Glucocorticoid sensitivity, functional polymorphism in glucocorticoid receptor gene and glucocorticoid receptor expression in tumorous tissue of patients with incidentally discovered adrenocortical adenomas

Uvod: Adrenalni incidentalomi su tumori otkriveni različitim vizualizacionim metodama učinjenim bez sumnje na adrenalnu patologiju. Većina ovih tumora su nefunkcijski adrenokortikalni adenomi. Izvestan broj AI se prezentuje subkliničkim hiperkorticizmom (SH), stanjem biohemijskog kortizolskog ekscesa, u odsustvu klasičnih znakova i simptoma hiperkorticizma. Prevalencija SH kod pacijenata sa AI se procenjuje između 5 i 24%. Polimorfizam gena za glukokortikoidni receptor (GR) menja glukokortikoidnu (GC) senzitivnost, što dovodi do promena u metaboličkom profilu. Cilj studije: je bio da se ispita prevalenca subkliničkog hiperkorticizma (SH) i pridruženih bolesti kod pacijenata sa unilateralnim (UAI) i bilateralnim AI (BAI). Ispitivali smo učestalost četiri polimorfizma gena za GR (BclI, ER22/23EK, N363S i 9β) i predispoziciju za nastanak AI. Analizirali smo ekspresiju GR u tumorskom, peritumorskom i normalnom adrenalnom tkivu. Materijal i metode: Ispitivanje je obuhvatilo 152 pacijenta, 105 (69.1%) sa UAI i 47 (30.9%) sa BAI. SH je definisan na osnovu serumskog kortizola >50nmol/L nakon prekonoćnog deksametazon supresivnog testa (DST) sa 1mg ili nakon dvodnevnog niskodoznog DST (LDDST) sa 2mg deksametazona, uz još jedan navedeni parametar (ponoćni serumski kortizol >208nmol/L, 24-h urinarni slobodni kortizol >245nmol/L ili ACTH<10ng/L). Telesni sastav i koštana gustina su mereni dual-energy X-ray absorptiometry metodom na nivou lumbalnog dela kičme, vrata femura i celog skeleta. DNK je dobijena iz leukocita periferne krvi. Prisustvo polimorfizama je detektovano metodama PCR, RFLP i sekvenciranjem DNK. Uzorci tkiva su analizirani Western blot metodom. Rezultati: Pacijenti sa BAI se nisu razlikovali u odnosu na pacijente sa UAI po starosti, BMI, obimu struka, učestalosti dijabetesa, hipertenzije i dislipidemije. Prevalencija SH u celoj grupi iznosila je 20.5% (prekonoćni DST), odnosno 20.0% (LDDST) i bila je značajno veća kod pacijenata sa BAI u odnosu na pacijente sa UAI (31.1% vs 15.2%, p=0.026). Nezavisni prediktor SH je bila veličina adrenalnog adenoma (odds ratio [OR]=1.055, 95% confidence interval [CI] 1.017-1.094, p=0.004), nakon korekcije za godine, obim struka, indeks telesne mase i prisustvo BAI. Kostna gustina na nivou kičme je bila značajno niža kod pacijenata sa BAI u odnosu na UAI (0.96±0.14 vs 0.87±0.15, p=0.002). Nije nađena razlika u kostnoj gustini na nivou vrata femura...Background: Adrenal incidentalomas (AIs) are clinically silent adrenal masses that are detected incidentally during imaging procedures performed for unrelated diseases. The vast majority of these lesions are non-hypersecreting cortical adenoma. However, some of them present with subclinical hypercortisolism (SH), the condition of biochemical cortisol excess without the classical signs or symptoms of overt hypercortisolism. The prevalence of SH in patients with AIs varies between 5 and 24%. Glucocorticoid receptor (GR) polymorphisms alter glucocorticoid (GC) sensitivity and have been associated with altered metabolic profiles. Objective: The aim of this study was to investigate the prevalence of SH and associated co-morbidities in patients with unilateral AIs (UAIs) and bilateral AIs (BAIs). We also evaluated the prevalence of the four GR polymorphism genes (BclI, ER22/23EK, N363S and 9β) and investigated whether they influence susceptibility for AIs. We analyzed GR expression in tumorous, peritumorous and normal adrenocortical tissue samples. Material and Methods: We enrolled 152 patients, 105 (69.1%) with UAIs and 47 (30.9%) with BAIs. SH was diagnosed in the presence of serum cortisol levels after 1mg dexamethasone suppression test (DST) or after 2-day low-dose DST (LDDST) >50nmol/l with at least one of the following parameter (midnight serum cortisol >208nmol/L, 24-h urinary free cortisol >245nmol/24h, or ACTH <10ng/L). Body composition and bone mineral density (BMD) were measured with dual-energy X-ray absorptiometry at lumbar spine (LS), femoral neck (FN), and total skeleton. DNA was obtained from peripheral blood leucocytes. The polymorphisms were detected using PCR, RFLP and DNA sequencing. Tissue samples were studied by Western blot. Results: Age, body mass index (BMI), and waist circumference (WC) were comparable, and diabetes, hypertension and dyslipidemia occurred with a similar frequency in BAI and UAI patients. The overall prevalence of SH was 20.5% based on post-1mg DST, and 20.0% based on post-LDDST cortisol levels, and was more prevalent in BAIs than UAIs patients (31.1% vs 15.2%, respectively, p=0.026). Factor related to SH was adrenal mass size (odds ratio [OR]=1.055, 95% confidence interval [CI] 1.017-1.094, p=0.004), after adjustment for age, WC, BMI, and presence of BAIs. LS BMD was lower in BAI than in UAIs patients (0.96±0.14 vs 0.87±0.15, p=0.002). There were no differences in FN BMD..

