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It is well known that the presence of different chemical groups in drug molecules influences their pharmacological properties. The aim of our study is to investigate whether newly synthesized derivatives of propafenone, with changes in benzyl moiety, have a different effect upon arrhythmia, compared to propafenone. 5OCl-PF and 5OF-PF are derivatives of propafenone with-Cl or –F substituent on the ortho position of the benzyl moiety. For verification of their antiarrhythmic effect, we used an in vivo rat model of aconitine-induced arrhythmia. 5OCl-PF speeded the appearance of supraventricular premature beats (SVPB) and death more than aconitine. All animals treated with 5OCl-PF developed ventricular premature beats in salvos (VPBS), bigeminies (VPBB) and paroxysmal ventricular tachycardia (PVT). 5OF-PF had a negative chronotropic effect and potentiated atrial excitability (more SVPB). It had a positive effect on the occurrence and onset time of supraventricular tachycardia, VPBS, and PVT. Based on the obtained results, it can be concluded that newly synthesized propafenone derivatives have no better antiarrhythmic effect than the parent compound. In the future, our research will be focused on the synthesis of different derivatives and examining their antiarrhythmic effects. Dobro je poznato da prisustvo različitih hemijskih grupa u molekulima leka utiče na njegova farmakološka svojstva. Cilj našeg istraživanja je ispitati da li novosintetisani derivati propafenona, s promenama u benzilnoj grupi, imaju drugačiji efekat na aritmiju u odnosu na propafenon. 5OCl-PF i 5OF-PF su derivati propafenona sa -Cl ili –F supstituentom na orto položaju benzilnog dela. Za proveru njihovog antiaritmičnog efekta koristili smo in vivo model na pacovima sa aritmijom izazvanom akonitinom. 5OCl-PF je ubrzao pojavu supraventrikularnih prevremenih otkucaja (SVPB) i smrt više nego akonitin. Sve životinje lečene sa 5OCl-PF razvile su ventrikularne prevremene otkucaje (VPBS i VPBB) i paroksizmalnu ventrikularnu tahikardiju (PVT). 5OF-PF je imao negativan hronotropni efekat i potencirao atrijalnu ekscitabilnost (više SVPB). Pozitivno je uticao na pojavu i vreme početka supraventrikularne tahikardije, VPBS i PVT. Na osnovu dobijenih rezultata se može zaključiti da novosintetisani derivati propafenona nemaju bolji antiaritmijski efekat od polaznog jedinjenja. U budućnosti, istraživanje će biti usmereno ka sintezi hemijski drugačijih derivata i ispitivanju njihovog antiaritmijskog efekta.

Doktorsku disertaciju čine tri celine kojima je zajednički element ispitivanje fenola, u pogledu njihovog sadržaja i dostupnosti u dijetetskim suplementima na bazi grožđa i japanskog troskota, ispitivanja uticaja suplementacije kod eksperimentalnih životinja, odnosno mogućnosti ekstrakcije fenolnih jedinjenja iz komine grožđa. Osnovni ciljevi prvog dela bili su ispitivanje bezbednosti i kvaliteta 14 suplemenata na bazi grožđa i japanskog troskota u pogledu ujednačenosti mase, sadržaja i brzine rastvorljivosti. U drugom delu je radi provere terapijske delotvornosti suplementa koji sadrži čist resveratrol ispitan njegov uticaj na glikemiju i lipidni status eksperimentalnih životinja. Treći deo je obuhvatio analizu pet različitih komina grožđa i razvoj metode ekstrakcije fenola iz odabrane komine kako bi se dobili ekstrakti sa što je moguće većim prinosom, koristeći rastvarače koji su bezbedni po ljudsko zdravlje. Rezultati su pokazali da dva od četrnaest suplemenata nisu ispunila zahteve farmakopeje za lekove u pogledu ujednačenosti mase i sadržaja aktivne komponente, dok ni jedan suplement nije ispunio zahteve u pogledu brzine rastvorljivosti aktivne supstance, zbog čega se postavlja pitanje delotvornosti ispitanih suplemenata. Ni kod jednog