State Carnitine Hospital, Hatay, Turkey

4Department of Biochemistry, Medical D. Bayram1, N. Gumral2, A. Gokcimen1, Faculty, Süleyman Demirel University, 2 Isparta, Turkey. S. Calıskan

30 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

1Department of Histology and 1Department of Biochemistry, Faculty of Embryology, Medical Faculty, Pharmacy, Istanbul University, 34116, Süleyman Demirel University, Isparta, Istanbul, TURKEY Turkey, 2Department of Chemistry, Faculty of 1 Department of Physiology, Medical Engineering, Istanbul University, 34320, Faculty, Süleyman Demirel Univesity, Istanbul, TURKEY Isparta, Turkey, Various substances are investigated nowadays The hazard of microwave electromagnetic fields for the prevention of diabetic complications. (EMF) to humans and animals has been During long-life diabetes, persistent high blood intensively studied during the last 20 years, glucose levels cause the destruction of small because large professional groups of people blood vessels in the nephrons, leading to are subjected to microwave exposure. Several diabetic nephropathy and chronic kidney studies have suggested that biological systems disease. Zinc is a trace element possessing a would exhibit sensitivity to 2.45 GHz of EMF wide range of functions and antioxidant emitted from wireless settings. This study was properties. This study was undertaken in order conducted to investigate the histopathological to illuminate the conflicting data on the status of effects of electromagnetic field at 2.45 GHz on zinc in diabetes, present in literature. Female rat skeletal muscle tissue and protection by Swiss albino rats were randomly divided into 4 Selenium (Se) and L-Carnitine (L-Car). groups: Group I, control; Group II control+zinc There were five study groups each consisting of sulfate; Group III, STZ-diabetic; Group IV, STZ- six animals. Group I: Controls, Group II: diabetic+zinc sulfate. Diabetes was induced by Shame- controls, Group III: 2.45 GHz EMF intraperitoneal injection of streptozotocin (STZ, exposed, Group IV: 2.45 GHz EMF exposed + 65 mg/kg body weight). Zinc sulfate was given Se treated (1.5 mg/kg/over day) and Group V: daily by gavage at a dose of 100mg/kg body 2.45 GHz EMF exposed + L-Car treated (100 weight every day for 60 days to groups II and mg/kg/day). 2.45 GHz EMF was applied to IV. At the last day of the experiment, rats were Groups III, IV and V for 28 days (1h/day). At the sacrificed, kidney tissue was taken, end of the 4th week, tissue samples were taken homogenized by means of a glass homogenizer for histological examination. in cold saline to make a 10% (w/v) Control and shame groups were observed as homogenate. Antioxidant enzymes, catalase, normal. There were slight changes in skeletal glutathione reductase, glutathione peroxidase, muscle tissues of L-Car and Se groups but glutathione-S-transferase, superoxide these differences were not statistically dismutase activities were determined in tissue meaningful (p>0.05). Some structural changes homogenates. The results were evaluated observed in 2.45 GHz exposed group, including statistically. It was shown that antioxidant skeletal muscle fibrillary degeneration and enzyme activities were significantly decreased increased of muscle fibers diameter when in zinc treated diabetic groups in comparison compared with other groups (p≤0.05). with untreated diabetic group, thus showing the Exposure to the 2.45 GHz EMF caused some beneficial effect of Zn treatment in diabetes. degeneration in skeletal muscle while these o changes were fixed in L-Car and Se given Poster N 59 groups in our study. Antioxidant effects of these agents may be useful for degeneration skeletal Activity paraoxonase and muscle. arylesterase in sportsmen and sedentary controls Poster No 58

Oral zinc supplementation protects E. Eren1, A. Yeğin2, A. Kurtoğlu2, rat kidney tissue from oxidative Nurullah Ay2, N. Şimşek2, N. Yılmaz2 stress in diabetic rats

1 N.Özsoy1, A. Can1, Ö. Mutlu2, Nuriye Antalya Atatürk Hospital, Biochemistry Akev1, R. Yanardağ2 Laboratory, Antalya, Turkey

31 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

2 Antalya Education and Research Süleyman Demirel University, Isparta, Hospital, Clinical Biochemistry, Antalya, Turkey. Turkey Forty female Wistar rats were randomly divided Physical activity aids the human body into four groups. Each group consisted of 10 antioxidant defense system via obscure rats. Control group received 1 ml/kg serum mechanisms. Paraoxonase (PON) and physiologic (SF) intraperitoneally (i.p.) once a arylesterase (ARE) are enzymes that own day, CAPE and GM+CAPE groups received 10 antioxidant characteristics. The studies μmol/kg/day CAPE i.p. for 12 days. GM group; investigate a possible beneficial relationship, injected i.p. with 1 ml/kg SF for 2 days before that increased levels of these enzymes due to GM treatment and after the GM and afterwards physical activity might help our understanding i.p with 100 mg/kg GM for 8 days, continued of the issue. In this study we focus on healthy only injected i.p. with 1 ml/kg SF for 2 days. young subjects unlike previous ones that have GM+ CAPE treated group; injected i.p with 10 prefered middle aged or older participants. μmol/kg CAPE for 2 days before GM treatment Serum paraoxonase and arylesterase activities and daily during GM treatment and after the GM were evaluated in 31 well-trained young treatment for 2 days. On the twelfth day of the sportsmen and 30 sedentary age-matched study all rats were sacrificed and then blood controls. Paraoxonase activity measurements samples and kidneys were taken. Left kidneys were performed in the absence of salt (basal were used for hystopathological evaluation. activity). The amount of generated p- Malonyldialdehyde (MDA) and nitric oxide (NO) nitrophenol was calculated from the molar levels, glutathione peroxidase (GSH-Px), absorptivity coefficient at pH 10.5, which was superoxide dismutase (SOD) and catalase 18,290 M−1cm−1. Phenylacetate was used as a (CAT) enzyme activities were determinated in substrate to measure the arylesterase activity. right kidneys of rats. Levels of BUN and Enzymatic activity was calculated from the creatinin were studied in serum. molar absorptivity coefficient of the produced Hystopathologic evaluation showed phenol, 1310·mol-1·L-1·cm-1. Although the nephrotoxicity findings in GM and GM+CAPE sportsmen were not receiving any special diet groups. Serum BUN and creatinin levels of GM or vitamin supplementation, they showed a group were significantly higher than control slightly improved antioxidant status, mainly group. In GM group, SOD, GSH-Px, CAT represented by increased paraoxonase enzyme activities were lower than the control (p<0.01), arylesterase levels (p<0.05), and in group. NO and MDA levels were higher than addition, elevated ratio of paraoxonase/HDL-C the control group. In GM+CAPE group, it has (p<0.01). In biological systems, cells respond to been observed that the oxidative stress amount mild oxidative stress by inducing their was decreased in renal tissue. According to antioxidant defenses and other protective these findings, it could be concluded that systems. Increased paraoxonase and oxidative stress may be a critical mechanism in arylesterase activities, in young sportsmen GM nephrotoxicity and using of CAPE maybe might indicate eliminated oxidative stress. It an effective treatment option in prevention of proves once again that physical activity oxidative stress due to GM nephrotoxicity. provides disease prevention. o Poster N 61 Poster No 60 The role of SAG as a prognostic Investigation of the role of oxidative factor on radio/chemotherapy in stress in experimental gentamycine patients with advanced colorectal induced nephrotoxicity and effect of cancer Caffeic Acid Phenethyl Ester on possible oxidative stress in rats Oya Orun1, S. Özden1, 2, H. Özyurt2, Z. Özgen2, M. Öncel2, O. Kılınç1, B. Kan3 Fadime Özge Aygün1, F.Z. Akçam1, O.Kaya1, B.M.Ceyhan2, R.Sütçü2 1Department of Biophysics, Marmara Department of Infectious Diseases1, Univ. School of Medicine, Department of Biochemistry and Haydarapasa, İstanbul. Clinical Biochemistry 2, Medical Faculty,

32 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

2Dr.Lütfi Kırdar Kartal Education and Research Hospital, Kartal, İstanbul. 3Department of Biophysics, Acıbadem Univ. School of Medicine, Maltepe, İstanbul.

Colorectal cancer is one of the most malignant disorders worldwide and show nearly 50% local recurrence posing a serious problem in treatment (Folkesson 2005, Adell 1999). Preoperative radiotherapy with or without chemotherapy is widely accepted to improve local control and overall survival rates. Resistance of cancer is one of the main causes for treatment failure. Predictive indicators will be highly valuable estimating the response of a patient. SAG (sensitive to apoptosis gene) protects cell from apoptosis induced by redox agents like hydroxyl radicals and irradiation. Prognostic value of apoptosis has been described at various reports. To assess the relation between SAG expression levels and sensitivity to radiation, 31 patients with rectal cancer were examined in this study before and after treatment. Our results indicate that expression levels of apoptosis related genes can be a regulatory factor for resistance against exposure and can considerably affect the therapeutic results, sensitivity against radiation and survival rates. This small group under study has revealed that SAG expression have correlated with 1 and 2 year survival rates and could have predictive value in progression of disease.

