Supplementary material for the article: Mihajlović, M.; Petrović, J.; Maletić, S.; Isakovski, M. K.; Stojanović, M.; Lopičić, Z.; Trifunović, S. Hydrothermal Carbonization of Miscanthus × giganteus: Structural and Fuel Properties of Hydrochars and Organic Profile with the Ecotoxicological Assessment of the Liquid Phase. Energy Convers. Manage. 2018, 159, 254–263. https://doi.org/10.1016/j.enconman.2018.01.003

Supplementary material for: [https://doi.org/10.1016/j.enconman.2018.01.003]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2100

Metabolism of sodium nitroprusside (SNP) in rat red blood cells

Sodium nitroprusside (SNP) is an established drug, clinically used in the treatment of hypertensive emergencies. The aim of this study is further elucidation of the molecular mechanism of the SNP action, particularly its metabolism in rat erythrocytes and reticulocytes. Rats erythrocyte and reticulocyte-rich suspensions were aerobically incubated without (control) or in the presence of SNP (0.1,0.25,0.5,1.0 and 1.5 mM). The concentrations of reactive nitrogen species (RNS) and reactive oxygen species (ROS) were determined after incubation. In rat erythrocytes, SNP did not alter nitrite level (NO+ ion indicator), while significantly increased concentrations of hydroxylamine (NO- ion indicator), S-nitrosothiols (SNO) and 3-nitrotyrosine (peroxynitrite indicator). Concentration of superoxide anion (O2-) decreased in the presence of low doses of SNP only, while level of hydrogen peroxide (H2O2) increased in dose-dependent manner in rat erythrocytes. On the other hand, SNP significantly increased nitrite, hydroxylamine and 3-nitrotyrosine concentrations in rat reticulocytes. In addition, low doses of SNP induced decrease of O2- level. Concentration of H2O2 did not alter in rat reticulocytes. On the basis of these data, we can conclude: SNP spontaneously liberated nitric oxide as NO- ion in rat erythrocytes and reticulocytes. In addition, applied experimental doses of SNP induced strong nitrosative and oxidative stress in these cells.Natrijum nitroprusid (SNP) je lek koji se klinički koristi u tretmanu hipertenzija. Cilj ovog rada je da se objasne molekularni mehanizmi delovanja SNPa, sa posebnim akcentom na metabolizam ovog leka u eritrocitima i retikulocitima pacova. Suspenzije eritrocita i crvenih krvnih ćelija bogate retikulocitima su aerobno inkubirane bez (kontrola) ili u prisustvu različitih koncentracija SNPa (0.1, 0.25, 0.5, 1.0 i 1.5 mM). Koncentracije reaktivnih vrsta azota (RNS) i kiseonika (ROS) su određivane nakon inkubacije. U eritrocitima pacova, SNP ne menja koncentraciju nitrita (indikator NO+ jona), dok značajno povećava koncentracije hidroksilamina (indicator NO- jona), S-nitrozotiola (SNO) and 3-nitrotirozina (indikator peroksinitrita). Koncentracija superoksid anjon radikala (O2-) je smanjena samo u prisustvu niskih doza SNP, dok je nivo vodonik peroksida (H2O2) povećan na dozno-zavisan način u eritrocitima pacova. S druge strane, SNP značajno povećava koncentracije nitrita, hidroksilamina i 3-nitrotirozina u retikulocitima pacova. Niske doze SNPa indukuju smanjenje O2- nivoa. Koncentracije H2O2 nisu promenjenje u retikulocitima pacova. Na osnovu iznetih podataka, možemo zaključiti: SNP spontano oslobađa azot monoksid kao NO" jon u eritrocitima i retikulocitima pacova. Primenjene eksperimentalne doze SNPa indukuju snažan nitrozacioni i oksidacioni stres u ovim ćelijama.nul