suplementa nisu kvantifikovani ostaci pesticida ni toksični metali, čime je potvrđena njihova bezbednost. Farmakodinamska ispitivanja na pacovima pokazala su antioksidantna, anti-dijabetska i hipolipidemijska svojstva dijetetskog suplementa na bazi resveratrola, čime je potvrđena njegova delotvornost. Za potvrdu delotvornosti suplementa na ljudima neophodno je sprovesti dobro dizajnirano kliničko ispitivanje na dovoljnom broju ispitanika. Analiza pet različitih komina grožđa pokazala je da ovaj nusprodukt proizvodnje vina sadrži različite fenole, od kojih je samo katehin bio prisutan u značajnoj količini. Kvalitativan i kvantitativan sastav komine značajno varira zavisno od sorte i berbe grožđa, kao i od primenjenog tehnološkog procesa proizvodnje vina. Imajući u vidu sveobuhvatne kriterijume u pogledu efikasnosti ekstrakcije, zaključeno je da su 55% etanol, odnos uzorak / rastvarač 1:40, pH 4,5, T 55°C i vreme od 30 min optimalni eksperimentalni uslovi za ekstrakciju fenola iz komine grožđa. U zavisnosti od osnovnog cilja procesa ekstrakcije ovi parametri se mogu lako modifikovati. Višestruke su mogućnosti za iskorišćenje komine grožđa, a jedan od njih bi mogla biti i da se koristi kao sirovina za izradu dijetetskih suplemenata. The dissertation consists of three parts with a common element – testing phenols in terms of content and availability in grape and Polygonum cuspidatum-based dietary supplements, testing the supplement’s impact on experimental animals and the possibility to extract phenolic compounds from grape pomace. The primary objectives of the first part were testing safety and quality of fourteen grape and Polygonum cuspidatum-based supplements in terms of uniformity of mass, content and dissolution profile. In the second part, to test the therapeutic efficacy of a supplement containing pure resveratrol, its effect on the glycemia and lipid profile of experimental animals was examined. The third part covers the analysis of five different grape pomaces and the development of a phenol extraction method from the selected grape pomace to obtain extracts with the highest possible yield, using solvents that are safe for human health. The results showed that two out of fourteen supplements have not met the Pharmacopoeia requirements in terms of uniformity of mass and active substance content, while no supplement has met requirements in terms of dissolution test of the active substance, which raises the question of the effectiveness of the tested supplements. No pesticide or heavy metal residues were quantified in any of the supplements which confirms their safety. Pharmacodynamic testing on rats has shown antioxidant, antidiabetic and hypolipidemic properties of the resveratrol-based dietary supplement which confirms its efficacy. In order to confirm the supplement efficacy on people, it is necessary to conduct a well-designed clinical trial on a sufficient number of human participants. The analysis of five different grape pomaces revealed that this byproduct of vine production contains different phenols, and only catechin was present in significant amounts. The qualitative and quantitative composition of grape pomace varies significantly depending on grape variety and harvesting as well as on the technological process that was applied in vine production. Considering the comprehensive criteria in terms of extraction efficacy, it was concluded that optimal experimental conditions for the extraction of phenol from grape pomace include 55% ethanol, sample/solvent ratio 1:40, pH 4.5, T 55°C for 30 minutes. Depending on the primary objective in the extraction process, these parameters can be easily modified. There are multiple possibilities for utilizing grape pomace and one of them could be as raw material in production of dietary supplements.

