Search

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.

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. 142076

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 ćelijske linije u kulturi: transformisane ćelije mišijih fibroblasta (L929) i maligne ćelije humane eritroleukemije (K562). Niske koncentracije deluju stimulativno na proliferaciju K562 ćelija u kulturi, dok je kod viših koncentracija situacija suprotna i uočava se dozno-zavisan uticaj SNP na proliferaciju K562 ćelija. Korišćenjem natrijum-nitroprusida i dve vrste superoksid dismutaza, Cu,Zn-SOD i Mn-SOD, stvoreni su uslovi generisanja više vrsta signalnih molekula i ispitan je odgovor obe ćelijske linije 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. null

Balans redoks potencijala u živoj ćeliji predstavlja imperativ održavanja zdravog fenotipa, i ukrajnjem, njenog preživljavanja. Nitrozativni stres može ozbiljno narušiti ćelijsku redokshomeostazu i, u kombinaciji sa oksidativnim stresom, uticati na ćelijsku proliferaciju idiferencijaciju, a u nekim slučajevima i na aktivaciju maligne transformacijeU ovom radu ispitivani su efekti donora NO natrijum-nitroprusida na dve ćelijske linije ukulturi: transformisane ćelije mišijih fibroblasta (L929) i maligne ćelije humaneeritroleukemije (K562). Natrijum- nitroprusid (SNP) je fotoreativan molekul sa veomakratkim poluživotom koji izaziva koncentraciono - zavisnu proliferaciju ili inhibicijućelijskog rasta in vitro.NO izaziva različite efekte u zavisnosti od eksperimentalnog modela,svoje relativne koncentracije kao i okruženja u kojem nastaje. Ispitivanja mogućnosti direktnetransformacije azot oksida u redoks aktivne vrste kao što su nitrozonijum katjon (NO+) initroksil anjon (NO-/HNO) i direktni efekti tih redoks potomaka u ćeliji tek su u začetku. Unašim eksperimentima, korišćenjem donora NO - natrijum nitroprusida (SNP) i dve vrstesuperoksid dismutaza, CuZn-SOD i Mn-SOD, stvorili smo uslove generisanja više vrstasignalnih molekula i ispitali odgovor transfomisanih (L929) i malignih (K562) ćelija na njih.Rezultati eksperimenata pokazuju da izabrani parametri (količina slobodnih tiolnih grupa iglutationa) mogu biti relevantni za praćenje efekata egzogenog azot oksida i njegovih redokspotomaka 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.

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. null

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. .

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.