Antioxidant effect of coenzyme Q10 in blood from cadmium-exposed rats

The effects of acute exposure to cadmium (Cd) on the blood antioxidant defense system (AOS), lipid peroxide (LP) concentration and hematological parameters, and the possible protective role of coenzyme Q10 (CoQ10) was studied. Male Wistar albino rats 3 months old were treated with cadmium as CdCl2 (0,4mg Cd/kg b.m., i.p., 24h before the sacrificing) or with coenzyme Q10 + Cd (20mg CoQ10/kg b.m., i.m., 48h + 0,4 mg Cd/kg b.m., i.p., 24h before the sacrificing). The hematological parameters: red blood cells count (RBCs), hemoglobin (Hb) concentration and hematocrite (Ht) value were significantly decreased in the blood of Cd treated rats. Intoxication with Cd was also followed by significantly increased of LP concentration. We also observed increased concentrations of non-enzymatic components of antioxidant defense system (AOS): reduced glutathione (GSH), vitamin C (Vit C) and vitamin E (Vit E). Pretreatment with CoQ10 exhibited a protective role on the toxic effects of Cd on the hematological values, LP concentration as well as on endogeneous antioxidant components.Cilj ovog rada je bio da se ispita uticaj kadmijuma (Cd) na antioksidacioni zaštitni sistem (AOS), koncentraciju lipidnih peroksida (LP) i hematološke parametre u krvi, kao i zaštitna uloga koenzima Q10 (CoQ10). Mužjaci pacova Wistar albino, stari tri meseca, akutno su tretirani kadmijumom (0,4mg Cd/kg t.m., i.p., 24h pre žrtvovanja) i koenzimom Q10 + Cd (20mg CoQ10/kg t.m., i.m., 48h + 0,4mg Cd/kg t.m., 24h pre žrtvovanja). Hematološki parametri: broj eritrocita (RBCs), koncentracija hemoglobina (Hb) i hematološka vrednost (Ht) su značajno smanjeni u krvi pacova posle tretmana kadmijumom. Cd značajno povećava i koncentraciju LP, kao i koncentracije neenzimskih komponenti AOS-a: redukovani glutation (GSH), vitamin C (Vit C) i vitamin E (Vit E). Eksperimenti sa pacovima koji su izazvanu anemiju i oksidaciona oštećenja (smanjuje koncentraciju LP), kao i značajno umanjuje toksične efekte Cd na komponente AOS-a.nul

Effect of SIN-1 on antioxidative defence system in red blood cells of rats

The main objective of this research is to establish the effect of 3-morpholino-sydnonimine-hydrochloride (SIN-1) on antioxidative defense system (AOS) in rat red blood cells. Rats erythrocyte and reticulocyte-rich suspensions were aerobically incubated without (control) or in the presence of SIN-1 (0.1, 0.25, 0.5, 1.0 and 1.5 mM). The concentrations of non-enzymatic components and activities of enzymatic component of AOS were determined after incubation. In rat erythrocytes SIN-1 did not alter vitamin C (Vit C), while induced increase of vitamin E (Vit E) concentrations. Unaltered concentration of reduced glutathione (GSH) following by decrease of oxidized glutathione (GSSG) resulted in decrease of reduction potential of this redox couple (decrease of GSSG/2GSH ratio). These data indicate that GSH redox couple is included in defense of rat erythrocytes from oxidative damage. Activities of all AOS enzyme did not altered in the presence of SIN-1 in rat erythrocytes. In reticulocytes a high doses of SIN-1 induced a decrease of Vit C and increase of Vit E concentrations, indicating defense role of antioxidative vitamins. Level of GSH redox couple is not changed. SIN-1 inhibited activity of manganese containing superoxide dismutase (Mn SOD) which was followed by stimulation of copper zinc containing superoxide dismutase (CuZn SOD) activity. SIN-1 did not alter activities of other AOS enzyme in rat reticulocytes.Cilj ovog istraživanja bio je da se objasni efekat 3-morfolino-sidnonimin-hidrohlorida (SIN-1) na antioksidacioni zaštitni sistem (AOS) u crvenim krvnim ćelijama pacova. Suspenzije eritrocita i crvenih krvnih ćelija bogate retikulocitima aerobno su inkubirane bez (kontrola) ili u prisustvu različitih koncentracija SIN-1 (0.1, 0.25, 0.5 1.0 i 1.5 mM). Koncentracije neenzimskih komponenti AOS i aktivnost enzima AOS određivane su nakon inkubacije crvenih krvnih ćelija. U eritrocitima pacova SIN-1 ne menja Vit C, dok povećava koncentraciju Vit E. Nepromenjena koncentracija redukovanog glutationa (GSH) praćena je smanjenjem nivoa oksidovanog glutaliona (GSSG) rezultujući smanjenjem redukcionog potencijala ovog redoks para (smanjenje GSSG/2GSH odnosa). Ovi podaci pokazuju da je GSH redoks par uključen u odbranu eritrocita od oksidacionih oštećenja. Aktivnost svih enzima AOS-a nisu bile promenjene u prisustvu SIN-1 u eritrocitima pacova. U retikulocitima pacova SIN-1 u visokim dozama izaziva smanjenje Vit C i povećanje Vit E koncentracije što ukazuje na zaštitnu ulogu antioksidacionih vitamina. Nivo GSH redoks para nije promenjen. SIN-1 inhibirá aktivnost Mn SOD što je praćeno stimulacijom aktivnosti CuZn SOD. SIN-1 ne menja aktivnost ostalih enzima AOS-a u retikulocitima pacova