Background/Aim. Recently, there has been an increased interest in novel dietary antioxidants, including omega-3 fatty acids and bioactive proteins present in milk. The aim of this study was to examine potential antioxidant effects of four-weeks long fish-based and milk-based diets in female Wistar rats. Methods. Four-months old rats were divided into three groups receiving either: control diet, diet enriched with fish meal, or diet enriched with milk. The activities of antioxidant enzymes: glutathione peroxidase (GPx), superoxide dismutase (SOD) and catalase (CAT), and concentration of thiobarbituric acid reactive substances (TBARS) were determined in liver homogenates obtained at the end of the treatment period. Results. Statistically significant higher activities of GPx (3.52 ± 0.73 U/mg) and CAT (147.25 ± 15.93 U/mg) were detected in rats fed with fishbased meal in comparison with both the control (GPx: 1.93 ± 0.11 U/mg; CAT: 99.37 ± 10.03 U/mg) and the group fed with milk-based diet (GPx: 1.72 ± 0.52 U/mg; CAT: 104.18 ± 37.49 U/mg). Despite somewhat lower concentration of TBARS in the milk-treated group (0.88 ± 0.23 nmoL/mg), no significant differences were detected in comparison with other groups (the control group: 1.00 ± 0.08 nmoL/mg; the fish-based diet group: 1.13 ± 0.15 nmoL/mg). Conclusion. Diet enriched with fish could improve one's oxidative status by enhancing activities of antioxidant enzymes in the liver tissue. On the contrary, we failed to obtain results suggesting that milk could serve as a source of dietary antioxidants. Uvod/Cilj. U poslednje vreme povećano je interesovanje za istraživanja novih antioksidanasa u ishrani, uključujući omega-3 masne kiseline i bioaktivne proteine prisutne u mleku. Cilj ove studije bio je ispitivanje potencijalnih antioksidativnih efekata hrane obogaćene ribljim brašnom i hrane obogaćene mlekom u prahu kod ženki Wistar pacova, u trajanju od četiri nedelje. Metode. Pacovi, starosti četiri meseca, podeljeni su u tri grupe koje su bile hranjene standardnom hranom (kontrolna grupa), hranom obogaćenom ribljim brašnom i hranom obogaćenom mlekom u prahu. U homogenatima jetre, posle četiri nedelje, određene su aktivnosti antioksidativnih enzima: glutation peroksidaze (GPx), superoksid dismutaze (SOD) i katalaze (CAT), kao i koncentracija reaktivnih supstanci tiobarbiturne kiseline (TBARS). Rezultati. Statistički značajno veće aktivnosti GPx (3,52 ± 0,73 U/mg) i CAT (147,25 ± 15,93 U/mg) nađene su kod pacova koji su dobijali hranu obogaćenu ribljim brašnom u odnosu na kontrolu (GPx: 1,93 ± 0,11 U/mg; CAT: 99,37 ± 10,03 U/mg) i grupu koja je hranjena hranom obogaćenom mlekom u prahu (GPx: 1,72 ± 0,52 U/mg; CAT: 104,18 ± 37,49 U/mg). Uprkos nešto nižoj koncentraciji TBARS u grupi koja je primala hranu obogaćenu mlekom u prahu (0,88 ± 0,23 nmoL/mg), nisu utvrđene statistički značajne razlike u poređenju sa drugim grupama (kontrola: 1,00 ± 0,08 nmoL/mg; grupa na ishrani obogaćenoj ribljim brašnom: 1,13 ± 0,15 nmoL/mg). Zaključak. Ishrana bogata ribom mogla bi delovati povoljno na oksidativni status preko poboljšanja aktivnosti antioksidativnih enzima jetre. Sa druge strane, rezultati ne pokazuju da bi mleko moglo biti dobar izvor dijetarnih antioksidanasa.

Gliklazid je jedan od najčešće korišćenih lekova u terapiji dijabetes melitusa tip 2. U poslednje vreme, utvrđeno je da gliklazid ispoljava i druge pozitivne farmakološke efekte kao što su imunomodulatorni i anti-koagulacioni efekti, ukazujući na njegovu potencijalnu primenu u terapiji dijabetes melitusa tip 1. Gliklazid se odlikuje varijabilnim stepenom apsorpcije nakon peroralne primene i iz tog razloga pretpostavlja se da bi tehnike njegove ciljane isporuke, kao što je mikroinkapsulacija, mogle da dovedu do poboljšanja njegove apsorpcije i njegove potencijalne primene u terapiji T1DM. Pokazano je da različite žučne kiseline, uključujući i holnu, imaju stabilizacione efekte u domenu primene mikrovezikula i kontrolisanog osobađanja lekova, te je moguće da bi njihov dodatak u mikrovezikularnu formulaciju gliklazida mogao dodatno da poboljša oslobađanje gliklazida, njegovu apsorpciju i antidijabetičke efekte. S tim u vezi, cilj ovog istraživanja je da se ispita hipoglikemijski efekat gliklazida primenjenog u obliku alginatnih mikrovezikula, sa ili