Effect of coenzyme Q10 on ascorbic acid, vitamin E, and coenzyme Q concentrations in testes of rats chronically exposed to cadmium

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The effect of cadmium on ascorbic acid, vitamin E and coenzyme Q concentrations in rat kidneys: A possible protective role of coenzyme Q10

Projekat ministarstva br. 03E18 i 03E2

Time course of hematological parameters in bleeding-induced anemia

In order to investigate daily changes of hematological parameters in bleeding-induced anemia, we treated Wistar albino male rats by daily bleeding (1.5-2 mL of blood from the tail vein for eight days). Blood samples were taken before (on day zero) and on the first to eighth days of bleeding. The values of hematocrit, hemoglobin, and erythrocyte count decreased significantly after the second, sixth, and second days of bleeding, respectively. The number of leuko­cytes and platelets, as well as Heinz body levels, increased significantly after the third and second days of treatment. The percentage of reticulocytes increased significantly from the second day and attained the maximum level (32.55 ± 0.96%) on the eighth day

Antioxidant Response in Gills of Eurasian Perch ( Perca fluviatilis ) to Cyanobacterial Bloom Exposure in the Gruža Reservoir

The aim of this study was to assess the impact of an Aphanizomenon flos-aquae bloom in the Gruža Reservoir on the antioxidant parameters measured in the gill cells of Eurasian perch (Perca fluviatilis). Effects of the bloom were evaluated through copper, zinc and manganese containing superoxide dismutase (CuZnSOD and MnSOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR) and biotransformation phase II enzyme glutathione-S-transferase (GST) activities and glutathione (GSH) and sulfydryl (–SH) groups concentrations. The activities of CuZn SOD and Mn SOD decreased during the cyanobacterial bloom compared to the period before the bloom. The opposite trend was observed for CAT and GSH-Px activities that increased. During the bloom GR and GST activities and GSH concentration decreased significantly. CuZn SOD, GR and GST activities are detected as the most important parameters for individual variations. Our results indicated that an A. flos-aquae bloom promotes oxidative stress in the gills of P. fluviatilis. The investigated parameters could be used in environmental risk assessment procedures to monitor the effects of A. flos-aquae blooms on fish. Observed changes in the antioxidant parameters in P. fluviatilis gills make them potential biomarkers in the research of ecological risk situations in freshwater ecosystems with frequent cyanobacterial blooms.Water Research and Management (2016), 6(2): 19-2

