GSSG Levels Research Articles

Isorhamnetin Reduces Glucose Level, Inflammation, and Oxidative Stress in High-Fat Diet/Streptozotocin Diabetic Mice Model.

Background: Isorhamnetin is a flavonoid that is found in medical plants. Several studies showed that isorhamnetin has anti-inflammatory and anti-obesity effects. This study aims to investigate the anti-diabetic effects of isorhamnetin in a high-fat diet and Streptozotocin-(HFD/STZ)-induced mice model of type 2 diabetes. Materials and Methods: Mice were fed with HFD followed by two consecutive low doses of STZ (40 mg/kg). HFD/STZ diabetic mice were treated orally with isorhamnetin (10 mg/kg) or (200 mg/kg) metformin for 10 days before sacrificing the mice and collecting plasma and soleus muscle for further analysis. Results: Isorhamnetin reduced the elevated levels of serum glucose compared to the vehicle control group (p < 0.001). Isorhamnetin abrogated the increase in serum insulin in the treated diabetic group compared to the vehicle control mice (p < 0.001). The homeostasis model assessment of insulin resistance (HOMA-IR) was decreased in diabetic mice treated with isorhamnetin compared to the vehicle controls. Fasting glucose level was significantly lower in diabetic mice treated with isorhamnetin during the intraperitoneal glucose tolerance test (IPGTT) (p < 0.001). The skeletal muscle protein contents of GLUT4 and p-AMPK-α were upregulated following treatment with isorhamnetin (p > 0.01). LDL, triglyceride, and cholesterol were reduced in diabetic mice treated with isorhamnetin compared to vehicle control (p < 0.001). Isorhamnetin reduced MDA, and IL-6 levels (p < 0.001), increased GSH levels (p < 0.001), and reduced GSSG levels (p < 0.05) in diabetic mice compared to vehicle control. Conclusions: Isorhamnetin ameliorates insulin resistance, oxidative stress, and inflammation. Isorhamnetin could represent a promising therapeutic agent to treat T2D.

Mitochondrial transplantation against gentamicin-induced toxicity on rat renal proximal tubular cells: the higher activity of female rat mitochondria.

Mitochondrial dysfunction is a fundamental mechanism leading to drug nephrotoxicity, such as gentamicin-induced nephrotoxicity. Mitochondrial therapy (mitotherapy) or exogenous mitochondria transplantation is a method that can be used to replace dysfunctional mitochondria with healthy mitochondria. This method can help in the treatment of diseases related to mitochondria. In this research, we studied the transplantation effect of freshly isolated mitochondria on the toxicity induced by gentamicin on renal proximal tubular cells (RPTCs). Furthermore, possible gender-related effects on supplying exogenous rat kidney mitochondria on gentamicin-induced RPTCs were investigated. At first, the normality and proper functioning of fresh mitochondria were assessed by measuring mitochondrial succinate dehydrogenase activity (SDH) and changes in mitochondrial membrane potential (MMP). Then, the protective effects of mitochondrial transplantation against gentamicin-induced mitochondrial toxicity were evaluated through parameters including lactate dehydrogenase (LDH) leakiness, reactive oxygen species (ROS) production, lipid peroxidation (LPO) content, reduced glutathione (GSH) level, extracellular oxidized glutathione (GSSG) level, ATP level, MMP collapse, and caspase-3 activity. According to the statistical analysis, transplanting the healthy mitochondria decreased the cytotoxicity, ROS production, MMP collapse, LPO content, GSSG levels, and caspase-3 activity caused by gentamicin in RPTCs. Also, it has caused an increase in the level of ATP and GSH in the RPTCs. Furthermore, higher preventive effects were observed for the female group. According to the current study, mitochondrial transplantation is a potent therapeutic method in xenobiotic-caused nephrotoxicity.

Simultaneous determination of intracellular reduced and oxidized glutathiones by the König reaction.

The König reaction is commonly used for the detection of cyanide and its derivatives, including thiocyanate and selenocyanate. We found that this reaction can be used to quantify glutathione fluorometrically, and applied it to the simultaneous determination of reduced and oxidized glutathiones (GSH and GSSG) using a conventional LC system with isocratic elution. The limits of detection were 6.04 nM and 9.84 nM for GSH and GSSG, respectively, and the limits of quantification were 18.3 nM and 29.8 nM, respectively. We also determined GSH and GSSG levels in PC12 cells exposed to paraquat, an oxidative stressor, and observed a decrease in GSH/GSSG ratio, as expected. Total GSH levels quantified by this method and by the conventional colorimetric method with 5,5'-dithiobis(2-nitrobenzoic acid) were comparable. Our new application of the König reaction offers a reliable and useful method to simultaneously quantify intracellular GSH and GSSG.

