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The aim of this study was to determine the effects of a specific mixture of metal nanoparticles (Ag, Cu, Fe and MnО2) orally administered to rats, on biochemical blood markers as indicators of chronic toxicity. Animals (Wistar line rats (n=20)) in experimental groups were given a solution of a mixture of metal salts or a mixture of nanoparticles with food for 90 days. On days 15, 30, 60 and 90 of the experiment, five rats from each group were anaesthetised with CO2 and decapitated, and blood samples were collected for biochemical studies and spectrophotometric measurement. The toxic effect of the chronic administration of the mixture at a dose of 4.0 mg/kg of body weight in white rats caused partial immunosuppression expressed as hypoproteinaemia, excessive formation of circulating immune complexes and acute phase serum mucoid proteins (P, Cilj je naše studije bio utvrditi učinke mješavine nanočestica određenih metala (Ag, Cu, Fe i MnО2) koje su oralno davane štakorima na biokemijske markere krvi kao indikatore kronične toksičnosti. Životinjama (štakorima soja Wistar (n=20)) je u eksperimentalnim skupinama hranom tijekom 90 dana davana je otopina mješavine metalnih soli ili mješavine nanočestica. Na 15., 60. i 90. dan eksperimenta, pet štakora je iz svake skupine pomoću CO2 anestezirano i dekapitirano te su prikupljeni uzorci krvi za biokemijske studije. Spektrofotometrijska mjerenja su provedena uporabom SHIMADZU UV-1800 instrumenta. Toksični je učinak kronične primjene NPMe mješavine pri dozi od 4,0 mg/kg tjelesne mase u bijelih štakora izazvao djelomičnu imunosupresiju izraženu kao hipoproteinemija, pretjerano formiranje cirkulirajućih imunokompleksa i mukoidne proteine u serumu (P

Rtanjski čaj, Satureja montana L. ima široku upotrebu u mnogim delovima sveta, koristi se kao kulinarska biljka i u tradicionalnoj medicini za lečenje ili ublažavanje različitih zdravstvenih simptoma. Međutim, brojne studije su pokazale da biljka ima farmakološki potencijal da se upotrebljava i kao antibakterijski, antidijabetesni, anti-HIV i antiproliferativni lek. Farmakološki efekti rtanjskog čaja mogu se pripisati prisustvu fitohemikalija poput flavonoida, sterola, tanina i etarskih ulja. Fenolna jedinjenja su uglavnom odgovorna za antioksidativnu aktivnost biljke Satureja montana L. U ovom istraživanju cilj je bio utvrđivanje uticaja ekstrakta biljke Satureja montana L. na parametre kao što su glikemija, status lipida i biohemijski parametri, zatim na indikatore koji ukazuju na funkciju jetre i bubrega u serumu zdravih, dijabetičnih pacova i pacova koji su bili na ishrani obogaćenoj mastima, kao i de se ispita uticaj na pokazatelje oksidativnog stresa kod životinja izloženih toksičnoj dozi paracetamola. Za određivanje hipoglikemijskog i antihiperlipidemijskog dejstva biljke Satureja montana L., laboratorijski pacovi soja WISTAR su podeljeni u 8 grupa, u grupama koje su korišćene za određivanje hipoglikemijskog potencijala dijabetes je indukovan streptozotocino nakon čega su životinje per os 28 dana, tretirane fiziološkim rastvorom, metforminom 100 mg/kg, ekstraktom rtanjskog čaja 500 mg/kg i njihovom kombinacijom. Za ispitivanje antihiperlipidemijskog efekta životinje su bile na ishrani obogaćenoj mastima i fiziološkim rastvorom, simvastatinom 10 mg/kg, ekstraktom rtanjskog čaja 500 mg/kg i njihovom kombinacijom. Za određivanje antioksidativnog potencijala biljke Satureja montana L. životinje su podeljene u četiri grupe i 7 dana su tretirane ekstraktom rtanjskog čaja i fiziološkim rastvorom. Nakon toga, pacovi su bili izloženi toksičnoj dozi paracetamola. U životinjskom serumu određivani su biohemijski parametri, koncentracija lipida i parametri funkcije bubrega i jetre. Određivanje aktivnosti lipidne peroksidacije i enzima antioksidativne zaštite u homogenatima jetre eksperimentalnih životinja, nakon primene toksične doze paracetamola, je ispitivano ex vivo. In vitro ispitivanja su vršena sa ekstraktima dobijenim rastvaranjem u različitim rastvaračima. Ispitivanja antioksidativne aktivnosti, kao i ukupni nivoi fenola i flavonoida, određeni su spektrofotometrijskom metodom. Kod životinja sa indukovanim dijabetesom 28-dnevni tretman ekstraktom Satureja montana L. u poređenju sa kontrolom grupom dijabetičnih životinja prouzrokovao je statistički značajno smanjenje glukoze. Na kraju studije, u grupi koja je primala ekstrakt Satureja montana L., uočene su statistički značajno niže vrednosti triglicerida u odnosu na kontrolnu dijabetičnu grupu. Na druge parametre lipidnog statusa, ispitivani ekstrakt nije pokazao značajne efekte. Kod životinja čiju smo ishranu obogatili mastima primenom ekstrakta Satureja montana L. došlo je do statistički značajnog snižavanja LDL holesterola u serumu, u odnosu na kontrolnu grupu koja je bila na isharni obogaćenoj mastima. Vrednosti parametara pokazatelja funkcije bubrega i jetre: AST, urea, kreatinin i mokraćna kiselina bile su značajno niže u grupi tretiranoj ekstraktom Satureja montana L. u odnosu na kontrolnu grupu koja je bila na ishrani obogaćenoj mastima. Pretretman životinja biljnim ekstraktom pre primene toksične doze paracetamola značajno je smanjio aktivnost alkalne fosfataze, koncentraciju direktnog bilirubina i kreatinina u serumu u odnosu na grupu tretiranu fiziološkim rastvorom i pojedinačnom dozom paracetamola. Aktivnost lipidne peroksidaze bila je značajno veća kod životinja koje su tretirane fiziološkim rastvorom u poređenju sa tretmanom ekstraktom Satureja montana L., pre toksične doza paracetamola. Pored toga, smanjen je broj inflamatornih ćelija zajedno sa aktivnošću enzima CIP2E1 kod životinja koje su prethodno tretirane ekstraktom Satureja montana L. Rezultati ovih istraživanja pokazali su hipoglikemijski potencijal ekstrakta Satureja montana L. kod životinja sa indukovanim dijabetesom. Pretretman ekstraktom Satureja montana L. pre primene toksične doze paracetamola, pokazao je potencijal hepatoprotektivnog, antiinflamatornog i antioksidativnog dejstva. Takođe, PCA analiza je pokazala da veliki uticaj na antioksidativni efekat imaju medijumi korišćeni za ekstrakciju. Satureja montana L. is widely used as culinary herb and in traditional medicine for the treatment or relief of various common health symptoms in many parts of the world. However, numerous studies have shown that it has pharmacological potential to be used as antibacterial drug, antidiabetic, anti-HIV and antiproliferative drug. Its pharmacological effects can be attributed to the presence of phytochemicals like flavonoids, sterols, tannins and essential oils. Phenolic compounds are mostly responsible for the anti-oxidative activity of Satureja montana L. In this research, the aim was to determine the influence of Satureja montana L. extract on glycemia, lipid status and biochemical parameters, indicators of liver and kidney function in the serum of healthy and diabetic rats and rats fed with high fat diet, as well as to examine it influence on the parameters of oxidative stress in animals exposed to the toxic dose of paracetamol. For determination of hypoglycemic and antihyperlipidemic effect of Satureja montana L., Wistar laboratory rats were divided into 8 groups, in groups used to determine hypoglycemic potential diabetes was induced by streptozotocin and after that, animals were treated for 28 days per os with saline solution, metformin 100 mg/kg, Satureja montana extract 500 mg/kg and their combination. For examination of hypolipidemic effect animals were treated with high fat diet and saline solution, simvastatin 10 mg/kg, Satureja montana L. extract 500 mg/kg and their combination. For determination of antioxidant potential of Satureja montana L. animals were divided into four groups and winter savory extract and saline solution were given them orally for 7 days. After that, rats were exposed to the toxic dose of paracetamol. Biochemical parameters, the lipids concentration and parameters of the kidney and liver function, were determined in animal serum. Ex vivo analyses included the determination of lipid peroxidation activity and the activity of antioxidant protection enzymes in liver homogenates of the test animals, after administration of a toxic dose of paracetamol. In vitro analyses were performed with extracts obtained with different solvents. Antioxidant activity tests as well as total phenolic and flavonoid content, were determined spectrophotometrically. In diabetic animals, 28 days treatment with Satureja montana L. extract brought to statistically significant decrease of glucose comparing to the control. Statistically significantly lower TG values were observed in the group that received Satureja montana L. extract compared to the control diabetic group, at the end of the study. On the other parameter of lipid status, the examined extract did not show significance effects. In animals fed with food enriched with fats, application of Satureja montana L. extract brought to statistically significant decrease of LDL cholesterol, relative to the control group fed with food enriched with fats. Concentration of parameters of the kidney and liver function: AST, urea, creatinine, and uric acid were significantly lower in the group treated with Satureja montana L. extract in comparison to the control group fed with food enriched with fats. Pre-treatment of animals with herbal extracts before application of the toxic dose of paracetamol significantly decreased alkaline phosphatase activity, concentration of direct bilirubin and creatinine in the serum relative to the group treated with saline solution and single dose of paracetamol. The activity of lipid peroxidation was significantly higher in animals that were treated with saline solution compared with Satureja montana L. extract, before the toxic doses of paracetamol. In addition, in animals pretreated with Satureja montana L. extract, the number of inflammatory cells is decreased along with the activity of CYP2E1 enzymes. The results of these studies showed hypoglycemic potential of Satureja montana L. extract in animals with induced diabetes. Pretreatment with Satureja montana L. extract prior to the administration of the toxic dose of paracetamol has shown potential of hepatoprotective, anti-inflammatory and antioxidative effect. Also, PCA analyses showed that the extraction mediums had a large influence on the antioxidative effect.

Elektromagnetsko se zračenje emitira iz prirodnog okruženja, ali i uporabom industrijskih i svakodnevnih uređaja za bežičnu komunikaciju, stoga su ljudski i životinjski organizmi stalno izloženi zračenju. Tijekom posljednjih godina, zbog brzog tehnološkog napretka, elektromagnetsko zračenje iz umjetnih izvora premašilo je vrijednosti zračenja prirodnog podrijetla. Opća zabrinutost svih nas, zbog sve većeg broja uređaja (mobilnih telefona, prijenosnih računala, Wi-Fi-ja i mikrovalnih pećnica), koji koriste radiofrekvencijsko elektromagnetsko zračenje (RF-EMZ) opravdana je zbog sve brojnijih dokaza o njihovoj štetnosti na žive organizme. Suvremeni uređaji moderne tehnologije emitiraju radiofrekvencijske elektromagnetske valove malih frekvencija koje ljudsko i životinjsko tijelo apsorbira što može imati potencijalne štetne učinke na: mozak, srce, endokrini sustav i reproduktivnu funkciju. Muški je reproduktivni sustav jedno od najosjetljivijih tkiva na RF-EMZ-e. Tako je primjerice, iz trenutno dostupnih studija provedenih in vitro i in vivo, jasno da RF-EMZ-e ima štetne učinke na spermatogenezu, odnosno kakvoću ejakulata ljudi i životinja – broj spermija u ejakulatu, preživljavanje, morfologiju i gibljivost spermija - utječe na stanični metabolizam i endokrini sustav i može prouzročiti genotoksičnost, genomsku nestabilnost i oksidativni stres, a to može prouzročiti neplodnost. Štetni učinci RFEMZ-a dijele se na toplinske i netoplinske. Većinanegativnih bioloških učinaka pripisuje se netoplinskim učincima, a toplinski se učinci nastali RF-EMZ mobilnog telefona, smatraju manje štetnima. Zbog stvaranja prevelike količine reaktivnih kisikovih spojeva u muškom spolnom sustavu hipertermija skrotuma i povećani oksidativni stres mogu biti ključni mehanizmi putem kojih RF-EMZ-e utječe na plodnost muškaraca. Navedeni su i negativni učinci povezani s vremenom korištenja, ponajprije mobilnog telefona. Stoga je cilj ovoga preglednog rada opisati neke od učinaka RF-EMZ-a na muški spolni sustav., Humans and animals are constantly exposed to electromagnetic radiation emitted from the natural environment, and through the use of industrial and everyday devices for wireless communication. In recent years, due to rapid technological progress, electromagnetic radiation from artificial sources has exceeded the values of radiation of natural origin. General concern about the increasing number of devices (mobile phones, laptops, Wi-Fi and microwave ovens) using radiofrequency electromagnetic radiation (RF-EMR) is justified due to increasing evidence of their harm to the living organism. Modern technology devices emit small frequency radiofrequency electromagnetic waves, which are then absorbed by the human and animal bodies and can potentially cause adverse effects on the brain, heart, endocrine system and reproductive function. It is believed that the male reproductive system is one of the most sensitive tissues to RF-EMR. It is clear from the literature that RF-EMR has harmful effects on ejaculate quality indicators (such as spermatozoa count in ejaculate and spermatozoa morphology and motility), affects cellular metabolism and the endocrine system, and causes genotoxicity, genomic instability and oxidative stress, which in turn may result in infertility. The adverse effects of RF-EMR are divided into thermal and non-thermal. Most negative biological effects are attributed to non-thermal effects, while the thermal effects from cell phone radiation are considered to be less harmful. However, scrotum hyperthermia and increased oxidative stress from the formation of excess reactive oxygen compounds in the male reproductive system can be key mechanisms by which RF-EMR affects male fertility. The negative effects associated with the duration of use of mobile phones are known and listed above. Therefore, the aim of this review article is to describe some of the effects of RF-EMR on the male reproductive system.

Литература даје ограничене податке о употреби матичњака код пацијената са инфекцијом вирусом хумане дефицијенције (енг. human immunodeficiency virus ‒ HIV), али подаци клиничких студија указују на бенефите примене ове биљке код поремећаја метаболизма угљених хидрата и липида. Циљ рада је био испитати деловање матичњака уз антиретровирусну терапију која садржи ефавиренз, дарунавир или ралтегравир, као и утврдити позитивне ефекте примене матичњака у регулацији метаболичког синдрома. Извршена је прелиминарна и детаљна хемијска анализа дијететског суплемента на бази екстракта листа матичњака. Испитано је антихипергликемично деловање екстракта у in vitro тестовима инхибиције ензима α-амилазе и α-глукозидазе. Поред тога, суплемент на бази екстракта матичњака и чиста розмаринска киселина су у in vitro условима подвргнути ферментацији помоћу пробиотских микроорганизама. Испитивање на пацијентима је укључило пацијенте Клинике за инфективне болести Клиничког центра Војводине који су инфицирани HIV-ом, са присутним метаболичким синдромом. Пацијенти су на основу лека који примају у терапији подељени на 3 групе, а поред прописане антиретровирусне терапије примали су и матичњак у дози од 600 m током 12 недеља. Пацијентима су на почетку и на крају периода испитивања мерени следећи параметри: концентрација розмаринске киселине у крви, концентрација антиретровирусног лека (ефавиренз, дарунавир или ралтегравир) у крви, број копија вируса HIV-а у крви, хематолошки, имунолошки и биохемијски параметри, показатељи оксидативног стреса, активност ензима антиоксидативне заштите, као и антропометријски параметри. Испитани суплемент на бази екстракта окарактерисан је као добар извор фенолних једињења, при чему је најзаступљенија компонента била розмаринска киселина. Утврђена је in vitro инхибиција оба ензима у релативно ниским концентрацијама, уз бољу активност у инхибицији α-глукозидазе. Путем in vitro ферментације показано је да суплемент на бази екстракта подлеже интензивном метаболизму од стране пробиотских сојева; розмаринска киселина у екстракту је стабилнија у поређењу са изолованом формом. Показано је и да ово једињење представља маркер употребе матичњака у плазми, при чему се розмаринска киселина у крви након 12 недеља употребе доминантно налази у облику глукуронида и сулфата. Нису пронађене статистички значајне разлике између концентрација испитиваних лекова одређених пре и после 12 недеља примене матичњака. Није забележено повећање броја копија вируса у крви. Хематолошки и имунолошки параметри који су праћени нису показали статистички значајну промену вредности. У оквиру биохемијских параметара није дошло до статистички значајних промена које се могу повезати са употребом матичњака, са изузетком испитиване групе ефавиренза, где је дошло до статистички значајног повећања аланин аминотрансферазе и аспартат аминотрансферазе, али је повећање наведених ензима било у оквиру физиолошких вредности. Показано је да након 12 недеља примене матичњака долази до статистички значајне редукције параметара оксидативног стреса у испитиваним групама пацијената који су примали ефавиренз и дарунавир, a у испитиваној групи са ефавирензом потврђено је и статистички значајно повећање активности глутатион-S-трансферазе. У оквиру антропометријски параметара уочено је повећање телесне масе након 12 недеља примене матичњака у испитиваној групи са ефавирензом. Овде представљено истраживање показало је да истовремена примена матичњака уз антиретровирусну терапију која укључује један од испитиваних лекова не утиче на концентрације ових лекова. Праћени параметри који су испитивани код пацијената показали су да је примена матичњака у дози од 600 mg током 12 недеља безбедна за пацијенте. Показани позитивни ефекти указују на повољно дејство код пацијената инфицираних HIV-ом са метаболичким синдромом. Literatura daje ograničene podatke o upotrebi matičnjaka kod pacijenata sa infekcijom virusom humane deficijencije (eng. human immunodeficiency virus ‒ HIV), ali podaci kliničkih studija ukazuju na benefite primene ove biljke kod poremećaja metabolizma ugljenih hidrata i lipida. Cilj rada je bio ispitati delovanje matičnjaka uz antiretrovirusnu terapiju koja sadrži efavirenz, darunavir ili raltegravir, kao i utvrditi pozitivne efekte primene matičnjaka u regulaciji metaboličkog sindroma. Izvršena je preliminarna i detaljna hemijska analiza dijetetskog suplementa na bazi ekstrakta lista matičnjaka. Ispitano je antihiperglikemično delovanje ekstrakta u in vitro testovima inhibicije enzima α-amilaze i α-glukozidaze. Pored toga, suplement na bazi ekstrakta matičnjaka i čista rozmarinska kiselina su u in vitro uslovima podvrgnuti fermentaciji pomoću probiotskih mikroorganizama. Ispitivanje na pacijentima je uključilo pacijente Klinike za infektivne bolesti Kliničkog centra Vojvodine koji su inficirani HIV-om, sa prisutnim metaboličkim sindromom. Pacijenti su na osnovu leka koji primaju u terapiji podeljeni na 3 grupe, a pored propisane antiretrovirusne terapije primali su i matičnjak u dozi od 600 m tokom 12 nedelja. Pacijentima su na početku i na kraju perioda ispitivanja mereni sledeći parametri: koncentracija rozmarinske kiseline u krvi, koncentracija antiretrovirusnog leka (efavirenz, darunavir ili raltegravir) u krvi, broj kopija virusa HIV-a u krvi, hematološki, imunološki i biohemijski parametri, pokazatelji oksidativnog stresa, aktivnost enzima antioksidativne zaštite, kao i antropometrijski parametri. Ispitani suplement na bazi ekstrakta okarakterisan je kao dobar izvor fenolnih jedinjenja, pri čemu je najzastupljenija komponenta bila rozmarinska kiselina. Utvrđena je in vitro inhibicija oba enzima u relativno niskim koncentracijama, uz bolju aktivnost u inhibiciji α-glukozidaze. Putem in vitro fermentacije pokazano je da suplement na bazi ekstrakta podleže intenzivnom metabolizmu od strane probiotskih sojeva; rozmarinska kiselina u ekstraktu je stabilnija u poređenju sa izolovanom formom. Pokazano je i da ovo jedinjenje predstavlja marker upotrebe matičnjaka u plazmi, pri čemu se rozmarinska kiselina u krvi nakon 12 nedelja upotrebe dominantno nalazi u obliku glukuronida i sulfata. Nisu pronađene statistički značajne razlike između koncentracija ispitivanih lekova određenih pre i posle 12 nedelja primene matičnjaka. Nije zabeleženo povećanje broja kopija virusa u krvi. Hematološki i imunološki parametri koji su praćeni nisu pokazali statistički značajnu promenu vrednosti. U okviru biohemijskih parametara nije došlo do statistički značajnih promena koje se mogu povezati sa upotrebom matičnjaka, sa izuzetkom ispitivane grupe efavirenza, gde je došlo do statistički značajnog povećanja alanin aminotransferaze i aspartat aminotransferaze, ali je povećanje navedenih enzima bilo u okviru fizioloških vrednosti. Pokazano je da nakon 12 nedelja primene matičnjaka dolazi do statistički značajne redukcije parametara oksidativnog stresa u ispitivanim grupama pacijenata koji su primali efavirenz i darunavir, a u ispitivanoj grupi sa efavirenzom potvrđeno je i statistički značajno povećanje aktivnosti glutation-S-transferaze. U okviru antropometrijski parametara uočeno je povećanje telesne mase nakon 12 nedelja primene matičnjaka u ispitivanoj grupi sa efavirenzom. Ovde predstavljeno istraživanje pokazalo je da istovremena primena matičnjaka uz antiretrovirusnu terapiju koja uključuje jedan od ispitivanih lekova ne utiče na koncentracije ovih lekova. Praćeni parametri koji su ispitivani kod pacijenata pokazali su da je primena matičnjaka u dozi od 600 mg tokom 12 nedelja bezbedna za pacijente. Pokazani pozitivni efekti ukazuju na povoljno dejstvo kod pacijenata inficiranih HIV-om sa metaboličkim sindromom. Literature contains limited data on the use of lemon balm in patients infected with the human immunodeficiency virus (HIV). However, data from clinical studies indicate the benefits of this plant’s use in carbohydrate and lipid metabolism disorders. The aim of this study was to examine the effect of lemon balm when used alongside antiretroviral therapy containing efavirenz, darunavir or raltegravir, as well as to determine the positive effects of the use of lemon balm in metabolic syndrome regulation. Both a preliminary and a detailed chemical analysis of lemon balm leaf extract were performed. The antihyperglycemic activity of the extract was investigated in in vitro α-amylase and α-glucosidase enzyme inhibition assays. Fermentation of the lemon balm extract, as well as of pure rosmarinic acid with probiotic microorganisms, was also performed in vitro. The investigation performed on patients included patients of the Clinic for Infectious Diseases of the Clinical Center of Vojvodina infected with HIV and with a co-occurring metabolic syndrome. The patients were divided into 3 groups based on the medication they received in antiretroviral therapy, and in addition to the prescribed therapy, they received lemon balm in a dose of 600 mg for 12 weeks. At the beginning and at the end of the study period, the following parameters were measured in patients: the concentration of rosmarinic acid in the patients’ blood, antiretroviral drug concentration (efavirenz, darunavir or raltegravir) in the patients’ blood, the number of copies of HIV in the patients’ blood, hematological, immunological and biochemical parameters, oxidative stress, the activity of enzymes of antioxidant protection, and anthropometric parameters. The tested extract was characterized as a good source of phenolic compounds, with rosemary acid being the most abundant compound. The extract showed in vitro inhibition of both enzymes at relatively low concentrations, with better activity in α-glucosidase inhibition. In in vitro fermentation, the extract has been shown to undergo intensive metabolism by probiotic strains; rosmarinic acid in the extract was more stable compared to the pure chemical compound. Blood analysis has shown that rosmarinic acid is a marker of the use of lemon balm in plasma, however, this compound is found in the form of a sulfoglucuronide in the blood after 12 weeks of use. No statistically significant differences were found between the concentrations of the tested drugs before and after 12 weeks of investigation. There was no increase in the number of copies of the virus in the blood. The monitored hematological and immunological parameters did not show a statistically significant change in values. Within the monitored biochemical parameters, there were no statistically significant changes that can be associated with the use of lemon balm, with the exception of the studied group of efavirenz, where there was a statistically significant increase in alanine aminotransferases and aspartate aminotransferases, but within reference values. It was shown that after 12 weeks of application of lemon balm, there was a statistically significant reduction in oxidative stress parameters in the studied groups of efavirenz and darunavir; in addition, a statistically significant increase in glutathione-S-transferase activity was shown in the efavirenz study group. Regarding the parameters of anthropometry, an increase in body weight was shown in the examined group of efavirenz after 12 weeks of application of lemon balm. The research presented here shows that the simultaneous use of lemon balm with antiretroviral therapy that includes one of the tested drugs does not affect the concentrations of these drugs. The monitored parameters in the patients showed that the use of lemon balm for 12 weeks in a dose of 600 mg was safe for patients. The presented positive results indicate a beneficial effect on HIV-infected patients with metabolic syndrome.