Effect of olive oil on antioxidative defense components in liver of cadmium intoxicated rats

The present study investigated the possible protective role of olive oil on antioxidative defense components in liver of cadmium (Cd) intoxicated animals. Male, Wistar albino 3 months old rats were injected with a single dose of: 1) CdCl2 (0.4 mg Cd/kg i.p. and sacrificed after 24h), 2) olive oil (0.4 ml olive oil/kg b.m., i.m., 48h before the sacrificing) and 3) olive oil + Cd (in above mentioned amounts). Treatment with Cd increased lipid peroxidation (LP) in blood and activities of alanine and aspartate transaminase (ALT and AST) in plasma. Concentrations of non enzymatic components of antioxidative defense system (AOS): vitamin C (Vit C) and vitamin E (Vit E) were significantly decreased in liver tissue of Cd treated rats. Pretreatment with olive oil reversed Cd induced alterations in LP content and transaminases activities, as well as on endogenous antioxidive defense components.U ovom radu ispitivana je moguća zaštitna uloga maslinovog ulja na antioksidacione zaštitne komponente u jetri pacova intoksiciranim kadmijumom (Cd). Mužjaci pacova Wistar albino, starosti tri meseca akutno su tretirani sa: 1) CdCl (0.4 mg Cd/kg t.m., i.p., 24h pre žrtvovanja), 2) maslinovim uljem (0.4 ml maslinovog ulja/kg t.m., i.m., 48h pre žrtvovanja) i 3) maslinovim uljem + Cd (u navedenim količinama). Posle tretmana kadmijumom značajno se povećava koncentracija lipidnih peroksida (LP) u krvi, kao i aktivnost alanin i aspartat transaminaza (ALT i AST) u plazmi. Koncentracije neenzimskih komponenti antioksidacionog zaštitnog sistema (AOS): vitamin C (Vit C) i vitamin E (Vit E) su značajno smanjene u jetri kod pacova tretiranih kadmijumom. Eksperimenti sa pacovima koji su u pred tretmanu dobijali maslinovo ulje pokazuju da maslinovo ulje sprečava ili ublažava kadmijumom izazvane promene u koncentraciji LP, aktivnosti, kao i koncentraciji endogenih antioksidacionih zaštitnih komponenti

Effect of olive oil on antioxidative defense components in liver of cadmium intoxicated rats

