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    Purification and characterization of nitric oxide synthase from bovine kidney and investigating drug-induced toxicities of some antibiotics on the enzyme activity
    (Bentham Science Publishers B.V., 2017) Işgör, Mehmet Mustafa; Ekinci, Deniz; Beydemir, Şükrü
    Introduction: Broad spectrums of antibiotics are widely used in the treatment of bacterial infections, particularly causing Staphylococcus aureus. These drugs have high acute renal injury (AKI) potency due to an increase in nitric oxide level. In this study, potency of nitric oxide synthase inhibition of some antibiotics was investigated according to vancomycin which was used as a reference antibiotic in renal injury. Methods and Results: Nitric oxide synthase (NOS) enzyme was purified by using 2'5'-ADP Sepharose 4B affinity chromatography from bovine kidney tissue in a single step with a yield of 3.58% and 1217-fold. Native and subunit molecular weights of the purified kidney NOS enzyme were calculated as 215 kDa and 114.8 kDa. Optimum pH, optimum ionic strength, optimum temperature and stable pH values for purified enzyme were determined as 7.5, 50 mM, 10°C and a pH range of 5.5-6.5, respectively. In vitro effects of antibiotics on purified enzyme activity were investigated by drawing Lineweaver-Burk plots. Enrofloxacin, kanamycin, chloramphenicol, gentamicin, vancomycin, cefazolin, streptomycin and ampicillin inhibited enzyme activity. IC50 values of these compounds were determined as 0.189 mM, 0.295 mM, 1.509 mM, 6.614 mM, 16.579 mM, 18.679 mM, 28.171 mM and 30.394 mM, respectively. Conclusion: Enrofloxacin and kanamycin were observed to have stronger NOS inhibitory effects as compared to vancomycin and may be more reliable antibiotics for use in renal infections as an alternative to vancomycin. © 2017 Bentham Science Publishers.
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    Role of epigallocatechin gallate on in vitro model of methylglyoxal-induced amyloidogenesis
    (BRNSS Publication Hub, 2016) Kucukgul, Altug; Işgör, Mehmet Mustafa; Cellat, Mustafa; Ata, Hayrettin
    Introduction: The study aimed to investigate the potential effects of epigallocatechin gallate (EG) on the reflux of methylglyoxal (MG)-induced amyloidogenesis in human glioblastoma (U87) cells. Materials and Methods: The effective concentrations of MG and EG were investigated via Trypan blue test. Glyoxalase-1 (GLO-1), ?-amyloid precursor protein (?APP), and Caspase 3 (Cas 3) expression levels were determined by quantitative real-time polymerase chain reaction and intracellular glutathione (GSH) contents of cells were measured. Results: MG at 250 ?M reduced viable cells by 33.4% as compared to control group. However, 5 ?M EG pre-treatment before MG prevented 22.4% of the cell loss caused by MG (P ? 0.05). MG stimulated ?APP and Caspase 3 levels by 4.13- and 3.46-fold; however, EG pre-treatment inhibited these increases by 1.76 and 3.09, respectively. In addition, EG pre-treatment increased GLO-1 levels by 3.71-fold and GSH levels by 2.30-fold according to MG group. Conclusion: EG demonstrated protective effect against cell death on U87 cells by suppressing amyloidogenic factors and apoptotic stimuli induced by MG.