The activity of superoxide-dismutase in animal cell culture CHO-K1 after treatment with fullerenol and mytomicine C

Eukaryotic cell survives in predominantly reduced conditions. Homeostasis of cellular redox system is an imperative of cell surviving and its normal metabolism. ROS are well recognized for playing a dual role as both deleterious and beneficial species, since they can be either harmful or beneficial to living systems. These species are mutagenic compounds known to lead to DNA damage, favor cell transformation, and contribute to the development of a variety of malignant diseases. All the effects of oxidants are influenced by the cellular antioxidant defenses. This multilayer system consists of low molecular weight components and several antioxidant enzymes. Superoxide dismutases (SODs) are the only enzymes dismuting superoxide radicals. Mitomycin C, a cross-linking agent, demonstrated genotoxicity in all in vitro and in vivo test systems in mammalian cells and animals. Water-soluble fullerenes are well known as cytotoxic agents for many cell lines in vitro. At the other side, fullerenols are good free radical scavengers and antioxidants both in vitro and in vivo. This paper investigates the effects of fullerenol on survival and fullerenol/ /mytomicine (MMC) treatment on superoxide-dismutase (SOD) activity in CHO-K1 cells. Samples were treated 3 and 24 h with fullerenol (C60(OH)24) at concentration range 0.01-0.5 mg/mL and survival was monitored with dye exclusion test (DET). The activity of total SOD was estimated in samples treated with chosen concentrations of fullerenol and MMC (0.5 and 0.1 mg/mL) after 3 and 24 h of cell incubation. Increasing of C60(OH)24 concentration leads to decreasing of percent of surviving cells 3 and 24 h after incubation. The activity of total SOD enhanced with higher concentration of fullerenol, while decreased in the highest concentration at both experimental points. In samples treated with MMC, as well as in samples treated with fullerenol (0.0625 mg/mL) + MMC was noticed boost in total SOD activity in comparison with controls. Treatment with fullerenol decreased SOD activity in rest of samples treated with MMC. Decreased activity of superoxide-dismutase in almost all samples treated with fullerenol and MMC might be contributed to antioxidative properties of fullerenol. Increased enzyme level at concentration of 0.0625 mg/mL may be due to its prooxidative activity.U ovom radu ispitivani su efekti fulerenola (C60(OH)24) na preživljavanje, kao i tretmana fulerenolom i mitomicinom c (MMC) na aktivnost ukupne superoksid-dismutaze u CHO-K1 (ovarijalnih ćelija hrčka) ćelijskoj liniji. U uzorcima ćelija tretiranim fulerenolom koncentracija 0,01-0,5 mg/mL, praćeno je preživljavanje testom odbacivanja boje (DET) u 3 i 24 h tretmanu. Aktivnost superoksid-dismutaze (SOD) merena je u uzorcima tretiranim fulerenolom izabranih koncentracija i mitomicinom c (0,5 i 0,1 mg/mL) nakon 3 i 24 h. Sa porastom koncentracije (C60(OH)24) opada procenat preživelih ćelija tokom 3 i 24 h. Aktivnost SOD raste sa porastom koncentracije fulerenola i u najvećoj koncentraciji opada u obe vremenske tačke eksperimenta. U uzorcima tretiranim fulerenolom i MMC došlo je do smanjenja aktivnosti SOD, izuzev pri koncentraciji fulerenola od 0,0625 mg/mL, kada je zapažen porast aktivnosti SOD u odnosu na kontrolne grupe.Projekat ministarstva br. 14207

Hypericum perforatum

St. John’s wort is a widely used medicinal plant. The quality of herbal drug, which is in most of the cases collected from nature, varies. Therefore, the aim of the present study was detailed chemical characterization of Hypericum perforatum subsp. perforatum samples collected in close time intervals during flowering and fruitification with the purpose to state the phenological stage characterized by maximum levels of active principles. The antioxidant potential and potential to inhibit biologically important enzymes, as well as the cytotoxicity and genotoxicity of the sample collected during the full flowering period, were evaluated. Data showed that the optimal period for the achieving of maximum level of active principles is the phenophase between floral budding and flowering stage. Significant antioxidant potential and the ability to inhibit biologically important enzymes (especially α-glucosidase) were recorded. The extract exhibited no genotoxicity in subcytotoxic concentrations, while increased cytotoxicity recorded in cotreatment with bleomycin on malignant cell lines was especially significant