bez dodatka holne kiseline na T1DM modelu pacova. Trideset šest pacova obolelih od T1DM indukovanog aloksanom i odgovarajuće zdrave kontrolne životinje su nasumično raspoređene u šest grupa (n=6) i tretirane jednokratnom dozom fiziološkog rastvora, suspenzijom gliklazida, gliklazidom u obliku alginatnih mikrovezikula, samo holnom kiselinom, i mikrovezikulama gliklazida sa ili baz dodatka holne kiseline. Uzorkovana je krv tokom 10 h nakon unete doze i merena je koncentracija glukoze u krvi I koncentracija gliklazida u serumu korišćenjem HPLC metode. Mikrovezikule gliklazida su ispoljile hipo-glikemijski efekat kod pacova obolelih od dijabetesa, uprkos njegovim smanjenim koncentracijama u serumu, dok je dodatak holne kiseline u mikrovezikularnu formulaciju smanjio hipoglikemijski efekat gliklazida. Ovo potvrđuje izostanak sinergističkog efekta između gliklazida i holne kiseline. Takođe, ni proces mikroinkapsulacije niti dodatak holne kiseline nisu doprineli poboljšanju apsorpcije gliklazida, što ukazuje na činjenicu da su njegovi hipoglikemijski efekti nezavisni od njegove apsorpcije i koncentracije u serumu. Stoga se može pretpostaviti da su hipoglikemijski efekti gliklazida pre pod uticajem crevno-metaboličke aktivacije nego ciljanog oslobađanja u digestivnom traktu sistemske apsorpcije. Mikrovezikule gliklazida ispoljavaju hipoglikemijski efekat kod pacova obolelih od T1DM nezavisno od insulina, te mogu imati potencijalnu primenu u terapiji T1DM. Ovaj rad su podržali: HORIZON 2020 MEDLEM projekat broj 690876; Projekat Sekretarijata naučnog i tehnološkog razvoja Vojvodine broj . 114-451-2072-/2016-02; Projekat Ministarstva obrazovanja, nauke i tehnološkog razvoja Republike Srbije broja 41012. Gliclazide is one of the most commonly prescribed drugs for the treatment of Type 2 diabetes. Recently, gliclazide has reported to have desirable pharmacological effects such as immunomodulatory and anti-clotting effects, suggesting its potential applications in Type 1 diabetes mellitus (T1D) therapy. Gliclazide has variable absorption after oral administration, and therefore use of targeted-delivery techniques, such as microencapsulation, may optimise gliclazide absorption and its potential applications in T1D. Different bile acids, including cholic acid, have demonstrated microcapsule-stabilising and controlled-release effects, and thus their incorporation into gliclazide microcapsules may further optimise gliclazide release, absorption and antidiabetic effects. Accordingly, this study aimed to investigate the hypoglycaemic effects of gliclazide alginate-based microcapsules, in combination with or without added cholic acid, in a T1D rat model. Thirty six alloxan-induced T1D rats and respective healthy controls were randomly assigned into six equally distributed groups (n=6) and gavaged with a single dose of normal saline solution, gliclazide suspension, gliclazide alginate-based microcapsules, cholic acid alone, combination of gliclazide and cholic acid or alginate-based microcapsules with combined gliclazide and cholic acid. Blood samples were collected over 10 h post-dose and analysed for blood glucose concentrations and gliclazide serum concentrations using HPLC. Gliclazide microcapsules exibited a hypoglycaemic effect in diabetic rats, despite detected lower serum concentrations, whereas cholic acid incorporation diminished the hypoglycaemic effects. This suggests a lack of synergistic effects between gliclazide and cholic acid. In addition, neither microencapsulation nor cholic acid incorporation optimised gliclazide absorption which suggests that hypoglycaemic effects of gliclazide are independent of its absorption and serum concentrations. This provides evidence that hypoglycaemic effects of gliclazide may be associated with gut-metabolic activation rather than gut-targeted delivery and systemic absorption. Conclusion Gliclazide microcapsules exibited hypoglycaemic effects in T1D rats independent of insulin and thus may have potentials in treatment of T1D. This research was supported by: HORIZON 2020 MEDLEM project Grant No. 690876; Project for Scientific and Technological Development of Vojvodina No. 114-451-2072-/2016-02; Project of Ministry of Education, Science and Technological Development of the Republic of Serbia, Grant No. 41012.

