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Purpose This study aims to examine the relationship between the chemical composition of juices obtained from fruits of autochthonous wild pomegranate (Punica granatum L.) grown in Montenegro and their cytotoxic effects on cancer cells. Design/methodology/approach To explore the potential value of wild pomegranate fruits, in vitro biological assays were carried out with juices whose composition was analyzed in detail for sugars, organic acids, vitamin C and phenolic compounds. The effect of juices on survival was determined in human lung A549, cervical HeLa and breast MCF-7 carcinoma cells by MTT assay. As a control, the cytotoxicity against normal fetal lung fibroblasts (MRC-5) was monitored. Findings Among cancer cell lines, considering the IC50 related to total phenolics, the lowest value – 13 µg/mL was found for the A549. The strongest effect on lung cells was assumed due to the favorable contribution of ellagitannins to total phenolics in juice as well as the given combination of anthocyanins and their synergistic action. For HeLa cells, the lowest IC50 value was obtained at 88 µg/mL, and the cytotoxicity could be matched with the effects of anthocyanins and catechin. For MCF-7 cells, the lowest IC50 was 504 µg/mL, and the elevated levels of vitamin C and ellagic acid derivatives should have a noticeable effect on these cells. Originality/value This study provides an important contribution to the knowledge on the effect of phytochemicals from wild pomegranate juice on lung, cervical and breast cancer cells, in vitro. The present observations suggest that the juice of wild pomegranate has the potential in the fight against cancer.

Radiotherapy-induced toxicity is a major dose-limiting factor in anti-cancer treatment. Ionizing radiation leads to the formation of reactive oxygen and nitrogen species (ROS/RNS) that are associated with radiation-induced cell death. Investigations of biological effects of fullerenol have provided evidence for its ROS/RNS scavenger properties in vitro and radioprotective efficiency in vivo. Therefore we were interested to evaluate its radioprotective properties in vitro in the human erythroleukemia cell line. Pretreatment of irradiated cells by fullerenol exerted statistically significant effects on cell numbers and the response of antioxidative enzymes to X-ray irradiation-induced oxidative stress in cells. Our study provides evidence that the pre-treatment with fullerenol enhanced the enzymatic activity of superoxide dismutase and glutathione peroxidase in irradiated K562 cells.

Polyhydroxy fulleren derivates have significant potential in nanomedical application. Research of polyanion nanoparticle fullerenol C60(OH)24 is of high significance for interpretation of biological mechanisms. This paper investigated the properties of polyanion nanoparticle fullerenol C60(OH)24 as a potential polydentat ligand. Fullerenol C60(OH)24 water solutions were added in solution of [Cu(NH3)4]2+ in order to form a dark brown complex. Absorbance of [Cu(NH3)4]2+ solution was decreasing with increasing concentration of polyanion nanoparticle nanoligand fullerenol. This relation was determined in all investigated concentrations of [Cu(NH3)4]2+. The ratio of Cu2+ complex composer to polyanion polydentat nanoligand fullerenol had an increase from 1.5 to 9, proportional to the increase of the complex composer concentration and decrease of polyanion polydentat nanoligand fullerenol in the alkali medium. The thermogram TGA-DTA of fullerenol and fullerenol and CuSO4 complex, clearly show endothermic effects (which are the result of dehydratation and dehydroxylation) and exothermic effects (as the result of degradation of C60(OH)24 molecules and processes of oxidation in CO, CO2. At the beginning of TGA-DTA fullerenol thermogram, there is a very well defined endothermic peak of water loss at 100?C, followed by mass decrease as a consequence of lost hydroxyl groups, covalent bounded for C60. The influence of the complex composer is manifested in the moving of thermal stability towards lower temperatures. The complex composer is a catalyst of the process of polyanion polydentat nanoligand fullerenol oxidation to CO and CO2. The temperature peak of fullerenol oxidation is at 490?C and in the case of complex oxidation two peaks were detected at 380 and 410?C. Polihidroksilovani fulerenski derivati imaju veliki potencijal u nanomedicinskoj primeni. Istrazivanja osobina polianjonske nanocestice fulerenola, C60(OH)24, od kljucnog su znacaja za tumacenje bioloskih mehanizama. U nasem radu je ispitana osobina polianjonske nanocestice fulerenola C60(OH)24 kao polidentatnog liganda. Vodeni rastvori fulerenola, C60(OH)24, dodavani su u rastvor [Cu(NH3)4]2+ do formiranja tamno smedjeg kompleksa. Apsorbancija rastvora [Cu(NH3)4]2+ smanjuje se sa povecanjem koncentracije polianjonskog polidentanog nanoliganda fulerenola u svim ispitanim koncentracijama [Cu(NH3)4]2+. U svim slucajevima, odnos graditelja kompleksa Cu2+ i polianjonskog polidentatnog nanoliganda povecava se priblizno 1,5 do 9 sa povecanjem koncentracije graditelja kompleksa i smanjenjem koncentracije polianjonskog polidentatnog nanoliganda u alkalnoj sredini. Graditelj kompleksa Cu2+ katalizuje proces oksidacije polidentatnog polianjonskog nanoliganda fulerenola do CO i CO2 na temperaturama od 380 i 410?C, dok se sam polidentani polianjonski ligand oksiduje u jednostepenom procesu na temperaturi od 490?C.

