Total Antioxidant Status Activity Research Articles

Efficiency of Combined Use of Fullerene C60 and Bovine Serum Albumin for Rehabilitation of Vitrified Fragments of Rat Immature Seminiferous Tubules

Background. Today, cryopreserved reproductive tissues are used to treat some forms of male infertility. However, after long-term preservation of fragments of seminiferous tubules of testes (FSTT) in a low-temperature bank (-196 °С) their morphological and functional characteristics decrease reversibly. To solve this problem after freezing-thawing, the use of rehabilitation media with special additives is promising. Due to the fact that serum albumin and fullerene C60 have powerful protective and antioxidant properties, their use allows to stabilize the plasma membrane, osmotic pressure, and reduce free radicals that make them promising candidates to use in the development of rehabilitation media for biological objects after cryopreservation. Objective. The efficacy of fullerene C60, bovine serum albumin (BSA), and their combination as components of rehabilitation medium of vitrified FSTT of immature rats was studied. Methods. Vitrified-thawed samples of FSTT were incubated (22 °C) for 30 minutes in Leibovitz's medium with addition of 15 mg/mL C60, 5 g/L BSA or their combination. Control samples were incubated in the medium without C60 or BSA addition. Metabolic activity (MTT test), histomorphological data, total antioxidant status (TAS), reactive oxygen species (ROS) production, activity of g-glutamyltransferase (gGGT), and glucose-6-phosphate dehydrogenase were determined in the samples after rehabilitation in the investigated media. Results. The use of C60 led to the increase of metabolic (by 1.26 times) and TAS (by 1.74 times) activities, to the decrease in the number of ROS+ cells (by 1.35 times) and to the improvement of the spermatogenic epithelium binding to the basement membrane versus control sample. Application of BSA did not significantly affect the studied biochemical indices but decreased the number of tubules with desquamation of spermatogenic epithelium in histological sections. The combined use of BSA and C60 had the best effect among investigated rehabilitation media that led to the increase of metabolic activity (by 1.51 times), TAS activity (by 1.78 times), gGGT activity (by 1.59 times), histostructure restoration and the decrease in the number of ROS+ cells (by 1.45 times) compared to the control samples. Conclusions. The use of C60 and BSA combination increases the metabolic and antioxidant activity of vitrified FSTT and also has a positive effect on their histostructural characteristics compared to control samples. It should be noted that the effect of С60 and BSA addition to rehabilitation medium exceeds the results of using the investigated additives separately (by the metabolic and gGGT activity as well as architectonics of vitrified FSTT). These data relate to reproductive medicine and can be used to develop an effective rehabilitation protocol for vitrified FSTT.

Elevated circulating nitric oxide levels correlates with enhanced oxidative stress in patients with hyperemesis gravidarum

Since the biochemical and molecular mechanisms responsible for ongoing oxidative stress in hyperemesis gravidarum (HEG) patients have not yet been fully elucidated, the aim of this study was to evaluate the possible role of nitric oxide (NO), malondialdehyde (MDA) and other oxidative stress markers in the disease pathophysiology. Moreover, the relation between oxidative stress markers and Helicobacter pylori (H. pylori) infection was also investigated. Women with pregnancies complicated by HEG (n = 33) were compared with pregnant women without HEG (n = 30) and with healthy non-pregnant women (n = 31). Serum NO, MDA, total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI) and H. pylori infection status were determined for each subject. Serum NO levels and OSI index were found to be increased (p = .001 and .013, respectively) and TAS levels were decreased (p < .001) in HEG patients compared with both controls regardless of H. pylori infection status. Serum MDA and TOS levels were not different between the study groups. Helicobacter pylori infection rates were similar in each group. The reduced antioxidant activities, as well as the increased OSI and NO levels in HEG patients indicate possible oxidative stress conditions in HEG patients. Moreover, serum NO levels may be used as an adjunctive marker to distinguish HEG patients from other causes of emesis during pregnancy.Impact statementWhat is already known on this subject? Current evidence suggests that oxidative stress is a significant factor responsible for a number of complications during pregnancy.What do the results of this study add? Hyperemesis gravidarum is an oxidative stress condition, as reflected by increased nitric oxide (NO) and decreased total antioxidant status activity, regardless of H. Pylori infection.What are the implications for clinical practice and/or further research? Full disclosure of the association between circulating NO and hyperemesis gravidarum would shed light on underlying biological mechanisms and could help clinical management of similar pregnancy-associated morbidity states.