Role of soy lecithin combined with soy isoflavone on cerebral blood flow in rats of cognitive impairment and the primary screening of its optimum combination.

Soy isoflavone (SIF) and soy lecithin (SL) have beneficial effects on many chronic diseases, including neurodegenerative diseases. Regretfully, there is little evidence to show the combined effects of these soy extractives on the impairment of cognition and abnormal cerebral blood flow (CBF). This study examined the optimal combination dose of SIF + SL to provide evidence for improving CBF and protecting cerebrovascular endothelial cells. In vivo study, SIF50 + SL40, SIF50 + SL80 and SIF50 + SL160 groups were obtained. Morris water maze, laser speckle contrast imaging (LSCI), and hematoxylin-eosin staining were used to detect learning and memory impairment, CBF, and damage to the cerebrovascular tissue in rat. The 8-hydroxy-2'-deoxyguanosine (8-OHdG) and the oxidized glutathione (GSSG) were detected. The anti-oxidative damage index of superoxide dismutase (SOD) and glutathione (GSH) in the serum of an animal model was also tested. In vitro study, an immortalized mouse brain endothelial cell line (bEND.3 cells) was used to confirm the cerebrovascular endothelial cell protection of SIF + SL. In this study, 50 μM of Gen were used, while the 25, 50, or 100 μM of SL for different incubation times were selected first. The intracellular levels of 8-OHdG, SOD, GSH, and GSSG were also detected in the cells. In vivo study, SIF + SL could increase the target crossing times significantly and shorten the total swimming distance of rats. The CBF in the rats of the SIF50 + SL40 group and SIF50 + SL160 group was enhanced. Pathological changes, such as attenuation of the endothelium in cerebral vessels were much less in the SIF50 + SL40 group and SIF50 + SL160 group. The 8-OHdG was reduced in the SIF50 + SL40 group. The GSSG showed a significant decrease in all SIF + SL pretreatment groups, but the GSH showed an opposite result. SOD was upregulated by SIF + SL pretreatment. Different combinations of Genistein (Gen)+SL, the secondary proof of health benefits found in vivo study, showed they have effective anti-oxidation and less side reaction on protecting cerebrovascular endothelial cell. SIF50 + SL40 in rats experiment and Gen50 + SL25 in cell test were the optimum joint doses on alleviating cognitive impairment and regulating CBF through protecting cerebrovascular tissue by its antioxidant activity. SIF+SL could significantly prevent cognitive defect induced by β-Amyloid through regulating CBF. This kind of effect might be attributed to its antioxidant activity on protecting cerebral vessels.

Serum levels of reduced glutathione, oxidized glutathione, and glutathione reductase activity in minor recurrent aphthous stomatitis patients

Background/purposeRecurrent aphthous stomatitis (RAS) is one of the most prevalent oral mucosa diseases with unknown etiology. Reduced glutathione (GSH) is a major intracellular non-protein physiological antioxidant, and it has been demonstrated that GSH deficiency may be related to cardiovascular, immune, and diabetes. The purpose of this investigation was to evaluate the potential roles of GSH, oxidized glutathione (GSSG), and glutathione reductase (GR) in the etiopathogenesis of minor recurrent aphthous stomatitis (MiRAS). Materials and methodsThe study comprised 87 patients with idiopathic MiRAS and 90 race-, age-, and gender-matched healthy individuals. The spectrophotometric method was used to determine serum GSH and GSSG concentrations as well as GR activity. The GSSG/GSH ratios were subsequently computed. For statistical evaluation, the independent sample t test, Pearson's chi-square test, Mann–Whitney U test, Kruskal–Wallis H test, and Binary logistic regression analysis were used. ResultsThe serum GSSG level, GR activity and GSSG/GSH ratio were statistically higher in MiRAS patients, whereas the concentration of serum GSH was significantly decreased. With the exception of GR, serum GSSG, GSH, and GSSG/GSH were all significantly associated with MiRAS. Serum GSSG can be regarded as a risk factor, whereas serum GSH and GSSG/GSH maybe considered as protective factors against the occurrence of MiRAS. ConclusionGSSG may be a potential danger factor to MiRAS and GSH may be a protective factor, while GR may not play an important role in the aetiopathogenesis of MiRAS.