Poster No 62

Serum vitamin levels in young basketball players and sedentary controls

E. Eren1, G. Aykal2, A. Yeğin2, N. Ay2, Özgür Aydın2, N. Yılmaz2

1 Antalya Atatürk Hospital, Biochemistry Laboratory, Antalya, Turkey 2 Antalya Education and Research Hospital, Clinical Biochemisty, Antalya, Turkey

33 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Physical activity is known to increase the effects on acetaminophen mediated damage antioxidant defense system and reduce on rat liver tissue.48 Wistar Albino female rats exercise-induced oxidative stress. were randomly divided into six equal groups. Nevertheless, the precise mechanisms by Groups planned as : Control. Acetaminophen which regular training is responsible for this (1 g/kg, ip, single dose), Melatonin (10 mg/kg, protection are not completely clear. The cohort ip, single dose), Carnozine (250 mg/kg, ip, studies that have indicated benefits of physical single dose), Acetaminophen plus Melatonin activity have all focused on middle aged or and Asetaminofen plus Carnozine, at the same older participants. The current study focuses doses. At the end of the study; Serum tolal on 10 year old young basketball players. oxidan status (TOS) and total antioxidant Antioxidant profiles α-tocopherol, retinol, status (TAS) levels measured and oxidative pyridoxal-5-phosphate, bilurubin, and uric acid stres index (OSI) values were calculated. Liver were evaluated in 31 well-trained young tissue samples examined histologically and basketball players and 30 sedentary age- liver tissue inducible nitric oxide sentase matched controls. Although the sportsmen (iNOS) receptor range examined were not receiving any special diet or vitamin immunohistochemically. In histological supplementation they showed a slightly examination; damage in the rat liver tissues improved antioxidant vitamin status, mainly and sentrolobuler necrozis caused by represented by an increased antioxidant Acetaminophen markedly decreased in the status. The latter was evidenced by an liver tissues of Melatonin and Carnosine plus increment in higher serum retinol, pyridoxal-5- groups (p<0.05). In the only Acetaminophen phosphate and α-tocopherol (p<0.05) given group, iNOS receptor range increased; concentrations. In biological systems, cells as seen more stained immunohistochemically. respond to mild oxidative stress by inducing In Acetaminophen plus Melatonin and their antioxidant defenses and other protective Acetaminophen plus Carnosine groups, a little systems. Increased vitamin levels in young more staining were present when compared to basketball players might indicate eliminated the control group. The increased values of oxidative stress. Physical activity once again TOS and OSI parameters in the promises a prolonged and healthier life. Acetaminophen group, significantly decreased in the Acetaminophen plus Carnosine and Poster No 63 Acetaminophen plus Melatonin groups, especially in the Acetaminophen plus Evaluation of the Effect of Carnosine group (p<0.05). We have concluded Carnosine and Melatonin on that; in preventing high doses of Acetaminophen’s toxic effects in the rat liver Acetaminophen Induced Acute Liver tissues, melatonin and especially carnosine Toxicity. has powerful effects.

Ozgur Fedakar1, M. Oncu1, F. Poster No 64 Gultekin2 Cellular telephone use and cancer 1Department of Histology and risk

Embryology, University of Suleyman 1 2 Demirel, Isparta, 32100, Turkey. Ozlem Coskun , S. Comlekci 2 Department of Biochemistry, 1 University of Suleyman Demirel, Department of Biomedical Device Isparta, 32100, Turkey. Technology, University of Suleyman Demirel University, Isparta, TURKEY. Acetaminofen is the most common using pain 2 and fever reducing medical agent worldwide. Department of Electronics and But there is a lot of study, suggesting this Communication Engineering, medicine makes suffering of liver via liver University of Suleyman Demirel damage. Melatonin, which is famous as a member of most powerful antioxidants, is a University, Isparta, TURKEY. hormone. Carnosine is a powerful antioxidant dipeptide. In our study, our goal was to reveal The cellular phones and base stations are to compare melatonin and carnosine for their getting more into our lives. The effects of this

34 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels fast development in technology have been with ionizing radiation sometimes may be creating a great concern and curiosity on required to diagnose. Ionizing radiation can public and science community. In the last two directly affect fetal development in several decades, many studies have been published in ways. Radiation dose and the time of exposure the media and in the scientific world on the are critically important in the development of effect of radiofrequency (RF) emissions these effects. Radiation can cause fetal death distributed from cellular phones on cancer. or congenital anomalies, growth deficiencies or There are contradictory results on this topic in several structural and/or functional the literature. The assessment of these studies development abnormalities. The purpose of from the biophysical point of view revealed that this study is to discuss the potential adverse RF energy in general did not cause a effects of radiologic imaging modalities on significant biological effect. However, there are pregnancy and fetus. also reports suggesting a direct or indirect effect of thermal RF energy on cancer Poster No 66 progression. Studies performed in humans mainly concentrated on brain cancer, leukemia The protective effects of ghrelin on and intraocular melanoma. Although no solid stomach tissue of neonatal diabetic association was found between cellular phone use and cancer risk, there are some studies rats showing an increased risk in acoustic neuroma 1 2 3 and glioma due to long term (≥ 10 years) Ozlem Sacan , N. Türk , A. Karatug , cellular phone use. Animal studies could not S. Bolkent3, R. Yanardag1, provide consistent evidence that RF energy S. Bolkent2 exposure at non-thermal intensities caused cancer. Therefore, planning of detailed 1 research on this subject, definition and Department of Chemistry, Faculty of application of legal limitations, pursue of Engineering, Istanbul University, studies are vital for this issue. Additionally, it is Avcilar, Istanbul, Turkey. important to inform public about the possible 2Department of Medical Biology, adverse effects of electromagnetic radiation and get them take the precautions about the Cerrahpasa Faculty of Medicine, use of the equipment generating radiation. Istanbul University Cerrahpasa, Istanbul, Turkey. 3Department of Biology, Faculty of o Poster N 65 Sciences, Istanbul University,

Vezneciler, Istanbul, Turkey. Health effects of radiation exposure during pregnancy Diabetes mellitus is associated with an 1 2 Ozlem Coskun , S. Comlekci increased production of reactive oxygen species and a reduction in antioxidant 1Department of Biomedical Device defenses. Ghrelin is a novel 28 amino acid gastric peptide, which controls the acid release Technology, University of Suleyman and is produced abundantly in the stomach. Demirel University, Isparta, TURKEY Effects of ghrelin treatment in diabetes are still unknown exactly. In the present study, we 2Department of Electronics and aimed to demonstrate the effect of ghrelin on Communication Engineering, stomach damage in streptozotocin (STZ)- induced diabetic rats. In this study, Wistar University of Suleyman Demirel albino newborn rats were divided into four University, Isparta, TURKEY groups. Group I: Saline was administered intraperitonally (i.p.). Group II: Ghrelin dissolved in saline was given to newborn rats Imaging modalities that do not use ionizing from day 3 after birth, 100 μg/kg daily, radiation (ie, ultrasonography and magnetic subcutaneously (s.c), during four weeks. resonance imaging) are preferred for diagnosis Group III: Second day after birth, 100 mg/kg of maternal and fetal illnees during pregnancy. STZ was administered i.p. as a single dose to However, radiographic imaging procedures newborn diabetic rats (n2-STZ) group. Group

35 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

IV: Diabetic animals were given ghrelin for four extender containing Curcumin (0,5 and 2 mM), weeks. Stomach tissue was taken from dithioerythritol (0,5 and 2 mM) and no additive animals, homogenized in 0.9% saline to make (control) was cooled to 5oC, and frozen in 0.25 up to 10% homogenate. The homogenate was ml French straws. French straws. Frozen used for protein level and enzyme activities. straws were thawed individually at 37oC for 20 Catalase (CAT), superoxide dismutase (SOD), s in a water bath for evaluation. glutathione peroxidase (GPx) and glutathione The extender supplemented with 0,5 mM dose reductase (GR) activities were determined. of curcumin led to lower percentage of total Stomach CAT, SOD, GPx and GR activities abnormality (20.40±2.36%), when compared to were decreased in diabetic rats. Administration the control (30.60±1.47%, p<0.05). Curcumin of ghrelin to diabetic rats increased stomach and dithioerythritol at 0,5 mM provided a CAT, SOD, GPx and GR activities. As a result, greater protective effect in the membrane we can propose that ghrelin could be a functional integrity (54.40±2.09% and potentially beneficial agent in reducing 50.00±2.68%), in comparison to control stomach damage in neonatal diabetic rats, (37.20±1.77%, p<0.001). While curcumin and probably by decreasing oxidative stress. dithioerythritol at 0,5 mM led to higher percentages of post-thaw motilities, when Poster No 67 compared to the control groups, these increases seemed to be insignificant. No Effects of Curcumin and significant differences were observed in sperm Dithioerythritol on Post-thawed Bull acrosome abnormalities among the groups (p>0.05). Supplementation with antioxidants Sperm did not significantly affect the LPO and AOP levels, compared to the control groups. The 1 2 N. Başpınar , Pınar PEKER AKALIN , maintenance of total GSH level in curcumin 0.5 M. N. Bucak3, P. B. Tuncer3, S. mM was demonstrated to be higher than that Sarıözkan4, K. Coyan5 of control, following the freeze-thawing (p<0.05). 1 Selcuk University, Faculty of Veterinary Medicine, Department of Poster No 68 Biochemistry, Konya, Turkey 2Ministry of Agriculture and Rural The Effect of Cysteine And Affairs, Central Veterinary Control and Glutathione on Sperm Parameters, Research Institude, Ankara, Turkey Malondialdehiyde and Glutathione 3Ministry of Agriculture and Rural Peroxidase Activities of Post- Affairs, Lalahan Livestock Central Thawed Bull Semen Research Institute, Lalahan, Ankara, Turkey Pürhan Barbaros Tuncer1, M. N. Bucak 4Erciyes University, Safiye Cikrikcioglu 1, S. Sarıözkan 2, S. Büyükleblebici1 Vocational College, Kayseri, Turkey , H. Kinet 1, O. Büyükleblebici 3 5 Selcuk University, Faculty of Veterinary Medicine, Department of 1 Ministry of Agriculture and Rural Reproduction and Artificial Affairs, Lalahan Livestock Central Insemination, Konya, Turkey Research Institute, Lalahan, Ankara, Türkiye The aim of this study was to determine the 2 , Safiye Cikrikcioglu effects of curcumin and dithioerythritol added Vocational Collage, 38039, Kayseri, into bull semen extender on sperm Türkiye parameters, LPO (lipid peroxidation), total 3 glutathione (total GSH) and antioxidant , Faculty of potential (AOP) levels of bull sperm following Veterinary Medicine, Department of the freeze-thawing process. Nine ejaculates Biochemistry, Ankara, Turkey obtained from three bulls were included in this study. Each ejaculate which was splitted into seven equal groups and diluted in a Tris-based