Serum malondialdehyde level and activity of total antioxidant status of dogs with age-related cataract.

determination of malondialdehyde (MDA) level and total antioxidant status (TAS) in the serum of dogs with age-related cataract. Forty dogs, 10-16 years of age, which were diagnosed with age-related cataract were examined. The control group consisted of 12 dogs, 8-13 years of age, without cataract. MDA was determined using a Perkin-Elmer LS 30 luminescence spectrometer using the method with thiobarbituric acid (TBA). TAS was determined using the colorimetric method with a set of Randox Total Antioxidant Status reagents. The results show a higher level of MDA with statistically significant difference and lower activity of TAS in the serum of dogs with age-related cataract. The results obtained indicate higher systemic lipid peroxidation and weakening of the general defense system in dogs with age-related cataract. These results suggest that such conditions may have an effect on the development of age-related cataract and are consistent with the theory of free radical age-related cataract development.

Effects of dietary Rhodiola rosea on growth, body composition and antioxidant capacity of white shrimp Litopenaeus vannamei under normal conditions and combined stress of low-salinity and nitrite

An 8-week feeding trial followed by an acute combined stress of low-salinity and nitrite were designed to examine the effects of Rhodiola rosea on growth, body composition and antioxidant capacity of the white shrimp Litopenaeus vannamei. Shrimp (3.60 ± 0.03 g) were randomly allocated to 4 groups, with 3 replicates per group and 30 shrimp per replicate. The dietary groups were given as follows: control and diets containing R. rosea (300, 1000 and 3000 mg kg−1). Antioxidant capacity including total antioxidant status (TAS), activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) as well as gene expression of GSH-Px and CAT in hepatopancreas of shrimp was analysed at the end of feeding trial and again during the combined stress. The results showed that supplemental R. rosea had no significant impacts on growth and whole body composition of shrimp. There was significant effect of R. rosea supplementation dose and times on the antioxidant status parameters of L. vannamei. R. rosea (3000 mg kg−1) significantly improved the resistance of L. vannamei against the combined stress of low-salinity and nitrite, as indicated by the significant higher activities of TAS, GSH-Px and CAT (P < 0.05), as well as by the higher transcript levels of GPx and CAT gene (P < 0.05).

Effects of Acute Administration of D,L-Homocysteine Thiolactone on the Antioxidative Status of Rat Intestine and Liver

Abstract Oxidative stress appears to play a role in the pathogenesis of several inflammatory gastrointestinal diseases. Increased homocysteine levels may play a role in the pathogenesis of Chron’s disease and ulcerative colitis. The aim of this study was to examine the influence of homocysteine on the antioxidant status of rat intestine and liver. The levels of thiobarbituric acid reactive substances (TBARS), activity of catalase (CAT) and total antioxidant status (TAS) were investigated in the isolated gut and liver of young male rats in the control group (8 rats) and after 3-hоur incubation in high doses of D, L-homocysteine thionolactone (Hcy) (10 μmol/L) (8 rats). Samples of duodenum, ileum, colon and liver were homogenized in sodium phosphate buffer (1:10). Homogenates were centrifuged at 10000 for 10 min at 4° C and the supernatant was taken for biochemical assays. Our results showed that high D, L-homocysteine thionolactone concentration reduced enzymatic catalase activity in homogenates of the isolated segments of duodenum (27.04%) p&lt;0.01; ileum (37.27%), colon (34.17%) and liver (67.46%) p&lt;0.001. Exposition to high D,L-homocysteine thiolactone concentration significantly increased TBARS levels in the duodenum (106.05%), ileum (47.24%), colon (112.75%) and liver (32.07%) (p&lt;0.01). Homocysteine also modifi ed the total antioxidant status of homogenates from the duodenum, ileum, colon and liver, increasing by 20.68% (duodenum), 24.74% (ileum), 14.88% (colon) and 19.35% (liver) (p&lt;0.001). Homocysteine induced a consistent oxidative stress in rat’s intestine and liver (reduced activity of catalase and increased level of TBARS), but the elevated activity of TAS in our experiments could be explained as an adaptive response to the generated free radicals which indicates the failure of the total antioxidant defense mechanism to protect the tissues from damage caused by homocysteine.

Effects of dietary chlorogenic acid on growth performance, antioxidant capacity of white shrimp Litopenaeus vannamei under normal condition and combined stress of low-salinity and nitrite

