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The 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/approach: Swiss 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. Findings: The 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/value: These 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.

Purpose: The 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/approach: Swiss 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. Findings: The 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/value: These 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.

The 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/approach: Swiss 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. Findings: The 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/value: These 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.

Purpose: The 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/approach: Swiss 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. Findings: The 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/value: These 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.

У овој дисертацији испитивана су антитуморска дејства антихипергликемијског лека метформина, вазодилататорног лека нитроглицерина, и комбинација ових лекова са дијагностичким средством 2-деокси-D-глукозом и/или радио и хемио сензибилизатором кофеином на хуманим културама аденокарцинома плућа (A549), колоректалног карцинома (HT29), аденокарцинома цервикса (HeLa), као и на контролној ћелијској култури нормалних фибробласта плућа (МRC 5). In vitro испитивање утицаја метформина, нитроглицерина, 2-деокси-D-глукозе и кофеина на проли- ферацију ћелија карцинома грлића материце (HeLa), ћелијској култури аденокарциномa плућа (A549) и ћелијској линији карцинома дебелог црева (HT29). Ћелије у експоненцијалној фази раста третиране су растућим концентрацијама метформина, нитроглицерина и 2-деокси-D-глукозе и утврдила се дозна зависност цитотоксичног ефекта. Метформин, кофеин и 2-деокси-D-глукоза су утицали на смањење процента преживљавања туморских ћелија, док је применом нитроглицерина овај ефекат изостао, иако у експериментима код истовремене примене нитроглицерина и кофеина постоји пад процента преживелих ћелија. Најпотентнији ефекат је постигнут код истовремене примене метформина и кофеина, док је разлог за одсуство снажног цитотоксичног ефекта метформина и 2-деокси-D-глукозе код комбиноване примене молекуларни механизам деловања појединачних супстанци. Снажан пролиферативни ефекат је евидентиран применом метформина и кофена на здравим фибробластима плућа., U ovoj disertaciji ispitivana su antitumorska dejstva antihiperglikemijskog leka metformina, vazodilatatornog leka nitroglicerina, i kombinacija ovih lekova sa dijagnostičkim sredstvom 2-deoksi-D-glukozom i/ili radio i hemio senzibilizatorom kofeinom na humanim kulturama adenokarcinoma pluća (A549), kolorektalnog karcinoma (HT29), adenokarcinoma cerviksa (HeLa), kao i na kontrolnoj ćelijskoj kulturi normalnih fibroblasta pluća (MRC 5). In vitro ispitivanje uticaja metformina, nitroglicerina, 2-deoksi-D-glukoze i kofeina na proli- feraciju ćelija karcinoma grlića materice (HeLa), ćelijskoj kulturi adenokarcinoma pluća (A549) i ćelijskoj liniji karcinoma debelog creva (HT29). Ćelije u eksponencijalnoj fazi rasta tretirane su rastućim koncentracijama metformina, nitroglicerina i 2-deoksi-D-glukoze i utvrdila se dozna zavisnost citotoksičnog efekta. Metformin, kofein i 2-deoksi-D-glukoza su uticali na smanjenje procenta preživljavanja tumorskih ćelija, dok je primenom nitroglicerina ovaj efekat izostao, iako u eksperimentima kod istovremene primene nitroglicerina i kofeina postoji pad procenta preživelih ćelija. Najpotentniji efekat je postignut kod istovremene primene metformina i kofeina, dok je razlog za odsustvo snažnog citotoksičnog efekta metformina i 2-deoksi-D-glukoze kod kombinovane primene molekularni mehanizam delovanja pojedinačnih supstanci. Snažan proliferativni efekat je evidentiran primenom metformina i kofena na zdravim fibroblastima pluća., In this dissertation, the anti-cancer effects of an antihyperglycaemic agent of metformin, a vasodilator drug nitroglycerin, and a combination of these drugs with a 2-deoxy-D-glucose diagnostic agent and / or radio and hemio sensitizer with caffeine on human cultures of adenocarcinoma of the lungs (A549), colorectal carcinoma (HT29), cervix adenocarcinoma (HeLa), as well as on the control cell culture of normal fibroblasts of the lungs (MRC 5). An in vitro study of the effects of metformin, nitroglycerin, 2-deoxy-D-glucose and caffeine on the proliferation of cervical cancer cells (HeLa), cell culture of the lung adenocarcinoma (A549), and colon cancer of the colon (HT29). The cells at the exponential growth stage were treated with rising concentrations of metformin, nitroglycerin and 2-deoxy-D-glucose, and the cytotoxic effect was determined. Metformin, caffeine, and 2-deoxy-D-glucose reduced the number of tumor cells, while nitroglycerin did not it could be concluded. Although there is a decrease in survival in experiments with the simultaneous administration of nitroglycerin and caffeine, the most effective effect is achieved in the simultaneous use of metformin and caffeine, while the reason for the absence of a potent cytotoxic effect of metformin and -deoxy-D-glucose is the molecular mechanism of the action of individual substances. The most significant effect was achieved with the simultaneous administration of metformin and caffeine to the cell culture of lung adenocarcinoma. A potent proliferative effect was recorded using metformin and 2-deoxy-Dglucose on healthy lung fibroblasts.