Serum and brain metabolomic study reveals the protective effects of Bai-Mi-Decoction on rats with ischemic stroke.

Bai-Mi-Decoction (BMD), which is composed of Eugenia caryophyllata, Myristica fragrans, Moschus berezovskii, and Crocus sativu, is a characteristic TCM multi-herb formula for brain disease. However, the mechanism of protective effects of BMD on ischemic stroke (IS) still has not been clarified. Our study is designed to elucidate the protective effects and underlying mechanisms of BMD on IS by employing pharmacodynamic and serum and brain metabolomic methods. In this experiment, 90 adult male Sprague-Dawley rats were randomly divided into the sham operation group (SHAM, vehicle), middle cerebral artery occlusion-reperfusion injury model group (MCAO/R, vehicle), positive control group (NMDP, 36mg/kg/day nimodipine), and low (BMDL, 0.805g/kg/day), moderate (BMDM, 1.61g/kg/day), and high (BMDH, 3.22g/kg/day) dosage of BMD prophylactic administration groups. The drugs were dissolved in 0.5% CMC-Na and orally administered to rats with equal volumes (100g/ml body weight) once a day for 14 consecutive days. Neurological deficit score, cerebral infarct volume, change in body weight, and serum NO, SOD, MDA, GSH, and GSSG levels were determined. Pathological abnormalities using hematoxylin and eosin staining and the expression of VEGF, caspase-3, and NF-κB were analyzed. Furthermore, serum and brain metabolic profiles were explored to reveal the underlying mechanism using UHPLC-QTOF-MS/MS technology. BMD exhibited significant neuroprotective effects on MCAO/R rats. As compared to the MCAO/R model group, it could reduce the neurological deficit score and cerebral infarct volume, increase body weight, enhance GSH, SOD, and GSSG activities, and decrease NO and MDA contents of MCAO/R rats. Meanwhile, BMD could ameliorate pathological abnormalities of MCAO/R rats through reducing neuronal loss, vacuolated spaces, shrunken neurons, and destructed neuron structure, as well as regulating the expression of VEGF, caspase-3, and NF-κB. UHPLC-QTOF-MS/MS-based serum and brain metabolomics analysis found a total of 53 differential metabolites between MCAO/R and SHAM groups, of which 30 were significantly regulated by BMD intervention, and further metabolic pathway analysis implied that the protective effects were mainly associated with amino acid and glycerophospholipid metabolisms. Our pharmacodynamic and metabolomic results revealed the neuroprotective effects of BMD on MCAO/R rats, and the underlying mechanisms were probably related to amino acid and glycerophospholipid metabolisms.

Protrusion of KCNJ13 Gene Knockout Retinal Pigment Epithelium Due to Oxidative Stress-Induced Cell Death.

This study was performed to elucidate the mechanisms of morphological abnormalities in a Leber congenital amaurosis 16 (LCA16) cell model using KCNJ13 knockout (KO) retinal pigment epithelial cells derived from human iPS cells (hiPSC-RPE). In KCNJ13 KO and wild-type hiPSC-RPE cells, ZO-1 immunofluorescence was performed, and confocal images were captured. The area and perimeter of each cell were measured. To detect cell death, ethidium homodimer III (EthD-III) staining and LDH assay were used. Scanning electron microscopy (SEM) was used to observe the cell surface. The expression levels of oxidative stress-related genes were examined by quantitative PCR. To explore the effects of oxidative stress, tert-butyl hydroperoxide (t-BHP) was administered to the hiPSC-RPE cells. Cell viability was tested by MTS assay, whereas oxidative damage was monitored by oxidized (GSSG) and reduced glutathione levels. The area and perimeter of KCNJ13-KO hiPSC-RPE cells were enlarged. EthD-III-positive cells were increased with more dead cells in the protruded region. The KO RPE had significantly higher LDH levels in the medium. SEM observations revealed aggregated cells having broken cell surfaces on the KO RPE sheet. The KCNJ13-deficient RPE showed increased expression levels of oxidative stress-related genes and total glutathione levels. Furthermore, t-BHP induced a significant increase in cell death and GSSG levels in the KO RPE. We suggest that in the absence of the Kir.7.1 potassium channel, human RPE cells are vulnerable to oxidative stress and ultimately die. The dying/dead cells form aggregates and protrude from the surviving KCNJ13-deficient RPE sheet.