The present study investigated the possible protective role of olive oil on antioxidative defense components in liver of cadmium (Cd) intoxicated animals. Male, Wistar albino 3 months old rats were injected with a single dose of: 1) CdCl2 (0.4 mg Cd/kg i.p. and sacrificed after 24h), 2) olive oil (0.4 ml olive oil/kg b.m., i.m., 48h before the sacrificing) and 3) olive oil + Cd (in above mentioned amounts). Treatment with Cd increased lipid peroxidation (LP) in blood and activities of alanine and aspartate transaminase (ALT and AST) in plasma. Concentrations of non enzymatic components of antioxidative defense system (AOS): vitamin C (Vit C) and vitamin E (Vit E) were significantly decreased in liver tissue of Cd treated rats. Pretreatment with olive oil reversed Cd induced alterations in LP content and transaminases activities, as well as on endogenous antioxidive defense components.U ovom radu ispitivana je moguća zaštitna uloga maslinovog ulja na antioksidacione zaštitne komponente u jetri pacova intoksiciranim kadmijumom (Cd). Mužjaci pacova Wistar albino, starosti tri meseca akutno su tretirani sa: 1) CdCl (0.4 mg Cd/kg t.m., i.p., 24h pre žrtvovanja), 2) maslinovim uljem (0.4 ml maslinovog ulja/kg t.m., i.m., 48h pre žrtvovanja) i 3) maslinovim uljem + Cd (u navedenim količinama). Posle tretmana kadmijumom značajno se povećava koncentracija lipidnih peroksida (LP) u krvi, kao i aktivnost alanin i aspartat transaminaza (ALT i AST) u plazmi. Koncentracije neenzimskih komponenti antioksidacionog zaštitnog sistema (AOS): vitamin C (Vit C) i vitamin E (Vit E) su značajno smanjene u jetri kod pacova tretiranih kadmijumom. Eksperimenti sa pacovima koji su u pred tretmanu dobijali maslinovo ulje pokazuju da maslinovo ulje sprečava ili ublažava kadmijumom izazvane promene u koncentraciji LP, aktivnosti, kao i koncentraciji endogenih antioksidacionih zaštitnih komponenti

Metabolism of SIN-1: Peroxynitrite donor in rat red blood cells

The aim of this study is elucidation of the molecular mechanism of the 3-morpholino-sydnonimine-hydrochloride (SIN-1) action, particularly its metabolism in rat erythrocytes and reticulocytes. SIN-1 is active metabolite of molsidomine, an established vasodilator drug, clinically used in the treatment of coronary artery disease. Rats erythrocyte and reticulocyte-rich suspensions were aerobically incubated without (control) or in the presence of SIN-1 (0.1, 0.25, 0.5, 1,0 and 1.5 mM). The concentrations of reactive nitrogen species (RNS), reactive oxygen species (ROS) and parameters that indicate oxidative damage of red blood cells were determined after incubation. In rat erythrocytes SIN-1 in high doses increased concentrations of nitrite (NO+ ion indicator), hydroxylamine (NO- ion indicator) and 3-nitrotyrosine (peroxynitrite indicator). Concentration of superoxide anion radical (O2) increased in the presence of SIN-1 on dose-dependent way (except in the presence of higher applied dose), while level of hydrogen peroxide (H2O2) did not alter. In rat reticulocytes SIN-1 significantly increased 3-nitrotyrosine concentrations only. In addition, low doses of SIN-1 induced decrease of O2.- level, while concentration of H2O2 increased dose-dependently. These data indicate that SIN-1-induced nitrosative and oxidative stress in rat red blood cells. The strong oxidative damage of erythrocytes and reticulocytes (increased level of methemoglobin, Heinz bodies and lipid peroxides concentrations) are appeared as consequence of SIN-1-induced oxidative stress.Cilj ovog rada je da se objasne molekularni mehanizmi delovanja 3-morfolino-sidnonimin-hidrohlorida (SIN-1) sa posebnim akcentom na metabolizam ovog leka u eritrocitima i retikulocitima pacova. SIN-1 je aktivni metabolit molsidomina, leka koji se klinički koristi u terapiji bolesti koronarnih arterija. Suspenzije eritrocita i crvenih krvnih ćelija bogate retikulocitima su aerobno inkubirane bez (kontrola) ili u prisustvu različitih koncentracija SIN-1 (0.1, 0.25, 0.5, 1.0 i 1.5 mM). Koncentracije reaktivnih vrsta azota (RNS) i reaktivnih vrsta kiseonika (ROS), kao i koncentracije parametara koji ukazuju na oksidaciona oštećenja crvenih krvnih ćelija, određivane su nakon inkubacije. U eritrocitima pacova, SIN-1 samo u visokim dozama povećava koncentracije nitrita (indikator NO+ jona) hidroksilamina (indicator NO- jona) i 3-nitrotirozina (indikator peroksinitrita). Koncentracija superoksid anjon radikala (O2.-) je povećana na dozno-zavisan način (osim u prisustvu maksimalne primenjene doze SIN-1) dok se nivo vodonik peroksida (H2O2) ne menja. U retikulocitima pacova SIN-1 značajno povećava samo koncentraciju 3-nitrotirozina. Niske doze SIN-1 indukuju smanjenje O2.- nivoa, dok koncentracije H2O2 rastu na dozno-zavisan način. Na osnovu iznetih podataka, možemo zaključiti da SIN-1 indukuje nitrozacioni i oksidacioni stres u crvenim krvnim ćelijama pacova. Ovakvo stanje je posledično praćeno snažnim oksidacionim oštećenjem eritrocita i retikulocita (povećanjem koncentracija methemoglobina, Heinz-ovih telašaca i lipidnih peroksida).nul