Primary Sjögren's syndrome (pSS) is a disease associated with the overexpression of proinflammatory cytokines, and oxidative stress is one of the factors responsible for its etiopathogenesis. This study aimed to investigate the thiol/disulphide homeostasis in pSS patients.The study included 68 pSS patients and 69 healthy controls. Thiol/disulphide homeostasis (total thiol, native thiol, and disulphide levels) was measured using the automatic spectrophotometric method developed by Erel and Neselioglu, and the results of the 2 groups were compared.The gender and age distributions of the pSS and control groups were similar (It was concluded that the thiol/disulphide balance shifted towards disulphide in patients with pSS.Primarni Sjögrenov sindrom (pSS) je bolest povezana sa prekomernom ekspresijom proinflamatornih citokina, dok je oksidativni stres jedan od faktora odgovornih za njegovu etiopatogenezu. Cilj ove studije je bio da ispita tioldisulfidnu homeostazu kod pacijenata sa pSS-om.Studija je obuhvatila 68 pacijenata sa pSS-om i 69 zdravih kontrolnih pojedinaca. Homeostaza tiol-disulfida (nivoi ukupnog tiola, nativnog tiola i disulfida) je izmerena pomoću automatske spektrofotometrijske metode koju su razvili Erel i Neselioglu. Rezultati dve grupe su upoređeni.Raspodela po polu i starosti pacijenata sa pSSom i pojedinaca u kontrolnoj grupi je bila slična (Zaključeno je da se ravnoteža tiol/disulfid pomera prema disulfidu kod pacijenata sa pSS-om.

The aim of the study was to investigate the association of paraoxonase 1 (PON1) polymorphism, PON1/arylesterase (ARE) activity and oxidative stress index (OSI) in breast cancer (BC) patients with type 2 diabetes (DM).Our study group consisted of 30 healthy women (HV group) and 66 female BC patients. The BC patients were divided into two groups: those with (n=37) and without DM (n=29) (BDM and NBDM group). Genotyping of PON1 Q192R and L55M polymorphisms were done by polymerase chain reaction (PCR) - restriction fragment length polymorphism (RFLP) method. Serum PON1/ARE enzyme activities, total oxidant status (TOS) and total antioxidant status (TAS) were analysed by spectrophotometric method. The ratio of TOS to TAS was accepted as the oxidative stress index (OSI).PON1 Q192R genotype frequency distribution was significantly different in the BDM group compared to the NBDM group (p=0.021). When alleles distribution was examined, R and L alleles were significantly lower, Q and M alleles were significantly higher in the BDM group than in the NBDM group (p0.001). TOS and OSI were statistically higher in BC patients than HV group (p0.001).Our results suggest that PON1 gene Q and M alleles may be the risk factors predisposing formation of BC due to increased oxidant damage seen in DM. However, these statements require further confirmation with screening PON1 polymorphism in a greater number of patients with DM, and also wide range follow-up studies are necessary for the same purpose.Cilj studije bio je istraživanje veze polimorfizma paraoksonaze 1 (PON1), aktivnosti PON1/arilesteraze (ARE) i indeksa oksidativnog stresa (OSI) kod pacijentkinja sa rakom dojke (BC) i dijabetesom tip 2 (DM).Naša studijska grupa se sastojala od 30 zdravih žena (HV grupa) i 66 žena sa rakom dojke (BC). Pacijentkinje iz BC grupe su podeljene u dve grupe: one sa (n = 37) i bez DM (n = 29) (BDM i NBDM grupa). Genotipizacija PON1 K192R i L55M polimorfizama vršena je polimeraznom lančanom reakcijom (PCR) – metodom polimorfizma dužine restrikcionih fragmenta (RFLP). Enzimske aktivnosti serumske PON1/ARE, ukupni oksidativni status (TOS) i ukupni antioksidativni status (TAS) analizirani su spektrofotometrijskom metodom. Odnos TOS-a prema TAS-u je prihvaćen kao indeks oksidativnog stresa (OSI).Frekvencija distribucije genotipova PON1 K192R bila je značajno drugačija u BDM grupi u odnosu na NBDM grupu (p = 0,021). Kada je ispitana alelna distribucija, R i L aleli su bili značajno niži, K i M aleli su bili značajno viši u BDM grupi nego u NBDM grupi (p0,001). TOS i OSI su bili statistički viši kod pacijentkinja sa rakom dojke odnosu na HV grupu (p0,001).Naši rezultati ukazuju na to da PON1 gen Q i M aleli mogu biti faktori rizika koji predisponiraju razvoj raka dojke zbog povećanog oksidativnog oštećenja kod DM. Ipak, ovakva tvrdnja zahteva dalju potvrdu skriningom PON1 polimorfizma kod većeg broja pacijentkinja sa DM, a u istu svrhu su potrebne i dodatne studije širokog spektra.

The accumulation of oxidative damage to DNA and other biomolecules plays an important role in the etiology of aging and age-related diseases such as type 2 diabetes mellitus (T2D), atherosclerosis, and neurodegenerative disorders. Mitochondrial DNA (mtDNA) is especially sensitive to oxidative stress. Mitochondrial dysfunction resulting from the accumulation of mtDNA damage impairs normal cellular function and leads to a bioenergetic crisis that accelerates aging and associated diseases. Age-related mitochondrial dysfunction decreases ATP production, which directly affects insulin secretion by pancreatic beta cells and triggers the gradual development of the chronic metabolic dysfunction that characterizes T2D. At the same time, decreased glucose oxidation in skeletal muscle due to mitochondrial damage leads to prolonged postprandial blood glucose rise, which further worsens glucose homeostasis. ROS are not only highly reactive by-products of mitochondrial respiration capable of oxidizing DNA, proteins, and lipids but can also function as signaling and effector molecules in cell membranes mediating signal transduction and inflammation. Mitochondrial uncoupling proteins (UCPs) located in the inner mitochondrial membrane of various tissues can be activated by ROS to protect cells from mitochondrial damage. Mitochondrial UCPs facilitate the reflux of protons from the mitochondrial intermembrane space into the matrix, thereby dissipating the proton gradient required for oxidative phosphorylation. There are five known isoforms (UCP1-UCP5) of mitochondrial UCPs. UCP1 can indirectly reduce ROS formation by increasing glutathione levels, thermogenesis, and energy expenditure. In contrast, UCP2 and UCP3 regulate fatty acid metabolism and insulin secretion by beta cells and modulate insulin sensitivity. Understanding the functions of UCPs may play a critical role in developing pharmacological strategies to combat T2D. This review summarizes the current knowledge on the protective role of various UCP homologs against age-related oxidative stress in T2D.

Compositions of stainless steel, nickel-titanium, cobalt-chromium and &beta, titanium orthodontic alloys were simulated with mixtures of Fe, Ni, Cr, Co, Ti and Mo metal ions as potential oxidative stress-triggering agents. Wild-type yeast Saccharomyces cerevisiae and two mutants &Delta, Sod1 and &Delta, Ctt1 were used as model organisms to assess the cytotoxicity and oxidative stress occurrence. Metal mixtures at concentrations of 1, 10, 100 and 1000 &micro, M were prepared out of metal chlorides and used to treat yeast cells for 24 h. Every simulated orthodontic alloy at 1000 &micro, M was cytotoxic, and, in the case of cobalt-chromium alloy, even 100 &micro, M was cytotoxic. Reactive oxygen species and oxidative damage were detected for stainless steel and both cobalt-chromium alloys at 1000 &micro, M in wild-type yeast and 100 &micro, M in the &Delta, Ctt1 mutants. Simulated nickel-titanium and &beta, titanium alloy did not induce oxidative stress in any of the tested strains.

Polycystic ovary syndrome (PCOS) is a complex endocrine and metabolic disorder of multifactorial etiopathology likely to involve the interactions between genetics and lifestyle. Chronic inflammation and oxidative stress (OS) may participate in the pathophysiology of the syndrome. The question of the extent to which OS and inflammation are causally related to the development of the syndrome and metabolic complications remains unanswered. By our knowledge, the role of the NLR family pyrin domain containing 3 (NLRP3) inflammasome as an important trigger of inflammatory pathways and NLRP3 and CARD8 polymorphisms has never been addressed in PCOS yet. We conducted a case-control study conducting of total 169 Slovenian PCOS patients and 83 healthy blood donors. They were genotyped for polymorphisms in antioxidative (SOD2 rs4880, CAT rs1001179, PON1 rs854560, and rs662) and inflammatory pathways genes (NLRP3 rs35829419, CARD8 rs2043211, TNF rs1800629, IL1B rs1143623, and rs16944, IL6 rs1800795) using competitive allele-specific polymerase chain reaction (PCR). Logistic regression and the Mann&ndash, Whitney test were used in the statistical analysis. SOD2 rs4880, CARD8 rs2043211, and IL1B rs16944 were associated with the risk of developing PCOS. Furthermore, the interactions between CARD8 rs2043211 and IL6 rs1800795 and between IL1B rs1143623 and IL6 rs1800795 also significantly affected the risk for PCOS. With regard to glucose homeostasis, CAT rs1001179, SOD2 rs4880, PON1 rs854560, NLRP3 rs35829419, and TNF rs1800629 were significantly associated with response to the glycemic load. Our data indicate that the genetic variability in the antioxidative and inflammatory pathways influences the development of PCOS and glucose homeostasis in PCOS patients.

Neurodegenerative diseases, namely Alzheimer’s disease (AD), Parkinson’s disease (PD), amyotrophic lateral sclerosis, Huntington’s disease, and multiple sclerosis are becoming one of the main health concerns due to the increasing aging of the world’s population. These diseases often share the same biological mechanisms, including neuroinflammation, oxidative stress, and/or protein fibrillation. Recently, there have been many studies published pointing out the possibilities to reduce and postpone the clinical manifestation of these deadly diseases through lifelong consumption of some crucial dietary substances, among which phytochemicals (e.g., polyphenols) and endogenous substances (e.g., acetyl-L-carnitine, coenzyme Q10, n-3 poysaturated fatty acids) showed the most promising results. Another important issue that has been pointed out recently is the availability of these substances to the central nervous system, where they have to be present in high enough concentrations in order to exhibit their neuroprotective properties. As so, such the aim of this review is to summarize the recent findings regarding neuroprotective substances, their mechanisms of action, as well as to point out therapeutic considerations, including their bioavailability and safety for humans.

Background: The environment in swimming pools, which contain chlorine, might interact with the airway epithelium, resulting in oxidative stress and/or inflammation during high intensity training periods. Methods: We evaluated pulmonary functional (metacholine challenge test, FEV1 and VC), cellular (eosinophils and neutrophils), inflammatory (FeNo, IL-5, IL-6, IL-8 and TNF-[alpha]), oxidative (8-isoprostanes) and angiogenesis factors (VEGF) in induced sputum and peripheral blood of 41 healthy non-asthmatic elite swimmers (median 16 years) during the period of high intensity training before a national championship. The second paired sampling was performed seven months later after training had been stopped for one month. Results: There was a ten-fold increase (median 82-924 pg/ml P < 0.001) in 8-isoprostanes in induced sputum and five-fold increase (median 82-924 pg/ml P < 0.001) in sera during training in comparison to the period of rest. However, there was no difference in FEV1 (113 vs 116%), VC (119 vs 118%), FeNo (median 34 vs 38 ppb), eosinophils (2.7 vs 2.9% in sputum 180 vs 165 cells/[micro]l in blood), neutrophils, different cytokines or VEGF in induced sputum or sera. The only exception was TNF-[alpha], which was moderately increased in sera (median 23 vs 40 pg/ml P=0.02) during the peak training period. Almost half (18 of 41) of swimmers showed bronchial hyperresponsiveness during the peak training period (PC20 cutoff was 4 mg/ml). There was no correlation between hyperresponsiveness and the markers of oxidative stress or inflammation. Conclusions: High intensity training in healthy, non-asthmatic competitive swimmers results in marked oxidative stress at the airway and systemic levels, but does not lead to airway inflammation. However, we could not confirm that oxidative stress is associated with bronchial hyperresponsiveness (AHR), which is often observed during the peak exercise training period.

Due to climate change, plants are being more adversely affected by heatwaves, floods, droughts, and increased temperatures and UV radiation. This review focuses on enhanced UV-B radiation and drought, and mitigation of their adverse effects through silicon addition. Studies on UV-B stress and addition of silicon or silicon nanoparticles have been reported for crop plants including rice, wheat, and soybean. These have shown that addition of silicon to plants under UV-B radiation stress increases the contents of chlorophyll, soluble sugars, anthocyanins, flavonoids, and UV-absorbing and antioxidant compounds. Silicon also affects photosynthesis rate, proline content, metal toxicity, and lipid peroxidation. Drought is a stress factor that affects normal plant growth and development. It has been frequently reported that silicon can reduce stress caused by different abiotic factors, including drought. For example, under drought stress, silicon increases ascorbate peroxidase activity, total soluble sugars content, relative water content, and photosynthetic rate. Silicon also decreases peroxidase, catalase, and superoxide dismutase activities, and malondialdehyde content. The effects of silicon on drought and concurrently UV-B stressed plants has not yet been studied in detail, but initial studies show some stress mitigation by silicon.

Štavelj (Rumex crispus, Polygonaceae) je višegodišnja zeljasta biljka, koja predstavlja bogat izvor fenolnih komponenti. Iako se smatra invazivnim korovom, mlado lišće štavelja je jestivo i često se koristi kao salata. Dalje, upotreba plodova štavelja opisana je u srpskoj i turskoj narodnoj medicini u lečenju gastrointestinalnih tegoba. Cilj ovog rada bio je procena in vitro i in vivo antioksidantne/prooksidantne i citotoksične aktivnosti, i određivanje eventualnog in vitro antiinflamatornog efekta vodenog ekstrakta ploda Rumex crispus. Ukupan sadržaj flavonoida određen je spektrofotometrijskom metodom. Kvalifikacija i kvantifikacija flavonoida potvrđena je visokoefikasnom tečnom hromatografijom (HPLC). Antioksidantna aktivnost vodenog ekstrakta ploda štavelja procenjena je na osnovu in vitro testova: Ferric-reducing antioxidant power (FRAP), sposobnosti ekstrakta da neutrališe slobodne radikale NO•, OH• i DPPH• i uticaja na lipidnu peroksidaciju u lipozomima. Citotoksičnost ispitivanog ekstrakta je određena in vitro na tumorskim ćelijskim linijama: humani karcinom cerviksa (HeLa), adenokarcinom (HT-29) i adenokarcinom dojke (MCF7). Takođe, moguća in vivo hepatoprotektivna i antioksidantna svojstva ekstrakta određena su kod oksidativnog stresa izazvanog CCl4 kod eksperimentalnih životinja. Pored toga, proverena je hipoteza u kojoj testiran ekstrakt pokazuje in vivo antiproliferativnu aktivnost kod Ehrlich-ovih (EAC) i Hepatoma AS30D ćelija, merenjem zapremine ascitesa, procenta vijabilnih ćelija i nivoa nekoliko antioksidantnih enzima. Optimizovan in vitro test za određivanje potencijala inhibicije ciklooksigenaze-1 (COX-1) i 12-lipooksigenaze (12-LOX) preduzet je u svrhu procene antiinflamatornog efekta vodenog ekstrakta ploda R. crispus. HPLC analiza otkrila je da je mikvelianin najdominantniji flavonoidni konstituent ekstrakta. Testirani ekstrakt pokazao je potencijalnu antioksidantnu aktivnost rezultujući velikom moći u neutralizaciji slobodnih radikala, i sposobnošću da smanji lipidnu peroksidaciju u lipozomima. Rezultati su ukazali na tkivno-selektivnu citotoksičnost ekstrakta ploda R. crispus in vitro. Najizraženija antitumorska aktivnost primećena je prema HeLa i MCF7 ćelijskim linijama. Podaci sugerišu da bi se ispitivani ekstrakt mogao smatrati potencijalnim in vivo hepatoprotektivnim i antioksidantnim agensom, sprečavajući oksidativna oštećenja jetre. S druge strane, pomenuti ekstrakt može pokazati in vivo prooksidantna svojstva, uzrokujući oksidativni stres u maligno transformisanim EAC i AS30D ćelijama i smanjujući zapreminu ascitesa i udeo vijabilnih ćelija, u poređenju sa kontrolnom grupom. Promene u aktivnosti antioksidantnih enzima su verovatno posledica indukovanog oksidativnog stresa u EAC i AS30D ćelijama, naročito kod pretretiranih životinja. Vodeni ekstrakt ploda štavelja pokazao je COX-1, kao i 12-LOX inhibitornu aktivnost, navodeći da bi ispitivani ekstrakt mogao biti antiinflamatorni agens. Vodeni ekstrakt ploda R. crispus ima potencijalnu antioksidantnu, citotoksičnu i antiinflamatornu aktivnost. Ispoljavanje prooksidantnih svojstava predstavlja mogući mehanizam antiproliferativnog efekta ekstrakta., Curly dock (Rumex crispus, Polygonaceae) is a wild perennial herbaceous plant, which products are described as a rich source of phenolic compounds. Apart from being considered a seriously invasive weed, young leaves of curly dock are edible and often used as salad. Furthermore, the use of its fruits has been described in Serbian and Turkish traditional medicine against stomach complaints. The objectives of this study were to evaluate in vitro and in vivo antioxidant/prooxidant and cytotoxic activities, and to determine an eventual in vitro anti-inflammatory effect of the aqueous extract of Rumex crispus fruits. Total flavonoid content was determined by spectrophotometric method. Qualification and quantification of flavonoids were confirmed using High performance liquid chromatography (HPLC). The aqueous extract of curly dock fruits was evaluated for its antioxidant activity by in vitro assays for Ferric-reducing antioxidant power (FRAP), NO•, OH• and DPPH•-free radical scavenging activities and the influence on lipid peroxidation in liposomes. The cytotoxicity of tested extract was examined in vitro in human cervix carcinoma (HeLa), colon adenocarcinoma (HT-29) and breast adenocarcinoma (MCF7). Also, the potential in vivo hepatoprotective and antioxidant properties of investigated extract were determined on CCl4-induced oxidative stress in experimental animals. Furthermore, the hypothesis that the examined extract might show in vivo antiproliferative activity in Ehrlich carcinoma (EAC) and Hepatoma AS30D cells was tested by measuring volume of ascites, percentage of viable cells and level of several antioxidant enzymes. The optimized in vitro test for determination of cyclooxygenase-1 (COX-1) and 12-lipoxygenase (12-LOX) inhibition potency was undertaken in order to estimate an anti-inflammatory effect of aqueous extract of R. crispus fruits. HPLC analysis revealed miquelianin as the most abundant flavonoid constituent of the extract. The tested extract might have an antioxidant activity resulting in scavenging of free radicals and ability to decrease lipid peroxidation in liposomes. The results could indicate tissue-selective cytotoxicity of R. crispus fruit extract in vitro. The most prominent antitumor activity was observed towards HeLa and MCF7 cell lines. The data suggested that investigated extract may be considered as potential in vivo hepatoprotective and antioxidant agent due to prevention of the liver injuries induced by oxidative damage. On the other hand, mentioned extract could exhibit in vivo prooxidant property, causing the oxidative stress in malignant transformed EAC and AS30D cells and reducing volume of ascites and percentage of viable cells, in comparison with control group. Changes in activities of antioxidant enzymes might be the results of induced oxidative stress in EAC and AS30D cells, especially in the pretreated animals. The aqueous extract of curly dock fruits showed COX-1, as well as 12-LOX inhibitory activity, suggesting that tested extract might be an anti-inflammatory agent. It could be concluded that aqueous fruit extract of R. crispus might have antioxidant, cytotoxic and anti-inflammatory activities. The prooxidant properties of examined extract could be the mechanism of potential antiproliferative effect of extract.

U organizmu se, pod fiziološkim uslovima, produkuju slobodni radikali. Iako se u organizmu nalaze u veoma niskoj koncentraciji, slobodni radikali mogu ispoljiti toksične efekte. Težeći da spare elektrone, u hemijskoj reakciji oksidacije, dolazi do brzog i nepredvidivog vezivanja za susedne molekule, proteine, lipide, ugljene hidrate i nukleinske kiseline od kojih su sačinjeni strukturni elementi ćelije, pokrećući unutrašnji put apoptoze. Antioksidansi su supstance koje sprečavaju ili značajno smanjuju oksidaciju biomolekula. Oksidativni stres je stanje koje nastaje kada produkcija slobodnih radikala premaši kapacitete antioksidativnih enzima da ih neutrališu. U antioksidativne enzime spadaju: superoksid dismutaza (SOD), katalaza (CAT), glutation peroksidaza (GPx), glutation reduktaza (GR) i glutation-S-transferaza (GST). Lipidna peroksidacija (LP) je proces oksidacije višestruko nezasićenih masnih kiselina od strane slobodnih radikala. Malondialdehid predstavlja biohemijski marker pomoću kog je moguće meriti stepen oksidativnog oštećenja ćelijskih membrana. Oksidativna modifikacija DNK dovodi do promene strukture DNK koje rezultuju genetskim oštećenjima. Najčešće korišćen marker oksidativnog stresa je urinarni 8-hidroksi-2-deoksiguanozin (8-OHdG). Oštećenja proteina, lipida, DNK čine važan osnov mnogih oboljenja kao što su ateroskleroza, neurodegenerativna oboljenja, dijabetes, gojaznost, proces starenja, retinopatija, hronične inflamatorne bolesti i karcinom. Polazeći od hipoteze da su ovi biomolekuli različiti u različitim stadijumima bolesti, oni bi mogli predstavljati prognostički marker proširenosti bolesti. Cilj istraživanja je bio da se ispita da li postoje razlike između kontrolne grupe (zdravih žena), pacijentkinja sa prekanceroznim lezijama na grliću materice (HSIL), pacijentkinja sa lokalno ograničenim (FIGO Ia-Ib) i pacijentkinja sa lokalno uznapredovalim karcinomom grlića materice (IIa-IV) u pokazateljima oštećenja DNK (određivanjem vrednosti 8-OHdG), pokazateljima oksidativnog stresa (određivanjem intenziteta lipidne peroksidacije (TBARS)), pokazateljima antioksidativne odbrane (određivanjem aktivnosti antioksidativnih enzima superoksid dismutaze (SOD), katalaze (CAT), glutation peroksidaze (GPx), glutation reduktaze (GR) i glutation-S-transferaze (GST)). Pored toga, cilj istraživanja je bio da se uporede vrednosti 8-OHdG, proizvoda lipidne peroksidacije (TBARS) i aktivnosti antioksidativnih enzima (SOD, CAT, GST, GPx, GR) unutar grupe pacijentkinja sa lokalno ograničenim karcinomom grlića materice podeljenih u dve podgrupe sa niskim i visokim rizikom u odnosu na relaps bolesti. Takođe, u radu je koreliran nivo 8-OHdG, MDA i antioksidativnih enzima sa relapsom bolesti. Istraživanje je izvedeno na Klinici za operativnu onkologiju, odeljenje za ginekologiju na Institutu za onkologiju Vojvodine, Zavodu za farmaciju Medicinskog fakulteta u Novom Sadu i Zavodu za zdravstvenu zaštitu radnika Novi Sad u periodu od 2013. godine do 2017. godine. Od ispitanica su prikupljani uzorci krvi i urina, pripremljeni na adekvatan način i čuvani na -80° do analiza. Aktivnost enzima antioksidativne zaštite, kao i intenzitet lipidne peroksidacije određivani su spektrofotometrijskim metodama, a koncentracija 8-OhdG određivana je gasnom hromatografijom uz masenu detekciju. Za sprovođenje istraživanja dobijena je saglasnost Etičkog odbora Instituta za onkologiju Vojvodine. Pokazano je da postoje statistički značajne razlike između kontrolne grupe (zdravih žena), pacijentkinja saprekanceroznim lezijama na grliću materice (HSIL), pacijentkinja sa lokalno ograničenim (FIGO Ia-Ib) u odnosu napacijentkinje sa lokalno uznapredovalim karcinomom grlića materice (IIa-IV) u pokazateljima oštećenja DNK (koncentracija 8-OHdG), pokazateljima oksidativnog stresa (intenziteta lipidne peroksidacije (TBARS)), pokazateljima antioksidativne odbrane (aktivnosti antioksidativnih enzima SOD, CAT i GST). Nisu pokazane razlike između ispitivanih grupa u aktivnosti enzima glutation peroksidaze (GPx) i glutation reduktaze (GR). Nisu pronađene razlike u koncentraciji 8-OHdG, proizvoda lipidne peroksidacije (TBARS) i aktivnosti antioksidativnih enzima (SOD, CAT, GST, GPx i GR) unutar grupe pacijentkinja sa lokalno ograničenim karcinomom grlića materice podeljenih u dve podgrupe sa niskim i visokim rizikom u odnosu na relaps bolesti. Aktivnosti CAT i GST bile najbolji prediktori rekurencije bolesti kod definisanih pacijentkinja. Na osnovu aktivnosti ova dva oksidativna enzima, separacija grupe pacijentkinja kod kojih nije došlo do rekurencije bolesti nakon perioda praćenja od ostale dve grupe kod kojih je došlo do rekurencije bolesti je bila moguća. Na osnovu dobijenih rezultata zaključuje se da je moguće koristiti navedene biomarkere kao dijagnostičke markere kod pacijentkinja sa karcinomom grlića materice. Ovi biomolekuli mogu pomoći lakšem svrstavanju pacijentkinja u određene grupe prema stadijumu bolesti, a sledstveno i bržem odabiru odgovarajućeg lečenja. Pored toga, pokazano je da su aktivnosti enzima CAT i GST prediktori rekurencije bolesti kod definisanih grupa pacijentkinja. Free radicals are produced in our body under physiological conditions. Although in very low concentrations, they can show some toxic effects. While trying to bind electrons, in the chemical reaction of oxidation, they rapidly and unpredictably bind to adjacent molecules- proteins, lipids, carbohydrates and nucleic acids from which the structural elements of the cell are made, triggering the internal pathway of apoptosis. Antioxidants are substances that prevent or significantly reduce the oxidation of biomolecules. Oxidative stress is a condition that occurs when the production of free radicals exceeds the capacity of antioxidant enzymes to neutralize them. The antioxidant enzymes include: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione-S-transferase (GST). Lipid Peroxidation (LP) is the process of oxidation of polyunsaturated fatty acids by free radicals. Malondialdehyde is a biochemical marker by which it is possible to measure the degree of oxidative damage of cell membranes. The oxidative modification of DNA leads to a change in DNA structure that results in genetic damage. The most commonly used marker of oxidative stress is urinary 8-hydroxy-2-deoxiguanosine (8-OHdG). The damage to proteins, lipids and DNA is an important basis for many diseases such as atherosclerosis, neurodegenerative diseases, diabetes, obesity, aging, retinopathy, chronic inflammatory disease and cancer. Starting from the hypothesis that these biomolecules are different at different stages of the disease, they could represent a prognostic marker of the progression of the disease. The aim of the study was to examine whether there were differences between the control group (healthy women), the patients with precancerous lesions on the cervix (HSIL), the patients with early stage cervical cancer (FIGO Ia-Ib) and the patient with locally advanced cervical cancer (IIa - IV) in the indicators of DNA damage (determining the value of 8-OHdG), indicators of oxidative stress (by determining the lipid peroxidation intensity (TBARS)), indicators of antioxidant defense (by determining the activity of antioxidative enzymes of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase GPx), glutathione reductase (GR), and glutathione-S-transferase (GST)). In addition, the aim of the study was to compare the values of 8- OHdG, lipid peroxidation products (TBARS) and the activity of antioxidant enzymes (SOD, CAT, GST, GPx, GR) within the group of patients with early stage cervical cancer divided into two subgroups- with low and high risk in relation to the relapse of the disease. The research was performed at the Clinic for operative oncology, Department of Gynecology at the Institute of Oncology of Vojvodina, Medical Faculty in Novi Sad, Department of Pharmacy and the Institute for Health Care of Novi Sad in the period from 2013 to 2017. Samples of blood and urine of the patients were collected, prepared adequately and stored at -80 ° until the analysis. The activity of the antioxidant enzymes as well as the lipid peroxidation were determined by spectrophotometric methods, and the concentration of 8-OHdG was determined by gas chromatography with mass detection. The approval of the Ethical Committee of the Institute for Oncology of Vojvodina was obtained before conducting the research. It has been shown that there are statistically significant differences between the control group (healthy women), patient with precancerous cervical lesions (HSIL), the patients with early stage cervical cancer (FIGO Ia-Ib) compared to a group of patients with locally advanced cervical cancer (IIa-IV) in indicators of damage to DNA (concentration of 8-OHdG), indicators of oxidative stress (lipid peroxidation (TBARS)), indicators of antioxidant defense (activities of antioxidant enzymes SOD, CAT and GST). There was no difference between the groups in activity of glutathione peroxidase enzyme (GPx) and glutathione reductase (GR). There were no differences in the concentration of 8-OHdG, lipid peroxidation products (TBARS) and the activity of antioxidant enzymes (SOD, CAT, GST, GPx and GR) within the group of patients with locally restricted cervical cancer divided into two subgroups with low and high risk in relation on relapses of the disease. CAT and GST activities were the best predictors of disease recurrence among defined groups. Based on the activities of these two oxidative enzymes, the separation of the group of patients who did not experience disease recurrence after a follow-up period from the other two groups in which recurrence of the disease occurred was possible. Based on the obtained results it is concluded that it is possible to use the studied biomarkers as diagnostic markers in patients with cervical cancer. These biomolecules can help in the patient's classification into certain groups according to the stage of the disease, and consequently the more efficient choice of appropriate treatment. In addition, CAT and GST enzyme activity have been shown to be predictors of disease recurrence in defined patient groups.