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.

PurposeThe purpose of this paper is to determine the polyphenol content, antioxidative potential and antiproliferative activity of Swiss chard from Montenegro, grown under different irrigation and fertilization regimes.Design/methodology/approachSwiss chard was grown in the open field (Lješkopolje, Montenegro) where it was subjected to different fertilization and irrigation regimes. Chard samples were analyzed for previously identified polyphenols and for antioxidant parameters. Additionally, in order to complete the biological activities, chard extracts were tested for antiproliferative activity against MFC-7 and HT-29 tumor cell lines.FindingsThe polyphenols identified in Swiss chard were flavonoids: vitexin-2?-O-xyloside, vitexin-6?-O-acetyl-2?-O-xyloside, vitexin-6?-O-malonyl-2?-O-xyloside and isorhamnetin-3,7-diglucoside. In the antioxidant tests, DPPH (2,2-diphenyl-1-picrylhydrazyl), and ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid)), chard extract had values of 7.00 and 8.50 (mean values) µmol Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid) equ/d.w., respectively. The tested chard extracts inhibited cell proliferation at different concentrations (3.125–50.0 µg/mL) against the MCF-7 cell line, after 24 and 48 h of incubation. The antiproliferative activity, expressed in terms of IC50, was 32.97 and 86.45 µg/mL after 24 h of incubation and 20.76 and 23.33 µg/mL after 48 h of incubation, for treated and untreated chard extracts, respectively.Originality/valueThese data suggest Montenegrin Swiss chard grown under different irrigation and fertilization treatments can be considered as a functional food and should be included in an everyday diet. The collected data could help in the growth improvement of chard with functional food properties.

There is inadequate published data referring to bioactivity of lemon balm tea and its Kombucha. The aim of this study, therefore, was to investigate antimicrobial, antiproliferative, genotoxic, and antigenotoxic potential of lemon balm tea and its Kombucha with consuming acidity. Antimicrobial activity was determined by agar-well diffusion method. Cell growth effects were determined in HeLa, MCF7, and HT-29 human tumor cell lines. Genotoxic and antigenotoxic effects were determined using chromosome aberration assay in Chinese hamster cell line CHO-K1. Differences between control and treated groups were evaluated using analysis of variance, at significance level of p  < 0.05. Kombucha from lemon balm tea ( Melissa officinalis L.) exibited antimicrobial activity against prokaryotic microorganisms independently of their cell wall structure (both Gram-positive and Gram-negative bacteria), while there was no observed activity against eukaryots (yeasts and moulds). There was absence of genotoxic effects while antigenotoxic effects of lemon balm Kombucha and tea were confirmed on MMC-damaged CHO-K1 cells. For the explanation of cell growth effects that were not concentration dependent, concept of hormesis was used. Antiproliferative activity was lower compared with traditional Kombucha and Satureja montana L. Kombucha, with lemon balm tea showing higher activity than its Kombucha.

Fullerenol (C60(OH)24) nanoparticles (FNP) have a significant role in biomedical research due to their numerous biological activities, some of which are cytoprotective and antioxidative properties. The aim of this study was to measure distribution of fullerenol nanoparticles and zeta potential in cell medium RPMI 1640 with 10% fetal bovine serum (FBS) and to investigate the influence of FNP on Chinese hamster ovary cells (CHO-K1) survival, as well as to determine the activity of three antioxidative enzymes: superoxide-dismutase, glutathione-reductase and glutathione-S-transferase in mitomycin C-treated cell line. Our investigation implies that FNP, as a strong antioxidant, influence the cellular redox state and enzyme activities and thus may reduce cell proliferation, which confirms that FNP could be exploited for its use as a cytoprotective agent.