36 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

The aim of this study was to determine the 4Department of Animal Nutrition and effects of antioxidants reduced glutathione Nutritional Diseases, Faculty of (GSH) and cysteine in Laiciphose® extender on semen parameters, malondialdehiyde (MDA) Veterinary, Dicle University, Diyarbakir and glutathione peroxidase (GPx) of post- thawed bull semen. Five groups, namely; GSH (0.5 and 2 mM), cysteine (5 and 10 mM) and This study was conducted to determine the control group, were conducted to test the effects of dietary natural clinoptilolite on serum effects of antioxidants in Laiciphose®. contents and feding performance. This Insemination doses were processed so that research was arranged according to each 0.25-ml straw contained 15 x 106 sperm. Randomized Plots Design by using 4 groups The addition of antioxidants did not provide with 3 replicates. Adult male Sprague–Dawley any significant effect on the percentages of rats (n=24) were randomly divided into 4 post-thaw sperm morphology (acrosome and groups with three replicates, including a control total abnormalities), subjective, computer group (without zeolite) and 3 doses of natural assisted sperm motility analysis (CASA) and zeolite (2, 4 and 6 %) in the diets. All rats were progressive motilities, as well as sperm motility fed the above concentrates during the characteristics (VAP, VSL, VCL, LIN and ALH), experimental period for 56 days. Blood compared to the control groups (p>0.05). samples were collected from each animal at However, cysteine at 10 mM dose gave rise to the end of the experiment. Dieatary a slight higher percentage of membrane clinoptilolite increased serum albumin, integrity assessed by HOST than those of the triglyceride and VLDL levels. However the other groups. With respect to fertility results differences among treatment groups were not based on 59-day non-returns, the significant for serum minerals (Ca, P, Mg, K, supplementation of GSH at 2 mM gave a lower Na, Cl, Fe), urea, Fe binding, LDL, ALP, rate (p<0.05). For MDA level, cysteine at 10 glucose, uric acid, total Fe, total protein, mM dose gave the highest level (4.99±0.44 globulin, cholesterol, HDL cholesterol, nmol/L) (p<0.001). GPX activity was creatinin; metabolizable energy and crude demonstrated to be higher upon the addition of protein consumption for 1 g live weight gain of 5 mM cysteine, when compared to the other rats. groups (p<0.05). According to diets, serum parameters were ranged for urea, Fe binding, LDL, alkaline fosfatase, glucose, uric acid, total Fe, total Poster No 69 protein, globulin, triglyceride, cholesterol, HDL cholesterol, albumin, VLDL, creatinine and Fe levels as follows (42.65 – 46.08, 314.17 – Effects of dietary zeolite on serum 325.50, 15.33 – 19.00, 662.90 – 816.44, contents and feeding performance 159.72 – 197.35, 1.56 – 1.73, 523.83 – 529.00, in rats. 6.53 – 6.82, 3.50 – 3.62, 40.78 – 58.40, 80.32 – 84.35, 54.44 – 57.32, 3.03 – 3.20, 8.16 – Ramazan Demirel, 1 Beran Yokuş,2 11.67, 0.54 – 0.58 and 198.33 – 214.33), Dilek Şentürk Demirel,1 M. Aydin respectively. Average serum macro mineral 3 4 contents were ranged between for Ca (10.28 – Ketani, M. Sedat Baran 10.70 mg/dl), K (5.56 – 6.25 mg/dl), Na (143.93 – 144.74 mg/dl), P (8.48 – 9.87 mg/dl), 1Department of Animal Science, Feeds Mg (2.62 – 2.83 mg/dl) and for Cl (101.87 – and Animal Nutrition, Faculty of 103.73mEq/L). Metabolizable energy and crude protein consumptions for 1 g live weight Agriculture, Dicle University, gain of rats were ranged between 59.91 - 21280, Diyarbakir – Turkey 69.34 (ME, kcal / LWG, g); 3.48 – 4.88 (CP, g / 2 Department of Biochemistry, Faculty LWG, g), respectively. of Veterinary, Dicle University, The highest blood serum urea, Fe binding and Diyarbakir LDL values were obtained from control group; 3 alkalin fosfatase, Ca and Na from group II Department of Histology and (2%); glucose, uric acid, total Fe, total protein, Embryology, Faculty of Veterinary, albumin, globulin, triglyceride, cholesterol, HDL Dicle University, Diyarbakir cholesterol, VLDL, P, Mg, Cl and protein consumption from group III (4%); creatinin, K, Fe levels and energy consumption were

37 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels obtained from group IV(6%). The results Ca (2+)- free and thapsigargin-added Krebs showed that the supplementation of solutions. It was found that both digoxin and clinoptilolite did not have positive effect on monensin caused apositive inotropic effect in serum concentrations of investigated normal and Ca (2+) - free Krebs solutions. parameters, however at the same time did not Although monensin produced a positive effect negatively health status of animals. inotropic effect in thapsigargin-treated papillary muscles, digoxin did not show a positive References inotropic effect. Because papillary muscles in Prvulović, D., A. Jovanović – Galović., B. Na (+) - free Krebs solution showed Stanić., M. Popović, and G. Grubor-Lajšić, spontaneous contractions, trusted results were 2007. Effects of Clinoptilolite Supplement in not obtained in Na (+) - free medium. It is Pig Diets on Performance and Serum concluded that monensin produces a positive Parameters. Czech Journal of Animal inotropic effect by depending on both extra Science, 52(6), 159-164. cellular Ca (2+) and Ca (2+) released from the Taş, M., R. Demirel., D. Şentürk Demirel., D. sarcoplasmic reticulum (SR). Digoxin, on the Kurt., S. Bacinoglu., Ü. Cirit, and M. A. Ketani, other hand, produces its effect by depending 2007. Effects Of Dietary Natural Zeolite On on the Ca (2+) released from SR. The Testicular Weight, Body Weight and Spermatological Characteristics In Rats. The Journal of Istanbul Univ. Faculty of Veterinary Med. 33 (3), 33 - 42. Abbès, S., J. B. Salah-Abbès., K. Nahdi., R. B. Younes., M. M. Hetta., A. A. El-Kady., M. A. Poster No 71 Abdel-Wahhab and R. Oueslati, 2007. Inactivation of Cadmium Induced Combined effects of niacin and Immunotoxicological Alterations in Rats by chromium on the heart tissue of Tunisian Montmorillonite Clay. International Immunopharmacology, 7, 750-760. hyperlipidemic rats

1 2 M. M. Doger , B. B. Sokmen , Refiye 1 Poster No 70 Yanardag

Investigation of the cellular 1Istanbul University, Faculty of mechanisms of actions of digoxin Engineering, Department of Chemistry, and monensin by using guinea-pig Istanbul, Turkey. papillary muscles 2Giresun University, Faculty of Arts Ramazan Üstün¹, İ. Meral² and Sciences, Department of Chemistry, Giresun, Turkey. ¹Department of Nurse, School of Healty, Karamanoğlu Mehmetbey Atherosclerotic vascular disease, including University, Karaman, 70100, TURKEY. hyperlipidemia is associated with endothelial dysfunction. Therefore, it is important to improve endothelial dysfunction for treatment ²Department of Physiology, School of of atherosclerotic disease. In this study, female Medicine, Yuzuncu Yıl University, Van, Swiss albino rats were used. They were 65200, TURKEY. divided into four groups. The animals of the first group (group I) were fed with pellet chow. The rats (group II) were fed with a lipogenic diet consisting of 2% cholesterol, 0.5% cholic This experiment was carried out to determine acid and 20% sunflower oil added to the pellet the possible mechanism of actions of chow, and given 3% alcoholic water for 60 monensin and digoxin by using guinea-pig days. The rats (group III) were fed with the papillary muscles. A total of 120 papillary same lipogenic diet and treated by gavage muscles obtained from 60 guinea-pigs were technique CrCl 6H O to a dose at 250 µg/kg used. Three treatment groups; control (% 0,1% 3 2 and 100 mg/kg niacin for 45 days, 15 days ethanol), monensin (10 μmol/l) and digoxin (3μ + after experimental animals were done mol/l) were compared in normal, Na ( ) - free,

38 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels hyperlipidemic. Group IV was fed with pellet smokers and in MPU than that of control chow and treated with 250 µg/kg CrCl3 6H2O subjects and also in MPU than that of smokers. and 100 mg/kg niacin for 45 days. On the 60th There was significant difference between MPU day, the heart tissue and blood samples were and control subjects. In Group II, important taken from animals. Heart lipid peroxidation correlation was found between consumed daily levels (LPO), serum gama glutamyl transferase MP amount and ENO levels. Also, there were (GGT) activity and serum protein carbonyl positive correlations between the number of levels were increased, serum paraoxonase cigarettes and duration of smoke or MP use (PON) activity and heart glutation (GSH) levels and ENO levels. Erythrocytes represent an were decreased in hyperlipidemic rats. important compartment for NO metabolism. As Treatment with combined niacin and chromium a free radical, NO mediates cellular toxicity and reversed these effects. In conclusion, the reacts with superoxide to form peroxynitrite present study revealed that the combined which an even more potent oxidant. It was treatment with niacin and chromium to concluded that smoke and ST use possibly hyperlipidemic rats might induce a protective affected oxidant and antioxidant balance of effect on heart GSH and LPO levels and serum erythrocytes and changes may be related with PON, GGT activities and protein carbonyl harmful effects of ST. levels. Poster No 73