Bor je bilo koji četinar roda Pinus koji pripada porodici Pinaceae. U Republici Srbiji nalaze se prirodne i veštačke šume crnog bora Pinus nigra Arnold. Kora bora se vekovima koristila u tradicionalnoj medicini, međutim tek u novije vreme su otkriveni i potvrđeni njeni pozitivni biološki efekti u medicini. Najpoznatiji komercijalni preparat kore bora je piknogenol, standardizovani ekstrakt kore francuskog primorskog bora Pinus pinaster Aiton, koji raste duţ obale jugozapadne Francuske. Pozitivni efekti koje ispoljava piknogenol, kao što su antioksidativno, hipoglikemijsko, hipolipidemijsko delovanje, zasnovani su na njegovom visokom sadržaju fenolnih jedinjenja. Danas, i pored saznanja da je kora bora bogata sekundarnim metabolitima ona najčešće završava kao nusproizvod drvne industrije. Ciljevi ovog rada bili su ispitivanje kvalitativnih i kvantitativnih karakteristika i biohemijskih aktivnosti ekstrakata kore crnog bora Pinus nigra sa Mokre gore i Tare, dobijenih različitim rastvaračima; upoređivanje glikemijskog i lipidnog delovanja ekstrakta kore bora Pinus nigra sa komercijalnim preparatom piknogenol; uticaj ekstrakta kore bora Pinus nigra i piknogenola na delovanje standardnih antidijabetika metformina i gliklazida; kao i uticaj na parametre oksidativnog stresa nakon izlaganja životinja toksičnoj dozi paracetamola. In vitro ispitivanja su uključivala analizu ekstrakata dobijenih različitim rastvaračima. Ukupni sadržaj fenola, tanina, flavonoida i proantocijanidna, kao i ispitivanje antioksidativne aktivnosti kroz pet različitih metoda, određivani su spektrofotometrijski. Vršena je analiza ekstrakata kore bora visokoefikasnom tečnom hromatografijom (HPLC). In vivo ispitivanje je rađeno na 156 albino laboratorijska pacova soja Wistar. Eksperimentalne ţivotinje su tokom 7 dana per os primale ekstrakt kore crnog bora poreklom sa Mokre gore 100 mg/kg, koji je pokazao najbolju aktivnost, piknogenol 50 mg/kg, metformin 100 mg/kg, gliklazid 10 mg/kg, kao i njihove kombinacije. Za ispitivanje uticaja primenjivanih supstanci na glikemiju ţivotinja korišćeni su test oralnog podnošenja glukoze i indukcija trajne hiperglikemije aloksanom. Od biohemijskih parametara u serumu je određivana koncentracija lipida, kao i parametric pokazatelji funkcije bubrega i jetre. Ex vivo ispitivanja uključivala su određivanje intenzitet lipidne peroksidacije i anktivnosti enzima antioksidativne zaštite u homogenatu jetre ispitivanih ţivotinja, nakon primene toksične doze paracetamola. HPLC analizom ekstrakata kore crnog bora utvrđeno je da ekstrakti poreklom sa Mokre gore i Tare u najvećoj meri sadrţe taksifolin, a zatim katehin, njegove derivate, kafenu kiselinu i epikatehin. PotvrĎen je visok antioksidativni kapacitet piknogenola, ali takođe i značajan antioksidativni kapacitet ekstrakata kore crnog bora, sa najboljim rezultatima kod ekstrakta poreklom sa Mokre gore. Sedmodnevni tretman ekstraktom kore bora 100 mg/kg, metforminom 100 mg/kg, gliklazidom 10 mg/kg i piknogenolom 50 mg/kg, kod ţivotinja sa aloksanski izazvanim dijabetesom, doveo je do sniženja glikemije u krvi. Uočeno je povećanje konentracije HDL holesterola i značajno sniženje serumskih triglicerida kod dijabetičnih i normoglikemičnih ţivotinja koje su primale piknogenol, ekstrakta kore bora, gliklazid i metformin u poređenju sa kontrolnom grupom ţivotinja. Zajednička primena ekstrakta kore bora i standardnih antidijabetika značajno je snizila vrednosti AST, ALP i direktni bilirubin u grupi dijabetičnih i normoglikemičnih ţivotinja u odnosu na kontrolnu grupu, čime se ukazuje na potencijalno hepatoprotektivno delovanje ekstrakta kore crnog bora. Oštećenje jetre izazvano toksičnom dozom paracetamola je potvrđeno ispitivanim parametrima, uključujući oksidativni status u homogenatu jetre i histološka ispitivanja. Rezultati našeg istraživanja su pokazali da piknogenol 50 mg/kg, kao i ekstrakt kore crnog bora 100 mg/kg značajno smanjuju nivo MDA u poređenju sa kontrolnom grupom i grupom koja je primala paracetamol. Na osnovu rezultata istraţivanja može se zaključiti da primena piknogenola i ekstrakta kore crnog bora: ublaţava poremećaj homeostaze glukoze, utiče povoljno na lipidni status, značajno potencira antihiperglikemijsko delovanje metformina i hipoglikemijski efekat gliklazida, sprečava poremećaj biohemijskih parametara pokazatelja funkcije jetre i bubrega u serumu ispitivanih životinja i ispoljava značajno in vitro antioksidativno delovanje i sprečava oštećenje jetre laboratorijskih životinja izazvano toksičnom dozom paracetamola., A pine is any conifer in the genus Pinus of the family Pinaceae. In the Republic of Serbia there are natural and artificial forests of black pine Pinus nigra Arnold. Pine bark has been used for centuries in traditional medicine, but only recently its positive biological effects has been revealed and confirmed in medicine. The most famous commercial pine bark product is Pycnogenol, a standardized extract of the French maritime pine bark (Pinus pinaster Aiton), which grows along the coast of southwestern France. The positive effects of Pycnogenol, such as antioxidant, hypoglycemic, hypolipidemic, are based on its high content of phenolic compounds. Nowdays, despite the knowledge that the pine bark is rich in secondary metabolites, it usually ends up as a by-product of the wood industry. The objectives of this dissertation were to investigate the qualitative and quantitative characteristics and biochemical activities of the black pine (Pinus nigra) extracts from Mokra gora and Tara, obtained by different solvents; comparison of glycemic and lipid action of Pinus nigra bark extract with commercial product Pycnogenol; the influence of black pine bark extract and Pycnogenol on the action of standard antidiabetics metformin and gliclazide; as well as the influence on the parameters of oxidative stress in animals exposed to the toxic dose of paracetamol. In vitro analyses included the analysis of extracts obtained with different solvents. The total phenolic, tannin, flavonoid and proanthocyanidin content, as well as the antioxidant activity tests, were determined spectrophotometrically. High-efficiency liquid chromatography (HPLC) was performed in order to determine black pine bark extract content. In vivo analyses was performed on 156 albino Wistar laboratory rats. Experimental animals received for 7 days per os: black pine bark extract obtained from Mokra gora 100 mg/kg, which showed the best activity; Pycnogenol 50 mg/kg; metformin 100 mg/kg; gliclazide 10 mg/kg; and their combinations. An oral glucose tolerance test and the aloxan-induced hyperglycaemia were used to examined the effects of the substances on experimental animals. Biochemical parameters, the lipids concentration and parameters of the kidney and liver function, were determined in animal serum. Ex vivo analyses included determination of lipid peroxidation activity and the activity of antioxidant protection enzymes in liver homogenates of the test animals, after administration of a toxic dose of paracetamol. HPLC analysis of black pine bark extracts obtained from Mokra gora and Tara showed the greatest content of taxifolin, then catechin, and its derivatives, caffeic acid and epicatechin. A high antioxidative capacity of Pycnogenol was confirmed, but also a significant antioxidant capacity of black pine bark extract, with the best results in the extract obtained from Mokra gora. Seven-day treatment in aloxan-induced diabetes animals with 100 mg/kg black pine bark extract, metformin 100 mg/kg, gliclazide 10 mg/kg and Pycnogenol 50 mg/kg led to a decrease of serum glycemic index. There was an increase in the HDL cholesterol concentration and a significant reduction in serum triglycerides in diabetic and normoglycemic animals which received Pycnogenol, black pine bark extract, gliclazide and metformin compared to the control group. Concomitant usage of the black pine bark extract and standard antidiabetics significantly reduced the levels of AST, ALP and direct bilirubin in the group of diabetic and normoglycemic animals compared to the control group, indicating the potential hepatoprotective action of the black pine bark extract. Damage to the liver caused by the toxic dose of paracetamol was confirmed by the liver homogenate oxidative status and histological examination. The results of our study have shown that Pycnogenol 50 mg/kg, as well as 100 mg/kg black pine bark extract significantly reduce the level of MDA in comparison with the control and the paracetamol treated group. Based on the results of the dissertation, it can be concluded that the use of Pycnogenol and black pine bark extract alleviate glucose homeostasis disorder; positively affects the lipid status; significantly increases the antihyperglycemic effect of metformin and the hypoglycemic effect of gliclazide; prevents impairment biochemical parameters of the liver and kidney function in the serum of the experimental animals; significantly exhibits in vitro antioxidant activity and prevents liver damage caused by toxic dose of paracetamol in experimental animals.

Током учесталог вежбања, нарочито професионалних спортиста, мишићи се привремено замарају, услед чега може доћи до пада функционалних способности. Замор мишића услед физичког вежбања постао је један од најважнијих предмета спортске науке. Циљ ове студије био је да се праћењем биохемијских параметара замора и оштећења мишића и оксидативног стреса у току једне појединачне тренажне епизоде утврди замор мишића и антиоксидативни статус кошаркашица након оптерећења изазваног са три различита врсте аеробног тренинга: континуирано ниско интензивни (НИК), континуирано високо интензивни (ВИК) и интервални аеробни тренинг (HIIT). У склопу укрштеног (crossover) дизајна изабран је узорак од 12 кошаркашица које су биле подвргнуте тестирању у предтакмичарској фази. Узорци крви узети су пре и одмах након тренажне епизоде у три наврата у размаку од по седам дана између сваког типа тренинга. Осим узимања узорака крви, проверан је пулс пре тренинга, одмах након тренинга, потом након првог, трећег и петог минута опоравка, затим, испитанице су радиле субјективну процену замора по Борговој скали одмах након тренинга, па затим у првом, трећем и петом минуту опоравка. Биохемијски параметри које смо анализирали били су маркери оштећења мишића, маркери целуларне биоенергетике, параметри оксидативно/антиоксидативног статуса. Резултати истраживања указују на то да маркери замора и оштећења мишића бележе варијабилност у резултатима с обзиром на врсту тренинга. Евидентиране су разлике у маркерима лактати, миоглобин и креатин киназа између НИК и HIIT. У мерењима срчане фреквенције у више временских тачака, статистички значајне разлике појавиле су се између ВИК и HIIT, што је у корелацији са добијеним налазима маркера оштећења мишића. Такође, субјективна процена замора прати добијене налазе и бележи значајно повећање после иста два тренинга одмах после, у првом и трећем минуту опоравка. Од маркера целуларне биоенергетике евидентиране су разлике у креатинину после НИК и HIIT тренинга. Индекс липидне пероксидације (TBARS), као један од параметара оксидативног стреса, бележи промене после НИК. Концентрација нитрита (NO2-) бележи такође статистички значајно повећање код НИК, али и код HIIT. Од испитиваних параметара антиоксидативне заштите једино каталаза (CAT) бележи значајно повећање код НИК. У процени стања замора мишића и деловања ензима оксидативно/антиоксидатине статуса, према добијеним резултатима, најсензитивнији маркери који су реаговали на различите тренажне епизоде, односно на различита аеробна оптерећења јесу миоглобин, лактати, креатин киназа и креатинин, а каталаза је показала могућност дискриминације различитих врста аеробног оптерећења. Tokom učestalog vežbanja, naročito profesionalnih sportista, mišići se privremeno zamaraju, usled čega može doći do pada funkcionalnih sposobnosti. Zamor mišića usled fizičkog vežbanja postao je jedan od najvažnijih predmeta sportske nauke. Cilj ove studije bio je da se praćenjem biohemijskih parametara zamora i oštećenja mišića i oksidativnog stresa u toku jedne pojedinačne trenažne epizode utvrdi zamor mišića i antioksidativni status košarkašica nakon opterećenja izazvanog sa tri različita vrste aerobnog treninga: kontinuirano nisko intenzivni (NIK), kontinuirano visoko intenzivni (VIK) i intervalni aerobni trening (HIIT). U sklopu ukrštenog (crossover) dizajna izabran je uzorak od 12 košarkašica koje su bile podvrgnute testiranju u predtakmičarskoj fazi. Uzorci krvi uzeti su pre i odmah nakon trenažne epizode u tri navrata u razmaku od po sedam dana između svakog tipa treninga. Osim uzimanja uzoraka krvi, proveran je puls pre treninga, odmah nakon treninga, potom nakon prvog, trećeg i petog minuta oporavka, zatim, ispitanice su radile subjektivnu procenu zamora po Borgovoj skali odmah nakon treninga, pa zatim u prvom, trećem i petom minutu oporavka. Biohemijski parametri koje smo analizirali bili su markeri oštećenja mišića, markeri celularne bioenergetike, parametri oksidativno/antioksidativnog statusa. Rezultati istraživanja ukazuju na to da markeri zamora i oštećenja mišića beleže varijabilnost u rezultatima s obzirom na vrstu treninga. Evidentirane su razlike u markerima laktati, mioglobin i kreatin kinaza između NIK i HIIT. U merenjima srčane frekvencije u više vremenskih tačaka, statistički značajne razlike pojavile su se između VIK i HIIT, što je u korelaciji sa dobijenim nalazima markera oštećenja mišića. Takođe, subjektivna procena zamora prati dobijene nalaze i beleži značajno povećanje posle ista dva treninga odmah posle, u prvom i trećem minutu oporavka. Od markera celularne bioenergetike evidentirane su razlike u kreatininu posle NIK i HIIT treninga. Indeks lipidne peroksidacije (TBARS), kao jedan od parametara oksidativnog stresa, beleži promene posle NIK. Koncentracija nitrita (NO2-) beleži takođe statistički značajno povećanje kod NIK, ali i kod HIIT. Od ispitivanih parametara antioksidativne zaštite jedino katalaza (CAT) beleži značajno povećanje kod NIK. U proceni stanja zamora mišića i delovanja enzima oksidativno/antioksidatine statusa, prema dobijenim rezultatima, najsenzitivniji markeri koji su reagovali na različite trenažne epizode, odnosno na različita aerobna opterećenja jesu mioglobin, laktati, kreatin kinaza i kreatinin, a katalaza je pokazala mogućnost diskriminacije različitih vrsta aerobnog opterećenja. During intense exercise, especially professional athletes, muscles are temporarily tired, resulting in a decline in functional abilities. Damage caused by muscle stiffness due to physical exercise has become one of the most important subjects of sports science. The aim of this study was to determine the muscle fatigue and the antioxidative status of basketball players following a load caused by three different types of aerobic training: continuous low intensity (LIC), continuous high intensity (HIC) and intermittent aerobic training (HIIT). As part of the crossover design, a sample of 12 basketball players that were tested at the pre-match phase was selected. Blood samples were taken before and immediately after the training episode on three occasions at intervals of seven days between each type of training. In addition to taking blood samples, pulse testing was performed before training, immediately after training, then after the first, third and fifth minutes of recovery, then the subjects were subjective assessment of fatigue on the Bragg scale immediately after the training, then in the first, third and fifth minutes recovery. The biochemical parameters we analyzed were markers of muscle damage, markers of cellular bioenergetics, parameters of oxidative stress and the activity of enzymes of protection against oxidative damage. The results of the study indicate that the markers of muscle damage record the variability in the results compared to the type of training. Differences in markers of lactates, myoglobin and creatine kinase were recorded between LIC and HIIT. In relation to the heart rate measured at several time points, statistically significant differences have occurred between HIC and HIIT, which is correlated with the findings of the markers of muscle damage. Also, a subjective estimate of fatigue follows the findings obtained and shows a significant increase after the same two trainings immediately after, in the first and third minutes of recovery. Differences in creatinine from LIC and HIIT training were recorded from cellular bioenergetic markers. The lipid peroxidation index (TBARS), as one of the parameters of oxidative stress, records changes after LIC. The concentration of nitrite (NO2-) also recorded a statistically significant increase in LIC, but also in HIIT. From the tested parameters of antioxidant protection, only catalase (CAT) showed a significant increase in NIK. In the assessment of muscular fatigue and oxidative / antioxidant status, according to the obtained results, the most sensitive markers that responded to different training episodes or different aerobic loads were myoglobin, lactates, creatine kinase and creatinine, while the catalase showed the possibility of discrimination of different species aerobic load.

Background Inflammation and oxidative stress are recognized as important contributors to Parkinson’s disease pathogenesis. As such, genetic variability in these pathways could have a role in susceptibility for the disease as well as in the treatment outcome. Dopaminergic treatment is effective in management of motor symptoms, but poses a risk for motor and non-motor adverse events. Our aim was to evaluate the impact of selected single-nucleotide polymorphisms in genes involved in inflammation and oxidative stress on Parkinson’s disease susceptibility and the occurrence of adverse events of dopaminergic treatment. Methods In total, 224 patients were enrolled, and their demographic and clinical data on the disease course were collected. Furthermore, a control group of 146 healthy Slovenian blood donors were included for Parkinson’s disease’ risk evaluation. Peripheral blood was obtained for DNA isolation. Genotyping was performed for NLRP3 rs35829419, CARD8 rs2043211, IL1β rs16944, IL1β rs1143623, IL6 rs1800795, CAT rs1001179, CAT rs10836235, SOD2 rs4880, NOS1 rs2293054, NOS1 rs2682826, TNF-α rs1800629, and GPX1 rs1050450. Logistic regression was used for analysis of possible associations. Results We observed a nominally significant association of the IL1β rs1143623 C allele with the risk for Parkinson’s disease (OR = 0.59; 95%CI = 0.38–0.92, p = 0.021). CAT rs1001179 A allele was significantly associated with peripheral edema (OR = 0.32; 95%CI = 0.15–0.68; p = 0.003). Other associations observed were only nominally significant after adjustments: NOS1 rs2682826 A allele and excessive daytime sleepiness and sleep attacks (OR = 1.75; 95%CI = 1.00–3.06, p = 0.048), SOD2 rs4880 T allele and nausea/vomiting (OR = 0.49, 95%CI = 0.25–0.94; p = 0.031), IL1β rs1143623 C allele and orthostatic hypotension (OR = 0.57, 95%CI = 0.32–1.00, p = 0.050), and NOS1 rs2682826 A allele and impulse control disorders (OR = 2.59; 95%CI = 1.09–6.19; p = 0.032). We did not find any associations between selected polymorphisms and motor adverse events. Conclusions Apart from some nominally significant associations, one significant association between CAT genetic variability and peripheral edema was observed as well. Therefore, the results of our study suggest some links between genetic variability in inflammation- and oxidative stress-related pathways and non-motor adverse events of dopaminergic treatment. However, the investigated polymorphisms do not play a major role in the occurrence of the disease and the adverse events of dopaminergic treatment.