Concentration of antioxidant compounds and lipid peroxidation in the liver and white muscle of hake (Merluccius merluccius L.) in the Adriatic sea

Specimens of a marine freshwater fish - hake (Merluccius merluccius L.) - were collected at the localities of Platamuni and Valdanos (Montenegro, South Adriatic) in the spring of 2003. Our results showed higher concentrations of LPO, Vit C, and Vit E in the liver in comparison with white muscle. The concentration of LPOin both tissues was higher, while that of Vit E was lower at Valdanos compared to Platamuni. These differences in parameters of oxidative stress are partly due to differences in temperature and the concentrations of nitrites, nitrates, and detergents in the waters of Valdanos compared to Platamuni.Tokom 2003. godine uzorkovane su jedinke morske ribe - oslića (Merluccius merluccius L.) iz voda sa lokaliteta Platamuni i Valdanos (Crna Gora, južni Jadran) u prolećnom periodu. Dobijeni rezultati su pokazali veće koncentracije LP, Vit Ci Vit E u jetri oslića u odnosu na belo mišićno tkivo. Koncentracija LP je bila veća, a Vit E manja u oba ispitivana tkiva oslića u vodama lokaliteta Valdanos u poređenju sa Platamuni. Ovakve promene parametara oksidacionog stresa posledica su promena temperature, kao i povećanih koncentracija nitrita, nitrata i deterdženata u vodama lokaliteta Valdanos u odnosu na Platamune.Projekat ministarstva br. 143035

Effects of cisplatin on lipid peroxidation and the glutathione redox status in the liver of male rats: The protective role of selenium

The role of oxidative stress in cisplatin (CP) toxicity and its prevention by pretreatment with selenium (Se) was investigated. Male Wistar albino rats were injected with a single dose of cisplatin (7.5 mg CP/kg b.m., i.p.) and selenium (6 mg Se/kg b.m, as Na2SeO3, i.p.) alone or in combination. The results suggest that CP intoxication induces oxidative stress and alters the glutathione redox status: reduced glutathione (GSH), oxidized glutathione (GSSG) and the GSH/GSSG ratio (GSH RI), resulting in increased lipid peroxidation (LPO) in rat liver. The pretreatment with selenium prior to CP treatment showed a protective effect against the toxic influence of CP on peroxidation of the membrane lipids and an altering of the glutathione redox status in the liver of rats. From our results we conclude that selenium functions as a potent antioxidant and suggest that it can control CP-induced hepatotoxicity in rats

Glutathione status in the blood of rats after reticulocytosis induced by phenylhydrazine and bleeding

In this experiment, we compared the in vivo effects of phenylhydrazine (PHZ) and bleeding treatment on the redox status and glutathione antioxidative mechanism parameters in the plasma and red blood cells (RBC) of rats. Results showed a lower level of reactive oxygen species (ROS), a higher level of lipid peroxidation and the effective antioxidative role of the glutathione system in the blood of bleeding rats. PHZ-treatment induced higher concentrations of ROS and an accumulation of oxidized glutathione in the plasma, while the glutathione system showed a satisfactory antioxidative capacity in the RBC of rats. When comparing the two anemic groups, the PHZ-treated rats showed marked oxidative stress in the plasma.Cilj ovog rada je bio da se isprate in vivo efekti fenilhidrazinom (PHZ) i krvarenjem indukovane retikulocitoze na parametre redoks i glutationskog antioksidativnog statusa u plazmi i crvenim krvnim ćelijama (RBC) pacova. Rezultati pokazuju niži nivo reaktivnih vrsta kiseonika (ROS), viši nivo lipidne peroksidacije i efikasnu antioksidacionu ulogu glutationskog sistema u krvi iskvavljenih pacova. Tretman PHZom prouzrokovao je više koncentracije ROSa i akumulaciju oksidovanog glutationa u plazmi, dok je glutationski sistem pokazao efikasan antioksidativni kapacitet u RBCu pacova. Kada se uporede dve anemične grupe, izraženiji oksidacioni stres se javlja u plazmi pacova tretiranih PHZom.Projekat ministarstva br. 143035