Effects of the exposed to biomass and cigarette smoke on antioxidant defense system, GST activity and GSH levels in rat liver Poster No 72

Serap Sahin Basak1, Yavuz Silig2, The effect of smoke and smokeless 3 tobacco use as Maraş powder on Omer Taner Dogan erythrocyte nitric oxide level 1Department of Chemistry, Faculty of 2 1 1 Science and Art, Department of N. Kurtul ,Saniye Böyükkara , E. 3 Gökpınar1 Biochemistry, Faculty of Medicine, Department of Respiratory Disease, 1University of Kahramanmaraş Sütçü Faculty of Medicine, University of İmam, Faculty of Science Department Cumhuriyet, Sivas, TURKEY. of Chemistry, Division of Biochemistry, The harmful effects of tobacco smoke have Kahramanmaraş been well-known for years. Moreover, these effects have been demonstrated by The use of smokeless tobacco (ST) is common experimental trials. Free radicals and reactive worldwide. Also, in Türkiye, especially in oxygen species (ROS) are produced during Kahramanmaraş and other Southern cities, a cigarette smoking. In this study, we kind of ST prepared from a tobacco of species investigated effects on antioxidant defense Nicotiana rustica L. and known as Maraş system, GST and GSH levels in liver of rats powder (MP) has been consumed widely as exposed to biomass (dried dung) and cigarette alternative to smoking. The habit is becoming smoke. Adult male Wistar-strain albino rats increasingly popular due to banning smoking in weighing about 300 g each and nourished our country. The ST is causatively associated under normal conditions were used in this with an increased risk for cardiovascular study. Rats were divided into five groups each diseases and cancers of the oral cavity, larynx, of which contains seven animals. Animals were and pharynx. In this study, erythrocyte nitric exposed to biomass smoke (group 1), to oxide (ENO) concentrations were measured by cigarette smoke (group 2) and to both cigarette Griess reaction in blood samples obtained from smoke and biomass together (group 3), for an smokers (Group I), MP users (MPU; Group II), hour daily for three months. Whereas the and healthy control subjects (Group III) who control group animals was not subjected to any were nonsmokers and nonusers of ST and the application. It has been found out GSH level, results were compared. The ENO the activities of GST and antioxidant enzymes concentrations were found to be lower in

39 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels in experimental groups were significantly albumin (BSA) on standard semen indicators elevated than control group. On the other (motility, acrosome and total abnormalities, hand, when compared to controls, MDA level HOST), endogenous antioxidant enzyme in all groups were significantly increased activities and fertilizing ability of frozen-thawed (group 1:1.51-fold, group 2:1.36 fold, and bull semen. Eighteen ejaculates from each of 3 group 3:1.41.fold). It has been determined that Holstein bulls were collected using an artificial antioxidant defense system was induced in all vagina and 9 replicates of the ejaculates were groups associated to an increase in free diluted in Bioxcell extender supplemented with radicals, however this induction could not antioxidants, including BSA (5 mg/ml), GSH (2 protect the liver cells from the damage caused mM), GGSG (2 mM), and an extender by free radical which was understate by containing no antioxidants (control). detecting the increase in lipid peroxidation. On Insemination doses (1.5 × 107 sperm/0.25 ml the other hand, it could be suggested that an straw) were prepared for the insemination of increase GST enzyme activity could be the cows at observed oestrus. Supplementation result of the induction of detoxification system. with antioxidants led to lower percentages of acrosome damage and total abnormalities, References compared to the controls (p< 0.01). Pregnancy Desai SR, Ryan SM, Colby TV. 2003. rate after insemination was highest (72.2%) in Smoking-related interstitial lung diseases: the group which was given BSA (p< 0.05). histopathological and imaging perspectives. There were no significant differences among Clin Radiol. 58:259-268. groups in GSH and glutathione peroxidase (GSH-PX) enzyme activities. Superoxide Pryor WA. 1997. Cigarette smoke radicals and dismutase (SOD) activities in all of the the role of free radicals in chemical experimental groups with antioxidants were carcinogenesis.Environ. Health Perspect. lower than the control group (p< 0.001). 105:875–882. Furthermore, BSA increased (p< 0.001) the Poster No 74 activity of catalase (CAT), following the freezing-thawing process. Influence of Various Antioxidants on Poster No 75 Microscopic-Oxidative Stress Indicators and Fertilizing Ability of The protective effects of N (1)-2,4- Frozen-Thawed Bull Semen dihydroxybenzylidene-N (4)-2- hydroxybenzylidene-S-methyl- Serpil Sarıözkan1, P. B. Tuncer2, M. N. thiosemicarbazidato-oxovanadium Bucak2, P. A. Ulutaş3 (IV) on brain injury in streptozotocin-induced diabetic rats 1Erciyes University, Faculty of Sevim Tunali1, R. Yanardag1, T. Bal- Medicine, Hakan Çetinsaya 2 2 Experimental and Clinical Research Demirci , B. Ülküseven 2 Center, Kayseri, Turkey Ministry of 1 Agriculture and Rural Affairs, Lalahan Istanbul University, Faculty of Livestock Central Research Institute, Engineering, Department of Chemistry, Lalahan, Ankara, Turkey Biochemistry Division, 34320, Avcilar, 3Adnan Menderes University, Faculty Istanbul, TURKEY of Veterinary Medicine, Department of 2 Biochemistry, Aydın, Turkey Istanbul University, Faculty of Engineering, Department of Chemistry, Cryopreservation is associated with the Inorganic Division, 34320, Avcilar, production of reactive oxygen substances Istanbul, TURKEY (ROS), which lead to lipid peroxidation of sperm membranes, resulting in a loss of sperm motility, viability and fertility. The aim of this Diabetes is usually accompanied by increased study was to determine effects of the production of free radicals or impared antioxidants of oxidized glutathione (GSSG), antioxidant defences. Oxidative damage reduced glutathione (GSH) and bovine serum contributes to biochemical abnormalities in

40 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels brain tissue. In the present study, we aimed to on deuterium contents of drinking water and demonstrate the effect of N (1)-2, 4 diet. Also D/H ratio is not constant in tissues Dihydroxybenzylidene-N (4)-2- and body fluids. The effect of the replacement hydroxybenzylidene-S-methyl- of H with D in biological systems is well thiosemicarbazidato-oxovanadium (IV) (VOL) documented, however, the possible role of on brain damage in male rats with naturally occurring D has not completely streptozotocin (STZ)-induced diabetes. Male, elucidated. Recent studies have shown that 3-3.5 months old, Swiss albino rats were reduction of D concentration in drinking water randomly divided into four groups. diminishes the growth rate of the tumors and Experimental diabetes was induced by induces apoptosis (1-3). In this study, the intraperitoneal injection of STZ in a single dose effect of deuterium depleted water (DDW) (85 65 mg/kg. Group I: control (intact) animals. ppm) consumption on the blood and liver Group II: control animals given VOL. Group III: parameters were investigated in healthy rats. STZ-induced diabetic animals. Group IV: STZ- Creatinine kinase, creatinine, glucose, protein, induced diabetic rats treated with VOL. VOL of bilirubin, urea, uric asid, total lipid, cholesterol, 0.2 mM/kg daily dose were administered by and triglyceride levels were determined in gavage technique to rats for 12 days, after the blood samples; lipid peroxidation (LPO) and experimental animals were made diabetic. On glutathione levels, tissue factor activity (DFa), day 12, the brain tissues were taken from carbonic anhydrase, sodium-potassium animals. Brain tissues were homogenized in ATPase, superoxide dismutase, catalase 0.9 % saline with a glass homogenizer to make (CAT), glutathione-S-transferase and up to 10 % homogenate (w/v). The glutathione peroxidase enzyme activities were homogenates were centrifuged and the clear also measured in liver tissue. No significant supernatants were used for protein levels and differences on these parameters were found enzyme activities. Brain xanthine oxidase, between either tap water or DDW consumption alanine transaminase, aspartate transaminase for 30 days. On the other hand, consumption of activities, nonenzymatic glycosylation and lipid DDW for 40 days decreased protein levels and peroxidation levels were increased in diabetic increased LPO, CAT and DFa in liver and rats. Also brain catalase, acetylcholinesterase increased creatinine kinase activity in blood activities and glutathione levels were significantly, compared to tap water drinking decreased in diabetic rats. Treatment with VOL group. As a result, the application duration and reversed these effects. As a result it might be probably the concentration of DDW concluded that treatment with VOL has a consumption are important factors. It is protective effect on damage of brain of STZ- necessary that more comprehensive research induced diabetic rats. has to be done to enlighten the mechanism of this issue. Poster No 76 Poster No 77 The effect of deuterium depleted 2+ water consumption on blood and Coupling of Ca microdomains to liver biochemical parameters in spatially and temporally distinct healthy rats cellular responses by the tyrosine kinase Syk. Sevinç Özgür1, A. Yarat1, T. Yardımcı2 1 1 1Department of Basic Medical Siaw W. Ng , C. Nelson , A.B. Parekh 1 Sciences, Biochemistry, Faculty of . Dentistry, Marmara University, 1 Istanbul, TURKEY Department of Physiology, Anatomy and Genetics, University of Oxford, 2Department of Biochemistry, Faculty Parks Road, Oxford OX1 3PT, United of Pharmacy, Marmara Kingdom. University, Istanbul, TURKEY Communication between the cell surface and Deuterium (D) is the only stable and non- the nucleus is essential for regulated gene radiating isotope of Hydrogen (H). D expression. In immune cells, excitation- concentration varies in human fluids depending transcription coupling is thought to involve