The activity of superoxide-dismutase in animal cell culture CHO-K1 after treatment with fullerenol and mytomicine C

Eukaryotic cell survives in predominantly reduced conditions. Homeostasis of cellular redox system is an imperative of cell surviving and its normal metabolism. ROS are well recognized for playing a dual role as both deleterious and beneficial species, since they can be either harmful or beneficial to living systems. These species are mutagenic compounds known to lead to DNA damage, favor cell transformation, and contribute to the development of a variety of malignant diseases. All the effects of oxidants are influenced by the cellular antioxidant defenses. This multilayer system consists of low molecular weight components and several antioxidant enzymes. Superoxide dismutases (SODs) are the only enzymes dismuting superoxide radicals. Mitomycin C, a cross-linking agent, demonstrated genotoxicity in all in vitro and in vivo test systems in mammalian cells and animals. Water-soluble fullerenes are well known as cytotoxic agents for many cell lines in vitro. At the other side, fullerenols are good free radical scavengers and antioxidants both in vitro and in vivo. This paper investigates the effects of fullerenol on survival and fullerenol/ /mytomicine (MMC) treatment on superoxide-dismutase (SOD) activity in CHO-K1 cells. Samples were treated 3 and 24 h with fullerenol (C60(OH)24) at concentration range 0.01-0.5 mg/mL and survival was monitored with dye exclusion test (DET). The activity of total SOD was estimated in samples treated with chosen concentrations of fullerenol and MMC (0.5 and 0.1 mg/mL) after 3 and 24 h of cell incubation. Increasing of C60(OH)24 concentration leads to decreasing of percent of surviving cells 3 and 24 h after incubation. The activity of total SOD enhanced with higher concentration of fullerenol, while decreased in the highest concentration at both experimental points. In samples treated with MMC, as well as in samples treated with fullerenol (0.0625 mg/mL) + MMC was noticed boost in total SOD activity in comparison with controls. Treatment with fullerenol decreased SOD activity in rest of samples treated with MMC. Decreased activity of superoxide-dismutase in almost all samples treated with fullerenol and MMC might be contributed to antioxidative properties of fullerenol. Increased enzyme level at concentration of 0.0625 mg/mL may be due to its prooxidative activity

The activity of superoxide-dismutase in animal cell culture CHO-K1 after treatment with fullerenol and mytomicine C

Eukaryotic cell survives in predominantly reduced conditions. Homeostasis of cellular redox system is an imperative of cell surviving and its normal metabolism. ROS are well recognized for playing a dual role as both deleterious and beneficial species, since they can be either harmful or beneficial to living systems. These species are mutagenic compounds known to lead to DNA damage, favor cell transformation, and contribute to the development of a variety of malignant diseases. All the effects of oxidants are influenced by the cellular antioxidant defenses. This multilayer system consists of low molecular weight components and several antioxidant enzymes. Superoxide dismutases (SODs) are the only enzymes dismuting superoxide radicals. Mitomycin C, a cross-linking agent, demonstrated genotoxicity in all in vitro and in vivo test systems in mammalian cells and animals. Water-soluble fullerenes are well known as cytotoxic agents for many cell lines in vitro. At the other side, fullerenols are good free radical scavengers and antioxidants both in vitro and in vivo. This paper investigates the effects of fullerenol on survival and fullerenol/ /mytomicine (MMC) treatment on superoxide-dismutase (SOD) activity in CHO-K1 cells. Samples were treated 3 and 24 h with fullerenol (C60(OH)24) at concentration range 0.01-0.5 mg/mL and survival was monitored with dye exclusion test (DET). The activity of total SOD was estimated in samples treated with chosen concentrations of fullerenol and MMC (0.5 and 0.1 mg/mL) after 3 and 24 h of cell incubation. Increasing of C60(OH)24 concentration leads to decreasing of percent of surviving cells 3 and 24 h after incubation. The activity of total SOD enhanced with higher concentration of fullerenol, while decreased in the highest concentration at both experimental points. In samples treated with MMC, as well as in samples treated with fullerenol (0.0625 mg/mL) + MMC was noticed boost in total SOD activity in comparison with controls. Treatment with fullerenol decreased SOD activity in rest of samples treated with MMC. Decreased activity of superoxide-dismutase in almost all samples treated with fullerenol and MMC might be contributed to antioxidative properties of fullerenol. Increased enzyme level at concentration of 0.0625 mg/mL may be due to its prooxidative activity.U ovom radu ispitivani su efekti fulerenola (C60(OH)24) na preživljavanje, kao i tretmana fulerenolom i mitomicinom c (MMC) na aktivnost ukupne superoksid-dismutaze u CHO-K1 (ovarijalnih ćelija hrčka) ćelijskoj liniji. U uzorcima ćelija tretiranim fulerenolom koncentracija 0,01-0,5 mg/mL, praćeno je preživljavanje testom odbacivanja boje (DET) u 3 i 24 h tretmanu. Aktivnost superoksid-dismutaze (SOD) merena je u uzorcima tretiranim fulerenolom izabranih koncentracija i mitomicinom c (0,5 i 0,1 mg/mL) nakon 3 i 24 h. Sa porastom koncentracije (C60(OH)24) opada procenat preživelih ćelija tokom 3 i 24 h. Aktivnost SOD raste sa porastom koncentracije fulerenola i u najvećoj koncentraciji opada u obe vremenske tačke eksperimenta. U uzorcima tretiranim fulerenolom i MMC došlo je do smanjenja aktivnosti SOD, izuzev pri koncentraciji fulerenola od 0,0625 mg/mL, kada je zapažen porast aktivnosti SOD u odnosu na kontrolne grupe.Projekat ministarstva br. 14207