IZ Kukuruzni plamenac (Ostrinia nubilalis Hbn.) predstavlja jednu od najznačajnijih štetočina kukuruza, kako u Srbiji, tako i u Evropi i Sjedinjenim Američkim Državama. Za suzbijanje kukuruznog plamenca u usevu kukuruza mogu se koristiti različite metode, ali za sada najrasprostranjenije je hemijsko suzbijanje. U tu svrhu postoje registrovani preparati, ali njihova efikasnost ne zavisi samo od aktivne supstance, već i od klimatskih faktora, ali i vremena i načina primene. U radu je ispitivana efikasnost insekticida na bazi: hlorantraniliprola, hlorantraniliprola + lambda-cihalotrina i indoksakarba, registrovanih za suzbijanje kukuruznog plamenca, kao i efikasnost različitih rokova primene hlorantraniliprola + lambda-cihalotrina. Takođe, ispitivan je uticaj larvi kukuruznog plamenca i intenziteta napada ove vrste na pojavu i intenzitet zaraze gljiva iz roda Fusarium, kao i uticaj različitih aktivnih supstanci na komponente antioksidativnog sistema zaštite gusenica kukuruznog plamenca. Nakon četiri godine ispitivanja efikasnosti insekticida i dve godine ispitivanja uticaja rokova hemijskog tretmana u suzbijanju kukuruznog plamenca, sprovedenih mikoloških i biohemijskih istraživanja, zaključeno je da su najveću efikasnost u suzbijanju gusenica i smanjenju oštećenja prouzrokovanih larvama kukuruznog plamenca, kako u stabljici kukuruza tako i u klipu, imali tretmani hlorantraniliprolom i hlorantraniliprolom + lambda-cihalotrinom. Tretman indoksakarbom je pokazao najmanju ali i dalje zadovoljavajuću efikasnost. Hemijski tretman imao je pozitivan efekat na prinos u svim godinama ispitivanja. Tretman hlorantraniliprolom je najviše uticao na prinos u 2013. i 2016. godini (35,3 % i 21 % viši u odnosu na kontrolnu grupu). U 2014. i 2015. godini, kada je brojnost gusenica kukuruznog plamenca bila značajno niža, najveći uticaj na prinos imao je tretman hlorantraniliprolom + lambda-cihalotrinom (11,3 % i 15,1 %). Tretmani hlorantraniliprolom i hlorantraniliprolom + lambda-cihalotrinom najviše su uticali na prinos u svim ispitivanim godinama. Ogledom ispitivanja rokova hemijskog tretmana, u obe godine istraživanja, utvrđeno je da je najveći uticaj na prinos ostvaren pri tretmanu kukuruza u periodu početka leta kukuruznog plamenca (19 i 40,6%). Ustanovljeno je da je u godini sa najvećom frekvencijom gusenica i oštećenja kukuruznog plamenca, tretman hlorantraniliprolom značajno uticao na smanjenje učestalosti oštećenih klipova i ujedno je u istom tretmanu zabeležena statistički značajno niža učestalost fuzarioznih klipova, ali pozitivna korelacija smanjenja oštećenosti klipova i fuzarioza nije potvrđena u svim oglednim godinama. Takođe je pokazano da je uticaj insekticida na smanjenje oštećenja klipa prouzrokovanih ishranom insekata i plesnivosti klipa evidentan u godinama sa visokom frekvencijom gusenica, iako se plesnivost klipa može javiti i nezavisno od oštećenja klipa u godinama sa povoljnim klimatskim uslovima za razvoj gljiva. Najzad, rezultati su pokazali da su sva tri tretmana insekticidom uticala na izmenu fizioloških pokazatelja larvi kukuruznog plamenca kao odgovor na oksidativni stres, različit potencijal za indukciju oksidativnog stresa, kao i da reakcija larvi kukuruznog plamenca na delovanje insekticida zavisi od tipa aktivne materije. Reakcija larvi kukuruznog plamenca na delovanje insekticida zavisi od tipa aktivne materije. Složeno delovanje korišćenih insekticida indukuje mehanizme antioksidativne odbrane i detoksifikacije u telu larvi i kod sva tri insekticida ogleda se u visokoj aktivnosti GST u odnosu na kontrolnu grupu. S druge strane, uočena je razlika u odgovoru na delovanje insekticida kod enzima SOD, CAT i GR. Ovim istraživanjem je dokazano da se druga generacija kukuruznog plamenca može uspešno suzbijati u usevu merkantilnog kukuruza sa samo jednim tretmanom, da efikasnost tretmana zavisi i od vremena primene, kao i da je najefikasnija aktivna supstanca za tu svrhu hlorantraniliprol. Potvrđen je uticaj tretmana insekticidom na smanjenje plesnivosti klipa posrednim putem kroz smanjenje broja oštećenih klipova, ali ne u svim godinama istraživanja. Larve koje su preživele hemijski tretman pokazale su izmenjene fiziološke pokazatelje oksidativnog stresa, ali za potpunije razumevanje mehanizama enzimske detoksifikacije potrebna su dalja istraživanja. AB The European Corn Borer (Ostrinia nubilalis Hbn.) is one of the most destructive pests of corn in Serbia and in other parts of its areal. In order to control its populations in maize crops a number of methods can be used, however, chemical control is the most commonly used. Several insecticides are registered for this purpose, yet their efficacy depends on a variety of factors, e.g. climatic conditions, time and mode of application. In this work the efficacy of several Insecticides (chlorantraniliprole, chlorantraniliprole + lambda-cyhalothrin and Indoxacarb) registered against the ECB was evaluated in a four year trial. In a second trial the timing of application of chlorantraniliprole + lambda-cyhalothrin was evaluated during a two year period. The possible effects of ECB larvae on the occurence and intensity of Fusarium Ear Rot in maize was also subject to evaluation in this paper. Finally, the effects of the aforementioned insecticides on the antioxidative defence system of ECB larvae was also part of this work. The results showed that the highest efficacy in reducing the number of ECB larvae and the damage they cause in maize stalks and ears were achieved in the chlorantraniliprole and chlorantraniliprole + lambda-cyhalothrin treatments, while the least efficacy values were obtained in the Indoxacarb treatment, which could still be defined as satisfactory. The chemical tretments showed positive effects on yield in all the trial years. Treatments with chlorantraniliprole had the most significant positive effects on the yields in years 2013. and 2016. with a 35.3% and 21% yield increase compared to the control. In years with the lowest pest pressure, 2014 and 2015, the highest positive impact on yield was achieved with the chlorantraniliprole + lambda-cyhalothrin treatments (11.3% and 15.1%). Treatments using chlorantraniliprole and chlorantraniliprole + lambda-cyhalothrin had the highest positive impact on yield in all trial years.Regarding the timing of application, the highest positive impact on yields were achieved when trials were undertaken at the beginning of the flight periodof the second ECB generation (19 and 40.6%). Treatments with chlorantraniliprole in trial years with the highest pest pressure considerably reduced the number of larvae and their damage to maize ears, and at the same time considerably reduced the incidence of Fusarium Ear Rot. However, this positive correlation was not confirmed in all trial years. It has been shown that the effects of insecticide treatments on the reduction of ear damage caused by insect feeding and Fusarium Ear Rot is evident in years with an increasedlarval frequency, even though FER can occur independently from ear damage in years with favourable climatic conditions for the development of the fungus. All three insecticide treatments affected the physiological parameters of ECB larvae, inducing a responce to oxidative stress. The different insecticides had a different potential for oxidative stress induction, which depends on the application of the active substance. All three insecticide treatments induced a higher activity of glutathione S-transferase compared to the control group, while different reaction towards superoxide dismutase, catalase and glutathione reductase were observed depending on the applied treatment. The results of the trials in this work showed that the second ECB generation can be managed with only one insecticide application, and that the efficacy of the treatment depends on the timing of the application. The highest results were obtained using chlorantraniliprole. The positive effects of insectide treatments on decreasing Fusarium Ear Rot incidence through the decrease of ear damage caused by ECB larvae was confirmed, however, this was not the case in all trial years. ECB larvae which survived the insecticide treatment showed clearly altered parameters of oxidative stress, however, a better understanding of the mechanisms of enzymatic detoxification should be looked for in further trials.

U radu je analiziran uticaj limunske kiseline na morfološke, fiziološke i biohemijskekarakteristike odabranih klonova vrba (Salix spp.) gajenih u zemljištu umereno zagađenom kadmijumom (Cd). Definisanjem parametara akumulacije, otpornosti i tolerantnosti različitih klonova vrba na prisustvo kadmijuma u podlozi, kao i uspešnosti primene limunske kiseline kao helatora u procesu asistirane fitoremedijacije, mogao bi se identifikovati klon (ili klonovi) pogodan za dekontaminaciju zemljišta zagađenog kadmijumom. Da bi se postigao cilj istraživanja biljke su gajene u polu-kontrolisanim uslovima tokom 3 meseca. Eksperiment je podeljen u dve faze (dve godine istraživanja),zasnovane na razlikama u metodološkom pristupu. Tokom prve godine limunska kiselina je dodata jednom, dok je tokom druge druge godine aplikacija ovog helatora vršena u tri ponavljanja. U radu je analizaran sadržaj i distribucija Cd u zemljištu irazličitim biljnim organima (koren, izdanak, mladi i stari listovi); uticaj Cd i limunske kiseline na morfometrijske parametre: visina i prečnik izdanka, masa i površina listova, broj listova, masa i zapremina korena; na fotosintetičke karakteristike: intenzitet fotosinteze, intenzitet transpiracije, stomatalnu provodljivost, intercelularnu koncentraciju CO2, efikasnost korišćenja vode, koncentraciju fotosintetičkih pigmenata; na mineralnu ishranu i distribuciju makronutrijenata u biljkama (sadržaj azota, fosfora i kalijuma); na sadržaj aminokiseline prolina i cisteina; na aktivnost antioksidativnih enzima (katalaza, askorbat-peroksidaza, gvajakol-peroksidaza), kao neezmiskih antioksidanata poput sadržaja redukovanog glutationa i tiola. Dobijeni rezultati su ukazali na genotipsku specifičnost analiziranih klonova vrba u procesu fitoremedijacije u zavisnosti od primenjenog tretmana, doze Cd i prisustva limunske kiseline. Toksično dejstvo nije bilo jasno uočljivo na morfološkom nivou, dok su primenjeni tretmani signifikantno uticali na fiziološko-biohemijske procese u biljkama. Sa povećanjem sadržaja Cd u biljkama je utvrđena tendencija smanjenja intenziteta fotosinteze, transpiracije, kao i efikasnosti koršćenja vode, dok je primena limunske kiseline ublažila štetan efekat ovog teškog metala, što je registrovano nakombinovanim tretmanima. Kao odgovor na akumulaciju Cd u biljnom tkivu utvrđenaje promena aktivnosti antioksidativnih enzima, kao i neezimskih komponentiodbrambenog sistema zaštite. Pored toga, u radu je utvrđeno da primena limunske kiseline utiče na usvajanje, akumulaciju i toleranciju vrba na prisustvo Cd, te rezultati ove studije imaju praktičan značaj u usavršavanju tehnika fitoremedijacije i dekontaminacije zagađenih područja., The role of citric acid on morphological, physiological and biochemical characteristics of selected willow clones (Salix spp.) grown in soil with moderately polluted cadmium (Cd) was presented. The aim of this study was to determine the Cd accumulation, translocation and tolerance of different willow clones, as well as the influence of citric acid as a chelator in the assisted phytoremediation process. In order to achieve this goal, plants were grown in semi-controlled conditions for 3 months. The experiment was divided into two phases (two years of research), based on differences in the methodological approach, regarding the application of citric acid. During the first-year experiment, citric acid was added once, while in the second year the application of this chelator was performed in three repetitions. The analyses included: the accumulation and distribution of Cd in soil and various plant organs (root, stem, young and old leaves); influence of Cd and citric acid on morphometric parameters: stem hight and diameter, number of leaves, biomass and area of leaves, biomass and volume of the root; on photosynthetic characteristics: net photosynthetic and transpiration rate, water use efficiency, stomatal conductance, intercellular concentration of CO2, concentration of photosynthetic pigments; on mineral nutrition and distribution of macronutrients in plants (nitrogen, phosphorus and potassium content); on the content of the amino acid proline and cysteine; on the activity of antioxidant enzymes (catalase, ascorbate peroxidase, guaiacol peroxidase), as non- enzymatic components such as reduced glutathione and thiol content.The obtained results indicated the genotypic specificity of the analyzed willow clones in the phytoremediation process, depending on the treatment applied, the Cd dose, and the presence of citric acid. The toxic effect was not evident on the morphological level, while the applied treatments showed a significant influence on the physiologicalbiochemical processes in the willows. The net photosynthetic rate, transpiration rate, as well as, water use eficiency showed decreasing with increasing Cd concentration in plant tissue, while the application of citric acid mitigated the harmful effect of this heavy metal, which was registered on combined treatments. In response to the accumulation of Cd in plant tissue, a change in the activity of anti-oxidant enzymes, as well as nonenzymatic components of the defense system has been established. In addition, theapplication of citric acid showed the promotion role on the absorption, accumulation and tolerance of willows grown in moderately polluted soil cadmium, which has a practical significance in the improvement of phytoremediation techniques and decontamination of polluted soil.

Kičica se u Srbiji tradicionalno koristi za lečenje različitih oboljenja, uključujući i dijabetes. Cilj ovog rada bio je rasvetljavanje mehanizama antioksidativnog i protektivnog dejstva ekstrakta kičice (EK) koji doprinose očuvanju strukture i funkcije β-ćelija pankreasa i ublažavanju komplikacija u dijabetesu. Efekti EK analizirani su na eksperimentalnom modelu dijabetesa pacova izazvanom streptozotocinom (STZ) kao i na Rin-5F β-ćelijama izloženim oksidativnom stresu primenom vodonik peroksida, natrijum nitroprusida ili STZ-a. EK je ispoljio antidijabetogeno dejstvo uočeno kao sniženje nivoa glukoze/glikohemoglobina, poboljšanje lipidnog statusa i povećanje nivoa insulina u cirkulaciji dijabetičnih pacova što se može pripisati detektovanom uticaju EK na oc uvanje strukture Langerhansovih ostrvaca i povec anju mase β-c elija koje produkuju insulin. ntioksidativni efekat EK na β-c elije ogledao se u redukovanju oksidativnih os tec enja, modulisanju aktivnosti i ekspresije enzima antioksidativne zaštite (C T, SOD, GPx i GR) i uravnotežavanju aktivnosti redoks-senzitivnih faktora (NFκB, Nrf-2, Sp1, FOXO3 ) uključenih u regulaciju transkripcije gena za antioksidativne enzime. Dodatni doprinos EK boljem preživljavanju β-ćelija i sekreciji/ekspresiji insulina ostvaren je finim promenama u aktivnosti Akt, ERK i p38 kinaza kao i PDX-1 i MafA proteina. Pored toga, antioksidativni efekat EK kod dijabetičnih životinja ogledao se u zaštiti eritrocita, jetre i bubrega od gliko-oksidativnih oštećenja što je doprinelo poboljšanju njihovih funkcionalnih parametara. Opisani efekti EK ukazuju na značaj daljeg razjašnjavanja mehanizama antioksidativnog i protektivnog dejstva EK i njegovih komponenti u cilju potencijalnog terapijskog delovanja u dijabetesu. Centaury is traditionally used in Serbia for treating different diseases, including diabetes. This study aimed to elucidate the mechanisms of the antioxidant and protective effects of the centaury extract (CE) that preserve the structure and function of pancreatic β-cells and alleviate diabetic complications. The effects of the CE were analyzed on the experimental model of streptozotocin (STZ)-induced diabetic rats and in Rin-5F β-cells exposed to oxidative stress after exposure to hydrogen peroxide, sodium nitroprusside or STZ. The CE exhibited an anti-diabetic effect, observed as a decrease in hyperglycemia and glycohemoglobin concentration, an improved lipid status, and as increased insulin level in the circulation of diabetic rats. These effects can be attributed to the detected impact of the CE on the preservation of the structure of the islets of Langerhans, and on the increase in insulin producing β-cell mass. The antioxidant effect of the CE on β-cells was manifested as a reduction in oxidative damage, changed activities and expression of antioxidant enzymes, CAT, SOD, GPx and GR, and as a readjustment of the activities of redox-sensitive factors, NFκB, Nrf-2, Sp1, FOXO3A, involved in the regulation of gene transcription of antioxidant enzymes. Additional contributions of the CE to improved β-cell survival and insulin secretion/expression were achieved by the fine-tuning of the activities of Akt, ERK and p38 kinases and transcription regulators, PDX-1 and MafA. The antioxidant effect of the CE in diabetic animals was also reflected on increased protection of erythrocytes, liver and kidneys from glyco-oxidative damage, which contributed to the improvement of their functional parameters in diabetic rats. The described effects of the CE point to the importance of further clarification of the mechanisms of the antioxidant and general protective effects of the CE and its components in view of its potential use for therapeutic intervention in diabetes.

The purpose of the study was to determine oxidative stress-related plasma thiol/disulphide, ischemia-modified albumin (IMA) levels and ferroxidase activity among women with obesity in insulin-resistant and non-insulin-resistant groups in comparison with an overweight group.We compared plasma thiol/disulphide, IMA levels, and ferroxidase activity between the study groups. We analyzed plasma thiol/disulphide homeostasis with a newly developed automated measurement method; IMA with Albumin Cobalt Binding Test and ferroxidase (ceruloplasmin) levels with an automated, colourimetric method.There were no significant differences between insulin-resistant and non-insulin-resistant women with obesity in terms of plasma native thiol, total thiol, disulphide, disulphide/native thiol ratio, disulphide/total thiol or native thiol/total thiol values. Ferroxidase activity was higher in insulin-resistant than in non-insulin-resistant women with obesity and higher in the total women with obesity group than in the overweight subjects (p0.001, and p=0.014, respectively). IMA was lower in the insulin-resistant group than in the non-insulin-resistant group and overweight groups (p=0.011, and p=0.042, respectively).The significantly greater increase in ferroxidase activity in insulin-resistant subjects with obesity may reflect its role as a positive acute phase protein. These findings may be related to the pathogenesis of the disease. Changes in oxidative status occur in women with obesity, and partially in overweight subjects. The ferroxidase activity of ceruloplasmin plays a crucial role in iron homeostasis and lowers oxidative stress by reducing the detrimental effects of iron.Svrha ove studije bila je da se odredi nivo oksidativnog stresa vezan za plazma tiol/disulfid, nivo ishemijski modifikovanog albumina (IMA) i aktivnost fer oksidaze kod gojaznih žena iz insulinski rezistentne i nerezistentne grupe u poređenju sa grupom žena sa prekomernom težinom.Uporedili smo plazma tiol/disulfid, nivoe IMA i aktivnost feroksidaze među ispitivanim grupama. Analizirali smo plazma tiol/disulfidnu homeostazu pomoću novorazvijene automatizovane metode merenja, IMA testom vezivanja albumin kobalta, a feroksidazu (ceruloplazmin) automatizovanom kolorimetrijskom metodom.Nije bilo značajnih razlika između insulinski rezistentnih i nerezistentnih žena i gojaznih žena u smislu vrednosti koje se odnose na prirodni tiol, ukupni tiol, disulfid, odnos disulfid/prirodni tiol, disulfid/ukupni tiol ili prirodni tiol/ukupni tiol. Aktivnost feroksidaze bila je viša kod žena rezistentnih na insulin nego kod gojaznih žena koje nisu insulinski rezistentne, a veća je u ukupnom broju gojaznih nego kod ispitanica sa prekomernom težinom (p0,001 i p = 0,014, po ispitivanju). IMA je bio niži u grupi koja je rezistentna na insulin nego u grupama koje nisu rezistentne na insulin i grupi sa prekomernom težinom (p = 0,011 i p = 0,042).Značajno veće povećanje aktivnosti feroksidaze kod gojaznih subjekata otpornih na insulin može odražavati njegovu ulogu kao pozitivnog proteina akutne faze. Ovi nalazi mogu biti povezani sa patogenezom bolesti. Promene u oksidativnom statusu javljaju se kod gojaznih žena i delimično kod ispitanica sa prekomernom težinom. Feroksidazna aktivnost ceruloplazmina igra ključnu ulogu u homeostazi gvožđa i smanjuje oksidativni stres smanjenjem štetnih efekata gvožđa.