The aim of this experimental study was to assess the effects of the acute administration of L-arginine alone and in combination with L-NAME (a non-selective NO synthase inhibitor) on the coronary flow and oxidative stress markers in isolated rat hearts. The experimental study was performed on hearts isolated from Wistar albino rats (n=12, male, 8 weeks old, body mass of 180-200 g). Retrograde perfusion of the isolated preparations was performed using a modified method according to the Langendorff technique with a gradual increase in the perfusion pressure (40–120 cmH2O). The following values were measured in the collected coronary effluents: coronary flow, released nitrites (NO production marker), superoxide anion radical and the index of lipid peroxidation (measured as thiobarbiturate reactive substances). The experimental protocol was performed under controlled conditions, followed by the administration of L-arginine alone (1 mmol) and L-arginine (1 mmol) + L-NAME (30 μmol). The results indicated that L-arginine did not significantly increase the coronary flow or the release of NO, TBARS and the superoxide anion radical. These effects were partially blocked by the joint administration of L-arginine + L-NAME, which indicated their competitive effect. Hence, the results of our study do not demonstrate significant effects of L-arginine administration on the coronary flow and oxidative stress markers in isolated rat hearts.

Background/Aim. Autogenous bone grafting has been the gold standard in clinical cases when bone grafts are required for bone defects in dentistry. The study was undertaken to evaluate multilevel designed carbonated hydroxyapatite (CHA) obtained by hydrothermal method, as a bone substitute in healing bone defects with or without corticosteroid treatment in rats as assessed by histopathologic methods. Methods. Bone defects were created in the alveolar bone by teeth extraction in 12 rats. The animals were initially divided into two groups. The experimental group was pretreated with corticosteroids: methylprednisolone and dexamethasone, intramuscularly, while the control group was without therapy. Posterior teeth extraction had been performed after the corticosteroid therapy. The extraction defects were fulfilled with hydroxyapatite with bimodal particle sizes in the range of 50-250 μm and the sample from postextocactional defect of the alveolar bone was analyzed pathohystologically. Results. The histopatological investigations confirmed the biologic properties of the applied material. The evident growth of new bone in the alveolar ridge was clearly noticed in both groups of rats. Carbonated HA obtained by hydrothermal method promoted bone formation in the preformed defects, confirming its efficacy for usage in bone defects. Complete resorption of the material's particles took place after 25 weeks. Conclusion. Hydroxyapatite completely meets the clinical requirements for a bone substitute material. Due to its microstructure, complete resorption took place during the observation period of the study. Corticosteroid treatment did not significantly affect new bone formation in the region of postextractional defects., Uvod/Cilj. Autogeni koštani graftovi predstavljaju zlatni standard u stomatologiji za popunjavanje koštanih defekata. Studija je sprovedena kako bi se ispitala efikasnost višefaznog karbonatnog hidroksiapatita (HA), dobijenog hidrotermalnam metodom, kao zamene za kost kod in vivo zarastanja koštanih defekata. Procena efikasnosti izvršena je patohistološkom analizom na pacovima (Sprague Dawley). Metode. Koštani defekti načinjeni su u alveolarnoj kosti ekstrakcijom bočnih zuba kod 12 pacova. Eksperimentalne životinje prvo su bile podeljene u dve grupe. Prva, kontrolna grupa, bila je bez terapije, dok je druga, eksperimentalna grupa intramuskularno dobijala kortikosteroidnu terapiju i to metilprednizolon i deksametazon. Ekstrakcija bočnih zuba izvršena je nakon resorpcije izazvane terapijom kortikosteroidima. Ekstrakcione rane ispunjene su hidroksiapatitom čestica veličine 50-250 μm, a uzorci uzeti iz postekstrakcionih defekata alveolarne kosti analizirane su patohistološki. Rezultati. Patohistološkom analizom potvrđena su biološka osteokonduktivna svojstva primenjenog materijala. Intenzivni rast nove kosti unutar alveolarnog grebena jasno je uočen u obe grupe eksperimentalnih životinja. Karbonatni HA dobijen hidrotermal nim metodom inicirao je formiranje kosti preko površine defekata, potvrđujući efikasnost njegove primene kod koštanih defekata. Do potpune resorpcije materijala došlo je posle 25 nedelja. Zaključak. Ispitivani hidroksiapatit u potpunosti zadovoljava kliničke zahteve kao zamena za kost, poštujući ograničenja eksperimentalne namene studije. Zbog mikrostrukture materijala došlo je do kompletne resorpcije tokom perioda posmatranja. Lečenje kortikosteroidima nije značajno uticalo na stvaranje nove kosti u predelu potekstrakcionih defekata. PR Projekat Ministarstva nauke Republike Srbije, br. 172026.