An eight-week feeding trial followed by an acute combined stress test of low-salinity and nitrite were performed to evaluate effects of chlorogenic acid (CGA) on growth performance and antioxidant capacity of white shrimp Litopenaeus vannamei. Shrimp were randomly allocated in 12 tanks (30 shrimp per tank) and triplicate tanks were fed with a control diet or diets containing different levels of CGA (100, 200 and 400 mg kg−1 feed) as treatment groups. Growth performance including weight gain (WG), biomass gain (BG), feed conversion ratio (FCR), and feed intake were determined after feeding for 56 days. Antioxidant capacity were evaluated by determining the activity of total antioxidant status (TAS), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) as well as the gene expression of GSH-Px and CAT in the hepatopancreas of shrimp at the end of feeding trial and again at the end of the combined stress test. The results indicated that supplemention of CGA had no significant effects on the growth performance and the activities of TAS, SOD, GSH-Px and CAT in hepatopancreas of shrimp cultured under normal conditions for 56 days. However, compared with the control group, CGA (200, 400 mg kg−1 feed) significantly improved the resistance of L. vannamei against the combined stress of low-salinity and nitrite, as indicated by the significant (P < 0.05) higher survival, higher activities of TAS, GSH-Px and CAT, as well as higher transcript levels of GPx and CAT gene in shrimp treated with CGA in the combined tress test. Our findings suggested that CGA possessed dual-modulatory effects on antioxidant capacity of L. vannamei and could be a potential feed additive that can enhance shrimp resistance against environmental stresses. The recommended application dosage is 200 mg kg−1 and further studies are needed to clarify the action model of CGA efficiency.

Antioxidant defence to ammonia stress of characins (Hyphessobrycon eques Steindachner) fed diets supplemented with carotenoids

This study aimed to find out whether dietary carotenoid (CD) supplement could influence the resistance of characins (Hyphessobrycon eques Steindachner) to ammonia stress. Two types of CD and its combination [astaxanthin (AX), β-carotene (BC), 1 : 1 combination of AX and BC (MX)] at three concentrations (10, 20 and 40 mg kg−1) were used resulting in nine pigmented diets. No differences in growth and survival of the fish among treatments were found after 8-week rearing. Experimental and control fish were then exposed to 15 mg total ammonia nitrogen L−1 (stress group) and 0.15 mg total ammonia nitrogen L−1 (normal group) for 72 h, and their blood was withdrawn. No mortality resulted under such TAN concentrations. Serum total antioxidant status (TAS), serum antioxidant enzymes [superoxide dismutase (SOD), glutathione peroxidases (GPx)] and serum transaminases [alanine aminotransferase (ALT), aspartate aminotransferase (AST)] were chosen as indices of fish antioxidant capacity or stress resistance. SOD, GPx and AST were affected by the interactions of dietary CD and ammonia stress. The activities of TAS, SOD, GPx and AST increased under the stress. Dietary CD reduced serum SOD, GPx, ALT and AST activities. In conclusion, dietary CD increased the resistance of characins to ammonia stress.

Relationship between Metabolic Syndrome Components and Oxidative Stress in Elderly Community-Dwelling Mexicans

Aim: To determine the relationship between metabolic syndrome (MetS) components and oxidative stress (OxS) in elderly community-dwelling Mexicans. Methods: We carried out a comparative cross-sectional study on 113 elderly subjects (≧60 years old); 50 of them did not have MetS and 63 did have MetS (based on Adult Treatment Panel III criteria). We measured total antioxidant status (TAS), plasma lipid peroxidation (LPO), antioxidant activity of superoxide dismutase (SOD) and glutathione peroxidase (GPx), and calculated the SOD/GPx ratio. High blood pressure (HBP) was defined as systolic blood pressure (SBP) ≧140 mm Hg and/or diastolic blood pressure (DBP) ≧90 mm Hg. Results: We found a statistically significant increase in the percentage of cases of severe OxS in elderly subjects with MetS in comparison to healthy elderly (17 vs. 8%, OR: 7.33, 95% CI: 1.38–42.39; p < 0.01). It was observed that subjects with 5 MetS components had a 10-fold higher risk of developing OxS than subjects with 1 component (OR: 11.00, 95% CI: 1.72–115.17; p < 0.01). A positive correlation between SBP and LPO (r = 0.202, p < 0.05), and a negative correlation of SBP and DBP with TAS activity (r = –0.232, p < 0.05; r = –0.211, p < 0.05) were also observed. Conclusion: Our findings suggest that MetS is linked to severe OxS, that the number of metabolic syndrome components is a significant risk factor in the development of OxS, and that HBP is the most important metabolic syndrome component linked to OxS in the elderly.

Band 3 Profile as a Marker of Erythrocyte Changes in Chronic Kidney Disease Patients