Melatonin Protects Against Titanium Oxide-Induced Neurotoxicity: Neurochemical, Neurobehavioral, and Histopathological Evidences.

titania (titanium dioxide, TiO2) is known to induce neurotoxicity and CNS dysfunctions. Numerous studies have explored the neuroprotective effects of melatonin against neurotoxicity. This study evaluates the potential of melatonin to protect against titania-induced neurotoxicity and the role of the Keap1/Nrf2/ARE signaling pathway. One group of animals were treated with Titania (0.045 and 0.075g/rat) alone while the other with added melatonin (1mg/kg and 3mg/kg) and behavioral alterations were assessed using OFT (open field test). Neurochemical and histopathological changes were also studied in the hippocampus by analyzing kelch ECH associating protein 1 (Keap1), nuclear factor erythroid 2-related factor 2 (Nrf2), and antioxidant response element (ARE). It was seen that the animals with added Melatonin had improved behavioral scores in the OFT, like anxiety and motor dysfunction triggered by TiO2. Melatonin also reduced lipid peroxidation, ROS, GSSG, IL1β, TNFα, Bax, and Keap1 levels, but boosted GSH, GPx, GR, SOD,IL10,IL4, Bcl2, Nrf2, and ARE levels and improved quadruple mitochondrial enzyme complex activity in titania-treated animals. Histopathological examination showed melatonin induced cytoprotection against vacuolization and necrosis in granular cells of DG and pyramidal cells of CA1 area of the hippocampus. In our study, pretreatment with melatonin reduced titania-induced neurotoxicity in the hippocampus through a mechanism potentially mediated by the Keap-1/Nrf2/ARE pathway.

Biodegradation and Decolorization of Textile Azo Dyes by Paramecium caudatum Isolated from Industrial Wastewater

Azo dyes in textile industry effluent are one of the major toxic contaminants causing a severe threat to life. Bioremediation is the most cost-effective and environmentally beneficial innovative biotechnologically technique used to mitigate dyes’ toxic effects in aquatic environments. The purpose of the present study was to determine the azo dyes degradation potential of a ciliate, Paramecium caudatum, isolated from industrial wastewater. Under optimum conditions, P. caudatum was found to possess a 90.86% decolorizing ability of RR2 (reactive red), 83.06% of RB5 (reactive blue) and 85.43% of LY (Levafix reactive yellow) dyes. The P. caudatum showed maximum growth at 25 °C and pH 7.5 in the presence of azo dyes at a concentration of 20 ppm (0.02 mg/mL). After being exposed to RR2, RB5, and LY azo dyes, the level of GSH in P. caudatum increased for the control i.e., 54, 43, and 23%, respectively. Sequentially, the GSSG level decreased by 26% after treatment with the LY azo dye, while exposure to RR2 and RB5 enhanced the value by more than twofold and by 0.86%. The results for the decolorization and biodegradation of azo dyes indicated that P. caudatum is a potential candidate for the treatment of textile industry effluents.

Blueberry Anthocyanins from Commercial Products: Structure Identification and Potential for Diabetic Retinopathy Amelioration.

The aim of the present study was to determine the major anthocyanins of blueberry extracts from northeast China and explore their vision health improvement effects. HPLC-Q-TOF-MS/MS results suggested that six different anthocyanins were accurately identified, among which the Cy-3-glu (C3G) was the most abundant, ranging from 376.91 ± 7.91 to 763.70 ± 4.99 μM. The blueberry extract contained a higher purity of anthocyanins, and the anthocyanosides reached 342.98 mg/kg. The anti-oxidative stress function of C3G on HG-treated ARPE-19 cells were evaluated, and showed that the GSSG level of HG-cells pretreated with 10 μM C3G was significantly decreased, while the Nrf2 and NQO1 gene expression levels were increased. Further molecular docking (MD) results indicated that the C3G displayed favorable binding affinity towards REDD1, and only the B-ring of the C3G molecule displayed binding interactions with the CYS-140 amino acids within the REDD1 protein. It implied that the oxidative stress amelioration effects of C3G on the ARPE-19 cells were related to the REDD1 protein, which was probably via the Nrf2 pathways, although further studies are needed to provide mechanism evidence. The present study provides novel insights into understanding the roles of blueberry anthocyanins in ameliorating oxidative stress-induced BRB damage in the retina.