41 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels global Ca2+ signals. The current study There are a lot of studies on vasodilatation of investigates whether excitation-transcription the effect of hyperglycemia, but there is a poor coupling can be driven by Ca2+ microdomains knowledge of the effect of hyperglycemia on arising from open CRAC channels. A vascular contractile responses. Furthermore combination of Ca2+ imaging, whole cell patch chronic exposure to high concentrations of has clamp and reverse transcription-PCR generally been studied. So the aim of this experiments were used to address this in rat study was to investigate whether short-term basophilic leukemia-1 cells, using thapsigargin hyperglycemia affected vascular contractility. as a stimulus. We find that in mast cells, Ca2+ Non-diabetic rat aorta rings were incubated in microdomains from store-operated Ca2+ normal (11 mM) (NG) and high (22 mM and 44 release-activated Ca2+ channels activate mM) (HG) glucose ambient for four hours. expression of the transcription factor c-fos. Responses of vasoconstriction to serotonin Local Ca2+ entry is sensed by the tyrosine (10-5 M), phenyleprinhe (10-6 M) and KCl (60 kinase Syk. Syk clusters at the cell periphery in mM) compared to the ambient including resting cells and is retained here after different glucose concentration. In addition, stimulation. Syk signals to the nucleus through MDA (malondialdehyde, a product of lipid the transcription factor STAT5 in a protein peroxidation) levels of aorta tissue was kinase C- and MEK/ERK-independent assessed by high-performance liquid pathway. Ca2+ microdomains following CRAC chromatography in different glucose channel activation also activate Ca2+- concentration (NG and HG).Responses of dependent phospholipase A2, followed by vasoconstriction to KCl were increased in the synthesis and secretion of cysteinyl presence of Krebs’ solution with high glucose leukotrienes. However, unlike c-fos expression, (p<0,05). Although there are slightly this is mediated via the MEK/ERK pathway. In increasement of respones observed with conclusion, Syk therefore couples Ca2+ serotonin, no statistically significant changes of microdomains to the activation of two spatially responses to phenyleprinhe and serotonin. and temporally distinct cellular responses, Besides, MDA levels increased significantly revealing the versatility of local Ca2+ signals in during the hyperglycemia (p<0,05).Short-term driving cell activation. hyperglycemia may lead to augment contractile response in aort rings through several mechanisms and our results showed that oxidative stress is most probably one of them.

Poster No 78 Poster No 79

Effects of short-term hyperglycemia The Effect of Raffinose And on the contractile responses of rat Methionine on Frozen/Thawed aorta Angora Buck Sperm Parameters, MDA and SOD Activities Soner Dönmez1, O. Gökalp2, M. Purhan Barbaros Tuncer 1 Mustafa Doğan3, H. Vural4, B. Yiğit4 Numan Bucak 1 Umut Taşdemir 1 Serpil Sarıözkan 2 Fatih Sakin 3 Department of Chemistry, Biochemistry Ahmet Ateşşahin 4 Olga Division, School of Art and Science, Büyükleblebici 5 University of Nevsehir, Nevsehir 50300, Turkey1 1 Ministry of Agriculture and Rural Department of Pharmacology, School Affairs, Lalahan Livestock Central of Medicine, Dicle University, Research Institute, Lalahan, Ankara, Diyarbakir 21100, Turkey2 Turkey Department of Pharmacology3, 2 Erciyes University, Safiye Cikrikcioglu Department of Biochemistry4, School Vocational College, Kayseri, Turkey. of Medicine, Suleyman Demirel 3 Mustafa Kemal University, Faculty of University, Isparta 32260, Turkey Veterinary Medicine, Department of Pharmacology and Toxicology, Hatay, Turkey

42 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

4 Fırat University, Faculty of Veterinary R.R.Costa, R.I. Reis, J.F. Aguiar and Medicine, Department of Wamberto A. Varanda Pharmacology and Toxicology, Elazığ, Department of Physiology, School of Turkey Medicine of Ribeirão Preto/USP Brazil 5 Ankara University, Faculty of Veterinary Medicine, Department of Luteinizing hormone (LH) is known to activate Biochemistry, Ankara, Turkey both adenyl cyclase and phospholipase C and also to increase intracellular Ca2+ 2+ concentration ([Ca ] ) in Leydig cells. Here we The aim of the present study was to determine i investigated the effects of modulators of PKA the effects of different doses of raffinose and 2+ and PKC on T-type Ca currents and on the methionine on post-thawed semen quality, 2+ overall [Ca ] transients induced by LH. MDA and SOD (superoxide dismutase) of i Treatment of the cells with 1 µg/ml LH Angora buck sperm following cryopreservation. increased the amplitude of the peak current at Ejaculates collected from three Angora bucks -20 mV, from -2.4 ± 0.7 to -3.8 ± 0.8 pA/pF (n = were evaluated and pooled at 37°C. Semen 22, p = 0.0381) and 1 µM PMA increased the samples, which were diluted with a Tris-based current to -3.6 ± 0.4 pA/pF, (n = 6, p = extender containing the additives raffinose 0.00033). Only minor changes were observed (2.5, 5 and 10 mM) and methionine (2.5, 5 and in the voltage dependence of activation, 10 mM) and an extender containing no inactivation or deactivation of the currents. antioxidants (control), were cooled to 5oC and Inhibition of both PKC and PKA with 400 nM frozen in 0.25 ml French straws to be stored in 2+ staurosporine blocks the [Ca ] changes liquid nitrogen. Frozen straws were thawed i induced by LH (n = 37 cells). A similar effect individually at 37°C for 20 sec in a water bath was seen with 10 µM H89, a specific inhibitor for evaluation. of PKA. Although PMA by itself slowly The freezing extender supplemented with 2.5 increased the fluorescence, the subsequent and 5 mM methionine led to higher addition of 1 µg/ml LH still triggers the typical percentages of computer-assisted sperm 2+ transients in [Ca ]i. Chelerythrine, a PKC motility (CASA) (63.6±7.0 and 63.4±3.1%, 2+ inhibitor, also does not avoid the Ca respectively), in comparison to the controls transients. (p<0.01) following the freeze-thawing process. Taken together our results show that PKA The freezing extender with raffinose (5 and 10 plays a leading role in mediating the LH mM) and methionine at three different doses 2+ induced changes in [Ca ] , a necessary step (2.5, 5 and 10 mM) led to lower percentages of i in the process of testosterone production and acrosome abnormalities, in comparison to the secretion by Leydig cells. controls (p<0.001). Malondialdehyde (MDA) formation was found to be lower (1.8 ± 0.1 Financial Support: FAPESP, CNPq, FAEPA, (nmol/L) in the group of 5 mM raffinose, CAPES compared to the control following the freeze- thawing process (P<0.01). The additives did not show any effectiveness on the o Poster N 81 maintenance of SOD activity, when compared to the controls (P>0.05). In conclusion, methionine and raffinose play a cryoprotective The effect of nicotine consumption role on CASA motilities and acrosome & deprivation on damage of multi- abnormalities. Raffinose 5 mM exhibited organ failure caused by sepsis: Role antioxidative properties, decreasing MDA of Neutrophils levels. Zarife Nigâr Özdemir1, D. Özbeyli1, A. 1 1 o F. Özdemir , M. B. Karaaslan , K. Poster N 80 1 1 2 Kaytaz , M. F. Kara ,O. E. Tok , F. 2 1 T-type calcium currents and Ercan , B. Ç. Yeğen intracellular calcium concentration 1 are modulated by PKA in mice Marmara University School of Leydig cells. Medicine, Department of Physiology

43 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

2Marmara University School of Medicine, Department of Histology and Evaluation of buccal mucosa Embryology epithelium in type I diabetic patients by an exfoliative cytology method Sepsis is a severe clinical condition causing and clinical findigs. multi-organ failure. Nicotine decreases the severity of many diseases by inhibiting pro- Zehra Safi Öz 1, F. Battal2, H. Atmaca inflammatory cytokines. In a sepsis model, we 3 2 aimed to show the protective effects of nicotine , B.Ermiş on two target organs. Male Wistar albino rats 1 received tap water with (50mg/ml) or without University of Zonguldak Karaelmas, Faculty of Medicine, Department of Medical Biology, nicotine for 14 days prior to surgical 2 University of Zonguldak Karaelmas, procedures. Under ketamine anesthesia, Faculty of Medicine, Department of sepsis (n=45) was induced by ligation and Pediatrics, punction of the cecum, while sham group (n=8) 3 had only laparotomy. In a subgroup of rats, University of Samsun Ondokuz nicotine drink was withdrawn for 5 days before Mayıs, Faculty of Medicine, sepsis induction, while in the acute nicotine Department of Endocrinology, Turkey group rats were administered a single injection of nicotine (30mg/kg, ip) before sepsis, but had Diabetes mellitus is one of the most common no oral intake. Rats were decapitated 24 hours endocrin metabolic disorders that causes after surgery to obtain lung, liver and kidney chronic hyperglisemia and disturbances in tissues to determine malondialdehyde (MDA) carbohydrate, lipid, and protein metabolism. and glutathione (GSH) levels, myeloperoxidase Although diabetes can cause important cellular (MPO) activities. Data were analyzed by changes. We also decided to evaluate cellular ANOVA and Tukey multiple comparison tests. changes in buccal epithelial cells using an Sepsis resulted in increased pulmonary and exfoliative cytology method and clinic findings hepatic MPO activities as compared to control in Type I diabetes. All women were asked to fill group (p<0.001), but in the chronic nicotine in an questionnaire focusing on detailed history group this increase was abolished (p<0.001). and symptoms. In 30 patients with Type I However, in the withdrawal and single-dose diabetes ( age mean 15,57 ± 7,9) and 30 nicotine groups pulmonary and hepatic MPO control individuals (age mean 15,3 ±6.5), activities were not different than the sepsis buccal smeras were obtained and stained with group. Histologically observed damages were Papanicolaou technique. Cellular changes ameliorated by all nicotine treatments at were evaluated in each slide by light varying degrees, but MDA and GSH levels microscopy. All buccal PAP smear and clinical were not different among experimental groups. data were documented in computer. All The findings of the present study raise the analyses were performed SPSS programme. possibility that long-term nicotine consumption The chi-square and Fisher’s exact tests were reduces sepsis-induced organ damage, which used for the statistical analysis. P values <0.05 appears to involve the inhibition of neutrophil were considered significant. The difference recruitment to the inflamed tissues. between of type I diabetes and binucleation, nuclear membrane irregularity, cytoplasmic polymorphism, karyorrhexis- karyopyknosis- References karyolysis and also clinical data such as sensitivity of buccal mucosa, halitosis and 1) Steiner AA, Oliveira DL, Roberts JL, xerostomia was statistically significant (p Petersen SR, Romanovsky AA. Nicotine <0.05). Type I Diabetes mellitus can cause administration and withdrawal affect survival in nuclear and cytoplasmic changes in the buccal systemic inflammation models. J Appl Physiol epithelium that are noticeable with this 105: 1028–1034, 2008. exfoliative cytology method and clinical 2) Lucidarme O, Seguin A, Daubin C, findings such as halitosis and xerostomia. Ramakers M, Terzi N, Beck P, Charbonneau P, du Cheyron D. Nicotine withdrawal and References agitation in ventilated critically ill patients. Crit Care. 9; 14(2):R58, 2010 Alberti S, Sapadella CT, Francischone TR,