Our aim was to study changes in red blood cell (RBC) membrane band 3 profile, as a cumulative marker of RBC changes, in chronic kidney disease (CKD) patients under haemodialysis and recombinant human erythropoietin (rhEPO) therapy and its linkage with resistance to this therapy. We studied 63 CKD patients, 32 responders and 31 non-responders to rhEPO therapy, and 26 healthy individuals. We evaluated the band 3 profile (% of band 3 monomer, high molecular weight aggregates (HMWAg), and proteolytic frag- ments (Pfrag)), membrane-bound haemoglobin (MBH), haematological data, total serum bilirubin, glutathione peroxidase (GPx) and superoxide dismutase activities, total antioxidant status (TAS) and plasma lipid peroxidation (TBA). Compared to controls, band 3 profile presented by CKD patients showed statistically significant lower HMWAg and Pfrag values and a significant higher value in band 3 monomer. GPx, TBA and TAS activities, and TBA/TAS ratio were also signifi- cantly higher in CKD patients. Comparing responders to non-responders CKD patients, significantly lower value in Pfrag and a trend for a higher value in MBH were found in non-responders. Our data suggest that CKD patients present younger RBC population, which could be related to the rhEPO therapy. The adverse plasma environment associated to CKD patients under hemodialysis imposes changes in band 3 profile, particu- larly in non-responders, suggesting that resistance to rhEPO therapy in CKD patients seems to be associated to an increase in RBC damage.

Oxidative stress may be a piece in the endometriosis puzzle

Objective To assess the total antioxidant potential of women with endometriosis-associated infertility, women with idiopathic infertility, and fertile controls. Design Retrospective study. Setting University medical hospital. Patient(s) Sixty-five women admitted for diagnostic laparoscopy. Main outcomes measure(s) Peritoneal fluid samples were analyzed for superoxide dismutase activity and total antioxidant status on spectrophotometry, glutathione peroxidase activity on enzyme-linked immunosorbent assay, and lipid peroxides levels on colorimetry. Result(s) Mean activity of superoxide dismutase, glutathione peroxidase, and total antioxidant status was lowest and lipid peroxide level was highest among infertile patients with endometriosis. Women with idiopathic infertility, in contrast, had the highest superoxide dismutase, glutathione peroxidase, and total antioxidant status activity and the lowest lipid peroxide level. Conclusion(s) High antioxidant potential is not a contributing factor in women with idiopathic infertility. Low total antioxidant status and low activity of antioxidant enzymes in the peritoneal fluid of infertile women with endometriosis probably do not influence fertility in these women, but these factors may play a role in the development of the disease.

Prognostic significance of free radicals: mediated injury occurring in the kidney donor.

Brain death is associated with hemodynamic disturbances in systemic circulation and metabolic storm, and, thus, free radical-mediated injury to donor tissues was hypothesized. An assessment of oxidative stress in the donor and its effect on posttransplant kidney graft function comprised the scope of the study. A prospective study was performed in 27 donors and 50 kidney transplant recipients. Sera from 27 brain-dead organ donors and preservation media were tested for malondialdehyde (MDA) and for total antioxidant status (TAS). Kidneys were preserved in University of Wisconsin-gluconate solution with machine perfusion. Mean ischemia time was 36.7+/-8 hours. Organs were transplanted to recipients on the Polish National Waiting List and posttransplant kidney function was monitored periodically. Posttransplant delayed graft function (DF) was diagnosed when a patient required at least one dialysis within first week after transplantation. Acute rejection was diagnosed clinically and confirmed with fine-needle biopsy if necessary. Thirty-two recipients had immediate graft function (IF), and 18 suffered from DF. MDA level in preservation solution at the end of machine perfusion was significantly higher in the DF group (52.6+/-31 vs. 25.3+/-19 micromol/L) whereas donor TAS activity was lower (1.14+/-0.2 vs. 0.97+/-0.3 mmol/mL). Patients who suffered from acute rejection received kidneys from donors with significantly higher serum MDA (66+/-73 micromol/ml vs. 23+/-49 for patients without rejection). Serum creatinine 12 to 48 months after transplantation correlated to donor- and preservation-solution MDA (P<0.006). Free-radical mediated injury occurring in the donor and during preservation is strictly correlated with immediate and long-term kidney function. It may also cause grafts to be prone to acute rejection.

Effects of rutin supplementation on antioxidant status and iron, copper, and zinc contents in mouse liver and brain.

The effect of rutin on total antioxidant status as well as on trace elements such as iron, copper, and zinc in mouse liver and brain were studied. Mice were administrated with 0.75 g/kg or 2.25 g/kg P. O. of rutin for 30 d consecutively. Following the treatment, the activity of total antioxidant status, catalase, Cu,Zn-superoxide dismutase, Mn-superoxide dismutase, zinc, copper, and iron were measured in mouse liver and brain. The results showed that rutin significantly increased the antioxidant status and Mn-superoxide dismutase activities in mouse liver, but it had no effect on these variables in the brain. Treatment with a higher concentration of rutin significantly decreased catalase activity and iron, zinc, and copper contents in mouse liver; it also resulted in a slower weight gain for the first 20 d. These results indicate that rutin taken in proper amount can effectively improve antioxidant status, whereas at an increased dosage, it may cause trace element (such as iron, zinc, and copper) deficiencies and a decrease in the activities of related metal-containing enzymes.