Metabolism of SIN-1: Peroxynitrite donor in rat red blood cells

The aim of this study is elucidation of the molecular mechanism of the 3-morpholino-sydnonimine-hydrochloride (SIN-1) action, particularly its metabolism in rat erythrocytes and reticulocytes. SIN-1 is active metabolite of molsidomine, an established vasodilator drug, clinically used in the treatment of coronary artery disease. Rats erythrocyte and reticulocyte-rich suspensions were aerobically incubated without (control) or in the presence of SIN-1 (0.1, 0.25, 0.5, 1,0 and 1.5 mM). The concentrations of reactive nitrogen species (RNS), reactive oxygen species (ROS) and parameters that indicate oxidative damage of red blood cells were determined after incubation. In rat erythrocytes SIN-1 in high doses increased concentrations of nitrite (NO+ ion indicator), hydroxylamine (NO- ion indicator) and 3-nitrotyrosine (peroxynitrite indicator). Concentration of superoxide anion radical (O2) increased in the presence of SIN-1 on dose-dependent way (except in the presence of higher applied dose), while level of hydrogen peroxide (H2O2) did not alter. In rat reticulocytes SIN-1 significantly increased 3-nitrotyrosine concentrations only. In addition, low doses of SIN-1 induced decrease of O2.- level, while concentration of H2O2 increased dose-dependently. These data indicate that SIN-1-induced nitrosative and oxidative stress in rat red blood cells. The strong oxidative damage of erythrocytes and reticulocytes (increased level of methemoglobin, Heinz bodies and lipid peroxides concentrations) are appeared as consequence of SIN-1-induced oxidative stress.Cilj ovog rada je da se objasne molekularni mehanizmi delovanja 3-morfolino-sidnonimin-hidrohlorida (SIN-1) sa posebnim akcentom na metabolizam ovog leka u eritrocitima i retikulocitima pacova. SIN-1 je aktivni metabolit molsidomina, leka koji se klinički koristi u terapiji bolesti koronarnih arterija. Suspenzije eritrocita i crvenih krvnih ćelija bogate retikulocitima su aerobno inkubirane bez (kontrola) ili u prisustvu različitih koncentracija SIN-1 (0.1, 0.25, 0.5, 1.0 i 1.5 mM). Koncentracije reaktivnih vrsta azota (RNS) i reaktivnih vrsta kiseonika (ROS), kao i koncentracije parametara koji ukazuju na oksidaciona oštećenja crvenih krvnih ćelija, određivane su nakon inkubacije. U eritrocitima pacova, SIN-1 samo u visokim dozama povećava koncentracije nitrita (indikator NO+ jona) hidroksilamina (indicator NO- jona) i 3-nitrotirozina (indikator peroksinitrita). Koncentracija superoksid anjon radikala (O2.-) je povećana na dozno-zavisan način (osim u prisustvu maksimalne primenjene doze SIN-1) dok se nivo vodonik peroksida (H2O2) ne menja. U retikulocitima pacova SIN-1 značajno povećava samo koncentraciju 3-nitrotirozina. Niske doze SIN-1 indukuju smanjenje O2.- nivoa, dok koncentracije H2O2 rastu na dozno-zavisan način. Na osnovu iznetih podataka, možemo zaključiti da SIN-1 indukuje nitrozacioni i oksidacioni stres u crvenim krvnim ćelijama pacova. Ovakvo stanje je posledično praćeno snažnim oksidacionim oštećenjem eritrocita i retikulocita (povećanjem koncentracija methemoglobina, Heinz-ovih telašaca i lipidnih peroksida).nul