Akrilamid je toksična hemijska supst anca koja je već dugi niz godina prisutna u životnoj sredini, jer se kao važan monomer koristi u različite industrijske i laboratorijske svrhe. U poslednjih petnaest godina, akrilamid je postao posebno zanimljiv za šire naučne krugove jer se pokazalo da se nalazi i u hrani biljnog porekla, posebno hrani bogatoj skrobom, koja se priprema pečenjem ili prženjem na temperaturama višim od 120°C. Do sada ustanovljeni negativni zdravstveni efekti akrilamida su veoma raznovrsni i mogu biti rezultat delovanja samog akrilamida ili delovanja njegovog metabolita glicidamida koji nastaje in vivo kada se jedan deo molekula akrilamida metaboliše oksigenacijom dvostruke veze pomoću enzima citohrom P450 2E1 (CYP2E1). Akrilamid je supstanca koja ima dokazan negativan efekat na organske sisteme kod ljudi i životinja, i koja je svrstana u moguće humane karcinogene. Negativan efekat akrilamida na egzokrini pankreas je poznat, ali o mogućim efektima akrilamida na endokrini pankreas se i dalje veoma malo zna. Ima puno dokaza koji ukazuju na to da akrilamid ima citotoksični efekat koji se manifestuje kroz uticaj na redoks-status ćelija i dovodi do promena u vrednostima biomarkera oksidativnog i nitrozativnog stresa, kao i u aktivnosti antioksidativnih enzima. Pankreas je jedan od ciljnih organa za delovanje akrilamida te je glavni predmet istraživanja doktorske teze bio proučavanje potencijalnog efekta akrilamida na endokrini pankreas pacova. Ispitivanje je vršeno na 3 eksperimentalne grupe juvenilnih mužjaka pacova soja Wistar, od kojih je jedna grupa bila kontrolna, dok su dve bile tretirane sa akrilamidom u dozama od 25 mg/kg tm i 50 mg/kg tm, 5 dana nedeljno, tokom 3 nedelje. Po isteku tretmana, nakon dekapitacije, kompletno tkivo pankreasa je fiksirano u 10% rastvoru formalina tokom 24 h i obrađeno prema standardnoj proceduri za kalupljenje u parafinu. Parafinski kalupi su sečeni na serijske preseke debljine 5 µm, nakon čega su bojeni histohemijskom i imunohistohemijskim metodama. Kod eksperimentalnih grupa posmatrane su histološke promene na endokrinom pankreasu, sa akcentom na α- i β-ćelije. Takođe, posmatrana je i ekspresija hormona insulina i glukagona, enzima inducibilne azot -oksi d sintetaze (iNOS) i CYP2E1, kao i ekspresija antioksidativnih enzima katalaza (CAT) i superoksid dismut aza 1 i 2 (SOD1 i SOD2) u ćelijama Langerhansovih ostrvaca. Potencijalna promena u funkcionalnosti β-ćelija je ispitana i kroz analizu nivoa glukoze u serumu pacova tretiranih sa akrilamidom. Budući da β-ćelije čine 80% ćelija koje grade Langerhansova ostrvca pankreasa, pored in vivo eksperimenata, ispitana je i toksičnost akrilamida na Rin-5F ćelijsku liniju insulinoma β-ćelija pacova u in vitro uslovima. Glavni cilj in vitro istraživanja je bio da se ispita uticaj rastućih koncentracija akrilamida na preživljavanje tretiranih Rin-5F ćelija, ali i efekat IC50 koncentracije ove supstance primenjene tokom različitih vremenskih intervala (0,5, 1, 3, 6, 12 i 24 h) na pojavu oksidativnog i nitrozativnog stresa. Redoks-status Rin-5F ćelija tretiranih sa akrilamidom je ispitan preko analize prisustva biomarkera oksidativnog i nitrozativnog stresa, akrivnosti CAT i ukupne SOD, kao i promene u ekspresiji gena za CAT, SOD1, SOD2 i iNOS. Pored toga, analiziran je i efekat istog tretmana na ekspresiju gena za insulin, CYP2E1, Bax i Bcl-2. U okviru teze je pokazano da akrilamid ne dovodi do značajnih promena u histološkoj građi, dijametru i broju Langerhansovih ostrvaca kod tretiranih životinja. Primena stereoloških metoda je ukazala na mikrostrukturne promene na endokrinom pankreasu na nivou α- i β-ćelija. U ovoj tezi je po prvi put pokazano da tretman akrilamidom negativno utiče na broj i površinu β-ćelija pankreasa. U tezi je, takođe, pokazan značajan dozno-zavisni pad u prisustvu insulina u β-ćelijama pankreasa. Uprkos tome, kod akrilamidom tretiranih životinja nije konstatovana promena u koncentraciji serumske glukoze. U ovoj tezi je pokazano da tretman akrilamidom dovodi do statistički značajnog porasta u broju α-ćelija kod životinja koje su primale nižu dozu tretmana, dok se broj α-ćelija kod životinja koje su primale višu dozu tretmana ne razlikuje značajno od kontrole. Tretman akrilamidom je doveo do značajnog porasta u količini prisutnog glukagona u α-ćelijama pankreasa. Tretman akrilamidom nije doveo do značajne promene u ekspresiji CAT, SOD1 i SOD2 u ćelijama Langerhansovih ostrvaca. Kod tretiranih životinja došlo do značajnog dozno-zavisnog porasta u ekspresiji enzima iNOS, dok je ekspresija CYP2E1 značajno dozno-zavisno opala nakon tretmana. U tezi je pokazano da tretman akrilamidom negativno utiče na vijabilnost Rin-5F ćelija, i utvrđeno je da IC50 koncentracija akrilamida za Rin-5F ćelije iznosi 10 mM. Rezultati teze pokazuju da tretman akrilamidom u IC50 koncentraciji u Rin-5F ćelijskoj liniji značajno povećava nivo malondialdehida (MDA) nakon tretmana u trajanju od 1, 12 i 24 h. Isti tretman značajno smanjuje nivo redukovanog GSH nakon tretmana od 1, 3, 6, 12 i 24 h, kao i nivo slobodnih –SH grupa nakon tretmana od 3 i 6 h. Tretman akrilamidom u IC50 koncentraciji signifikantno pojačava aktivnost CAT nakon tretmana od 1 h, dok tretman u trajanju od 12 h značajno smanjuje aktivnost ovog enzima. Ovaj tretman smanjuje aktivnost SOD nakon 1, 12 i 24 h, dok tretman u trajanju od 6 h značajno pojačava aktivnost enzima SOD. U tezi je, takođe, pokazan i veoma značajan porast u nivou prisutnih nitrita, koji je direktno proporcionalan sa nivoom azot-oksida i nivoom akivnosti enzima iNOS. Ovaj nalaz ukazuje na potencijalnu pojavu nitrozati vnog stresa u akrilamidom-tretiranim Rin-5F ćelijama. U tezi je po prvi put pokazano da tretman akrilamidom dovodi do značajnih varijacija u transkripciji gena za iNOS, SOD1, SOD2, CAT, CYP2E1, Bax i Bcl-2 u tretiranim Rin-5F ćelijama, dok isti tretman ne dovodi do promene nivoa transkripcije gena za insulin. Tretman akrilamidom u koncentraciji od 10 mM tokom rastućih vremenskih perioda dovodi do porasta u relativnoj količini iRNK gena za iNOS u svim tačkama tretmana, do porasta nivoa iRNK za SOD1 i SOD2 nakon tretmana od 12 i 24 h, kao i do porasta količine iRNK za CAT nakon tretmana od 3 h. U tezi je pokazano i da akrilamid izaziva promene u sintezi iRNK za enzim CYP2E1 koji je posebno značajan u kontekstu detoksikacije ove toksične supstance. Porast u transkripciji gena za CYP2E1 je uočen nakon tretmana u trajanju od 0,5 i 1 h, dok je do smanjenja transkripcije došlo nakon tretmana od 12 i 24 h. Tretman akrilamidom u koncentraciji od 10 mM tokom rastućih vremenskih perioda dovodi do porasta u relativnoj količini iRNK gena za Bax u svim tačkama tretmana, i do porasta u transkripciji gena za Bcl-2 nakon tretmana od 0,5, 1 i 3 h. Sumirajući sve rezultate ove teze, moze se zaključiti da je endokrini pankreas jedno od ciljnih tkiva, na koje akrilamid ostvaruje višestruki negativni uticaj. Acrylamide is a toxic chemical used as an important monomer for various industrial and laboratory purposes, which makes it highly present in the environment. In the last fifteen years, acrylamide has become especially interesting for wider scientific circles when it was found in staple foodstuff rich in starch, prepared at temperatures higher than 120°C. The established negative health effects of acrylamide are very diverse and can be the result of the acrylamide action itself or the action of its metabolite glycidamide that occurs in vivo, when acrylamide molecule is metabolized via oxygenation of the double bond by the cytochrome P450 2E1 (CYP2E1). Acrylamide is a substance with a proven adverse effect on humans and animals, and it is classified as a possible human carcinogen. The negative effect of acrylamide on the exocrine pancreas has already been recognized, but the possible effects of acrylamide on endocrine pancreas are still mostly undetermined. There is a significant amount of evidence to suggest that acrylamide exerts a cytotoxic effect which manifests through the changes in level of oxidative and nitrosative stress biomarkers, as well as in the activity of antioxidant enzymes. Since, pancreas is one of the target organs for acrylamide, the main subject of doctoral thesis was to investigate the potential effect of acrylamide on the rat endocrine pancreas. The investigation was conducted on 3 experimental groups of juvenile male Wistar rats, of which one group was the control group, while two groups were treated with acrylamide at doses of 25 mg/kg bw and 50 mg/kg bw, 5 days a week, during 3 weeks. After termination of the treatment, decapitation was performed, and the complete pancreatic tissue was fixed in a 10% formalin solution for 24 h and treated according to the standard paraffin embedding procedure. Paraffin molds were cut into 5 μm thick serial sections, after which they were stained with histochemical and immunohistochemical methods. Histological changes ofthe endocrine pancreas, with the emphasis on α- and β-cells, were examined in three experimental groups of rats. In addition, the expression of insulin and glucagon hormone, the inducible nitric oxide synthase (iNOS) and CYP2E1 enzymes, and the expression of antioxidative enzymes catalase (CAT) and superoxide dismutases 1 and 2 (SOD1 and SOD2) in the islets of Langerhans were also investigated. A potential change in the functionality of β-cells was also examined by analyzing glucose level in the serum of rats treated with acrylamide. In pancreatic islets of Langerhans the majority of cells (>80%) are β-cells. Therefore, in addition to in vivo experiments, the toxicity of acrylamide was examined in vitro on rat insulinoma Rin-5F cell line.The main goal of in vitro research was to investigate the impact of increasing acrylamide concentrations on the viability of treated Rin-5F cells, and also to examine whether IC50 concentration of this substance, applied at different intervals of time (0.5, 1, 3, 6, 12 and 24 h), induce oxidative and nitrosative stress. Redox-status of Rin-5F cells treated with acrylamide was examined by analyzing oxidative and nitrosative stress biomarkers, CAT and total SOD activity, as well as changes in the expression of the CAT, SOD1, SOD2 and iNOS. In addition, the effect of the same treatment on the transcription of the insulin, CYP2E1, Bax and Bcl-2 gene was analyzed.The results of the thesis showed that acrylamide treatment does not lead to significant changes in the histological structure, diameter and number of islets of Langerhans of treated animals. Application of stereological methods indicated microstructural changes of α- and β-cells ofendocrine pancreas. It has been shown for the first time that treatment with acrylamide negatively affects the number and surface area of pancreatic β-cells. In addition, a significant dose-dependent decline in the amount of insulin in pancreatic β-cells was also demonstrated. However, no change in serum glucose level was observed in treated animals. Acrylamide treatment led to a statistically significant increase in the number of α-cells in animals receiving a lower dose of treatment, while the number of α-cells in animals receiving a higher dose of treatment did not differ significantly from the control. Treatment with acrylamide led to a significant increase in the amount of the glucagon in α-cells. Treatment with acrylamide did not cause a significant change in the expression of CAT, SOD1 and SOD2 in islets of Langerhans. However, there was a significant dosedependent increase in the expression of iNOS enzyme, whereas expression of CYP2E1 significantly decreased in dose-dependent manner in treated animals. Results of the thesis showed that acrylamide exerts a negative effect on the viability of Rin-5F cell line. It has been established that the IC50 concentration of acrylamide for the Rin-5F cell line is 10 mM. The results of the thesis indicate that treatment of Rin-5F cell line with IC50 concentration of acrylamide for 1, 12, and 24 h significantly increased the level of malondialdehyde (MDA). Exposure to acrylamide for 1, 3, 6, 12 and 24 h significantly decreased the level of reduced GSH, while the level of free -SH groups was reduced after 3 and 6 h of acrylamide treatments. Treatment with IC50 concentration of acrylamide significantly enhanced CAT activity after 1 h of acrylamide exposure, while 12 h exposure significantly reduced the activity of this enzyme. Application of acrylamide reduced SOD activity after 1, 12, and 24 h exposure, while 6 h exposure significantly increased the activity of SOD enzymes. Results of the thesis also showed a very significant increase of the nitrite level, which is directly proportional to the level of nitrogen oxide (NO) and the level of the iNOS activity. This finding points to the potential occurrence of nitrosative stress in acrylamide-treated Rin-5F cells. It has been shown for the first time that acrylamide treatment leads to significant variations in transcription of iNOS, SOD1, SOD2, CAT, CYP2E1, Bax and Bcl-2 genes in treated Rin-5F cells, while the same treatment does not affect transcription of the insulin gene. Treatment with acrylamide at a concentration of 10 mM for increasing periods of time leads to an increase in the relative amount of the iNOS gene iRNA at all treatment points. Twelve and and 24 h of acrylamide exposure increased the transcription of the SOD1 and SOD2 genes. Transcription of CAT gene was increased after 3 h ofacrylamide exposure. Furthermore, it has been shown that acrylamide treatment leads to variations in the mRNA synthesis of CYP2E1 gene, which is particularly significant in the context of detoxification of this toxic substance. An increase in the transcription ofthe CYP2E1 gene was observed after 0.5 and 1 h of acrylamide exposure, while the reduction of transcription occurred after 12 and 24 h of acrylamide exposure. The treatment with 10 mM acrylamide has led to an increase of the transcription of the Bax gene at all treatment points, and also to an increase of transcription of the Bcl-2 gene after of 0.5, 1, and 3 h of acrylamide exposure. Summarizing all the results of this thesis, it can be concluded that the endocrine pancreas is one of the target tissues of acrylamide, to which this substance exerts a multiple adverse effects.

Epigenetic mechanisms including posttranslational histone modifications and DNA methylation are emerging as important determinants of bone homeostasis. With our case-control study we aimed to identify which chromatin-modifying enzymes could be involved in the pathology of postmenopausal osteoporosis and osteoarthritis while co-regulated by estrogens, oxidative stress and hypoxia. Gene expression of HAT1, KAT5, HDAC6, MBD1 and DNMT3A affected by oxidative stress and hypoxia in an in vitro qPCR screening step performed on an osteoblast cell line was analysed in trabecular bone tissue samples from 96 patients. Their expression was significantly reduced in patients with postmenopausal osteoporosis and osteoarthritis as compared to autopsy controls and significantly correlated with bone mineral density and several bone histomorphometry-derived parameters of bone quality and quantity as well as indicators of oxidative stress, RANK/RANKL/OPG system and angiogenesis. Furthermore, oxidative stress increased DNA methylation levels at the RANKL and OPG promoters while decreasing histone acetylation levels at these two genes. Our study is the first to show that higher expression of HAT1, HDAC6 and MBD1 is associated with superior quantity as well as quality of the bone tissue having a more favourable trabecular structure.

Актуелни трендови употребе биљних производа у заштити здравља људи и животиња довели су до широке примене природних полифенола присутних у биљкама, као што су зелени чај, боровница, аронија, итд. На основу досадашњих испитивања настанка оксидативног стреса као и начина превенције, можемо закључити да су грожђе, семенке грожђа, као и комина показали одређене протективне ефекте. In vivo студије ефикасности полифенола су изузетно комплексне и до сада су дале контроверзне резултате. У овде представљеном раду дат је преглед до сада постигнутих сазнања ефеката употребе полифенола грожђа у заштити од оксидативног стреса домаћих животиња. Current trends on the use of plant products in the protection of human and animal health have resulted in the widespread use of natural polyphenols present in plants such as green tea, chokeberries, blackberries, etc. According to a wide array of trials on oxidative stress and its prevention it can be concluded that grapes, grapeseeds and pomace have some protective effects. In vivo polyphenol efficacy studies are extremely complex and up to date have raised a number of controversial results. In the here presented paper we have offered a review on the knowledge on grape polyphenols in oxidative stress protection of domestic animals.

Overproduction of free radicals accompanied with their insufficient removal/neutralization by antioxidative defense system impairs redox hemostasis in living organisms. Oxidative stress has been shown to be involved in all the stages of carcinogenesis and malignant melanocyte transformation. The aim of this study was to examine association between oxidative stress development and different stages of melanoma.The measured oxidative stress parameters included: superoxide anion radical, total and manganese superoxide dismutase, catalase and malondialdehyde. Oxidative stress parameters were measured spectrophotometrically in serum samples from melanoma patients (n=72) and healthy control subjects (n=30). Patients were classified according to AJCC clinical stage.Average superoxide anion and malondialdehyde concentrations were significantly higher in melanoma patients than in control group, with the highest value of superoxide anion in stage III, while malondialdehyde highest value was in stage IV. The activity of total and manganese superoxide dismutase was insignificantly higher in melanoma patients than in control group, while catalase activity was significantly higher. The highest activity of total activity of manganese superoxide dismutase was in stage IV. Catalase activity was increasing with the disease progression achieving the maximum in stage III.Results of our study suggest that melanoma is oxidative stress associated disease, as well as deteriorated cell functioning at mitochondrial level.Prekomerna produkcija slobodnih radikala i njihova nedovoljna eliminacija/neutralizacija putem sistema antioksidativne odbrane, narušava redoks homeostazu u organizmu. Oksidativni stres je uključen u sve faze razvoja karcinoma i maligne transformacije melanocita. Cilj ove studije bio je da se ispita razvoj oksidativnog stresa u razlićitim stadijumima melanoma.Parametri za procenu oksidativnog stresa su: superoksil anjon radikal, ukupna i mangan superoksidna dizmutaza, katalaza i malondialdehid. Parametri oksidativnog stresa su mereni metodom spektrofotometrije u serumu bo lesnika sa melanomom (n=72) i zdravih kontrolnih oso ba (n=30). Bolesnici su klasifikovani prema AJCC kriterijumu.Prosečne koncentracije superoksil anjon radikala i malonaldehida bile su značajno veće kod bolesnika sa melanomom u odnosu na kontrolnu grupu, najveća vrednost superoksil anjon radikala bila je u III stadijumu, a malondialdehida u IV stadijumu. Aktivnost ukupne i mangan superoksidne dizmutaze bila je neznačajno povećana kod obolelih od melanoma u odnosu na kontrolnu grupu, dok je aktivnost katalaze bila statistički značajno veća. Najveća aktivnost ukupne superoksidne dizmutaze bila je u III stadijumu, a mangan superoksidne dizmutaze u IV stadijumu.Zaključak Aktivnost katalaze je rasla sa napredovanjem bolesti i dostigla maksimum u III stadijumu.Rezultati našem studije ukazuju na povezanost melanoma i oksidativnog stresa, kao i na pogoršanu funkciju ćelija na nivou mitohondrija.

Oksidativni stres i hronična inflamacija smatraju se glavnim uzrocima pojave dijabetičnih komplikacija, među kojima su i oštećenja jetre. Važnu ulogu medijatora ovih procesa može imati endogeni protein HMGB1, koji u vanćelijsku sredinu dospeva iz nekrotičnih, oštećenih i aktiviranih ćelija. Kako je u dijabetesu ponašanje HMGB1 proteina slabo izučavano, u ovoj doktorskoj disertaciji ispitivan je doprinos HMGB1 oštećenjima jetre pacova sa streptozotocinom-izazvanim DT1. Pokazano je da nivo oštećenja jetre tokom dijabetesa koreliše sa prisustvom vanćelijskog HMGB1. Ovaj protein, u dijabetičnoj jetri, biva strukturno modifikovan acetilacijom, fosforilacijom i O-GlcNAc glikozilacijom što koreliše sa njegovim izlaskom iz jedra ćelija u citoplazmu i povećanjem njegovog prisustva u jetri i serumu. Rezultati u vezi sa snižavanjem nivoa vanćelijskog HMGB1 tretmanom dijabetičnih pacova melatoninom ili etil piruvatom, ukazuju da HMGB1 doprinosi oštećenju jetre u dijabetesu održavanjem stanja hronične inflamacije, stišavanjem antioksidativne odbrane i stišavanjem regeneracije. Vanćelijski HMGB1 kroz interakcije sa TLR4 receptorom aktivira MAPK/NF-κB p65 i JAK1/STAT3 signalne puteve, doprinoseći povećanju produkcije proinflamacijskih citokina TNF-α i IL-6 i akutno-faznog proteina haptoglobina. Podsticanjem NF-κB p65 inflamacijskog puta, HMGB1 deluje negativno na citoprotektivni odgovor u dijabetičnoj jetri tako što onemogućava aktivnost Nrf2 proteina, odgovornog za stišavanje inflamacije i produkciju antioksidativnih enzima. Na stišavanje regenerativnog potencijala jetre, aktivirana HMGB1/TLR4 osa utiče preko uvećanja prisustva negativnih regulatora ćelijskog ciklusa - proteina p53 i p21, i smanjenjem nivoa ciklina D1. Dobijeni rezultati ukazuju na složenost delovanja HMGB1 proteina u dijabetesu i na značaj sprečavanja oslobađanja HMGB1 ili blokade HMGB1/TLR4 ose u cilju odlaganja nastanka oštećenja jetre. Oxidative stress and chronic inflammation are considered to be the main causes of diabetic complications, one of which is liver damage. An important mediator of these processes may be the endogenous HMGB1 protein, when released into the extracellular environment from the necrotic, damaged or activated cells. As the HMGB1 role in diabetes was insufficiently studied, in this doctoral dissertation the contribution of HMGB1 to liver damage of streptozotocin-induced diabetic rats was investigated. It has been shown that the level of liver damage in diabetes correlates with the presence of extracellular HMGB1. In diabetic liver, this protein is structurally modified by acetylation, phosphorylation, and O-GlcNAc glycosylation, which correlates with its translocation from the nucleus to the cytoplasm and an increase in its presence in the liver and serum. Reduction of the level of extracellular HMGB1 by melatonin or ethyl pyruvate treatment of diabetic rats, shows that HMGB1 contributes to diabetic liver damage by maintaining a chronic inflammation, by lowering antioxidant defense and by reducing regeneration. Extracellular HMGB1 activates MAPK/NF-κB p65 and JAK1/STAT3 signaling pathways through interactions with the TLR4 receptor, thus contributing increased production of proinflammatory cytokines TNF-α and IL-6 and the acute-phase protein, haptoglobin. By stimulating the NF-κB p65 inflammatory pathway, HMGB1 acts negatively on the cytoprotective response of the diabetic liver, by disabling Nrf2 protein activity, which is responsible for reduction of inflammation and antioxidant enzymes production. Activated HMGB1/TLR4 axis reduces regenerative potential of the liver by increasing the presence of negative cell cycle regulators - proteins p53 and p21, and also by decreasing the level of cyclin D1. The obtained results indicate the complexity of HMGB1 protein action in diabetes and underlines the importance of preventing the release of HMGB1 or blockage of HMGB1/TLR4 axis in order to delay the occurrence of liver damage. 1

SummaryOxidative stress plays a role in the pathogenesis of many chronic diseases. It is recognized in overt hypothyroidism while its existence in subclinical hypothyroidism (SCH) is not well established. The aim of this study was to determine whether there was increased oxidation of lipids and proteins in SCH, and examine their association with lipids and thyroid hormones.Methods: Male adults (35-59 years) with SCH (n=467) and euthyroid controls (n=190) were studied. Anthropometric measurements, plasma lipids, thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3), total antioxidant capacity (T-AOC), lipid peroxidation products, malondialdehyde (MDA), advanced oxidation protein products (AOPP) and dityrosine concentrations were measured.Results: Plasma concentrations of MDA were significantly higher (pConclusion: Oxidative stress was increased in subclinical hypothyroidism as evidenced by the elevated lipid peroxidation product, malondialdehyde, while protein oxidation was absent. Thus, reduction of oxidative stress may be beneficial in patients with subclinical hypothyroidism

SummaryBackground: This study investigated the effects of a nutritionally relevant intake of eicosapentaenoic (EPA) and docosahexaenoic (DHA) fatty acids derived from oily fish or a fish oil supplement on selected cardiovascular risk factors in average middle-aged individuals.Methods: Thirty-three participants were randomized to receive salmon (oily fish) providing 274 mg EPA + 671 mg DHA/day or a commercial fish oil supplement providing 396 mg EPA + 250 mg DHA/day in a cross-over trial over an 8-week period separated by a 6-month washout period. Blood samples were collected before and after each intervention and lipids, inflammatory and oxidative stress parameters were determined.Results: Plasma levels of EPA, DHA and total n-3 fatty acids significantly increased after both interventions. A decreasing trend in triglycerides was more pronounced with salmon than with the fish oil supplement, but the changes noticed were not significant. Although there were no relevant changes in inflammatory marker concentrations at the end of both interventions, significant negative correlations were noticed between total plasma n-3 fatty acids and soluble intercellular adhesion molecule and Creactive proteinconcenthroughout the whole intervention period (pConclusion: Following two sets of recommendations for n- 3 fatty acids intake aimed at the general public had only a moderate effect on the selected cardiovascular risk factors in average healthy middle-aged subjects over a short-term period

SummaryBackground: The aim of the study was to evaluate parameters of oxidative and nitrosative stress as well as antioxidative parameters in a group of renal transplant recipients with stable graft function and no clinical signs of cardiovascular disease. We also aimed to determine the correlations among these parameters and to evaluate potential differences in all the biomarkers with regard to the immunosuppression protocol.Methods: We enrolled 57 renal transplant recipients and 31 controls who were age and sex matched with the renal transplant recipients. All of the patients included in this study had post-renal transplant surgery at least 12 months earlier and were on standard immunosuppressive therapy. In this study, we determined thiobarbituric acid-reactive substances in plasma and red blood cells and advanced oxidation protein products, nitrosative stress parameters (asymmetric and symmetric dimethylarginine - ADMA and SDMA), and antioxidative parameters (total SH groups and catalase activity).Results: The results of our study demonstrated that the levels of oxidative and nitrosative stress were significantly increased compared to the healthy population (pConclusion: There was no significant difference in oxidative and nitrosative stress parameters with respect to the immunosuppressive protocol.

The aim of the present study was to assess the effects of two metabolically different exercise programs on the redox state of women who were physically inactive before the beginning of the study. For this purpose, participants (women 25±5 years old) chose one of two popular fitness programs, Pilates or Tae Bo, and attended it 3 times a week for 12 weeks. At the beginning and end of the study, body composition analysis and venous blood sampling were performed. The levels of superoxide anion radical, hydrogen peroxide, nitric oxide and lipid peroxidation were measured in plasma, and the levels of reduced glutathione and the activity of superoxide dismutase and catalase were measured in erythrocytes. Only the Tae Bo program induced changes (positive) in body composition, whereas both exercise programs induced slight oxidative stress in exercisers. In the Tae Bo group, the levels of hydrogen peroxide were significantly increased, whereas the levels of reduced glutathione were decreased after three months of training. In the Pilates group, hydrogen peroxide and catalase activity were increased, and nitrites decreased. However, at the end of the study, those two groups had no significantly different values for any pro/antioxidant compared with the subjects who served as controls. This finding suggests that moderate physical activity, such as recreational fitness programs, may induce the increased production of reactive oxygen species but do not lead to a serious disturbance of the redox homeostasis of exercisers.

Ispitivanja primene vodonika u kliničkim uslovima su prilično nova, međutim efikasnost molekularnog vodonika u dosadašnjim kliničkim ispitivanjima je evidentna. Cilj ovog istraživanja bio je da se ispita uticaj suplementacije sa vodom obogaćenom molekularnim vodonikom (HRW) na bazalne vrednosti parametara oksidativnog stresa kod vrhunskih džudista. Eksperimentalni tretman odvijao se u dve faze. U prvoj fazi ispitanici eksperimentalne grupe (N=6) su bili suplementirani sa HRW (4g) u trajanju od 28 dana, dok je kontrolna grupa (N=6) koristila placebo. Nakon suplementacije ispitanici su imali „Washout“ period u trajanju od 28 dana, a zatim je ponovljen identičan protokol na ispitanicima u trajanju od 28 dana. U drugoj fazi eksperimentalnog tretmana ispitivani su akutni efekti suplementacije molekularnim vodonikom na acido baznu ravnotežu vrhunskih džudiskinja. U ovoj fazi studije ispitanice eksperimentalne grupe (N=4) su suplementirane sa HRW (4g) , dok je kontrolna grupa (N=4) koristila placebo. Nakon suplementacije ispitanice su imale „Washout“ period u trajanju od 4 dana, a zatim je ponovljen identičan protokol na ispitanicama pri čemu su ispitanice eksperimentalne grupe (N=4) koristile placebo, dok je kontrolna grupa (N=4) suplementirana sa HRW. Rezultati ispitivanja aktivnosti antioksidativnih enzima kod Placebo i HRW grupe ukazuju na pojavu statistički značajnih razlika u antioksidativnom statusu kod vrhunskih džudista. Odgovor organizma usled unosa HRW ogleda se u povećanju aktivnosti enzima SOD i sadržaja GSH usled sprovođenja programirane fizičke aktivnosti. Takođe, trend opadanja sadržaja MDA u HRW grupi je bio detektabilan. Primena HRW nije izazvala statistički značajne promene u biohemijsko hematološkim parametrima. Akutni efekti suplementacije sa HRW uticali su statistički značajno na redukciju akumulacije laktata kao i na povećanje pH krvi nakon primene specifičnog džudo fitnes testa (SJFT) kod vrhunskih džudistkinja. Ipak, primena suplementa nije značajno uticala na poboljšanje performansi džudistkinja. Dobijeni rezultati ukazuju na ergogenu antioksidantnu moć molekularnog vodonika, pri čemu su dalja istraživanja neophodna kako bi se podesili uslovi u kojima bi molekularni vodonik ispoljio svoje maksimalno delovanje i u sportu. Research of hydrogen application are quite new in clinical conditions, however the efficiency of molecular hydrogen in previous clinical studies is evident. The aim of this study was to investigate the effect of supplementation with hydrogen rich water (HRW) on baseline oxidative stress parameters in elite male judokas. Experimental treatment was carried out in two phases. In the first phase, experimental group (N=6) were supplemented daily with HRW (4g) for 28 days, while the control group (N=6) was used placebo. After supplementation intake all subjects had "washout" period for 28 days, after which the same protocol was repeated for a period of 28 days. In the second stage of experimental treatment, acute effects of supplementation with molecular hydrogen on acid base balance of elite female judokas were examined. In this phase of the study, subjects from experimental group (N=4) were supplemented daily with HRW (4g), while the control group (N=4) used placebo. After supplementation all subjects had a "washout" period for a 4 days, after which the same protocol was repeated wherein the members of the experimental group (N=4) used placebo, while the control group (N=4) was supplemented daily with HRW. The results of analyzed antioxidant enzymes activity in placebo and HRW group indicated on the occurrence of statistically significant differences in antioxidant status of elite judo athletes. Obtained results indicate that response of the body due to intake HRW is reflected in increasing levels of SOD and GSH due to the implementation of programmed physical activity. Also, trend of MDA decreasing in HRW group was detectable. In addition, application of HRW did not cause negative changes in the biochemical and hematological parameters. Acute effects of supplementation were affected to a statistically significant reduction in lactate accumulation and pH increase after applied special judo fitness test (SJFT) on the top judoka. However, use of supplements had no significant effect on improving performance in elite judokas. Obtained results confirm ergogenic antioxidant power of molecular hydrogen, wherein further studies are necessary in order to adjust the conditions in which molecular hydrogen will exerted its maximum in sport.