o Assis GF, Tania M. 2003. Exfoliative cytology Poster N 82 of the oral mucasa in type II diabetic patients:

44 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels morphology and cytomorphometry. Journal of significant increase in BP as compared with Oral Pathology& Medicine. 32 (9):538-543 control group. Propolis administration to L- NAME treated rats reduced BP but this was Shareef BT, Ang KT, Naik VR. 2008. not statistically significant. These results Qualitative and Quantitative exfiliative cytology suggest that propolis decrease TH activity and of normal oral mucosa in type 2 diabetic thereby may modulate the synthesis of patients. Med. Oral. Patol Oral Cir Bucal. 1; 13 catecholamine and BP. (11) - E693-6

Poster No 84 Poster No 83 Changes in tyrosine hydroxylase Effects of propolis on tyrosine activity and adrenomedullin level by hydroxylase activity and blood treatment of organoselenium pressure in nitric oxide synthase compounds in rat hypothalamus inhibited hypertensive rats exposed to 7,12- dimethylbenzanthracene A. Gogebakan1, Zeliha Selamoglu Talas1, I. Ozdemir2, E. Sahna3 I. Ozdemir1, Zeliha Selamoglu Talas2, Y. Gok1, B. Ates1, I. Yilmaz1 1Department of Biology, Faculty of Arts and Science, Nigde University, Nigde, 1Department of Chemistry, Faculty of Turkey Arts and Sciences, Inonu University, 2Department of Chemistry, Faculty of Malatya, Turkey Arts and Sciences, Inonu University, 2Department of Biology, Faculty of Arts Malatya, Turkey and Science, Nigde University, Nigde, 3Department of Pharmacology, Faculty Turkey of Medicine, Firat University, Elazig, Turkey The effects of synthetic organoselenium compounds (Se I and Se II) on the activity of tyrosine hydroxylase (TH), the rate-limiting Reduction in the synthesis or bioavailability of enzyme in catecholamine synthesis as well as nitric oxide plays a significant role in the adrenomedullin (ADM) level were determined development of hypertension. Propolis is a in the hypothalamus of adult rats exposed to resinous product collected by honeybees from DMBA (7,12-dimethylbenz[a]anthracene). various plant sources. Tyrosine hydroxylase DMBA, an organic environmental pollutant, is a (TH) is the rate limiting enzyme in the polycyclic aromatic hydrocarbon that can biosynthesis of the catecholamines. The aim of induce a range of toxic effects and stress in this study was to examine the effect of propolis rats. Selenium is an essential trace element, on blood pressure (BP), TH and total RNA which interacts with antioxidants, and has levels in the adrenal medulla, heart and anticancer and antihypertensive properties. TH hypothalamus tissues in chronic nitric oxide is an aromatic amino acid hydroxylase whose synthase (NOS) inhibited rats by NG-nitro-L- activity is elevated in response to a range of arginine methyl ester (L-NAME). Rats received stress inducers. TH activity is normally NOS inhibitor (L-NAME, 40 mg/kg, regulated by negative feedback in intraperitoneally) for 15 days to produce catecholamine biosynthesis. ADM is an hypertension and propolis (200mg/kg, by abundantly present peptide in a broad range of gavage) the last 5 days. TH activity and total normal tissues including adrenal medulla, RNA levels significantly increased in adrenal lungs, kidneys and brain. Plasma ADM levels medulla, heart and hypothalamus tissues in L- are elevated in a number of diseases including NAME-treated groups (P<0.05). TH activity essential hyptertension and chronic renal and total RNA levels of L-NAME + propolis- failure. The antioxidant properties of ADM offer treated rats significantly reduced (P<0.05) as protection against organ damage induced by compared with L-NAME-treated groups. TH high blood pressure, ischemia and aging. activity in propolis-treated rats were reduced to DMBA treatment increased the TH activity and the control group values. L-NAME led to a ADM level in the hypothalamus. These

45 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels increases were found to be inhibited by Se I References and Se II treatments.These studies demonstrate that synthetic organoselenium 1. Nazıroğlu M. Role of selenium on calcium compounds can suppress DMBA-induced signaling and oxidative stress- induced stress related changes in the rat molecular pathways in epilepsy. Neurochem hypothalamus. Therefore, the antioxidant and Res 2009; 34:2181–2191. antihypertensive effects of Se I and Se II may have important effects in the maintainance of 2. Uğuz AC, Nazıroğlu M, Espino J, Bejarano homeostasis. I, González D, Rodríguez AB, Pariente JA. Selenium modulates oxidative stress-induced cell apoptosis in human myeloid HL-60 cells Poster No 85 via regulation of caspase-3, -9 and calcium influx. J Memb Biol 2009; 232:15–23.

Effects of Magnetic Fields on Blood o Fields on Blood Biochemistry of Poster N 86

Diabetic Rats Determination of GST activity with CDNB of Orthrias angorae Ayşe Demirkazık1, S. Gültürk2, 2 2 2 E.Özdemir , T. Demir , S.Uğur , Cem Ozic1,2, S.Gül2

1 Department of Biophysics, Medical 1,2 Department of Biology, University of Faculty, Cumhuriyet University, Sivas, Anadolu, Eskisehir 26470, TURKEY Turkey. 2 Department of Biology, University of

2 Kafkas, Kars 36100, TURKEY Department of Physiology, Medical Faculty, Cumhuriyet University, Sivas, Glutathione S-transferases, GSTs (EC Turkey 2.5.1.18) are a family of diverse multifunctional proteins mainly involved in detoxification of We investigated the effects of a long-term xenobiotics and antioxidant defense .GSTs treatment with insulin, or/and magnetic field on have been found in virtually all plants, the blood biochemistry parameters in vertebrates, insects, yeasts, and bacteria. streptozotocin (STZ)-induced diabetic rats. GSTs detoxify herbicides, organic pollutants, Fifty three adult Wistar albino male rats and naturaltoxins, in addition to less (Cumhuriyet University Animal Center, Sivas, characterized role in oxidative stress tolerance. Turkey), weighing 250-300 g, were used. In most of the organisms, GST expression is Before study procedure, rats were randomly modulated in response to exposure to assigned in four groups: sham, exposed to no prooxidant xenobiotics. Overall, it is believed MF; MF, exposed to MF; DM, induced with that the induction of GSTs is an evolutionary streptozotocin; DMMF, induced with conserved response of the organisms towards streptozotocin and exposed to MF. Then, five exposure to prooxidants . Under certain mL blood was collected from intracardiac non- conditions, the interaction between glutathione fasting rats final experiment before the rats and CDNB is totally dependent on the were sacrificed by decapitation. Pre- and post- presence of active GST. The GST-catalyzed glucose levels, TG, cholesterol, HDL, LDH, Na, formation of GS-DNB produces a dinitrophenyl K, Ca, Mg and Fe electrolits were elevated thioether which can be detected by analyzer devices. There were statically spectrophotometer at 340 nm. One unit of GST differences on the mean values of pre- and activity is defined as the amount of enzyme post- glucose levels, TG, cholesterol, HDL, producing 1 μmol of GS-DNB conjugate/min LDH between sham and DMMF. In conclusion, under the conditions of the assay. In brief, the the states of improved glucose metabolism incubation mixture of GST toward CDNB may be prevented blood biochemistry activity assay consisted of 0.1M phosphate parameters and the hypoglycemic effect of buffer, 1 mM of GSH and 1 mM of CDNB magnetic field on the function of β cells may be solution. After preincubation at assay able to help increase in insulin concentration temperature (25 °C), the reaction was initiated and sensitivity to glucose metabolism. by the addition of 10 μL of liver subcellular fraction. GST activity (toward CDNB) (μmol/mg

46 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels protein/min) in homogenate was found to be exposure groups were found significant 0.71±0.04. CDNB in GST activity was compared to sham group (p<0.05).However, determined be used Orthrias angorae. GST no significant alterations were found in other activity with 1-chloro-2,4-dinitrobenzene endpoints such as MDA, MPO, TAC, TOS and (CDNB) was determined by the method of Habig et al. OSI levels. In conclusion, CAT levels were affected by long-term exposure of both 100 Poster No 87 and 500 μT ELF-MF in rats’ eye. However, further and more researches need to be Oxidative and Antioxidative performed to explain interaction mechanisms Parameters In The Eyes Of Rats between ELF-MF exposure and eyes.