In the health care of animals phytoteraphy is being applied very intensively, mostly for the prophylactic purpose and with the aim of treating weaker forms of diseases and chronic course of a disease. It is well known from a traditional folk medicine that Helleborus L. can induce the functions of defensive cell effectors and increase the activity of rapid, unspecific mechanisms of immune response. The plants of the genus Helleborus L. contain saponosides which increase the permeability of the erythrocytes membrane and that leads to the haemolysis and loss of haemoglobin. In this research paper we have studied the influence which the rhisome and root extract of H. odorus Waldst. et Kit., may have on the value of the erythrocytes count, concentration of haemoglobin and haematocrit value. In the rats in the trial group the extract of underground plant organs of Helleborus odorus Walds. et Kit. was applied intramuscularly in the concentration of 0,5, 5 or 20mg/100g TM. In the rats in a control group a sterile physiological solution in the quantity of 0,25ml/100g TM was applied in the same way. Fitoterapija se u zdravstvenoj zaštiti životinja veoma intenzivno primenjuje u profilaktičke svrhe i u cilju lečenja blažih oblika bolesti i bolesti hroničnog toka. Iz tradicionalne, narodne medicine poznato je da kukurek podstiče funkcije efektorskih odbrambenih ćelija i povećava aktivnost brzih, nespecifičnih mehanizama imunskog odgovora. Biljke roda Helleborus L. sadrže saponozide koji povećavaju permeabilnosti membrane eritrocita što dovodi do hemolize i gubitka hemoglobina. U ovom radu ispitivali smo uticaj ekstrakta rizoma i korena H. odorus Waldst. et Kit. na vrednost broja eritrocita, koncentraciju hemoglobina i hematokritsku vrednost. Pacovima oglednih grupa ekstrakt kukureka je aplikovan intramuskularno u koncentraciji 0,5, 5 ili 20mg/100g TM. Pacovima kontrolne grupe na isti način je aplikovan sterilan fiziološki rastvor u količini od 0,25ml/100g TM.

Objective. Although it is known that nitric oxide modulates the activity of hypothalamic-pituitary-adrenal axis, its functional significance has not been elucidated. Additionally, there is no information on the effect of nitric oxide on cortical expression of glucocorticoid receptor. The purpose of this study was to investigate the influence of endogenous nitric oxide on structure and function of rat adrenal cortex and adrenocortical expression of glucocorticoid receptor. Methods. Adult female Wistar rats showing diestrus day 1 were treated with Nω-nitro-L-arginine-methyl ester (LNAME), an inhibitor of all three isoforms of nitric oxide synthase, at a dose of 30 mg/kg, subcutaneously. The concentrations of adrenocorticotropic hormone (ACTH) and corticosterone were determined by radioimmunoassay. Stereological and immunohistochemical analyses were performed on paraffin sections. Results. Blockade of nitric oxide production significantly increased blood levels of ACTH and corticosterone. Stereological analysis showed that treatment with L-NAME significantly decreased numerical density of the cells in all cortical zones. Consistent with the decreased numerical density, L-NAME significantly increased the volume of cells in cortical zones. Inhibition of nitric oxide synthesis decreased expression of glucocorticoid receptor in zona fasciculata and zona reticularis. Conclusion. Obtained results indicate that endogenous nitric oxide inhibits activity of adrenal cortex and modulates expression of glucocorticoid receptor. Cilj. Iako je poznato da azot-monoksid moduliše aktivnost hipotalamo-hipofizno-nadbubrežne osovine, funkcionalni značaj tog delovanja nije rasvetljen. Pored toga, dejstvo azot-monoksida na kortikalnu ekspresiju glukokortikoidnog receptora nije istraženo. Cilj ovog rada bio je da se ispita uticaj endogenog azot-monoksida na strukturu i funkciju kore nadbubrežne žlezde pacova i adrenokortikalnu ekspresiju glukokortikoidnog receptora. Metode. Odrasle ženke pacova Wistar soja u metestrusnoj fazi ciklusa tretirane su Nω-nitro-L-arginin metil esterom (L-NAME), inhibitorom sve tri izoforme azotmonoksid sintaze, u dozi 30 mg/kg, subkutano. Koncentracije adrenokortikotropnog hormona (ACTH) i kortikosterona u krvi određene su radioimunološkom metodom. Na parafinskim presecima nadbubrežne žlezde izvršena je stereološka i imunohistohemijska analiza korteksa. Rezultati. Blokada sinteze azot-monoksida značajno povećava koncentracije ACTH i kortikosterona u krvi. Stereološka analiza je pokazala da tretman sa L-NAME značajno smanjuje numeričku gustinu ćelija u svim kortikalnim zonama. U skladu sa smanjenom numeričkom gustinom, L-NAME značajno povećava volumen ćelija u sve tri zone korteksa. Inhibicija sinteze azot-monoksida smanjuje ekspresiju glukokortikoidnog receptora u zoni fascikulati i zoni retikularis. Zaključak. Dobijeni rezultati ukazuju na mogućnost da endogeni azot-monoksid inhibiše aktivnost kore nadbubrežne žlezde i da moduliše ekspresiju glukokortikoidnog receptora.