Raloksifen je predstavnik selektivnih modulatora estrogenih receptora koji se koristi u terapiji osteoporoze i invazivnog oblika raka dojke u postmenopauzi. Raloksifen se relativno dobro resorbuje iz gastrointestinalnog trakta, ali pri prvom prolasku kroz jetru podleže biotransformaciji u značajnom procentu, što je uzrok njegove niske biološke raspoloživosti. Bioraspoloživost kod ljudi iznosi 2%, a kod Wistar pacova 39%. Različite supstance se koriste da bi se poboljšala bioraspoloživost lekova. Žučne kiseline, kao što je deoksiholna kiselina, omogućavaju bolji prolazak kroz biološke membrane drugim supstancama, te mogu povećati bioraspoloživost lekova. Apigenin je široko rasprostranjeni flavonoid koji inhibiše različite metaboličke puteve i na taj način može usporiti metabolizam i eliminaciju i povećati koncentraciju lekova u krvi. Ciljevi ovog istraživanja su bili da se ispita da li apigenin i natrijum-deoksiholat mogu povećati bioraspoloživost raloksifena, njihov uticaj na biohemijske parametre i parametre hemostaze, kao i da se ispita antioksidativni potencijal apigenina. Ispitan je i uticaj apigenina na akutno oštećenje jetre usled primene toksične doze paracetamola. U istraživanju su korišćeni zdravi, beli pacovi muškog roda, soja Wistar. U ogledu su ukupno korišćene 84 eksperimentalne životinje. Sva ispitivanja na životinjama je odobrila Etička komisija Univerziteta u Novom Sadu. Raloksifen je primenjen intravenski i per os, dok su natrijum-deoksiholat i apigenin aplikovani peroralno. Uzorci krvi, urina i fecesa su korišćeni za određivanje farmakokinetskih parametara, dok su za određivanje biohemijskih, hemostatskih i parametara oksidativnog stresa korišćeni serum i uzorci jetre laboratorijskih životinja. Pretretman natrijum-deoksiholatom je doveo do smanjenja koncentracije raloksifena u krvi zbog olakšanog i brzog prodora raloksifena u periferne kompartmane. Time je značajno produženo poluvreme eliminacije i srednje vreme zadržavanja raloksifena i značajno je povećan volumen distribucije raloksifena. Apigenin je doveo do manjeg pada koncentracije raloksifena u prvim satima nakon intravenske primene raloksifena, dok su koncentracije raloksifena bile značajno više nakon osmog časa od primene leka. Uticaj raloksifena na biohemijske parametre je bio značajno veći nakon intravenske nego nakon peroralne primene. Nakon intravenske primene raloksifena je značajno povećana aktivnost enzima jetre, ALP, ALT, AST i GGT, dok su pokazatelji funkcije bubrega, urea, mokraćna kiselina i kreatinin bili sniženi. U grupama koje su pretretirane natrijum-deoksiholatom i apigeninom vrednosti ovih parametara bile su niže u odnosu na grupu tretiranu samo raloksifenom. Statistički najznačajniji uticaj je imala primena trojne kombinacije, raloksifena, natrijum-deosiholata i apigenina, koja je dovela do značajnog pada aktivnosti enzima jetre, i u odnosu na grupu tretiranu raloksifenom i u odnosu na kontrolnu grupu. Kod životinja tretiranih kombinacijom apigenina i paracetamola pokazatelji toksičnosti su bili značajno niži, naročito vrednosti ALT i ALP, u odnosu na grupu koja je dobijala samo paracetamol. Hepatotoksičnost izazvana toksičnom dozom paracetamola je potvrđena i histopatološkim promenama na jetri, koje nisu primećene u grupi životinja tretiranih kombinacijom apigenina i paracetamola. Ispitivanjem je utvrđeno da apigenin može da spreči paracetamolom indukovano povećanje nivoa MDA, što ukazuje da apigenin pozitivno utiče na očuvanje integriteta ćelije. Aktivnost enzima CAT i GR u homogenatima jetre je bila značajno povećana nakon primene toksične doze paracetamola u odnosu na kontrolnu grupu. Aktivnost enzima CAT i GR u grupi tretiranoj kombinacijom apigenina i paracetamola je bila približna vrednostima u kontrolnoj grupi. Na osnovu rezultata istraživanja može se zaključiti da natrijum-deoksiholat i apigenin značajno utiču na farmakokinetiku raloksifena. Primena natrijum-deoksiholata dovela je do pada koncentracije raloksifena u krvi, značajnog prelaska raloksifena iz krvi u periferne kompartmane i povećanja njegovog volumena distribucije, dok je apigenin značajno usporio metabolizam i eliminaciju raloksifena i doveo do njegovog produženog zadržavanja u krvi. Natrijum-deoksiholat i apigenin su pokazali pozitivan uticaj na biohemijske parametre, parametre hemostaze i smanjenje nivoa oksidativnog stresa. Kombinacija natrijum-deoksiholata i apigenina je pokazala sinergistički uticaj na navedene parametre, odnosno dovela je do značajnih promena u odnosu na pojedinačnu primenu ovih supstanci. Rezultati ispitivanja ukazuju na to da apigenin smanjuje stepen lipidne peroksidacije i da dovodi do značajnog povećanja enzimskih antioksidantnih mehanizama odbrane kod pacova kod kojih je hepatotoksičnost indukovana paracetamolom., Raloxifene is selective estrogen receptor modulator used in treatment of osteoporosis and invasive breast cancer in postmenopausal women. Raloxifene is well absorbed from the gastrointestinal tract, but undergoes extensive first-pass metabolism, which results in very low bioavailability of raloxifene, 2% in humans, and 39% in Wistar rats. Various supstances are used for increasing bioavailability of other drugs. Bile acids, such as deoxycholic acid, promote transport of other supstances through biological membranes, and consequently, may increase their bioavailability. Apigenin is a widespread flavonoid, which inhibits different metabolic pathways. Thus, apigenin can slow down metabolism and elimination of drugs, and raise drug concentration in blood. Aims of this study were to investigate if apigenin and sodium deoxycholate could increase bioavailability of raloxifene, their influence on biochemical and hemostasis parameters, and to investigate antioxidative potential of apigenin. Furthermore, influence of apigenin on acute liver damage after toxic dose of paracetamol was examined. In vivo experiments were performed on 84 laboratory healthy male Wistar rats. All experiments were approved by Ethics Committee of University of Novi Sad. Raloxifene was applied intravenously and per os, while sodium deoxycholate and apigenin were given perorally. Blood, urine and feces samples were used for pharmacokinetic parameters measurement, whereas serum and liver samples were used for evaluation of biochemical, hemostasis and oxidative stress parameters. Pretreatment of sodium deoxycholate led to raloxifene blood concentration decrease due to easier penetration of raloxifene in peripher compartments. As a result, raloxifene half-life and mean residence time were significantly longer and volume of distribution was increased. Apigenin caused lower decrease in raloxifene concentration in first few hours after raloxifene intravenous application, while raloxifene concentrations after apigenin pretreatment were significantlny higher 8 hours after raloxifene application. Influence of raloxifene on biochemical parameters was more significant after intravenous than after per os application. Intravenous application of raloxifene led to increased activity of liver enzymes, ALP, ALT, AST and GGT, while parameters of kidney function, urea, uric acid and creatinine were decreased in comparison to the control group. In experimental groups pretreated with sodium deoxycholate and apigenin these parameters were lower than in the group treated only with raloxifene. Statistically the most significant effects were in the group treated with combination of raloxifene, sodium deoxycholate and apigenin, which caused significant decrease in activity of liver enzymes compared both with raloxifene and control group of animals. In experimental animals treated with combination of apigenin and paracetamol bioindicators of paracetamol toxicity were significantly lower, especially activity of ALT and ALP, in comparison to the group treated only with paracetamol. Hepatotoxicity induced by toxic dose of paracetamol was also confirmed by histopathological alterations in liver, which were not observed in the experimental group treated with combination of apigenin and paracetamol. In this study it was confirmed that apigenin could prevent paracetamol-induced MDA level increase, which suggests that apigenin have positive effects on cell integrity. Activity of CAT and GR in liver homogenates was significantly increased after toxic dose of paracetamol in comparison to the control group, while activity of these enzymes in the group treated with apigenin and paracetamol was similar to values in the control group. Results of this study showed that sodium deoxycholate and apigenin can significantly change pharmacokinetic parameters of raloxifene. Sodium deoxycholate caused signicant decrease in raloxifene blood concentration, extensive distribution from blood to peripheral compartments and increase of raloxifene volume of distribution. Apigenin inhibited metabolism and elimination of raloxifene and thus prolonged half-life and mean residence time of raloxifene. Sodium deoxycholate and apigenin showed positive effects on biochemical and hemostasis parameters and decreased the level oxidative stress. Combination of sodium deoxycholate and apigenin showed synergistic effects on these parameters in comparison to effects of separate application of sodium deoxycholate and apigenin. The result of our study indicates that apigenin inhibits the level of lipid peroxidation and significantly increase the enzyme antioxidant defence mehanisms in paracetamol induced hepatotoxicity in rats.

Bosiljak (Ocimum basilicum L.) pripada familiji Lamiaceae i jedna je od najrasprostranjenijih i najčešće gajenih biljaka širom sveta. Njegova upotreba je široko rasprostranjena zbog bogatog sadržaja i skladnog odnosa vitamina, minerala kao i različitih fenolnih jedinjenja kao glavnih nosilaca antioksidativne aktivnosti. Poslednjih godina velika pažnja je usmerena ka fenolnim fitokomponentama kao potencijalnim promoterima zdravlja. Ciljevi ovog rada bili su ispitivanje kvalitativnih i kvantitativnih karakteristika, kao i biohemijskih aktivnosti ekstrakata bosiljka dobijenih različitim rastvaračima, kao i ispitivanje uticaja farmaceutsko-tehnološke formulacije u obliku mikrovezikula ekstrakta bosiljka na glikemijski, lipidni i oksido-redukcioni status kod oglednih životinja, u odnosu na vodeni ekstrakt. In vitro ispitivanja su uključivala analizu 100 ekstrakata dobijenih različitim rastvaračima i stepenom usitnjenosti. Ukupni fenolni i flavonoidni sadržaj, kao i antioksidativna aktivnost određena je spektrofotometrijskom metodom. Takođe, kvantifikovane i kvalifikovane su fenolne komponente (fenolne kiseline i flavonoidi) primenom visokoefikasne tečne hromatografije (HPLC). In vivo ispitivanje je rađeno na 84 albino laboratorijska pacova soja Wistar. Per os su primenjivani vodeni ekstrakt bosiljka, rastvor natrijumove soli monoketoholne kiseline, njihove kombinacije, kao i ekstrakt u obliku farmaceutsko-tehnološke formulacije, mikrovezikula. Posle sedmodnevnog tretmana, merene su koncentracije glukoze u krvi, a nakon žrtvovanja, u serumu su određeni parametri lipidnog statusa i antioksidativnog stresa. Ex vivo analizama procenjivani su efekti ekstrakta bosiljka na enzimske i neenzimske parametre antioksidativnog odbrambenog sistema i oksidativne modifikacije lipida. Praćen je i uticaj tretmana na akutno oštećenje jetre usled primene paracetamola. Svi analizirani ekstrakti bosiljka su pokazali prisustvo velikog broja fenolnih jedinjenja iz klase fenolnih kiselina i flavonoida. Detektovane i kvantifikovane aktivne komponente bile su fenolne kiseline: hlorogenska, p-hidroksibenzoeva, kafena, vanilinska, ferulna, rozmarinska i cimetna, kao i apigenin, kvercetin, naringenin i rutin kao predstavnici flavonoidnih jedinjenja. Primena ekstrakta bosiljka i natrijumove soli monoketoholne kiseline, same ili u kombinaciji, a takođe i u farmaceutsko-tehnološkoj formulaciji, pokazale su smanjenja oštećenja tokom oksidativnog stresa, značajno antidijabetesno delovanje, povoljan uticaj na lipidni status i protektivni efekat na funkcije jetre i bubrega. Na osnovu dobijenih rezultata može se zaključiti da ekstrakt bosiljka sadrži značajnu količinu fenolnih jedinjenja odgovornih za antioksidativnu aktivnost. Vodeni ekstrakt bosiljka, sam ili u kombinaciji sa monoketoholnom kiselinom, i u obliku farmaceutsko-tehnološke formulacije snižava nivo glukoze u krvi, ubrzava obnovu β ćelija, ima povoljan efekat na lipidni status i ne dovodi do toksičnih promena na jetri i bubrezima kod eksperimentalnih životinja., Basil (Ocimum basilicum L.) belongs to the family Lamiaceae and represents one of the most widespread and most commonly cultivated plant worldwide. Its use is widespread due to its rich content and harmonious relationship of vitamins, minerals and various phenolic compounds which are responsible for the antioxidant activity. In recent years, great attention is directed to phenolic phytocomponents as potential promoters of health. The objectives of this study were to test the qualitative and quantitative characteristics, as well as the biochemical activity of basil extracts obtained by various solvents and investigate the impact of pharmaceutical technological formulation of basil extract on glycemic, lipid and oxidation-reduction status of the aqueous extract in the experimental animals. In vitro studies have included 100 extracts obtained by various solvents and degrees of fragmentation. Total phenolic and flavonoid content, as well as antioxidant activity was determined by spectrophotometric method. Also, phenolic compounds (phenolic acids and flavonoids) were qualified and quantified using high performance liquid chromatography (HPLC). In vivo testing was performed on 84 laboratory albino Wistar rats. The aqueous extract of basil, a solution of sodium salt of monoketocholic acid, their combinations, as well as an extract in the form of the pharmaceutical technological formulation were administered per os. After seven-day-treatment, the concentrations of glucose were measured in the blood and after sacrificing of animals, the lipid and antioxidative parameters were determined in serum. Ex vivo analysis assessed the effect of the extract of basil on the parameters of enzymatic and non-enzymatic antioxidant defense system and the oxidative modification of lipids. The effect of paracetamol on acute liver injury was also monitored. All analyzed basil extracts showed the presence of the large number of phenolic compounds from the class of phenolic acids and flavonoids. Chlorogenic, p-hydroxybenzoic, caffeic, vanillic, ferulic, rosmarinic, and cinnamic acid, as well as apigenin, quercetin, naringenin and rutin were detected and quantified. The application of basil extracts and the sodium salt of monoketocholic acid, either alone or in combination, and also in pharmaceutical technological formulation of microvesicles has shown a reduction in damage during oxidative stress, then a significant antidiabetic activity, favorable effect on the lipid profile and protective effect on the liver and kidney function. Based on these results it can be concluded that basil extract contains significant amounts of phenolic compounds responsible for antioxidant activity. The aqueous extract of basil, alone or in combination with monoketocholic acid, and in the form of the pharmaceutical technological formulations lowers glucose levels in the blood, accelerates the restoration of β cells, has a favorable effect on the lipid status and does not lead to toxic changes in the liver and kidneys in experimental animals.

Ćelijska smrt je jedan od uzroka patoloških promena u jetri tokom dijabetesa. Protein koji može imati važnu ulogu u ovim procesima, bilo kao marker nekroze ili aktivator signalnih puteva koji vode ćelijskoj smrti ili preživljavanju, je HMGB1. U vanćelijsku sredinu HMGB1 dospeva iz nekrotičnih, oštećenih ćelija i aktiviranih imunskih ćelija. U ovoj doktorskoj disertaciji ispitivana je uloga ovog proteina u određivanju profila ćelijske smrti u jetri pacova sa dijabetesom kroz njegov doprinos u modulaciji procesa apoptoze i autofagije. Rezultati ovog istraživanja pokazali su da je nivo HMGB1 proteina u serumu i jetri pacova sa dijabetesom u korelaciji sa nivoom oštećenja jetre i nekroze, što ga svrstava među potencijalne biološke markere za praćenje dijabetičnih komplikacija u jetri. Na aktivaciju procesa apoptoze i autofagije HMGB1 protein deluje dvojako, kroz aktivaciju nizvodnih signalnih puteva pokrenutih interakcijama vanćelijskog HMGB1 sa TLR4 i RAGE receptorima, i kroz uticaj citoplazmatskog HMGB1 na formiranje autofagozoma kroz interakcije sa Beklinom1. Pokazano je da nivo vanćelijskog HMGB1 proteina i njegov doprinos profilu ćelijske smrti u jetri zavisi od intenziteta prisutnog oksidativnog stresa. U uslovima oksidativnog stresa prisutnog u nelečenom dijabetesu, HMGB1 favorizuje puteve apoptoze i doprinosi ćelijskoj smrti i oštećenju jetre. Smanjenjem oksidativnog stresa i nivoa vanćelijskog HMGB1 melatoninom ili etil piruvatom, favorizuje se delovanje citoplazmatskog HMGB1 u pravcu protektivne autofagije i uklanjanja oštećenih unutarćelijskih struktura. Ovi podaci ukazuju na terapeutski značaj melatonina i etil piruvata u regulaciji aktivnosti HMGB1 u dijabetesu i na ulogu HMGB1 proteina u regulaciji i povezivanju apoptoze i autofagije. Cell death underlies hepatic pathological changes in diabetes. HMGB1 is a protein with important roles in these changes, whether as a necrotic marker or a pathway activator determining cell death or survival. HMGB1 is released into the extracellular space by damaged or necrotic cells and activated immune cells. The role of HMGB1 in determining the profile of cell death in the liver of diabetic rats was studied by examining its role in the modulation of apoptosis and autophagy. The levels of HMGB1 protein in the serum and liver of diabetic rats correlate with the degree of necrosis and liver damage, thus rendering HMGB1 as a potential biological marker of diabetic complications. To activate apoptosis and autophagy, HMGB1 protein acts by stimulating downstream signaling pathways triggered by interactions of extracellular HMGB1 with TLR4 and RAGE receptors, and through the activity of cytoplasmic HMGB1 on the formation of autophagosomes via interaction with Beclin1, which has a central role in autophagy. Extracellular expression of HMGB1 and its contribution to the cell death profile in the liver depends on the level of oxidative stress. In oxidative stress accompanying untreated diabetes, HMGB1 promotes apoptosis and resulting liver damage. The reduction of oxidative stress by melatonin and HMGB1 release by ethyl pyruvate stimulates the cytoprotective role of cytosolic HMGB1 in autophagy activation These results points to the therapeutic significance of melatonin and ethyl pyruvate in the regulation of HMGB1 activity in diabetes, and the role of HMGB1 in the regulation and interplay between apoptosis and autophagy. 1

Uvod:Apendicitis je oboljenje, koje se najčešće javlja u dečjem uzrastu. Poslednjih godina se laparoskopska apendektomija sprovodi sve češće u ovom uzrastu, međutim ne postoji jasan konsenzus o optimalnom izboru anestetika za održavanje opšte anestezije u toku ove hirurške metode u dečjem uzrastu. Cilj istraživanja: Utvrditi uticaj vrste anestezije i vrste hirurške procedure na odgovor organizma na hirurški stres tokom operacije crvuljka. Metodologija: Klinički prospektivno istraživanje je sprovedeno na Klinici za dečiju hirurgiju, na Institutu za zdravstvenu zaštitu dece i omladine Vojvodine. Istraživanjem je obuhvaćeno 120 dece, uzrasta od 7 do 17 godina, bez postojećih komorbiditeta, koji su operisani zbog zapaljenja crvuljka. U zavisnosti od vrste operativnog zahvata i vrste primenjene anestezije deca su podeljena u četiri grupe bolesnika. Kod sve četiri ispitivane grupe uzimana je venska i kapilarna krv, nekoliko minuta nakon uvoda u anesteziju, u momentu vađenja crvuljka iz trbuha i 12 časova nakon kraja hirurške intervencije. Laboratorijski su određeni markeri oksidativnog stresa (TBARS), metaboličkog odgovora na hirurški stres (laktat, glikemija), inflamatornog odgovora organizma (IL-6, leukociti), gasne analize, parametri oksigenacije i ventilacije, i hemodinamski parametri ispitanika. Rezultati:U istraživanju je dobijen rezultat da je zapaljenje crvuljka oboljenje koje se češće javlja kod dečaka. Tokom apendektomije u dečjem uzrastu, sevofluran je bolje kontrolisao arterijsku tenziju, dok na srčanu frekvencu vrsta anestezije nije imala uticaja. Sevofluran je anestetik tokom čije primene je manji inflamatorni odgovor tokom laparoskospske apendektomije. Propofol deluje suprimirajuće na oksidativni stres, ali nije nađena statistička značajnost u odnosu na vrednosti dobijene analizom uticaja sevoflurana na parametre oksidativnog stresa. Zaključak: Laparoskopska apendektomija u odnosu na laparotomiju nije praćena većim stepenom hirurškog stresa, a sevofluran je anestetik koji tokom anestezije za laparoskopsku apendektomiju u dečijem uzrastu daje bolju kontrolu kliničkog, metaboličkog i inflamatornog odgovora., Introduction: Appendicitis is a disease which appears most commonly in children. In recent years appendectomy in children is performed by laparoscopy, but there is no consensus yet on the optimal choice of anesthetics during general anesthesia for this procedure. Aim: To determine the influence of type of anesthesia and type of surgical procedure for appendectomy, on surgical stress in children. Methodology: A prospective clinical trial in Clinic of pediatric surgery in Novi Sad, Vojvodina. The study included 120 children aged from 7 to 17 years, with no commorbidities, who underwent appendectomy. Children were divided in four groups based on the type of anesthesia and type of surgery they received. In all participants, venous and capillary blood was sampled for analyzis 10 minutes after induction of anesthesia, at the moment of appendix removal and 12 hours after the procedure. The laboratory analysis included markers of oxidative stress (TBARS), metabolic response to surgical stress (lactate, blood glucose), inflammatory response (IL-6, leucocites), bloodgas analyses, parameters of oxygentation and ventilation and haemodynamic parameters of the participants. Results: In the study appendicitis was more common in boys. During laparoscopic appendectomy sevoflurane controlled better the blood pressure, but not the heart rate. Sevoflurane maintained a better control of parameters of the inflammatory response. Propofol decreased the oxidative stress, but there was no statistical difference compared to the effects of sevoflurane on oxidative stress. Conclusion: Laparoscopic appendectomy shoved no difference in the level of surgical stress compared to laparotomy, and sevoflurane appeared as an anaesthetic which had a better control of the metabolic, clinical and inflammatory response.