The influence of fullerenol on the cell number, cell area and colony forming unit ability in irradiated human erythroleukemic cell line

DET (dye exclusion test) cell count and cell area by computer analysis of the images were determined in cell lines of human eritroleukemia (K562), which were irradiated with X-rays in one dose of 24 Gy and pretreated with 10 nmol/mL fullerenol (Cgo(OH)24). Cell samples obtained using a citocentrifuge and May-Grünvald Giemsi (MGG) during, were analyzed. The cell colony formation ability was monitored using quantative CFU (colony forming unit) test. Irradiation decreases the number of K562 cells, but fullerenol significantly increases cell number on 24th and 48th hour of the experiment. Cell area is larger, and the number of formed cell colonies after irradiation is significantly smaller compared to pretreated groups during the whole experiment. Pretreatment with fullerenol maintains a smaller cell area, and the number of colony formed units was larger compared to the irradiated cells

The influence of fullerenol on antioxidative enzyme activity in irradiated human erythroleukemic cell line (K562)

Cell culture K562 samples were treated with fullerenol (C6o(OH)24) at a concentration of 10 nmol/mL and thereafter irradiated with X-rays (24Gy). The activity of gamma-glutamyltransfrease (γ-GT), total superoxide-dismutase (SOD) and glutathion-peroxidase (GSH-Px) was determined 1, 24 and 48 hours after irradiation. Irradiation induces an increase in the activity of all the investigated enzymes. Fullerenol in the applied dose decreased the γ-GT activity 24 and 48 h after irradiation. The total SOD activity is increased in both pretreated groups except in the irradiated group at the 48th hour. Treatment with fullerenol before irradiation increased GSH-Px activity in irradiated groups and decreased it in the control groups.U uzorcima ćelijske kulture humane eritroleukemije (K562) ozračenim X-zracima doze 24 Gy i predtretiranim fulerenolom (C60(OH)24) koncentracije 10 nmol/mL određivana je aktivnost gama-glutamiltransferaze (γ-GT), ukupne superoksid-dismutaze (SOD) i glutation-peroksidaze (GSH-Px) 1, 24 i 48 sati nakon zračenja. Iradijacija povećava aktivnost sva tri ispitivana enzima. U 24-om i 48-om satu nakon zračenja zapaža se sniženje aktivnosti γ-GT u grupama predtretiranim fulerenolom. Fulerenol utiče na povišenje aktivnosti SOD u obe tretirane grupe, osim u grupi ozračenih uzoraka u 48-om satu gde je došlo do sniženja aktivnosti. Predtretman fulerenolom u zračenim grupama povećava, a u neozračenim eksperimentalnim grupama snižava aktivnost GSH-Px.nul