This paper presents the results of investigations of parameter values of the red blood cell count in Wistar rats, after intramuscular application of rhizome and root extract of Helleborus odorus W. et K (HE). The trial was conducted in experimental conditions on 49 Wistar rats divided into 7 groups with 7 animals per unit (control and six experimental groups). The control group was treated by an intramuscular injection of sterile saline (0.25 ml/100 g BM). For the purpose of determing the duration of the HE effects, the animals were injected with a dose of 10 mg/100 g BMand blood was collected by cardial taps after 24, 48 and 72 hrs. Moreover, we investigated the influence of elevated temperatures on the noted HE extract effects, 24 hrs after the application of the inactivated (56oC 30 min) or denaturized (100oC 5 min) extract, as well as the influence of dialyzed HE. The obtained results of haematological analyses have pointed out the weak hemolyses activity of the extract. In all experimental groups, the erythrocyte count was decreased, but remained within reference values. Haemoglobin concentration and haematocrit values in the experimental and the control groups of rats were decreased in comparison to the physiological values, while the applied extract did not significantly affect the examined parameters. . U ovom radu su prikazani rezultati ispitivanja vrednosti parametara crvene krvne slike pacova soja Wistar, nakon intramuskularne aplikacije ekstrakta rizoma i korena Helleborus odorus W. et K. Ogled je izveden u eksperimentalnim uslovima na 49 pacova soja Wistar, podeljenih u 7 grupa sa po 7 jedinki (jedna kontrolna i šest oglednih grupa). Kontrolnoj grupi pacova je intramuskularno aplikovan fiziološki rastvor (0,25 ml/100 g t.m.). U cilju određivanja dužine trajanja efekata ekstrakta, pacovima je aplikovan tečni ekstrakt podzemnih organa kukureka u koncentraciji od 10 mg/100 g t.m., a krv je uzorkovana kardijalnom punkcijom nakon 24h, 48h i 72h. Ispitivan je i uticaj ekstrakta izlaganog povišenoj temperaturi na uočene efekte, 24h posle aplikacije inaktivisanog (56oC 30 min) ili denaturisanog ekstrakta (100oC 5 min), kao i efekat dijalizata ekstrakta. Dobijeni rezultati hematoloških analiza su ukazali na slabu hemolitičku aktivnost ekstrakta. Kod svih oglednih grupa, prosečan broj eritrocita bio je smanjen, ali se održao u granicama referentnih vrednosti. Zabeležene koncentracije hemoglobina i hematokrita u krvi pacova kontrolne i oglednih grupa bile su niže u odnosu na fiziološke vrednosti, pri čemu korišćeni ekstrakt nije značajno uticao na ispitivane parametre.