Exposed to Extremely Low o Frequency Magnetic Fields. Poster N 88 Mehmet Zulkuf Akdag1, S. Dasdag1, H. 2 3 Colchicine modulates oxidative Cakir , O. Erel 2+ stress and Ca release in *Department of Biophysics, Medical polymorph nuclear leucocytes of Faculty, Dicle University, Diyarbakir, remission (but not unremission) of Turkey. patients with family Mediterranean 2 Department of Biochemistry, Medical fever Faculty, Harran University, Urfa, 1 1 Turkey. H. Demirkan , Mehmet Şahin , A. C. 3 Department of Biochemistry, Ataturk Uğuz2, İ.S. Övey2, M. Nazıroğlu2, İ. Education Hospital, Ankara, Turkey. Erturan3, E. Tunç1

Most of the scientific concerns arise from several studies, which suggest possible 1Department of Internal Medicine, interaction mechanisms between extremely Division of Rheumatology, Faculty of low frequency magnetic fields (ELF-MFs) and living organisms. However, the molecular Medicine, Suleyman Demirel University, mechanism through which ELF-MFs can Isparta, Turkey. influence cellular behavior is still unclear. On 2 the other hand, it has been suggested that the Department of Biophysics, Faculty of interaction between living organisms and ELF- Medicine, Suleyman Demirel University, MFs could involve interferences with reactive Isparta, Turkey. oxygen species (ROS) and free radical production and half-life extensions. Therefore, 3Department of Dermatology, Faculty of the present study aimed to investigate the Medicine, Suleyman Demirel University, effect of ELF-MF on oxidative and antioxidative parameters in rat eyes. In this study, 30 male Isparta, Turkey. Sprague–Dawley adult male rats were used. It has been suggested that there is an Rats in two experimental groups were exposed abnormality in the oxidative stress of chronic to 100 and 500 μT ELF-MF, which are the inflammatory patient with familial safety standards of public and occupational Mediterranean fever (FMF) (1). A exposure for 2 h/day for 10 months. Same pharmacological characterization of Ca2+ influx procedure applied to the rats in sham group pathways in polymorph nuclear leucocytes except ELF-MF exposure. The levels of (PMN) is problematic because of lack of specific inhibitors. It was reported that catalase (CAT), malondialdehyde (MDA), colchicine induced antioxidant and cation myeloperoxidase (MPO), total antioxidative channel blocker effects in heart cells (2, 3). We capacity (TAC), total oxidant status (TOS), and aimed to investigate effects of colchicine on oxidative stress index (OSI) were measured in oxidative stress and Ca2+ release in plasma rat eye. The decrease of CAT levels in and polymorph nuclear leucocytes of attack,

47 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels remission and unremission patients with family Ameliorating Effects of the Caffeic Mediterranean fever. Eighteen FMF patients Asid Phenethyl Ester on Diazinon and six age-matched health control within four groups were used in the study. First group Poisoning used was control. Second group was patients 1 2 with active FMF. Third and fourth groups were E. Oğuzhanoğlu , Osman Gökalp , L. patients with patients with remission and Yavuz1, C. A. Andaç2 remission, respectively. Colcichine (1,5 mg/day) was given to consisting second, third 1 and fourth groups for one month. Lipid Medical School, Department of peroxidation (LP) and cytosolic Ca2+ release Anesthesiology and Reanimation, levels in PMN of FMF patients within attack Suleyman Demirel University Isparta, and unremission periods were higher than in Turkey. control although their levels were higher in 2Medical School, Department of FMF attack patients than in remission group. Vitamin E and -carotene levels were higher in Pharmacology, Dicle University, patients with remission period than in control Diyarbakır, Turkey. and patients with attack period. LP levels were also higher in serum of patients with attack The caffeic acid phenethyl ester period than in control although serum LP levels (CAPE) is a compound structurely similar to were lower in patients with remission period the flavonoids and is found in the bee gums than in patients with attack period. However, produced by honeybees. CAPE possesses PMN and serum LP levels and Ca2+ release antiinflammatory, antioxidant and immune concentrations were further increased in regulatory effects. We aimed to investigate patients with unremission period. In conclusion whether CAPE has an ameliorating effect on we observed that colcichine induced protective the diazinon poisoning on rats. Rats were effects on oxidative stress by modulating divided into 4 groups. The control (the first) vitamin E, -carotene and Ca2+ release levels group of the rats were not treated with any in FMF patients with remission period. substance. The second group of the rats were Colcichine did not affect on the values in orally given 200 µg/kg subletal doze of patients with unremission period. diazinon. The third group of the rats were given interperitoneally (ip) 10 µmol/kg of CAPE one

day before 200 µg/kg of diazinon was applied References orally. The fourth group of the rats were ip injected 10 µmol/kg of CAPE half an hour after 1. Gurbuz L, Yamanel L, Bulucu F, Inal V, 200 µg/kg of diazinon was orally administered. Aydin A. Oxidative status in familial Blood samples were obtained from the aorta of Mediterranean fever with or without proteinuria. the rats under anesthesia at 48th hour. After Free Rad Biol Med 2005;38:271-275. analysis of the blood samples, it was found 2. Modrianský M, Dvorák Z. Microtubule both in the third and fourth groups of the rats disruptors and their interaction with that CAPE diminished the inhibiting effect of biotransformation enzymes. Biomed Pap Med diazinon on the butyrylcholine esterase Fac Univ Palacky Olomouc Czech Repub. enzyme by half. The mechanism of the action 2005;149:213-215. of the ameliorating effect of CAPE was further computationally investigated by 3. Iribe G, Ward CW, Camelliti P, Bollensdorff Autodock_Vina.2 Three binding conformations C, Mason F, Burton RA, Garny A, Morphew of CAPE with binding energies ΔGo1= -8.2 MK, Hoenger A, Lederer WJ, Kohl P. Axial kcal/mol, ΔGo2= -7.9 kcal/mol and ΔGo3= -7.2 stretch of rat single ventricular cardiomyocytes 2+ kcal/mol, respectively, and one binding causes an acute and transient increase in Ca conformation of diazinon with a binding energy spark rate. Circ Res. 2009;104:787-795. of ΔGo= -6.7 kcal/mol were determined in the crystal structure of the active site of butyryl choline esterase (PDB ID: 2WID).1 Therefore, it was concluded that CAPE binds to butyryl choline esterase with roughly 2-10 times greater affinity than that of diazinon.

Poster No 89 References

48 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

1) Gokalp O, Buyukvanli B, Cicek E, et al.The hours later. In all groups the superoxide effects of diazinon on pancreatic damage and dismutase (SOD), catalase (CAT), glucose-6- ameliorating role of vitamin E and vitamin C. phosphate dehydrogenase (G6PD) activities Pesticide Biochemistry and Physiology 2005; and the malondialdehyde (MDA) levels were 81(2): 123-128. determined. 2) Carletti E, Aurbek N, Gillon E, Loiodice M, Nicolet Y, Fontecilla J, Masson P, Thiermann SOD and CAT activities decreased and G6PD H, Nachon F & Worek. F, Structure-activity activity increased in the group treated hexane analysis of aging and reactivation of human extract (p<0.05). SOD and CAT activities butyrylcholinesterase inhibited by analogues of increased while there is not a significant tabun. Biochem Journal 2009, 421(1): 97-106. difference at the G6PD activity in the group 3) Trott O, Olson A.J., AutoDock Vina: treated Ethanol extract. MDA levels were not improving the speed and accuracy of docking observed in all groups administered extract with a new scoring function, efficient (p>0.05).It is known that the organisms are optimization and multithreading, Journal of adapted to the high free radical levels by Computational Chemistry 2010, 31: 455-461. increasing the antioxidant enzyme activities http://vina.scripps.edu. (Stephensen et al..2002). However, it is Poster No 90 suggested that the decrease at the SOD and CAT enzymes is caused by the accumulation The Effects of Extracts Obtained of free superoxide radical and the H2O2 by From Nepeta Italica L. Species on inhibiting the protein synthesis (Husain et the Antioxidant Enzymes al..2001; Santhakumarı et al, 2003).It was reported that the extract administration caused Sercan Özbeka, F. Özdemirb, Ş. increasing at the SOD and CAT enzyme Konaka, İ. Özmenb, T. İ. Büyükoğlua, H. activities and was suggested that it was related Gençd to the increased antioxidant state (Karuna et al..2009). In our study, MDA levels were not a Health Vocation Training School, alter for all groups. This situation may be Mehmet Akif Ersoy University, Burdur, explained the bad effects of free radicals may Turkey be eliminated without causing any cell damage. b Faculty of Arts and Sciences, Süleyman Demirel University, Isparta, References Turkey 1. Stephensen E, Sturve J,. Förlin L, 2002. c Faculty of Veterinary Medicine, Effects of redox cycling compounds on Mehmet Akif Ersoy University, Burdur, glutathione content and activity of glutathione- Turkey related enzymes in rainbow trout liver, Comp. Biochem. Physiol. C 133:435–442. d Faculty of Education, Mehmet Akif Ersoy University, Burdur, Turkey 2.Lenartova V, Holovska K, Pedrajas JR, Martınezlara E, Peinado J, Lopez-Barea, J, We aimed to examine the effects on the Rosival I, Kosuth P, 1997. Antioxidant and antioxidant enzymes of the extracts obtained detoxifying fish enzymes as biomarkers of river from the Nepeta italica L.. pollution. Biomarkers 2:247-252.