The aim of this study was to characterize volatile and non-volatile compounds of rosemary from the North Adriatic region and to determine its antiproliferative activity, alone or in combination with radiomimetic bleomycin (BLM) on three malignant and one non-transformed human cell lines. Chemi-cal analysis of the volatile compounds revealed the presence of monoterpenes (93.8%), among which 1.8-cineol (32.9%) and camphor (15.5%) were the dominant compounds. Also, obtained results showed that the major polyphenolic constituents in rosemary extract were phenolic acids (rosmarinic acid and its derivatives up to 69.2 mg 100 g -1), as well as flavones and flavonols in the following order: lute-olin>isorhamnetin>quercetin>kaempferol>apigenin. Cell growth tests showed that rosemary extract alone exerted moderate antiproliferative activity, as well as a synergistic antiproliferative effect with bleomycin (EC 50 344.3-461.5 µg mL -1 and 58.6-292 µg mL -1 , respectively). The anti-tumor effect of rosemary extract in combination with BLM was much stronger, compared to BLM itself on the breast cancer cells. Through its proposed sensitizing effect, rosemary extract, in combination with the standard chemotherapeutics, could be used for the investigations of possible therapeutic modalities.

Fullerenol (C-60(OH)(24)) nanoparticles (FNP) have a significant role in biomedical research due to their numerous biological activities, some of which have cytoprotective and anti-oxidative 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.

Fullerenol (C-60(OH)(24)) nanoparticles (FNP) have a significant role in biomedical research due to their numerous biological activities, some of which have cytoprotective and anti-oxidative 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.

Fullerenol C-60(OH)(24) nano particles (FNP) show various biological activities in different experimental models. We evaluated genotoxic and antigenotoxic effects of FNP on human peripheral blood lymphocytes (PBL) using two cytogenetic assays. The effects of FNP were evaluated on the frequency of chromosomal aberrations (CA) and micronuclei (MN) on undamaged and mitomycin C (MMC)-damaged PBL. FNP number distribution in a culture medium with serum showed that predominant particles were about 180 nm and 90 nm respectively. Cytogenetic assays showed that FNP decreased chromosomal aberrations and micronucleus frequency on the undamaged and the MMC- damaged human PBL at concentration range from 5.54 mu M to 221.60 mu M. Our research confirmed that FNP did not exhibit genotoxic but induced antigenotoxic effects at subcytotoxic concentrations on human lymphocytes.

In this study we performed the dynamic and static scintigraphy of the domestic dog, using the newly synthesized radiopharmaceutical, [Tc-99m(CO)(3)(H2O)(3)]-C-60(OH)(22-24). In the current study, an advanced one-step method for the functionalization of fullerenol by Tc-99m is described. Optical properties of as-prepared samples and the mechanism responsible for the functionalization were investigated using UV-VIS and FTIR spectroscopy, respectively. Also, the presence of the Tc complex on fullerenol was confirmed by using the energy dispersive X-ray spectroscopy, HPLC and MALDI TOF techniques. This simple and effective method of producing a new radiopharmaceutical is of interest not only for its application in various areas of technology and biology, but also for investigating its potential use in radiation technology for nanoengineering of materials. With dynamic scintigraphy, performed during 30 minutes (120 frames, 15 sec per frame), we obtained ratios of heart, liver and spleen counts: 222/249/168; 178/320/217; 120/348/239 respectively. By static scintigraphy after 1 hour, we detected the activity in heart, liver, spleen and intestines. After 4 hours, the radiopharmaceutical activity was detected in salivary glands. The detection after 21 hours showed the activity in kidneys and urinary bladder, while the activity in intestines was absent. After 24 hours, we detected the activity in liver, spleen, kidneys and urinary bladder. Pharmacokinetic investigations performed in this study are of key interest for the further fullerenol in vivo research.

In this study we performed the dynamic and static scintigraphy of the domestic dog, using the newly synthesized radiopharmaceutical, [Tc-99m(CO)(3)(H2O)(3)]-C-60(OH)(22-24). In the current study, an advanced one-step method for the functionalization of fullerenol by Tc-99m is described. Optical properties of as-prepared samples and the mechanism responsible for the functionalization were investigated using UV-VIS and FTIR spectroscopy, respectively. Also, the presence of the Tc complex on fullerenol was confirmed by using the energy dispersive X-ray spectroscopy, HPLC and MALDI TOF techniques. This simple and effective method of producing a new radiopharmaceutical is of interest not only for its application in various areas of technology and biology, but also for investigating its potential use in radiation technology for nanoengineering of materials. With dynamic scintigraphy, performed during 30 minutes (120 frames, 15 sec per frame), we obtained ratios of heart, liver and spleen counts: 222/249/168; 178/320/217; 120/348/239 respectively. By static scintigraphy after 1 hour, we detected the activity in heart, liver, spleen and intestines. After 4 hours, the radiopharmaceutical activity was detected in salivary glands. The detection after 21 hours showed the activity in kidneys and urinary bladder, while the activity in intestines was absent. After 24 hours, we detected the activity in liver, spleen, kidneys and urinary bladder. Pharmacokinetic investigations performed in this study are of key interest for the further fullerenol in vivo research.