Fullerenol ([C.sub.60][(OH).sub.24]) nanoparticles (FNP) have a significant role in biomedical research due to their numerous biological activities, some of which have cytoprotective and antioxidative properties. The aim of this study was to measure distribution of fullerenol nanoparticles and zeta potential in cell medium RPMI 1640 with 10% fetal bovine serum (FBS) and to investigate the influence of FNP on Chinese hamster ovary cells (CHO-K1) survival, as well as to determine the activity of three antioxidative enzymes: superoxide-dismutase, glutathione-reductase and glutathione-S-transferase in mitomycin C-treated cell line. Our investigation implies that FNP, as a strong antioxidant, influences the cellular redox state and enzyme activities and thus may reduce cell proliferation, which confirms that FNP could be exploited for its use as a cytoprotective agent. Keywords: fullerenol, mytomocine C, antioxidative enzyme, CHO K1 cell line. Available online at the Journal website: Zahvaljujuci sirokoj bioloskoj aktivnosti, npr. citoprotektivnom i antiksidativnom svojstvu, fulerenol ima znacajnu ulogu u biomedicinskim istrazivanjima, na sta ukazuju i rezultati brojnih istrazivanja. Cilj ovog istrazivanja je bio merenje zeta potencijala i raspodele fulerenol nanocestica u medijumu RPMI 1640 + 10% FBS (Fetal Bovine Serum), kao i ispitivanje uticaja fulerenola na prezivljavanje celija i aktivnost tri antioksidativna enzima: superoksid-dismutaze, glutation-reduktaze i glutation-S-transferaze, u mitomicinom C tretiranoj celijskoj liniji. Istrazivanje je obavljeno na celijama jajnika kineskog hrcka CHO-K1 (Chinese Hamster Ovary cells) koristeci DET test (Due Exclusion Test) za brojanje celija, kao i set spektrofotometrijskih metoda za odredivanje antioksidativne aktivnosti. Celije su pre tretmana mitomicinom C tretirane fulerenolom u dve razlicite koncentracije, a potom inkubirane i analizirane nakon 3 i 24 h. Dodavanje fulerenolskih nanocestica u medijum sa 10% FBS nije izazvalo promene u raspodeli velicina cestica po broju ili zapremini, dok se vrednost zeta potencijala medijuma (θ) promenila sa -3,6 na -7,9 mV. Fulerenol ispoljava protektivni efekat na ?elije CHO-K1 koje su tretirane mitomicinom C. Mitomicin C povecava aktivnost sva tri ispitana enzima, dok sam fulerenol u veoma maloj meri utice na aktivnost pomenutih enzima. Pretretman sa fulerenolom smanjuje stres indukovan mitomicinom C po vremenski i dozno zavisnom obrascu. Nase istrazivanje potvrduje da nanocestice fulerenola uticu na redoks stanje i enzimsku aktivnost ispitivane celijske linije, sto ukazuje na to da mogu sniziti nivo celijske proliferacije i naci primenu kao citoprotektivni agens. Kljucne reci: Fulerenol * Mitomicin C * Antioksidativni enzimi * CHO K1 celijska linija

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 prezivljavanje, kao i tretmana fulerenolom i mitomicinom c (MMC) na aktivnost ukupne superoksid-dismutaze u CHO-K1 (ovarijalnih celija hrcka) celijskoj liniji. U uzorcima celija tretiranim fulerenolom koncentracija 0,01-0,5 mg/mL, praceno je prezivljavanje 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 prezivelih celija tokom 3 i 24 h. Aktivnost SOD raste sa porastom koncentracije fulerenola i u najvecoj koncentraciji opada u obe vremenske tacke eksperimenta. U uzorcima tretiranim fulerenolom i MMC doslo je do smanjenja aktivnosti SOD, izuzev pri koncentraciji fulerenola od 0,0625 mg/mL, kada je zapazen porast aktivnosti SOD u odnosu na kontrolne grupe.

This paper investigates the effects of sodium nitroprusside as NO donor on two cell lines in culture: transformed cells of mice fibroblasts (L929) and malignant cells of human eritroleukemia (K562). Low concentrations of NO have stimulative effect, while high concentrations have inhibitive effects on proliferation of K562 cells in a dose-dependent manner. In our experiments, by using sodium nitroprusside (SNP) as NO donor and two kinds of superoxide dismutase, Cu,Zn-SOD and Mn-SOD, we created conditions to generate several kinds of signal molecules and investigated reaction of transformed (L929) and malignant (K562) cells to dose. Results of experiments are showing that chosen parameters (amount of free thiol groups and glutathione) may be relevant in monitoring the effect of exogenous nitrate oxide and its redox descendants in different, both transformed and malignant cell lines. U ovom radu ispitivani su efekti donora NO natrijum-nitroprusida (SNP) na dve celijske linije u kulturi: transformisane celije misijih fibroblasta (L929) i maligne celije humane eritroleukemije (K562). Niske koncentracije deluju stimulativno na proliferaciju K562 celija u kulturi, dok je kod visih koncentracija situacija suprotna i uocava se dozno-zavisan uticaj SNP na proliferaciju K562 celija. Koriscenjem natrijum-nitroprusida i dve vrste superoksid dismutaza, Cu,Zn-SOD i Mn-SOD, stvoreni su uslovi generisanja vise vrsta signalnih molekula i ispitan je odgovor obe celijske linije na njih. Rezultati eksperimenata pokazuju da izabrani parametri (kolicina slobodnih tiolnih grupa i glutationa) mogu biti relevantni za pracenje efekata egzogenog azot oksida i njegovih redoks potomaka kod razlicitih, transformisanih i malignih celijskih linija.