ODP389 Lipid Peroxidation System Reactivity in Adolescents During Constitutional Obesity Non-drug Therapy

Abstract Background In recent years, there has been a worldwide trend toward preventing and controlling childhood obesity by non-drug therapy. The success of this trend largely depends on better understanding the various metabolic and regulatory systems functions. Lipid peroxidation (LPO) - antioxidant defense (AOD) nonspecific system reactivity should be of special attention. The aim of the study was to reveal the lipid peroxidation and antioxidant defense parameters changes in different sex's adolescents during constitutional obesity non-drug therapy. Materials and methods Adolescents of 12-17 years old with constitutional obesity (CO) (22 boys and 24 girls) before and after constitutional obesity non-drug therapy were examined. The therapy included routine recommendations for food consumption modification, motivational education in the School of Rational Eating, a steady diet, a low-calorie diet, aerobic physical exercises (running track), impulse physiotherapy on the rectus abdominis. Healthy teenager's data (20 boys and 16 girls) were used as a control. We analyzed the content of LPO products - diene conjugates (DC), ketodienes and conjugated trienes (KD and CT), TBA-active products and AOD factors activity: total antioxidant activity (TAA), α-tocopherol, retinol, reduced (GSH) and oxidized (GSSG) glutathione, superoxide dismutase activity (SOD). Spectrophotometric and fluorometric methods were used. Results The boys with CO had decreased values of primary LPO products – DC (by 1.35 times lower, p=0. 001) and AOD factors: TAA (by 1.16 times lower, p=0. 036) and SOD activity (by 1.49 times lower, p&amp;lt;0. 0001), against the increased background values of secondary products - KD and CT (by 1.42 times higher, p=0. 046) in comparison with control results. Boys with CO therapy did not lead to normalization of the studied parameters. The girls with CO had decreased values of TAA values (by 1.4 times lower, p=0. 0001), SOD (by 1.42 times lower, p&amp;lt;0. 0001) and α-tocopherol (by 1.33 times lower, p=0. 012) compared to the control group. After the course of therapy, there was an increase in GSSG levels (by 1.14 times higher, p=0. 029). Conclusions This study showed the presence of negative changes in the pro- and antioxidant status in adolescents with CO, which did not normalize after a course of non-drug therapy. Additional pharmacological correction by prescription of antioxidant drugs becomes relevant in addition to the course of non-drug therapy. Presentation: No date and time listed

DIET SUPPLEMENTATION WITH YEAST BETA-GLUCANS, DIETARY ANTIOXIDANTS AND VITAMIN K IN BROILER CHICKENS MITIGATED AFLATOXIN-INDUCED GROWTH RETARDATION

Mitigation of inevitably absorbed aflatoxins following the addition of toxin binders are insufficiently reported. Therefore, effect of yeast beta-glucans, antioxidants such as vitamins C (VC) and E (VE), selenium (Se) and anti-haemorrhagic agent (vitamin K- VK) in the diet on changes in biochemical processes that characterised aflatoxins poisoning in broiler chickens was investigated. Aspergillus flavus 3228 inoculated maize was used to formulate a basal diet (BD) containing 270 ± 16µg.kg-1 total aflatoxins (AFB1 and AFB2). Unsexed one-day-old Arbor-Acres broiler chicks (n = 180) were randomly assigned into six dietary treatments, comprising Negative Control (NC- aflatoxin-free diet), BD and BD containing beta-glucans, VC, VE, VK and selenium. Two levels of beta-glucans: 250 and 375 mg.kg-1 (BD250 and BD375) and two combinations of vitamins with or without selenium: [(VE + VC) +VK] = ECK and [(VE + VC) + VK + Se] = ECKSe were combined to give BD250 + ECK; BD250 + ECKSe; BD375 + ECK and BD375 + ECKSe. Selenium, VC, VE and VK, were included in the diets at 0.3, 250, 200 and 3.0 mg.kg-1 of feed, respectively in augmented (2 x 2) +2 factorial arrangement in completely randomised design. The diets were fed to the chicken ad libitum for seven weeks. Serum malondialdehyde, reduced (GSH) and oxidised (GSSG) glutathione levels and ratio, body weight gain (BWG) were measured and feed conversion ratio (FCR) and percentage mortality were determined. Data were analysed using ANOVA at α0.05. Serum malondialdehyde and GSSG levels were significantly reduced (P &lt; 0.05) from 159.41 ± 23.68 nM.mL-1 and 5.96 ± 5.20 µM.mL-1 in BD, to 69.68 ± 26.97 nM.mL-1 and 2.21 ± 0.88 µM.mL-1 in birds on BD375 + ECKSe. Birds fed BD375 + ECKSe had GSH:GSSG (3.58 ± 1.71), BWG (1,903.98 ± 32.56 g.bird-1) significantly higher and FCR (1.88 ± 0.04) was reduced (P &lt; 0.05) compared to 1.06 ± 0.81; 956.27 ± 19.34 g.bird-1 and 2.38 ± 0.04, respectively, in birds on BD. Birds fed BD375 + ECKSe had no variation (P &gt; 0.05) in performance when compared to NC. Mortality decreased significantly (P &lt; 0.05) from 39.39 ± 5.25 % in BD to 9.09 ± 5.25 % in BD375 + ECK. Combinations of 375 mg.kg-1 beta-glucans, vitamins E, C, K and selenium reduced lipid peroxidation activity, feed conversion ratio and oxidative stress, with increased bodyweight gain. Finally, combinations of beta-glucans, antioxidants and vitamin K demonstrated effectiveness in preventing changes in biochemical processes leading to aflatoxins deleterious effects in broiler chickens.