Žučne kiseline (ŽK) su strukturno raznoliki molekuli, koji pored uloge koju ostvaruju putem žuči, deluju i kao signalni molekuli i ostvaruju kako endokrina tako i parakrina dejstva. Činjenica da je do sada u terapijske svrhe primenjivana samo ursodeoksiholna kiselina (UDK), posledica je brojnih ograničenja u mogućnosti primene ostalih prirodnih ŽK, i ističe potrebu za otkrivanjem novih sintetskih ŽK i liganda. Cilj istraživanja bio je ispitivanje sintetske 12-monoketoholne kiseline (MK) i prirodne UDK u modelu holestaze i aloksanom izazvanog dijabetesa. Ispitivanja su vršena na pacovima soja Wistar. Analizirana je telesna masa, glikemija, pokazatelji jetrene funkcije (AST; ALT, γ-GT, ukupni i direktni bilirubin), a iz homogenate jetre određen je intenzitet lipidne peroksidacije i aktivnost antioksidativnih enzima (CAT, GSH-Px, GSH-R, GSH-ST). Isečci tkiva jetre su histološki obrađeni i bojeni hematoksilin-eozin metodom i histohemijskim metodama (retikulin, Mallory, Periodic Acid Schiff- Alcian Blue (PAS/AB)). Imunohistohemijski je ispitana proliferacija hepatocita (Ki-67), markeri apoptoze (p53, Bcl-2, Bcl-X, Bax) i ekspresija nuklearnog farnesoid X receptora (FXR). Rezultati istraživanja pokazuju da ispitivane ŽK pomažu očuvanje telesne mase u holestazi i dijabetesu, i značajno snižavaju glikemiju kod dijabetičnih jedinki. Parametri jetrene funkcije u holestazi i dijabetesu su regulisani primenom MK i UDK. Obe ŽK u značajnoj meri smanjuju intenzitet lipidne peroksidacije i pojačavaju enzimsku antioksidativnu odbranu hepatocita u holestazi i dijabetesu. Ekspresija markera apoptoze nije značajno promenjena izazvanjem modela holestaze i dijabetesa, kao ni primenom ispitivanih ŽK. Nasuprot tome, izazivanje holestaze i dijabetesa značajno smanjuje proliferaciju hepatocita, dok primena MK i UDK poništava ovaj efekat i značajno povećava proliferaciju hepatocita. Hiperglikemija u aloksanskom dijabetesu nije dovela do pojačane ekspresije FXR. Izazivanje holestaze kod zdravih i dijabetičnih životinja dovelo je do porasta ekspresije FXR, koja je redukovana primenom MK i UDK. Na osnovu dobijenih rezultata može se zaključiti da sintetska 12-monoketoholna kiselina pokazuje slična hipoglikemijska, hepatoprotektivna i antioksidativna dejstva kao i prirodna ursodeoksiholna kiselina. Bile acids (BAs) are structurally diverse molecules, which have theroles in the digestive system, which are exercised through the bile. Beside those, BAs act as a signaling molecules and achieve endocrine and paracrine effects. In addition to its own metabolism, bile acids modulate the metabolism of lipids and glucose. The fact that so far only ursodeoxycholic acid (UDC) is used for therapeutic purposes, speak clearly about of numerous limitations on the application of other natural BAs, and highlights the need to develop new synthetic Bas and ligands. The aim of this study was to investigate the influence of synthetic 12-monoketocholic acid (MC) and natural bila acid UDC in the model of cholestasis and alloxan-induced diabetes. Tests were performed on male Wistar rats. We analyzed the body mass, glucose, liver function tests (AST, ALT, γ-GT, total and direct bilirubin). Using liver tissue homogenates we determined intensity of lipid peroxidation (by concentration of malondilaldehyde) and the activity of antioxidant enzymes (CAT, GSH-Px, GSH -R, GSH-ST). Liver tissue were histologically processed and stained with hematoxylin-eosin method and histochemical methods (reticulin, Mallory, Periodic Acid Schiff- Alcian Blue (PAS / AB)). Imunohistochemical examination included hepatocyte proliferation (Ki-67), markers of apoptosis (p53, Bcl-2, Bcl-X, Bax), and expression of the nuclear farnesoid X receptor (FXR). Results of the research show that MC prevented decrease in body mass during cholestasis and diabetes, and significantly reduced glycemia in diabetic animals. The liver function tests in cholestasis and diabetes are normalised by MC and UDC aplication. Both BAs significantly reduce lipid peroxidation and enhance enzymatic antioxidant defense of hepatocytes in cholestasis and diabetes. The expression of markers of apoptosis was not significantly changed in models of cholestasis and diabetes, as well as the application of the tested BAs. In contrast, in cholestasis and diabetes model, the proliferation of hepatocytes was significantly reduced, while the use of MC and UDC reversed this effect and significantly increased the proliferation of hepatocytes. Hyperglycemia in alloxan-induced diabetes did not lead to overexpression of FXR. Induction of cholestasis in healthy and diabetic animals resulted in an increase in the expression of FXR, which is reduced by using the MK and the UDC. Based on these results we can conclude that a synthetic 12-monoketocholic acid shows similar hypoglycemic, hepatoprotective and antioxidant effects as natural ursodeoxycholic acid.

It is known that kidneys are target tissue for Cd toxicity but there is not enough literature data on kidney effects of BDE-209. Moreover, Cd as toxic metal and BDE-209 as organic halogenated pollutant can share similar mechanism of action like, oxidative stress which is already assumed. However, how will Cd act as prooxidant in presence of BDE-209 is still not investigated, therefore, the objective of this study was to assess this BDE-209 influence. Results of this study indicated slight decrease in MDA in rat's kidney homogenates after subacute exposure to Cd and/or BDE-209, while SOD activity was increased and content of -SH groups decreased. Based on dose-response assessment, BDE-209 did not influence Cd toxicity mediated by chosen oxidative stress parameters, namely the derived CEDL5 values were the same for single Cd and for Cd given in combination with the BDE-209. In conclusion, dynamic of oxidative process has influence on the parameters levels. Co-exposure with polybrominated organic pollutant did not influence Cd effects on oxidative stress parameters. Having in mind that this is pilot study, performed with only one dose of BDE-209, further investigations are necessary, including changes in dose ranges, duration of exposure etc. Moreover, results contributes to the issue of mixture toxicology even BDE-209 did not influenced oxidative stress as mechanism of Cd toxicity. Poznato je da su bubrezi ciljni organ toksičnosti kadmijuma, ali je mnogo manje literaturnih podataka o toksičnim efektima BDE-209 na bubrege. Kadmijum, kao toksičan metal i BDE-209, kao organska halogenovana zagađujuća supstanca mogu imati sličan mehanizam dejstva posredovan oksidativnim stresom. Međutim, kako će Cd da deluje kao prooksidant u prisustvu BDE-209 još uvek nije proučavano, te je cilj ovog rada bio da se proceni uticaj BDE-209 na oksidativni stres posredovan kadmijumom. Rezultati ovog istraživanja ukazuju na blagi pad koncentracije MDA u homogenatima bubrega pacova nakon subakutne ekspozicije Cd i/ili BDE-209, dok se aktivnost SOD povećava i sadržaj - SH grupa smanjuje. Na osnovu procene odnosa doza-odgovor, BDE-209 nije imao uticaja na toksičnost Cd izazvanu posredstvom oksidativnog stresa (posmatrajući odabrane parametre). Potvrda je dobijena i iz izračunatih donjih granica pouzdanosti doze od 5% - CEDL5 jer su vrednosti su bile iste za sam Cd i Cd primenjen u kombinaciji sa BDE-209. Može se zaključiti da dinamika oksidativnih procesa ima uticaj na vrednosti parametara oksidativnog stresa, da stoga istovremena ekspozicija Cd i polibromovanom organskom polutantu nije promenila uticaj samog Cd na parametre oksidativnog stresa. Imajući u vidu da je ovo pilot studija, izvršena sa samo jednom dozom BDE-209, potrebna su dalja istraživanja, uključujući i promene opsega doza, trajanje ekspozicije itd. Pored toga, rezultati ove studije doprinose tumačenju toksikologija smeša, iako BDE-209 nije imao uticaja na oksidativni stres kao mehanizam Cd toksičnosti za date uslove ekspozicije.

Biološki fakultet Ključna pretpostavka evolucione teorije starenja jeste da opadanje uzrasno-specifičnog preživljavanja organizama predstavlja rezultat smanjenja intenziteta prirodne selekcije nakon početka reprodukcije. U osnovi evolucije starenja nalaze se dva populaciono-genetička mehanizma: akumulacija mutacija (AM) i antagonistička plejotropija (AP). U skladu sa pretpostavkama evolucione teorije starenja, kod eksperimentalnih populacija pasuljevog žiška (Acanthoscelides obtectus) selektovanih više od 170 generacija za ranu (E režim) ili kasnu (L režim) reprodukciju, uočena je divergencija u dužini života i drugim preadultnim i adultnim osobinama životne istorije. Evolucija veće dužine života u L režimu u saglasnosti je sa pretpostavkama AP modela, dok u evoluciji kraćeg života E žižaka važnu ulogu ima akumulacija mutacija. Tretiranjem E i L žižaka pesticidima (parakvat i tebufenpirad) koji generišu slobodne radikale, potvrđena je pozitivna korelacija između dužine života i otpornosti na oksidativni stres. Budući da mitohondrija predstavlja glavno mesto sinteze energije i slobodnih radikala, u ovoj tezi je istraženo da li je uzrasno-specifična selekcija oblikovala varijabilnost mtDNK, nDNK i njihovih epistatičkih interakcija. Genotipizacijom mitohondrijskog COI gena i mikrosatelitskih lokusa pokazana je efikasnost uzrasno-specifične selekcije u međupopulacionoj divergenciji mitohondrijskog i jedarnog genskog pula kod pasuljevog žiška. Odvijanje procesa mito-jedarne koevolucije analizirano je korišćenjem mito-jedarnih linija u kojima su E i L mitohondrijski genomi kombinovani sa E i L jedarnim genomima. Pokazano je da su linije sa narušenim mito-jedarnim interakcijama imale niže aktivnosti mitohondrijskih kompleksa elektron transportnog lanca i lošiju preadultnu performansu. According to the evolutionary theory of ageing, age-specific decline in survival is the result of decreasing intensity of natural selection after the onset of reproduction. The evolution of ageing may be explained by two different, but not mutually exclusive, genetic mechanisms: antagonistic pleiotropy (AP) and mutation accumulation (MA). In accordance with evolutionary theory of ageing, two sets of the seed beetle (Acanthoscelides obtectus) experimental lines selected for more than 170 generations for early (E regime) or late (L regime) reproduction, show divergence in longevity and ageing patterns, as well as in other preadult and adult life history traits. Evolution of long life in L regime is predominantly based on antagonistic pleiotropy gene effects, while mutation accumulation plays important role in evolution of short life in E regime. By exposure of E and L beetles to two pesticides (paraquat and tebufenpyrad) that generate free radicals, it was confirmed that extended longevity in L beetles (especially in females) was associated with higher oxidative stress resistance. Additionally, since mitochondria are main site of free radical and energy production, effects of age-specific laboratory selection on mtDNA, nDNA and their epistatic interaction were assessed. Genotyping of mitochondrial COI gene and nuclear microsatellite loci showed that selection in E and L regimes led to divergent evolution of mtDNA and nDNA gene pools. Mitonuclear coevolution was investigated using mitonuclear introgression lines in which E and L mitochondrial genomes were expressed in both E and L nuclear background. It was found that lines with disrupted mitonuclear interactions had lower activity of the mitochondrial electron transport chain complexes and lower preadult performance.

The unceasing need for oxygen is in contradiction to the fact that it is in fact toxic to mammals. Namely, its monovalent reduction can have as a consequence the production of short-living, chemically very active free radicals and certain non-radical agents (nitrogen-oxide, superoxide-anion-radicals, hydroxyl radicals, peroxyl radicals, singlet oxygen, peroxynitrite, hydrogen peroxide, hypochlorous acid, and others). There is no doubt that they have numerous positive roles, but when their production is stepped up to such an extent that the organism cannot eliminate them with its antioxidants (superoxide-dismutase, glutathione-peroxidase, catalase, transferrin, ceruloplasmin, reduced glutathion, and others), a series of disorders is developed that are jointly called 'oxidative stress.' The reactive oxygen species which characterize oxidative stress are capable of attacking all main classes of biological macromolecules, actually proteins, DNA and RNA molecules, and in particular lipids. The free radicals influence lipid peroxidation in cellular membranes, oxidative damage to DNA and RNA molecules, the development of genetic mutations, fragmentation, and the altered function of various protein molecules. All of this results in the following consequences: disrupted permeability of cellular membranes, disrupted cellular signalization and ion homeostasis, reduced or loss of function of damaged proteins, and similar. That is why the free radicals that are released during oxidative stress are considered pathogenic agents of numerous diseases and ageing. The type of damage that will occur, and when it will take place, depends on the nature of the free radicals, their site of action and their source. Neprekidna potreba za kiseonikom je kontradiktorna sa činjenicom da je on toksičan po sisare. Naime, njegova jednovalentna redukcija može imati za posledicu proizvodnju kratkoživećih, hemijski vrlo aktivnih, slobodnih radikala i nekih neradikalskih agenasa (azot-oksida, superoksid anjon-radikala, hidroksilnog radikala, peroksilnog radikala, singletnog kiseonika, peroksinitrita, vodonik-peroksida, hipohloraste kiseline i dr.). Neosporno je da oni imaju brojne poželjne uloge, ali kada im se proizvodnja pojača toliko da organizam ne može svojim antioksidansima (superoksid dismutaza, glutation peroksidaza, katalaza, transferin, ceruloplazmin, redukovani glutation i sl.) da ih ukloni, razvija se serija poremećaja koji se jednim imenom nazivaju 'oksidativni stres'. Reaktivne kiseonične vrste koje karakterišu oksidativni stres su sposobne da napadnu sve glavne klase bioloških makromolekula, odnosno proteine, DNK i RNK molekule, a naročito lipide. Pod uticajem slobodnih radikala dolazi do peroksidacije lipida ćelijskih membrana, oksidativnih oštećenja DNK i RNK molekula, razvoja genetskih mutacija, fragmentacije i promene funkcije najrazličitijih proteinskih molekula. Sve to ima za posledicu: narušavanje propustljivosti ćelijskih membrana, poremećaja ćelijske signalizacije i homeostaze jona, smanjenja ili gubitka funkcija oštećenih proteina i sl. Zato se slobodni radikali, koji se oslobađaju tokom oksidativnog stresa, smatraju patogenim agensima mnogih bolesti i starenja. Koja vrsta oštećenja će se desiti i kada, zavisi od prirode slobodnih radikala, mesta delovanja i njihovog izvora.

Objectives: Peroxynitrite is species claimed to propagate ischemia/reperfusion damage. In this report levels of serum uric acid (UA), a peroxynitrite scavenger, are compared with creatine phosphokinase (CPK) in male patients before and after open-heart surgery in order to asses if increased levels of UA may protect heart from biochemical damage induced by peroxynitrite during the coronary by-pass grafting (CABG) intervention. Methods: 45 male patients (16 carvedilol pretreated (6.25 mg/ daily, during 6 weeks before surgery, mean age 55.3±1.7 years, range 50-71) and 29 patients without carvedilol pretreatment (mean age 58.3±1.4 years, range 47-73) underwent elective CABG were examined. Study inclusion criteria were CABG performed on two and more coronary-vessels with aortic crossclamp during 30-40 minutes. For assessment of patients objective health status before operations Euro- SCORE were used. Serum uric acid (UA) levels and creatine phosphokinase (CPK) were measured spectrophotometricaly by using a quantitative enzymatic assay. Results: Carvedilol pretreated patients had higher amount of serum UA (p

Cilj: Malondialdehid (MDA) je jedan od toksičnih produkata lipidne peroksidacije, a koristi se u evaluaciji oksidativnog stresa tijekom šećerne bolesti. Cilj ovog rada bio je ispitati učinak akarboze (inhibitora α-glukozidaza) na koncentraciju glukoze u serumu i MDA u homogenatu jetre NOD (engl. non-obese diabetic) miševa. Materijali i metode: U NOD miševa šećerna bolest inducirana je i.v. aplikacijom aloksan-monohidrata (75 mg/kg t. mase). NOD miševi podijeljeni su u 4 skupine (n=6): Kontrolni, zdravi NOD miševi (K), Kontrolni, zdravi NOD miševi tretirani 7 dana akarbozom (K/A), dijabetični NOD miševi (D) te dijabetični NOD miševi tretirani 7 dana akarbozom (D/A). Koncentracija glukoze u krvi izmjerena je glukoza oksidaza-peroksidaza metodom, a koncentracija MDA u homogenatu jetre određena je upotrebom metode s tiobarbiturnom kiselinom. Rezultati: Nakon sedmodnevnog tretmana akarbozom zabilježeno je statistički značajno smanjenje koncentracije glukoze u krvi u skupini D/A u odnosu na skupinu D (p, Aim. Malondialdehyde (MDA) is one of the toxic product of the lipid peroxida-tion, and that plays an important role in the evaluation of oxidative stress in the diabetes mellitus. The aim of this study was to study the effect of acarbo-se (α-glucosidase inhibitor) on serum glucose concentration and MDA in liver homogenate of NOD (non-obese diabetic) mice. Material and methods. Diabetes was induced in mice by i.v. administration of alloxan-monohydrate (75 mg/kg b.w.). The mice were divided into 4 groups (N=6): control, healthy mice (C), control healthy mice on 7-day treatment with acarbose (C/A), NOD mice (D), and NOD mice on 7-day acarbose treatment (D/A). The levels of blood glucose and of MDA in liver homogenate were determined by glucose oxidase-peroxidase method and a method with thiobarbituric acid, respectively. Results. Significant reduction in blood glucose levels was observed after 7-day administration of acarbose in D/A group compared to D group (p

The research was focused on biotic stress caused by Macrophomina phaseolina (Tassi) Goid., a plant pathogen that causes charcoal rot by infecting the root and lower stem of soybean. Differences in pathogenicity of fungi cultures from five isolates (two from sugar beet, one from maize, one from soybean and one from sunflower) have been investigated in seedlings of two soybean cultivars (Meli and Balkan). Comparison of antioxidant systems of soybean seedlings showed different reactions of plants infected with M. phaseolina isolates of different origin. The highest pathogenicity showed isolate from sugar beet ŠR62/4 (2.80), while no significant difference was found for isolates ŠR55(3)/09 (2.58), MphSo (2.32) and MphSu (2.26). The isolate from maize, MphK, showed lowest pathogenicity (1.94). For the most of the biochemical parameters tested (lipid peroxidation, phenylalanine ammonium-lyase activity, total polyphenols, tannins and proanthocyanidins contents) the isolate from sugar beet (ŠR55(3)/09) induced the most significant reaction in soybean seedlings among all isolates investigated. Presence of this pathogen in the field conditions should be further monitored and the crop rotation should be seriously taken into account. U radu je ispitivan biotički stres u klijancima soje izazvan gljivom Macrophomina phaseolina (Tassi Goid.), patogenom biljaka i uzročnikom ugljenaste truleži korena i donjeg dela stabla. Različite karakteristike izolata gljive (dobijenih sa šećerne repe (2), kukuruza (1), soje (1) i suncokreta (1)), ispitane su u dve sorte soje (Meli i Balkan). Poređenje antioksidantnih sistema u biljkama soje ukazalo je na različite odgovore između sorti soje inficiranih izolatima M. phaseolina različitog porekla. Izolati su izazvali intenzivnu reakciju u inficiranim klijancima: najveća patogenost zabeležena je kod izolata sa šećerne repe ŠR62/4 (2.80). Izolati ŠR55(3)/09 (2.58), MphSo (2.32) i MphSu (2.26) nisu pokazali statistički značajnu razliku u patogenosti. Izolat sa kukuruza, MphK, pokazao je najnižu patogenost (1.94). Za većinu ispitivanih biohemijskih parametara (lipidna peroksidacija, aktivnost fenilalanin amonijum-liaze, sadržaji ukupnih polifenola, tanina i proantocijanidina), izolat ŠR55(3)/09 sa šećerne repe, izazvao je najintenzivnije i najznačajnije reakcije u klijancima soje u poređenju sa svim ostalim ispitanim izolatima. Njegovo prisustvo u poljskim uslovima se mora podvrgnuti monitoringu, a o rotaciji useva na takvim oranicama se mora povesti više pažnje.

Aktivacija hipolamo-hipofizo-adrenalne (HHA) ose predstavlja jedan od odgovora organizma na izlaganje stresnim uslovima. Dok aktivacija HHA ose uzrokovana kratkotrajnim stresorima ima adaptivni karakter, hroničan stres može dovesti do brojnih patoloških promena među kojima su i poremećaji raspoloženja. Uzrok deregulacije HHA ose u stresnim uslovima još uvek nije u potpunosti utvrđen, a smatra se da može biti povezan sa i pojavom oksidativnog stresa. Stoga je ispitana i okarakterisana aktivnost HHA ose, merenjem koncentracije kortikosterona (CORT) kao markera stresnog odgovora, kao i mehanizmi regulacije antioksidativnih enzima CuZnSOD i MnSOD, kao biomarkera oksidativnog stresa, u uslovima akutnog, hroničnog i kombinovanog stresa u hipokampusu i prečeonoj kori mozga i jetri mužjaka pacova Wistar soja. Cilj ove teze je bio bolje razumevanje biohemijskih i molekularnih mehanizama delovanja antioksidativnih enzima u moždanim strukturama i jetri pacova nakon izlaganja 21-dnevnoj socijalnoj izolaciji, koja predstavlja jedan od životinjskih modela depresije. Akutno izlaganje imobilizaciji ili niskoj temperaturi u trajanju od 2 sata korišćeno je kao pokazatelj “normalnog“ odgovora na stres, dok je kombinacija hroničnog i akutnog stresa služila kao potvrda maladaptivnog efekta hroničnog stresa. Rezultati istraživanja su pokazali povećanu koncentraciju CORT-a u serumu životinja izloženih akutnim stresorima, dok su kod životinja izlaganih hroničnoj socijalnoj izolaciji vrednosti ostale nepromenjene. Međutim, značajno niža koncentracija CORT-a između životinja koje su izlagane kombinovanom i odgovarajućem akutnom stresu ukazala je na poremećaj u regulaciji HHA ose koji je izazvana hroničnom socijalnom izolacijom, a što je rezultovalo u neadekvatnm odgovoru na nove stresne situacije. Akutni stres imobilizacije doveo je do povećanja ekspresije MnSOD proteina u mitohondrijama obe moždane strukture, dok je kod životinja akutno izlaganih niskoj temperaturi ili hroničnom stresu izolacije nivo ovog proteina ostao nepromenjen. Međutim, aktivnosti CuZnSOD i MnSOD-a u prečeonoj kori nisu pratile detektovanu proteinsku ekspresiju ukazujući time na moguće stanje oksidativnog stresa. Dodatni akutni stres doveo je do opadanja nivoa MnSOD proteina u mitohondrijama prečeone kore, što uz istovremeno povećanje u citosolnoj frakciji kao i oslobađanje citohroma c može ukazati na narušavanje integriteta mitohondrijske membrane... The response of the organisms to stressors involves activation of hypothalamic-pituitary-adrenal (HPA) axis. HPA axis activation is adaptive in the short term, but chronic exposure to stressful stimuli may lead to pathological changes, such as mood disorders. Although precise cause of deregulation of HPA axis due to stress exposure is still unknown, it is thought that may be associated with increased oxidative stress. Therefore, we examined and characterized HPA axis activity, by measuring the concentration of serum corticosterone (CORT) as marker of stress response, and mechanisms of regulations of antioxidative enzymes CuZnSOD and MnSOD, as biomarkers of oxidative stress, in the prefrontal cortex, hippocampus and liver of male Wistar rats exposed to acute, chronic and combined stress. The aim of this thesis is better understanding the biochemical and molecular mechanisms of action of antioxidative enzymes in the brain structures and liver of rats subjected to 21-day social isolation, that represent an animal model for depresion. Acute stressors of 2h immobilization or cold are used to indicate ’normal’ stress response, while combination of chronic and acute stress served as confirmation of maladaptive effect of chronic stress. Results showed increased concentration of serum CORT level in the animals exposed to acute immobilization or cold stress, while in animals expsosed to chronic isolation these values remain unchanged. However, significantly lower CORT level detected between animals exposed to combined and appropriate acute stress showed compromised HPA axis funcioning, caused by chronic social isolation, which has resulted in inadequate response to the novel stressful situations. Acute immobilization stress resulted in increased expression of MnSOD protein in mitochondria of both brain structures, while in animals acutely exposed to cold or chronic social isolation level of this protein remained unchanged. However, detected discrepancy between CuZnSOD and MnSOD activies and protein expression in the prefrontal cortex suggesting the possible state of oxidative stress. Additional acute stress decreased mitochondrial MnSOD protein level, which along with the increase in citosolnoj fraction and the release of cytochrome c indicate compromised integrity of the mitochondrial membrane in the prefrontal cortx...