The Nepeta italica extracts were obtained by 3. Husain K,Scott BR, Reddy SK, Somani SM, using ethanol and hexane solutions and 2001. Chronic ethanol and nicotine interaction administered orally to rats. The rats were on rat tissue antioxidant defense system. separated into 3 groups (Group I; normal Alcohol 25:89–97. saline, Group II; ethanol, group III; hexane). The extracts were administered single dose 4. Santhakumarı P, Prakasam A, Pugalendi KV, 2003. Modulation of oxidative stress (1.0 g/kg). The blood samples were taken 3

49 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels parameters by treatment with Piper betle leaf MDA levels in all groups. in streptozotocin induced diabetic rats. Indian Journal of Pharmacology 35:373-378. NADPHs produced by G6PD are necessary for nucleotides in the formation of reduced 5.Karuna R, Sreenivasa Reddy S, Baskar R, glutathione in erythrocytes. In our study, G6PD Saralakumari D, 2009. Antioxidant potential of activity was increased insignificant in group aqueous extract of Phyllanthus amarus in rats, given hexane extract, this situation might be Indian Journal of Pharmacology 41;2:64-67. explained by the high value of reduced glutathione against the oxidative stress. o Poster N 91 G6PD enzyme activities decreased in the group given ethanol extract. It is reported that The Effects of The Extracts the decrease of G6PD activity shows a Obtained from The Nepeta Cilicia decrease at NADPH production (Özmen et al., Boiss on the Antioxidant Enzymes 2005). Also the increase at CAT activity is related to the antioxidant activity (Kocabaş, Sercan Özbeka, Fevziye Özdemirb, 2008). It is known the MDA level is one of the İsmail Özmenb, Şevkinaz Konaka, most sensitive indicators of lipid peroxidation Tülay İleri Büyükoğluc, Hasan Özçelikb also oxidative stress (Knight, 1988), therefore our the finding shows that the bad effects of a Health Vocation Training School, free radicals may be decreased. Mehmet Akif Ersoy University, Burdur, References Turkey b Faculty of Arts and Sciences, 1. Özmen İ, Ercişli S, Hizarcı Y, Orhan E, Süleyman Demirel University, Isparta, 2005. Investigation of antioxidant enzyme Turkey activities and lipid peroxidation of Rosa canina c Faculty of Veterinary Medicine, and Rosa dumalis fruits. Acta Hort.690:245- Mehmet Akif Ersoy University, Burdur, 248. Turkey 2. Kocabaş N, 2008. Homosisteinin indüklediği

oksidatif stres üzerinde quercetinin koruyucu It was to determine the effects on some etkisi, AKÜ Sağlık Bilimleri Ens., Afyon. antioxidant enzymes of the ethanol and 3. Knight JA, Pleper RK, Macellon L, 1988. hexane extracts obtained from the Nepeta Specificity of the thiobarbituric acid reaction: Its cilicia. use in studies of lipid peroxidation. Clin Chem. The N. cilicia extacts obtained with ethanol and 34:2433-2438. hexane organic solvents were given to the rats. Poster No 92 Rats were separated into 3 groups of 8 animals each. The extracts were given orally Effect of memantine on brain injury and single dose (1,0 g/kg). The blood samples and oedema after cerebral ischemia were taken from at 3 h after the administration.

Some antioxidant enzyme activities and MDA 1 1 levels were determined. Siğnem Eyuboğlu , G. Büyükgüçlü , M. Uğur1, T. Gençol1, E. Piranen1, Ü. 2 1 1 The ethanol extract caused an important Kılıç , B. Yılmaz , E. Kılıç decrease (p<0.05). The hexane extract increased the glucose-6-phosphate 1 Department of Physiology, Faculty of dehydrogenase (G6PD) activity, but not Medicine, University of Yeditepe, significantly (P>0,05). Catalase (CAT) activities Istanbul, Turkey increased in the group given ethanol extracts 2 Department of Medical Biology, (p<0.05). It was not observed any change at Faculty of Medicine, University of superoxide dismutase (SOD) activities and

50 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Yeditepe, Istanbul, Turkey *Department of Biophysics, Medical Faculty, Dicle University, Diyarbakir, Turkey. In vitro data suggested that rt-PA (recombinant tissue plasminogen activator) may aggravate ** Department of Biochemistry, Ataturk neuronal injury by enhancing N-methyl- D- Education Hospital, Ankara, Turkey. aspartate (NMDA) receptor signalling, possibly due to cleavage of its NR1 receptor subunit. It was subsequently shown that rt-PA treatment also potentiated NMDA toxicity in vivo when In the last decade, public opinion has been focused on hazards of radiofrequency (RF) NMDA was directly applied into the brain. radiation emitted from mobile phones. Based on these data, it was hypothesized that Recently, most of studies, which are increased NMDA receptor transmission may investigate the effects of mobile phone be responsible for the rt-PA induced exposure concentrated to the oxidative aggravation of injury after stroke (Kilic et al., process in the cells. Although many of studies 2001; 2005). We hypothesized therapeutically performed on effects on mobile phone delivered t-PA might increase ischemic injury, which may be reversed by an NMDA exposure on brain, the studies on the effects of antagonist memantine. mobile phone exposure on eyes relatively are limited. Whereas, the eyes also one of the In this study, male C57/BL6 mice were used. most important organs exposed to The animals were divided into 4 groups, radiofrequency radiation emitted from mobile submitted to 90 mins of focal cerebral ischemia phones because of the position of them in the and 24 hours reperfusion. The animals were head. However, the studies on the effects of treated with (1) vehicle, (2) rt-PA (10 mg/kg, mobile phones recently focused on the IV), (3) memantine (20 mg/kg, IP) and (4) oxidative process in cells. Therefore, the aim of memantine add on to rt-PA, just after stroke this study was to investigate the effect of 900 onset. Twenty-four hours after ischemia, brain MHz radiofrequency radiation on the oxidative injury was evaluated.rt-PA delivered parameters in the eyes of rats. The study was immediately after reperfusion onset increased carried out on 19 Wistar Albino adult male rats. infarct volume and brain oedema at 24 hours The rat heads in a carousel exposed to after focalcerebral ischemia. Memantine did 900MHz radiofrequency radiation emitted from not decrease infarct volume significantly, when a generator, which simulate mobile phones. administrated alone. In addition, memantine For the study group (n: 12), rats exposed to the did not reverse rt-PA induced brain injury. radiation 2 h per day (7 days in a week) for 10 months. For the sham group (n: 7), rats were We provide evidence that rt-PA increases brain placed into the carousel and the same injury. However, the NMDA antagonist procedure was applied except that the memantine does not reverse rt-PA induced generator was turned off. The levels of toxicity, indicating memantine is not attractive catalase (CAT), malondialdehyde (MDA), as an add-on treatment with thrombolytics. myeloperoxidase (MPO), total antioxidative

o capacity (TAC), total oxidant status (TOS), and Poster N 93 oxidative stress index (OSI) were measured in rat eye. TAC, TOS and OSI levels were found The effect of long-term 900 MHz RF higher in the eyes of rats exposed to 900 MHz exposure on Rat Eyes in Terms of radiofrequency radiation (p<0.05). In Oxidative and Antioxidative conclusion 900 MHz radiofrequency radiation parameters. emitted from mobile phone can induce oxidative process in the eyes of rats. More Suleyman Dasdag*, M.Z. Akdag*, O. performance is necessary. Erel**

51 3rd International Congress on Cell Membranes and Oxidative Stress: Focus 2010 on Calcium Signaling and TRP Channels

Poster No 94 Intraperitonel NAC (10 mg/kg BW) for 10 days was given to consisting third group after giving N-acetylcysteine modulates one dose of DOX. After ten days, liver tissues doxorubicin-induced liver Damage removed to study antioxidant values and in Rats histologically examined. Lipid peroxidation levels were higher in DOX M. Cem Koçkar1, M. Nazıroğlu2, 2 3 4 group than in control whereas lipid O.Celik , H. T. Tola , D. Bayram , peroxidation levels were lower in NAC group 5 A.Koyu than in control. Vitamin C and E levels were lower in DOX group than in control whereas 1 Department of Internal Medicine, Division vitamin C and E levels were higher in NAC of Gastroenterology, Medical Faculty, group than in DOX group. Glutathione Süleyman Demirel University, Isparta, peroxidase, reduced glutathione, vitamin A, - Turkey carotene values did not change by DOX and NAC administrations. In liver tissue samples of 2Department of Biophysics, Medical DOX group, there were mononuclear cell Faculty, Süleyman Demirel University, infiltrations, vacuolar degeneration, Isparta, Turkey. hepatocytes with basophilic nucleus and sinusoidal dilatations. The histological findings 3Department of Pediatrics, Şanlıurfa State were decreased by NAC administration. Hospital, Şanlıurfa, Turkey. In conclusion, DOX caused increase in lipid 4Department of Histology and Embryology, peroxidation levels of liver in rat although Medical Faculty, Süleyman Demirel antioxidant values decreased. The changes were recovered by NAC administration. University, Isparta, Turkey.

5 Department of Physiology, Medical Faculty, Süleyman Demirel University, Isparta, Turkey.

Doxorubicin (DOX) is a chemotherapeutic agent, which is widely used in cancer treatment. The most common side effect of DOX is on cardiovascular system. There are some studies suggesting oxidative stress- induced toxic changes on liver related to DOX administration. Various methods have been tried to prevent DOX-induced the toxic effects. In this study, we aimed to investigate whether N-acetylcysteine (NAC), a fatty acid co-factor, has a protective effect on hepatic tissue against a possible DOX-induced damage.

Twenty-four male Wistar albino rats weighing between 200-250 grams were included in this study. All animals were divided into three equal groups. First group was used as control. Second group received single dose of intraperitoneal DOX (2.5 mg/kg BW).

52