Glutathione Component of Antioxidant Status in Menopausal Women with Insomnia.

The study involved 138 women aged 45-60 years in perimenopause (n=55) and postmenopause (n=83) with insomnia (main groups) and without it (control). The levels of reduced (GSH) and oxidized (GSSG) glutathione and glutathione peroxidase activity were determined in erythrocytes; activities of glutathione S-transferase and glutathione reductase were measured in blood serum. The differences with the control groups were found only in perimenopause: higher glutathione reductase activity and reduced GSSG level and GSH/GSSG ratio in women with insomnia (p<0.05). The results of the comparative analysis between the main groups showed lower glutathione reductase activity, increased GSSG level, and a decrease the GSH/GSSG ratio in the postmenopausal period compared with the perimenopause (p<0.05). These results can be used in choosing the tactics for complex therapy of insomnia in menopausal women to correct free radical homeostasis and prevention of oxidative stress.

Haematological and Chosen Biochemical Parameter Assessment of the Antioxidant System in Red Deer (Cervus Elaphus) Blood in Early and Late Pregnancy.

The aim of this study was to investigate changes in haematological parameters and the antioxidant system in the early and late pregnancy of red deer (Cervus elaphus). Blood samples were collected from 30 red deer females 50 days after impregnation and 40 days before calving. Complete blood counts and stained blood smears were assessed. Paraoxonase 1 (PON1), superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) activities, glutathione disulphide (GSSG), total glutathione, total bilirubin, total protein, albumin, uric acid, malondialdehyde (MDA), beta-hydroxybutyrate (BHB), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides, cholesterol levels and total antioxidant status (TAS) were measured. The haematological characteristics of blood collected in the third trimester showed higher haemoglobin levels, haematocrit, mean corpuscular volume (P < 0.05) and a lower mean cell haemoglobin concentration (P < 0.05) in comparison to samples collected on the 50th day after mating. Activity of CAT and MDA, LDL, and triglyceride levels were lower while GR, GSSG, total glutathione, total protein and BHB levels were increased in blood samples drawn in the third trimester compared with the first trimester blood samples. There were no changes in SOD, PON1 activities, TAS, bilirubin, albumin, HDL, cholesterol or uric acid concentrations. Red deer's compensatory mechanisms facilitate its optimal adaptation to seasonal changes evidenced by mild pronounced haematological disturbances and an effective antioxidant system during pregnancy.

Dietary Intervention with Blackcurrant Pomace Protects Rats from Testicular Oxidative Stress Induced by Exposition to Biodiesel Exhaust.