The influence of fullerenol on antioxidative enzyme activity in irradiated human erythroleukemic cell line (K562)

Cell culture K562 samples were treated with fullerenol (C6o(OH)24) at a concentration of 10 nmol/mL and thereafter irradiated with X-rays (24Gy). The activity of gamma-glutamyltransfrease (γ-GT), total superoxide-dismutase (SOD) and glutathion-peroxidase (GSH-Px) was determined 1, 24 and 48 hours after irradiation. Irradiation induces an increase in the activity of all the investigated enzymes. Fullerenol in the applied dose decreased the γ-GT activity 24 and 48 h after irradiation. The total SOD activity is increased in both pretreated groups except in the iradiated group at the 48th hour. Treatment with fullerenol before irradiation increased GSH-Px activity in irradiated groups and decreased it in the control groups

The Oxidative Stress Parameters as Useful Tools in Evaluating the DNA Damage and Changes in the Complete Blood Count in Hospital Workers Exposed to Low Doses of Antineoplastic Drugs and Ionizing Radiation

Hospital workers at the Oncology Department are occupationally exposed to antineoplastic drugs (ANTNP) or low doses of ionizing radiation (Irrad). Therefore, the aim of this study was to evaluate the level of DNA damage, the oxidative stress parameters and complete blood count (CBC) of hospital workers in order to analyze the negative health effects of ANTNP and low dose Irrad. The frequency of micronuclei (MN) and proliferation index (PI) were analyzed by cytokinesis-block test. The oxidative stress biomarkers evaluated were the level of lipid peroxidation in plasma and catalase activity (CAT) in erythrocytes. A group of 86 hospital workers (35 exposed to ANTPN and 51 to Irrad) had increased MN frequency, CAT activity and level of lipid peroxidation compared to the control group, which consisted of 24 volunteers. The hemoglobin level was lower in the ANTNP group compared to thecontrol group, while a significant difference in RBC was recorded between thecontrol and Irrad groups, and in platelet count betweentheIrrad and ANTNP group. The results showed increased DNA damage, oxidative stress parameters, as well as impairment on complete blood count in hospital workers occupationally exposed to antineoplastic drugs and low-dose ionizing radiation. As this research has shown the importance of oxidative stress, we suggest that in addition to routine methods in periodic medical evaluation, the possibility of applying oxidative stress parameters is considered. Moreover, hospital workers exposed to ANTNP and Irrad in the workplace should undergo not only a more complete health prevention procedure but also have a more appropriate health promotion

The influence of fullerenol on antioxidative enzyme activity in irradiated human erythroleukemic cell line (K562)

Cell culture K562 samples were treated with fullerenol (C6o(OH)24) at a concentration of 10 nmol/mL and thereafter irradiated with X-rays (24Gy). The activity of gamma-glutamyltransfrease (γ-GT), total superoxide-dismutase (SOD) and glutathion-peroxidase (GSH-Px) was determined 1, 24 and 48 hours after irradiation. Irradiation induces an increase in the activity of all the investigated enzymes. Fullerenol in the applied dose decreased the γ-GT activity 24 and 48 h after irradiation. The total SOD activity is increased in both pretreated groups except in the irradiated group at the 48th hour. Treatment with fullerenol before irradiation increased GSH-Px activity in irradiated groups and decreased it in the control groups.U uzorcima ćelijske kulture humane eritroleukemije (K562) ozračenim X-zracima doze 24 Gy i predtretiranim fulerenolom (C60(OH)24) koncentracije 10 nmol/mL određivana je aktivnost gama-glutamiltransferaze (γ-GT), ukupne superoksid-dismutaze (SOD) i glutation-peroksidaze (GSH-Px) 1, 24 i 48 sati nakon zračenja. Iradijacija povećava aktivnost sva tri ispitivana enzima. U 24-om i 48-om satu nakon zračenja zapaža se sniženje aktivnosti γ-GT u grupama predtretiranim fulerenolom. Fulerenol utiče na povišenje aktivnosti SOD u obe tretirane grupe, osim u grupi ozračenih uzoraka u 48-om satu gde je došlo do sniženja aktivnosti. Predtretman fulerenolom u zračenim grupama povećava, a u neozračenim eksperimentalnim grupama snižava aktivnost GSH-Px.nul