Etiopathogenesis of the most frequent endocrinopathy, Hashimoto thyroiditis (HT) is not clear. Our investigations dating from 1990 on an experimental model for examination of thyroiditis in Wistar rats show morphological changes in the rat thyroid evoked by potassium iodine (KJ) administration. KJ is used as a drug in therapy of numerous diseases such as Erythema Nodosum, Vasculitis Nodularis, Sweet’s Syndrome, as well as tuberculosis and granulomatosis. This study examined the effect of the maximal dose of potassium iodine on the thyroid gland and status of T3, T4 hormones and TSH in Wistar rats. We have groups of Wistar rats: treated with low iodine dose (LKI = 22,5 µg/g b.w.), and with high iodine dose (HKI = 225 µg/g b.w.) of KI, solutions (n=10). Untreated nonimmunised animals served as controls. Solutions were administered daily intraperitonealy during a period of 26 consecutive days. Histological changes within the gland in HKI treated group were the infiltration of the mononuclear cells associated with destruction of tissue architecture, formation of macrofolicules and proliferation of connective tissue significantly higher than in LKI treated rats. Rats injected with LKI had depletion of the serum amount of T3 and T4, as HID injected rats. The early iodide induced cell necrosis and inflammation in the nonimunised animals not carrying a genetic susceptibility, is in fact a new experimental model of the LT. Etiopatogeneza najčešće endokrinopatije, Hašimotovog tiroiditisa (HT) nije razjašnjena. Naša istraživanja imunomodulirajućeg efekta kalijum jodida (KJ), započeta 1990. godine, jasno pokazuju da KJ izaziva eksperimentalni tiroiditis (ET) na dozno zavistan način u funkciji vremena, bez prethodne imunizacije. KJ se primenjuje u ljudskoj ishrani za jodiranje soli, za terapiju vaskulitisa malih krvnih sudova, erythema nodosum, vaskulitisa nodularisa, Sweet-ovog sindroma, kao i tuberkuloze i granulomatoze. Pacovima Wistar soja ubrizgavan je KJ intraperitonealno u niskoj i visokoj koncentraciji (NKJ - 225 µg/g i VKJ - 675 µg/g) tokom 26 dana. Kontrolna grupa životinje tretirana je fiziološkim rastvorom. Na histološkom preparatu štitnjače pacova tretiranih NKJ naziru se ostaci folikula bez koloida i tireocita, infiltracija mononuklearnim ćelijama i proliferacija vezivnog tkiva, a u onih tretiranih VKJ vidi se proliferacije veziva, gubitka folikularne građe i infiltraciju mononuklearnim ćelijama. Zapaljenjske promene su znatno izraženije kod pacova tretiranih VKJ. Praćenjem hormonskog statusa (T4, T3,TSH ) i morfoloških promena štitaste žlezde, ustanovljeno je da terapijske doze KJ koji se primenjuje za lečenje, indukuju nastanak eksperimentalnog tiroiditisa kod genetski neosetljivih životinja, po tipu hroničnog destruktivnog Hašimoto tiroiditisa kod ljudi. Nekroza i zapaljenje pokrenute jodidom u neimunzovanih životinja, bez genske osetljivosti nov je, originalan, eksperimentalni model tiroiditisa.

Tungstates and polyoxotungstates have been known to be bioactive compounds for a long time. Based on our previous work about sodium tungstate (ST) and 12-tungstophosphoric acid (WPA) as hepatoprotective agents, we performed a seven week long experiment in rats, using ST and WPA pretreatments in thioacetamide (TAA) induced acute liver necrosis. The possible influences of dimethyl sulfoxide (DMSO), given orally for 3 consecutive days before inducing hepatic lesions, were also investigated. The effects were evaluated by the activity of serum enzymes, oxidative stress parameters, antioxidative defense markers, and histopathology in Wistar rats. The obtained results suggest that oral pretreatment with tungstates, especially ST, in combination with DMSO, before the TAA inducted liver necrosis, could be useful for the prevention of hepatic injury in rats. Volframati i polioksovolframati su decenijama poznati kao biološki aktivna jedinjenja. Kao nastavak prethodnog istraživanja hepatoprotektivnog efekta natrijum volframata (ST) i 12-volframfosforne kiseline (WPA), izveden je eksperiment na Wistar pacovima, koji su pretretirani sa ST i WPA tokom sedam nedelja a nakon tog perioda im je intraperitonealno ubrizgan tioacetamid (TAA) u cilju indukovanja akutne nekroze jetre. Ispitivan je uticaj dimetil sulfoksida (DMSO), koji je primenjivan per os u toku 3 konsekutativna dana pre indukcije hepatične nekroze. Efekti su evaluirani praćenjem aktivnosti serumskih enzima, parametara oksidativnog stresa, antioksidativnog odbrambenog markera, kao i histopatološkom analizom tkiva jetre. Dobijeni rezultati ukazuju da oralni pretretman pacova sa volframatima, pre svega ST, u kombinaciji sa DMSO, može biti koristan u prevenciji nekroze jetre indukovane tioacetamidom.