Effects of acute hypoxia on the energy status and antioxidant defense system in the blood of carp - Cyprinvs carpio L.

The influence of acute hypoxia on glucose, pyruvate, lipid peroxide (LP) reduced glutathione (GSH) concentrations and lactate level in the whole blood of carp (Cyprinus carpio L) under aquarium conditions were studied. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), the concentrations of ATP and ADP and ATP/ADP ratio in the red blood cells (RBCs) were analyzed. Glutathione-S-transferase (GST) activity was determined in the plasma. Our experiments showed that short-term and long-term hypoxia causes significant changes of all examined haema-tological parameters. Increased concentration of LP and increased SOD CAT and GST activities, as well as a decreased GSH-Px activity showed that under hypoxic conditions oxidative stress and RBCs damage were produced.Uticaj akutne hipoksije na koncentraciju glukoze, piruvata, lipidnih peroksida (LP), redukovanog glutationa (GSH) i na nivo laktata u krvi šarana (Cyprinu carpio L) je analiziran u akvarijumskim uslovima. U crvenim krvnim zrncima (RBCs) je analizirana aktivnost enzima superoksid-peroksidaze (GSH-Px) kao i koncentracije ATP, ADP i odnos ATP/ADP. U plazmi je analizirana aktivnost glutation S-transferaze (GST). U našim eksperimentima je utvrđeno da kratkotrajna progresivna hipoksija izaziva značajne promene svih ispitivanih hematoloških parametara. Povećanje koncentracije LP i povećanje aktivnosti enzima SOD, CAT i GST uz istovremeno smanjenje aktivnosti GSH-Px potvrđuju nastanak oksidacionog stresa i oštećenje RBCs pod hipoksičnim uslovima.nul

Effects of acute hypoxia on the energy status and antioxidant defense system in the blood of carp - Cyprinvs carpio L.

The influence of acute hypoxia on glucose, pyruvate, lipid peroxide (LP) reduced glutathione (GSH) concentrations and lactate level in the whole blood of carp (Cyprinus carpio L) under aquarium conditions were studied. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), the concentrations of ATP and ADP and ATP/ADP ratio in the red blood cells (RBCs) were analyzed. Glutathione-S-transferase (GST) activity was determined in the plasma. Our experiments showed that short-term and long-term hypoxia causes significant changes of all examined haema-tological parameters. Increased concentration of LP and increased SOD CAT and GST activities, as well as a decreased GSH-Px activity showed that under hypoxic conditions oxidative stress and RBCs damage were produced.Uticaj akutne hipoksije na koncentraciju glukoze, piruvata, lipidnih peroksida (LP), redukovanog glutationa (GSH) i na nivo laktata u krvi šarana (Cyprinu carpio L) je analiziran u akvarijumskim uslovima. U crvenim krvnim zrncima (RBCs) je analizirana aktivnost enzima superoksid-peroksidaze (GSH-Px) kao i koncentracije ATP, ADP i odnos ATP/ADP. U plazmi je analizirana aktivnost glutation S-transferaze (GST). U našim eksperimentima je utvrđeno da kratkotrajna progresivna hipoksija izaziva značajne promene svih ispitivanih hematoloških parametara. Povećanje koncentracije LP i povećanje aktivnosti enzima SOD, CAT i GST uz istovremeno smanjenje aktivnosti GSH-Px potvrđuju nastanak oksidacionog stresa i oštećenje RBCs pod hipoksičnim uslovima.nul