Oksidativni stres nastao usled nesklada u produkciji slobodnih radikala i antioksidativnoj zaštiti sve češće se prepoznaje kao biohemijski mehanizam potencijalno uključen u patogenetska zbivanja u velikom broju sistemskih bolesti, pa tako i parodontopatije. Sa druge strane, parodontopatija kao inflamatorno destruktivno oboljenje i sama dovodi do pojačanog stvaranja slobodnih radikala. Ispitivanja pokazuju da parodontopatiju ne prati samo poremećaj u redoks ravnoteži lokalno u usnoj duplji, već da reaktivni oblici kiseonika proizvedeni tokom parodontopatije difunduju u krv dovodeći do oksidacije biomolekula krvi i nastanka cirkulatornog oksidativnog stresa. Cilj ovog istraživanja bio je da uporedimo stepen oksidativnog stresa u pljuvački i krvi u grupi pacijenata sa parodontopatijom i grupi pacijenata sa zdravim parodoncijumom kao i da ispitamo uticaj kauzalne terapije parodontopatije na stepen oksidativnog stresa u pljuvački i krvi kod pacijenata sa parodontopatijom.U istraživanje je bilo uključeno 30 pacijenata sa hroničnom parodontopatijom i 20 ispitanika sa zdravim parodoncijumom.Za procenu stanja parodoncijuma korišćeni su: Plak indeks (PI), Gingivalni indeks (GI), Indeks krvarenja gingive (IK), Dubina sondiranja (DS) i Nivo pripojnog epitela (NPE). Laboratorijsko ispitivanje se sastojalo od određivanja markera oksidativnog oštećenja DNK-8-hidroksideoksiguanozina (8-OHdG), markera peroksidacije lipida-malondialdehida (MDA) i antioksidativnog enzima-superoksid-dismutaze (SOD) u pljuvački i krvi. Kod pacijenata sa parodontopatijom bila je preduzeta kauzalna terapija. Klinička procena stanja parodoncijuma i laboratorijska ispitivanja su vršena na početku istraživanja i 3 meseca nakon završetka kauzalne terapije kod pacijenata sa parodontopatijom, odnosno samo na početku istraživanja kod pacijenata sa zdravim parodoncijumom. Vrednosti svih ispitivanih markera oksidativnog stresa u pljuvački pacijanata sa parodontopatijom bile su statistički značajno više u odnosu na pacijente sa zdravim parodoncijumom (8-OHdG p=0,043, MDA p=0,000 i SOD p=0,002). Pacijenti sa parodontopatijom imali su i statistički značajno višu vrednost MDA u krvi u odnosu na zdrave pacijente (p=0,013), dok se vrednosti 8-OHdG i SOD nisu razlikovale među ispitivanim grupama. Nakon sprovedene kauzalne terapije parodontopatije došlo je do značajnog smanjenja svih ispitivanih markera oksidativnog stresa u pljuvački, dok su njihove vrednosti u krvi ostale gotovo nepromenjene. Pacijenti sa parodontopatijom imali su značajno više vrednosti markera oksidativnog stresa u pljuvački i značajno višu vrednost MDA u krvi u odnosu na pacijente sa zdravim parodoncijumom. Kauzalna terapija parodontopatije dovela je do značajnog smanjenja markera oksidativnog stresa u pljuvački, dok na vrednost ovih markera u krvi nije imala uticaj., Oxidative stress arises due to the imbalance between the free radical production and the antioxidant defenses and is increasingly being recognized as a biochemical mechanism potentially present in pathogenic processes in a significant number of systemic illnesses, including periodontitis. On the other hand, periodontitis, as a destructive inflammatory disease, also contributes to the increased free radical production. Research indicates that periodontitis is accompanied not only by the disruption in bedox balance, localized in the oral cavity, but that reactive oxygen species produced during the periodontal disease are diffused into the blood, leading to the oxidation of blood biomolecules and the development of circulating oxidative stress. Aim: The aim of this study was to compare the level of oxidative stress in a group of patients with periodontitis to that of a group of patients with healthy periodontium and to evaluate the impact of non-surgical periodontal therapy on the level of oxidative stress in patients with periodontitis. Materials and methods: Study participians were 30 patients with chronic periodontitis and 20 periodontally healthy individuals. Their periodontal condition was assessed: Plaque index (PI), Gingival index (GI), Papilla bleeding index (PBI), Probing depth (PD) and Clinical attachment level (CAL). Laboratory assessment consisted of determination of DNA oxidative damage marker-8-hydroxydeoxyguanosine (8 OHdG), marker of lipid peroxidation- malondialdehyde (MDA) and antioxidant enzyme-superoxid dismutase (SOD) in saliva and blood. Patients with periodontitis were subjected to the non-surgical periodontal therapy. Clinical evaluation of periodontal conditions and laboratory tests were performed at baseline and 3 months after the non-surgical periodontal therapy. Results: The values of all studied oxidative stress markers in saliva were significantly higher in patients with periodontitis, compared to those with healthy periodontium (8-OHdG p =0,043, MDA p = 0,000 and SOD p = 0,002). The value of MDA in blood was also statistically significantly higher in patients with periodontitis relative to healthy individuals (p = 0,013), while the 8-OHdG and SOD values did not differ between the studied groups. Following the non-surgical periodontal therapy there was a ignificant reduction in the oxidative stress markers in saliva, while their values in the blood remained virtually unchanged. Conclusion: The values of oxidative stress markers in saliva and MDA in blood were significantly higher in patients with periodontitis, compared to those with a healthy periodontium. While non-surgical periodontal therapy led to a significant reduction in the oxidative stress markers in saliva, it had no effect on the value of these markers in the blood

There is ever increasing data indicating the vmast contribution of oxidative stress to the pathogenesis of numerous diseases (atherosclerosis, hypertension, heart failure, diabetes mellitus, stroke, rheumatoid arthritis, and others). Thus, in the pathogenesis of atherosclerosis the primary role is held by reactive oxygen species that are synthetized by endothelial cells of arterial blood vessels, leukocytes and macrophages. Furthermore, native particles of lipoproteins of small density become atherogenic through oxidation caused by reactive oxygen species. The oxidation of small-density lipoproteins stimulates the inflammatory process, and it in turn steps up adhesion and the inflow of monocytes and affects the synthesis and release of numerous proinflammatory cytokines involved in the further course of the process. One of the reasons for the development of arterial hypertension is the simultaneous activation of NAD(P)H oxidase and 12/15-lipoxygenase, since it results in the stepped up production of reactive oxygen species. These stimulate the production of matrix metalloproteinase 2, which lead to vascular remodelling and to increased apoptosis of heart muscle cells. Stepped up apoptosis is linked with myocardial infarction, cardiomyopathies and the development of heart failure. The sensitivity of β-cells of the endocrine part of the pancreas to reactive oxygen species favor the naturally low concentrations of the collectors of free radicals in them, as well as an increase in the concentration of proinflammatory cytokines, glucosis and lipids that induce a reduction in the mass and function of β-cells. Hyperglycemia in diabetes mellitus causes tissue damage through non-enzyme glycosylation of intracellular and extracellular proteins, which results in: reduced enzyme activity, damaged nucleic acid, disrupted natural decomposition of proteins, and activation of cytotoxic pathways. These processes are the basis of the pathogenesis of numerous complications of diabetes mellitus. Since inducible nitrogen-oxide synthesis launches processes that stimulate apoptosis of cerebral endothelial cells, and superoxide-anion radicals, hypochloric acid and hydrogen peroxide damage the parenchyma of an ischemic brain and biomacromolecules (causing lipid peroxidation, oxidation of proteins and deoxyribonucleic acid), brain damage occurs during cerebral ischemia and reperfusion. Sve više podataka ukazuje na veliki doprinos oksidativnog stresa patogenezi brojnih bolesti (aterosklerozi, hipertenziji, srčanoj insuficijenciji, šećernoj bolesti, moždanom udaru, reumatoidnom artritisu i dr.). Tako, u patogenezi ateroskleroze primarno mesto zauzimaju reaktivne kiseonične vrste koje sintetišu endotelne ćelije arterijskih krvnih sudova, leukociti i makrofagi. Uz to, nativne čestice lipoproteina male gustine postaju aterogene, oksidacijom izazvanom reaktivnim kiseoničnim vrstama. Oksidacija lipoproteina male gustine stimuliše zapaljenjski proces, a on pojačava adheziju i dotok monocita i utiče na sintezu i oslobađanje brojnih proinflamatornih citokina uključenih u dalji tok procesa. Jedan od razloga za razvoj arterijske hipertenzije je istovremena aktivacija NAD(P)H oksidaza i 12/15-lipoksigenaze, jer rezultira pojačanom proizvodnjom reaktivnih kiseoničnih vrsta. One podstiču proizvodnju metaloproteinaza-2 matriksa, koje dovode do vaskularnog remodeliranja i do pojačane apoptoze srčanih mišićnih ćelija. Pojačana apoptoza je povezana sa infarktom miokarda, kardiomiopatijama i razvojem srčane insuficijencije. Osetljivosti β-ćelija endokrinog dela pankreasa na reaktivne kiseonične vrste pogoduju prirodno niske koncentracije skupljača slobodnih radikala u njima, kao i porast koncentracije proinflamatornih citokina, glukoze i lipida koji indukuju smanjenje mase i funkcije β-ćelija. Hiperglikemija kod šećerne bolesti izaziva oštećenja tkiva neenzimskim glikozilovanjem intracelularnih i ekstracelularnih proteina, usled čega se smanjuje enzimska aktivnost, oštećuju nukleinske kiseline, re meti prirodna razgradnja proteina i aktiviraju citotoksični putevi. Ovi procesi su i osnova patogeneze mnogih komplikacija šećerne bolesti. S obzirom na to da inducibilna azot-oksid sintaza pokreće procese koji podstiču apoptozu cerebralnih endotelnih ćelija, a superoksid-anjon radikal, hipohlorna kiselina i vodonik-peroksid oštećuju parenhim ishemičnog mozga i biomakromolekule (izazivaju peroksidaciju lipida, oksidaciju proteina i dezoksiribonukleinske kiseline), tokom cerebralne ishemije i reperfuzije nastaju oštećenja mozga.

Introduction. Paraoxonase 1 (PON1) is a multifunctional enzyme associated with high-density lipoprotein particles (HDL). It is a cellular antioxidant that hydrolyses oxidized macromolecules, especially low-density lipoproteins (ox-LDL). Because increased oxidative stress is believed to play a crucial role in the initiation and propagation of atherosclerosis, coding (Q192R and L55M) and promoter (C(-107)T) region polymorphisms of pon1 gene, that are responsible for catalytic efficiency, activity and the level of the enzyme, have been of great interest as a potential markers of susceptibility for atherogenesis. Objective. The aim of the study was to assess possible association between these pon1 gene variants and clinical manifestations of the atherosclerosis and oxidative stress. Methods. A total of 60 angiographically documented patients with manifested atherosclerotic disease and 100 control individuals were analyzed. Genomic DNA was isolated from the peripheral blood cells and genotyping was performed using polymerase chain reaction followed by the restriction fragment length polymorphism (PCR-RFLP) analysis. Results No significant difference in allele and genotype frequencies of all three examined polymorphisms was found between the atherosclerotic patients and healthy controls. The obtained results could not support an association of pon1 gene variants with the oxidative stress and atherogenesis. Conclusion. These polymorphisms cannot be considered risk factors of atherosclerosis in Serbian population. A larger study is required in order to establish possible contribution of pon1 variants to atherosclerosis-related cardiovascular diseases., Uvod. Paraoksonaza 1 (PON1) je multifunkcionalni enzim koji je vezan za lipoproteine visoke gustine (HDL). To je ćelijski antioksidans koji hidrolizuje oksidovane makromolekule, naročito oksidovane lipoproteine niske gustine (ox-LDL). Smatra se da povišeni oksidativni stres igra ključnu ulogu u inicijaciji i propagaciji ateroskleroze, pa su polimorfizmi u kodirajućem (Q192R i L55M) i promotorskom (C(-107)T) regionu gena pon1, koji su odgovorni za katalitičku efikasnost, aktivnost i nivo enzima, od velikog interesa kao potencijalni markeri osetljivosti na aterogenezu. Cilj rada. Cilj ove studije je bio da se ispita moguća povezanost varijanti gena pon1 i kliničkih manifestacija ateroskleroze i oksidativnog stresa. Metode rada. Analizirano je 60 bolesnika s angiografski dokumentovanim manifestacijama ateroskleroze i 100 zdravih ispitanika. Genomska DNK je izolovana iz ćelija periferne krvi, a genotipizacija je urađena primenom reakcije lančane polimeraze, posle koje je urađena analiza dužine restrikcionih fragmenata (tzv. PCR-RFLP analiza). Rezultati. Učestalosti alela i genotipova tri ispitivana polimorfizma nisu pokazale značajne razlike između ispitanika obolelih od ateroskleroze i zdravih osoba. Dobijeni rezultati ne ukazuju na povezanost analiziranih varijanti gena pon1 i oksidativnog stresa i aterogeneze. Zaključak. Ovi polimorfizmi se ne mogu smatrati faktorima rizika za razvoj ateroskleroze u srpskoj populaciji. Potrebna je studija sa većim brojem ispitanika, kako bi se utvrdio mogući doprinos varijanti gena pon1 na nastanak kardiovaskularnih oboljenja u čijoj osnovi je ateroskleroza.

U ovom radu je ispitivan uticaj Antitoksičnognutritiva(ATN), preparata na bazi prirodnih aluminosilikata,na biohemijske, fiziološke, tehnološke i proizvodne parametre uzgoja pilića. ATN je smeša zeolita klinoptilolita, gline monmorilonita i male količine aktivnog uglja. Dodatak ovog preparata u hranivo za piliće u količini od 5 g/kg nije izazvao promene u normalnoj biohemijskoj i fiziološkoj homeostazi životinja. Hematološki parametri, koncentracija metabolita, elektrolita i aktivnost enzima seruma i jetre je bila u granicama referentnih vrednosti. ATN ne utiče na prirast životinja, ni na konverziju hrane, ali dovodi do povećanja relativnih masa pojedinih organa digestivnog trakta.Uočava se da dodatkom ATN-a u hranivo dolazi do smanjenja količine masti a povećanja količine proteina u belom mesu. ATN takođe povećava i sadržaj pepela ubelom i crvenom mesu.Akutni ili hronični tronedeljni oralni unos pojedinih toksikanata (mikotoksina aflatoksina B1i ohratoksina A, herbicida parakvata, jona olova ili toksinacijanobakterija-mikrocistisa) dovodi do poremećaja normalne biohemijske i fiziološke homeostaze pojedinih organa i tkiva pilića, što je utvrđeno na osnovu rezultata hematoloških i biohemijskih analiza, određivanja enzima antioksidativne zaštite i lipidneperoksidacije, kao i odredjivanja parametara uzgoja i težine organa. ATN, dodat u hranu u količini od 5 g/kg, mogao da bude dobar protektivni agens za delovanje aflatoksina, parakvata, jona teških metala i mikrocistisa, ali ne i ohratoksina., This study investigated the effects of dietary supplementation with hydrated aluminosilicate (Antitoxic Nutrient-ATN), based on zeolitic ore (clinoptilolite), clay bentonite (montmorillonite),and small amounts of activated charcoal, on performance, hematological, serum, and liver biochemical parameters, as well as organ weights and meat quality in broiler chickens. The dietary addition of ATN has no adverse effects on serum and liver biochemical parameters and does not affect the normal physiological homeostasis of animals. However, the results of this study demonstrate that supplementation with 5 g/kg of ATN influenced organ weights, and chemical composition of broiler chicken meat.This study also evaluated the effectiveness of ATN to protect broilers from adverse effects of five different toxic substances (mycotoxins aflatoxin B1, and ochratoxin A, herbicide paraquat, heavy metal ions supplied as lead-acetate, and microcystis, toxin of cyanobacteria). Toxic substances induced oxidative stress and disturb normal biochemical and physiological homeostasis of different tissues and organs in poultry. The results from this study demonstrate that the biochemical variables of serum, liver, kidney, lung and other organs were negatively affected by all five toxic substances. The additionof 5 g/kg of ATN was protective against aflatoxin B1, lead-acetate, paraquat and microcystis, but not against ochratoxin A.

Uloga vodikova peroksida (H2O2) u atmosferskoj kemiji i njegov doprinos u nastanku slobodnih radikala počeli su se proučavati tek posljednjih nekoliko desetljeća. Fotokemijskim reakcijama s ozonom i H2O2 nastaju oksidansi (slobodni radikali) koji mogu oksidirati biomolekule unutar stanica te dovesti do smrti stanica i ozljeda tkiva. Zbog toga se slobodni radikali smatraju uzrokom više od sto bolesti. H2O2 smatra se boljim indikatorom za atmosferski oksidacijski kapacitet od ozona. U atmosferi može biti prisutan u plinovitoj i tekućoj fazi te pokazuje tipične dnevne i sezonske varijacije. Me|utim, zbog skupe i slo`ene opreme, mjerenja H2O2 su rijetka i ograničena na samo nekoliko mjesta u svijetu. Mjerenja u slojevima leda na Grenlandu pokazala su da koncentracije H2O2 rastu posljednjih 200 godina. Značajan porast primijećen je upravo posljednjih dvaju desetljeća, a procjene pokazuju da će i dalje rasti zbog smanjene emisije sumporova dioksida. Mjerenja H2O2 u Hrvatskoj do sada još nisu bila provedena te će uporedo s već postojećim dugogodišnjim rezultatima mjerenja ozona i dušikovih oksida dati uvid u stanje i utjecaj na oksidativni stres., The past few decades saw a rising interest in the role of hydrogen peroxide (H2O2) in atmospheric chemistry and its contribution to the formation of free radicals. Free radicals (oxidants) are formed by photochemical reactions between ozone and H2O2. Free radicals formed within cells can oxidise biomolecules, and this may lead to cell death and tissue injury. For this reason, free radicals are believed to cause more than 100 diseases. H2O2 has been suggested as a better indicator of atmospheric oxidation capacity than ozone. Atmospheric H2O2 can appear in the gas phase or in the aqueous phase. It shows typical diurnal and seasonal variations. However, measurements of H2O2 with expensive and sophisticated equipment are rare and limited to but a few sites in the world. Measurements in Greenland ice cores showed that H2O2 concentrations increased over the last 200 years and most of the increase has occurred over the last 20 years. Evaluations show that concentrations will still rise as a result of decreasing SO2 emission. H2O2 measurements have not been carried out in Croatia until now, and, accompanied by the existing longterm measurements of ozone and nitrogen oxides, they will provide an idea of the oxidative capacity of the atmosphere and its influence on oxidative stress.

U radu su prikazani rezultati ispitivanja uticaja niskih doza γ-zračenja (do 200 Gy) na antioksidantne osobine i parametre oksidativnog stresa u različitim fazama razvoja soje i uticaja srednjih doza γ-zračenja (1-10 kGy) na neke antioksidantne osobine semena soje. Vršena su određivanja aktivnosti antioksidantnih enzima (SOD,CAT,GPx,GSH-Px), količine hidroksil radikala, malonildialdehida i redukovanog glutationa, sadržaja rastvorljivih proteina, ukupnih fenola i tanina, intenziteta oksidacije proteina, ukupne antioksidantne aktivnosti FRAP metodom, DPPH-skevindžer aktivnosti i inhibicije NO radikala. ESR spektroskopijom praćen je uticaj ekstrakata ozračene i neozračene soje na količinu hidroksil radikala u Fentonovom model sistemu. Pomoću HPLC analize određen je sadržaj izoflavona u semenu ozračene i neozračene soje.Upoređivanjem antioksidantnih statusa ozračenih i neozračenih genotipova soje može se zaključiti da je γ-zračenje, kojim je bilo izloženo seme, dovelo do stimulacije antioksidantnog sistema soje u različitim fazama razvoja kroz dve generacije. Utvrđeno je da promene aktivnosti pojedinih komponenti antioksidantnog sistema zavise od doze primenjenog zračenja, genotipa kao i od faze razvoja biljke.Na osnovu lipidne peroksidacije kao kriterijuma (biomarkera) osetljivosti na radijacioni ioksidativni stress, ispitivani genotipovi se mogu svrstati u dve grupe: radiosenzitivni genotipovi Venera i Ana i radiorezistentni genotipovi Meli i Novosađanka. Rezultati ESR spektroskopije su potvrdili da su genotipovi Ana i Venera osetljiviji na radijacioni i oksidativni stres od genotipova Meli i Novosađanka. Naši rezultati ukazuju da srednje doze γ-zračenja poseduju izvesno „antioksidantno dejstvo“ koje se manifestuje povećanjem DPPH skevindžer aktivnosti i smanjenjem intenziteta oksidacije proteina usemenu soje. Nije utvrđena ni jedna negativna posledica na ispitivane nutritivne karakteristike semena soje., The results obtained through the investigation about the influence of low level doses of γ-radiation (up to 200 Gy) on the antioxidant properties and oxidative stress parameters in different phases of soy development and the influence of the middle level doses of γ-radiation (from 1 kGy to 10 kGy) on some antioxidant parameters of soybean seed were presented in this paper. The investigation of the antioxidant enzymes (SOD,CAT,GPx,GSH-Px), hydroxyl-radical, malonyldialdehide and reduced glutathione quantities, soluble protein and total phenol and tannin content, protein oxidation intensity, total antioxidant activity by FRAP-method, DPPH scavenger ability and NO radical inhibition was performed. ESR spectroscopy was used to follow the change of hydroxyl radical quantities in the Phenton model system. HPLC was used to analyse the isoflavone content in the irradiated and nonirradiated soybean.By comparison of antioxidant statuses of irradiated and nonirradiated soy, it was established that the γ-irradiation of soybean stimulated the antioxidant system of soy during the development through two generations. It was observed that the changes in antioxidant activities of different components of antioxidant system of soybean depend on dose of radiation, genotype and phase of the development.On the basis of lipid peroxidation as criterion (biomarker) of sensitivity on radiation and oxidative stress, investigated genotypes can be classified intotwo groups: radiosensitive genotypes Venera and Ana and radioresistant genotypes Meli and Novosađanka. Results of ESR spectroscopy confirmed that genotypes Ana and Venera are more sensitive to radiation and oxidative stress than genotypes Meli andNovosađanka.Our results indicate that middle doses of γ-radiation possess such „antioxidant activity“ which is manifestated with the increase of DPPH scavenger activity and decrease of protein oxidation intensity in soybean seed. We did not establish any negative implication on investigated nutritive characteristics of soybean seed.

Considering free radicals (FR) involvement in mechanisms of numerous xenobiotics' adverse effects (UV radiation, poisonings, some drugs, smoking, environmental pollutions, etc.), the goal of this paper is to elucidate principles of oxidative stress (OS), modes of antioxidanase (AO) actions, as well as, to pont at numerous brand new AO pharmaceutical products/drugs, their rationale usage and to emphasize expansion of activities regarding this subject. Free radicals (FR) produced in high concentration initiate development of OS. Partly, FR are involved in numerous pathologies (degenerative, malignant, cardiovascular diseases, diabetes mellitus, poisonings, etc.), aging, as well in physical exercises. Fundamental scientific studies in this field trigger complex machinery of pharmaceutical-management activities, such as: conferences, congresses, work shops, educative programs and projects, publishing of the science-popular books and brochures. Moreover, strategic program leads to the manufacture of the new product, what implies innovative and modern technology processing, and its realization on the market. Usually, wide range of human population consumes AO. ICT marketing (information-communication-technology) and concept of social marketing (rational usage of the recent AO products/drugs, prevention of diseases, cost benefit effect, etc.) are very well recognized. Marketing survey has shown increased consumption of AO. Importantly, diagnostic kits for determination of OS in presymptomatic phase of diseases are also available on the market, but yet unattractive for daily routine in our country, unlike in developed countries. Imajući u vidu uključenost slobodnih radikala (SR) u mehanizme štetnog delovanja brojnih ksenobiotika (UV zračenje, trovanja, neki lekovi, pušenje, zagađenje čovekove okoline itd.) cilj ovog rada je da približimo i objasnimo osnovne principe nastanka oksidativnog stresa (OS) i način delovanja antioksidanasa (AO), kao i da ukažemo na racionalno korišćenje preparata sa AO delovanjem koji su na tržištu i na porast aktivnosti koji se tiču ove problematike. Nastali u prekomernoj količini, SR iniciraju nastanak i razvoj OS. Jednim delom su uključeni u brojna patološka zbivanja (degenerativne bolesti, maligne bolesti, šećerne bolesti, kardiovaskularne bolesti, trovanja, starenje i td.), starenju, kao i pri fizičkoj aktivnosti. Fundamentalna naučna istraživanja u ovoj oblasti su pokrenula kompleksnu mrežu farmaceutsko-menadžerskih aktivnosti, usmerenih na organizovanje konferencija, kongresa, radionica, edukativnih programa i publikovanje naučnopopularne literature. Takođe, strateška planiranja idu ka proizvodnji novog proizvoda, što podrazumeva primenu inovativnih tehnoloških postupaka/procesa kod razvoja i proizvodnje, kao i njegovu realizaciju na tržištu. Preparati sa antioksidantnim dejstvom su najčešće namenjeni širokoj populaciji. U promotivnim akcijama ICT marketinga (information-communicatontehnology) prepoznajemo koncept socijalnog marketinga (pravilna i racionalna upotreba novog proizvoda/leka, preventiva bolesti, cena leka, itd.). Istraživanje tržišta proizvoda sa AO dejstvom ukazuje na porast prodaje ovih proizvoda. Značajno je napomenuti da ovo tržište obuhvata i niz proizvoda za laboratorijsku dijagnostiku OS u presimptomatskoj fazi bolesti, što je nedovoljno prepoznato i prihvaćeno na domaćem tržištu.