The exposure to diesel exhaust emissions (DEE) contributes to negative health outcomes and premature mortality. At the same time, the health effects of the exposure to biodiesel exhaust emission are still in scientific debate. The aim of presented study was to investigate in an animal study the effects of exposure to DEE from two types of biodiesel fuels, 1st generation B7 biodiesel containing 7% of fatty acid methyl esters (FAME) or 2nd generation biodiesel (SHB20) containing 7% of FAME and 13% of hydrotreated vegetable oil (HVO), on the oxidative stress in testes and possible protective effects of dietary intervention with blackcurrant pomace (BC). Adult Fisher344/DuCrl rats were exposed by inhalation (6 h/day, 5 days/week for 4 weeks) to 2% of DEE from B7 or SHB20 fuel mixed with air. The animals from B7 (n = 14) and SHB20 (n = 14) groups subjected to filtered by a diesel particulate filter (DPF) or unfiltered DEE were maintained on standard feed. The rats from B7+BC (n = 12) or SHB20+BC (n = 12), exposed to DEE in the same way, were fed with feed supplemented containing 2% (m/m) of BC. The exposure to exhaust emissions from 1st and 2nd generation biodiesel resulted in induction of oxidative stress in the testes. Higher concentration of the oxidative stress markers thiobarbituric acid-reactive substances (TBARS), lipid hydroperoxides (LOOHs), 25-dihydroxycholesterols (25(OH)2Ch), and 7-ketocholesterol (7-KCh) level), as well as decreased level of antioxidant defense systems such as reduced glutathione (GSH), GSH/GSSG ratio, and increased level of oxidized glutathione (GSSG)) were found. Dietary intervention reduced the concentration of TBARS, 7-KCh, LOOHs, and the GSSG level, and elevated the GSH level in testes. In conclusion, DEE-induced oxidative stress in the testes was related to the biodiesel feedstock and the application of DPF. The SHB20 DEE without DPF technology exerted the most pronounced toxic effects. Dietary intervention with BC in rats exposed to DEE reduced oxidative stress in testes and improved antioxidative defense parameters, however the redox balance in the testes was not completely restored.

Indication of the impact of environmental stress on the responses of the bivalve mollusk Unio tumidus to ibuprofen and microplastics based on biomarkers of reductive stress and apoptosis

The vulnerability of bivalve mollusks to micropollutants is estimated mainly in single model exposures. However, chronic environmental stress and complex exposures can modulate their responses. To evaluate the impact of population-dependent adaptations on the ability to react to common micropollutants, we compared freshwater bivalves Unio tumidus from two distinct populations, pure (Pr) and contaminated (Ct), in their exposures to microplastics (MP, 1 mg L−1, size 0.1–0.5 mm), pharmaceutical ibuprofen (IBU, 0.8 μg L−1), or their combination (Mix) for 14 days. Control groups from both sites showed remarkable differences, with lower levels of total antioxidant capacity (TAC), metallothionein protein (MTSH), NADH and NAD+, cytochrome P450-related EROD, glutathione-S transferase (GST), and citrate synthase (CS) but higher levels of GSH, GSSG, caspase-3 and cathepsin D (CTD) in the Ct-control group. These data indicate a chronic stress impact in the Ct population. Under exposures, we found an almost common strategy in both populations for NAD+/NADH and MTSH suppression and CTD induction. Additionally, Mix exposure caused an increase in CS, and IBU did not change GSH in both populations. However, the expected response to IBU – the suppression of caspase-3 – was indicated only in PrIBU- and PrMix-mollusks. CTD efflux increased dramatically only in PrMP- and PrMix- groups, and suppression of EROD and GST was detected in the PrMix-group. According to discriminant analysis, exposed Pr-groups were highly differentiated from control, whereas Ct-control and exposed groups had common localization demonstrating high resistance to environmental stress. Thus, the same exposures resulted in different adverse outcome pathways depending on the population.

Determination of glutathione in blood via capillary electrophoresis with pH-mediated stacking.

A new approach has been developed for the direct determination of reduced (glutathione [GSH]) and oxidized (glutathione disulfide [GSSG]) GSH in whole blood by means of capillary electrophoresis. Its features include GSH-stabilizing sample preparation, the use of an internal standard, and pH-mediated stacking. Blood stabilized with acid citrate and K3 EDTA was treated with acetonitrile with N-ethylmaleimide, and then the analytes were extracted with diethyl ether. The total analysis time was 8min using a 50-µm (i.d.) by 32.5-cm (eff. length) silica capillary. The background electrolyte was 0.075-M citrate Na pH 5.8 with 200-µM cetyltrimethylammonium bromide and 5-µM sodium dodecyl sulfate, and the separation voltage was -14kV. The quantification limit (S/N=15) of the method was 1.5µM for GSSG. The accuracy levels of GSH and GSSG analysis were 104% and 103%, respectively, and between-run precision levels were 2.6% and 3.2%, respectively. Analysis of blood samples from healthy volunteers (N=24) showed that the levels of GSH and GSSG and the GSH/GSSG ratio in the whole blood were 1.05±0.14mM, 3.9±1.25µM, and 256±94, respectively. Thus, the presented approach can be used in clinical and laboratory practice.