We examined effect of α-tocopherol addition and influence of nitric oxide from sodium nitroprusside (SNP) on free thiol content and SOD activity in exponentially growing K562 cells (human erythroleukaemia) in RPMI 1640 (Sigma) supplemented with 2 mM glutamine, 10 % heat-inactivated fetal calf serum and antibiotics at 37 oC in a humidified 5 % carbon dioxide atmosphere. Cells concentration was of 0.85 x 106 per flask. Our results indicate potential dual role of α-tocopherol: induction of more reduced cell state (increased free -SH) and increased resistance of cancer cells to treatment which induce oxidative stress (increased SOD). α-Tocopherol pretreated K562 cells do not expected respond on the SNP treatment. ., Ispitivali smo efekat dodatka α-tokoferola i uticaj azot-oksida iz natrijum-nitroprusida (NNP) na sadržaj slobodnih tiol grupa i SOD aktivnost kod K562 ćelija (humana eritroleukemija) u eksponencijalnoj fazi rasta, koje su gajene u RPMI 1640 (Sigma) sa dodatkom 2 mM glutamina, 10 % toplotom-inaktiviranog fetalnog govedjeg seruma i antibiotika, na 37 oC u vlažnoj atmosferi sa 5 % ugljen-dioksida. Koncentracija ćelija bila je 0,85 x 106 po posudi. Naši rezultati ukazuju na moguću dvostruku ulogu α-tokoferola kod ovih ćelija: indukciju više redukovanog stanja (porast slobodnih -SH grupa), ali i porast njihove otpornosti na tretmane koji indukuju oksidacioni stres (porast SOD aktivnosti). α-Tokoferolom pretretirane K562 ćelije ne daju očekivani odgovor na tretman sa NNP. .

Balans redoks potencijala u živoj ćeliji predstavlja imperativ održavanja zdravog fenotipa, i u krajnjem, njenog preživljavanja. Nitrozativni stres može ozbiljno narušiti ćelijsku redoks homeostazu i, u kombinaciji sa oksidativnim stresom, uticati na ćelijsku proliferaciju i diferencijaciju, a u nekim slučajevima i na aktivaciju maligne transformacije U ovom radu ispitivani su efekti donora NO natrijum-nitroprusida na dve ćelijske linije u kulturi: transformisane ćelije mišijih fibroblasta (L929) i maligne ćelije humane eritroleukemije (K562). Natrijum- nitroprusid (SNP) je fotoreativan molekul sa veoma kratkim poluživotom koji izaziva koncentraciono - zavisnu proliferaciju ili inhibiciju ćelijskog rasta in vitro.NO izaziva različite efekte u zavisnosti od eksperimentalnog modela, svoje relativne koncentracije kao i okruženja u kojem nastaje. Ispitivanja mogućnosti direktne transformacije azot oksida u redoks aktivne vrste kao što su nitrozonijum katjon (NO+) i nitroksil anjon (NO-/HNO) i direktni efekti tih redoks potomaka u ćeliji tek su u začetku. U našim eksperimentima, korišćenjem donora NO - natrijum nitroprusida (SNP) i dve vrste superoksid dismutaza, CuZn-SOD i Mn-SOD, stvorili smo uslove generisanja više vrsta signalnih molekula i ispitali odgovor transfomisanih (L929) i malignih (K562) ćelija na njih. Rezultati eksperimenata pokazuju da izabrani parametri (količina slobodnih tiolnih grupa i glutationa) mogu biti relevantni za praćenje efekata egzogenog azot oksida i njegovih redoks potomaka kod različitih, transformisanih i malignih ćelijskih linija., The redox potential balance in the living cell isthe imperative of continuation of healthy phenotype, and subsequently of its survival. Nitrosative stress may seriously damage cell's redox homeostasis, and in combination with oxidative stress may influence cell proliferation and differentiation, in some cases even activation of malignant transformation. This paper investigates 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). The sodium nitroprusside(SNP) is a photo reactive molecule with very short half-life, causing concentration- dependant proliferation or inhibition of cell growth in vitro.The NO causes different effects depending on experimental model, its relative concentration and environment where it is formed. Investigations of possibility of direct transformation from nitrogen oxide to redox-active species as nitrosonium cation (NO+) and nitroxyl anion (NO −/HNO), as well as direct effects ofthose redox descendants within the cell are only in beginning. In our experiments,by using sodium nitroprusside (SNP) as NO donor and two kind of superoxide dismutase, CuZn-SOD and Mn-SOD, we created conditions to generate several kinds of signal molecules and investigated reaction of transformed (L929) and malignant (K562) cells tothose. Results of experiments are showing the parameters chosen (amount of free thiol groups and glutathione) may be relevant in measuring the effect of exogenous nitrate oxideand its redox descendants in different, both transformed and malignant cell lines.