Paracetamol-induced changes of haemato-biochemical and oxidative stress parameters in rat blood: protective role of vitamin C and betaglucan

Paracetamol (acetaminophen) is widely used as an ov er-the-counter analgesic and antipyretic drug. The aim of this stu dy was to investigate the possible protective effects of vitamin C (100mg/kg/day i.p.) and β -glucan (40 mg/kg/day i.p.) on altered haematological, biochemical and oxidative s tress parameters in the blood of rats treated with paracetamol (100 mg/kg/day i.p.) for 3 days. Exposure of rats to paracetamol caused changes of some haematological parameters (R BCs count, Hb concentration, Ht value and WBCs count), suggesting that the paraceta mol induced haematotoxicity. Paracetamol reduced serum total protein (TP), album in and globulin, while increased alanine aminotransferase (ALT), aspartate aminotran sferase (AST) and lactate dehydrogenase (LDH) activities compared to the cont rol. The results indicate that paracetamol is led to significant decrease in the c oncentration of Na + and K + and increase of Ca 2+ in the serum compared to the control. Coadministra tion of vitamin C and β -glucan with paracetamol reversed these changes of haematol ogical and biochemical parameters and diminished the toxic effects of paracetamol. Th e obtained results indicated that the concentration of LPO in erythrocytes significantly increased in, while the concentration of GSH significantly decreased in the group treated with paracetamol compared to control group. Coadministration of vitamin C and β -glucan with paracetamol reversed paracetamol-induced alterations in these oxidative stress parameters. This study suggests that paracetamol has significant prooxidative effec ts and may disrupt oxidant/antioxidant balance in erythrocytes. Furthermore, coadministrat ion with vitamin C and β -glucan have protective effects on paracetamol-induced oxidative damage and haematotoxicity.Kragujevac Journal of Science (2016), 38: 135-14

Effects of cisplatin on lipid peroxidation and the glutathione redox status in the liver of male rats: The protective role of selenium

The role of oxidative stress in cisplatin (CP) toxicity and its prevention by pretreatment with selenium (Se) was investigated. Male Wistar albino rats were injected with a single dose of cisplatin (7.5 mg CP/kg b.m., i.p.) and selenium (6 mg Se/kg b.m, as Na2SeO3, i.p.) alone or in combination. The results suggest that CP intoxication induces oxidative stress and alters the glutathione redox status: reduced glutathione (GSH), oxidized glutathione (GSSG) and the GSH/GSSG ratio (GSH RI), resulting in increased lipid peroxidation (LPO) in rat liver. The pretreatment with selenium prior to CP treatment showed a protective effect against the toxic influence of CP on peroxidation of the membrane lipids and an altering of the glutathione redox status in the liver of rats. From our results we conclude that selenium functions as a potent antioxidant and suggest that it can control CP-induced hepatotoxicity in rats.U našoj studiji ispitivana je uloga oksidacionog stresa u toksičnosti cisplatina (SR) i njegova prevencija pretretmanom selenom (Se). Mužjaci Wistar albino pacova su inicirani jednom dozom cisplatina (7.5 mg CP/kg t.m., i.p.) i selena (6 mg Se/kg t.m, kao Na2SeO3, i.p.) pojedinačno ili u kombinaciji. Rezultati pokazuju da intoksikacija SR uzrokuje oksidacioni stres i promenu glutation redoks statusa: redukovanog (GSH), oksidovanog (GSSG) i GSH/GSSG indeksa (GSH RI), kao i povećanje lipidne peroksidacije (LPO) u jetri pacova. Tretman Se koji je prethodio tretmanu SR pokazao je zaštitne efekte protiv toksičnog delovanja SR na peroksidaciju lipida membrane i promenu glutation redoks statusa u jetri pacova. Na osnovu naših rezultata zaključujemo da Se, deluje kao snažan antioksidans i da može imati ulogu u kontroli SR indukovane hepatotoksičnosti kod pacova.Projekat ministarstva br. 143035