Differential Sensitivity of Two Leukemia Cell Lines towards Two Major Gas Plasma Products Hydrogen Peroxide and Hypochlorous Acid

Oxidative stress has major implications for health and disease. At the same time, the term collectively describes the reactions to different types of reactive oxygen species (ROS) and oxidants, including hydrogen peroxide (H2O2) and hypochlorous acid (HOCl). However, how both compare in terms of cytotoxicity and mechanism of action is less known. Using two leukemia cell lines, Jurkat and THP-1, as model systems at similar cell concentrations, we found an 8-fold greater sensitivity of the former over the latter for H2O2 exposure. Unexpectantly, this was not the case with HOCl exposure. Jurkat cells were 2-fold more resistant to HOCl-induced cytotoxicity than THP-1 cells. In each cell type, the relatively more toxic oxidant also induced activation of caspases 3 and 7 at earlier time points, as time-lapse fluorescence microscopy revealed. The effects observed did not markedly correlate with changes in intracellular GSH and GSSG levels. In addition, siRNA-mediated knockdown of the Nrf2 target HMOX-1 encoding for HO-1 protein and the growth and survival factor IL-8 revealed Jurkat cells to become more sensitive to HOCl, while HO-1 and IL-8 siRNA-mediated knockdown in THP-1 cells produced greater sensitivity towards H2O2. siRNA-mediated knockdown of catalase increased oxidant sensitivity only negligibly. Collectively, the data suggest striking HOCl-resistance of Jurkat and H2O2 resistance of THP-1 cells, showing similar protective roles of HO-1 and IL-8, while caspase activation kinetics differ.

Hypersensitivity to ferroptosis in chromophobe RCC is mediated by a glutathione metabolic dependency and cystine import via solute carrier family 7 member 11

Chromophobe (Ch) renal cell carcinoma (RCC) arises from the intercalated cell in the distal nephron. There are no proven treatments for metastatic ChRCC. A distinguishing characteristic of ChRCC is strikingly high levels of reduced (GSH) and oxidized (GSSG) glutathione. Here, we demonstrate that ChRCC-derived cells exhibit higher sensitivity to ferroptotic inducers compared with clear-cell RCC. ChRCC-derived cells are critically dependent on cystine via the cystine/glutamate antiporter xCT to maintain high levels of glutathione, making them sensitive to inhibitors of cystine uptake and cyst(e)inase. Gamma-glutamyl transferase 1 (GGT1), a key enzyme in glutathione homeostasis, is markedly suppressed in ChRCC relative to normal kidney. Importantly, GGT1 overexpression inhibits the proliferation of ChRCC cells invitro and invivo, suppresses cystine uptake, and decreases levels of GSH and GSSG. Collectively, these data identify ferroptosis as a metabolic vulnerability in ChRCC, providing a potential avenue for targeted therapy for these distinctive tumors.

Physiological and Biochemical Adaptations to High Altitude in Tibetan Frogs, Nanorana parkeri.

The Xizang plateau frog, N. parkeri (Anura: Dicroglossidae), is endemic to the Tibetan Plateau, ranging from 2,850 to 5,100 m above sea level. The present study explores physiological and biochemical adaptations to high altitude in this species with a particular emphasis on parameters of hematology, oxidative stress, and antioxidant defense in adult and juvenile N. parkeri collected from high (4,600 m a.s.l) and low (3,400 m a.s.l) altitudes. Hematological results showed that hemoglobin concentration ([Hb]), hematocrit (Hct), and red blood cell (RBC) counts were significantly higher in high-altitude N. parkeri. High-altitude juveniles had lower RBC sizes than low-altitude juveniles. Higher levels of GSH and GSSG were indicated only in juveniles from high altitude, not in adults. High-altitude individuals also showed lower oxidative damage, assessed as malondialdehyde (MDA) and carbonyl groups (CG) in the liver. High-altitude adults also showed higher activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX), and glutathione-S-transferase (GST) as well as total antioxidant capacity (T-AOC) in the liver as compared to low-altitude adults. Moreover, higher GPX activity and T-AOC were observed in the heart and brain of high-altitude adults. Liver CAT, GPX, and T-AOC showed significant increases in high-altitude juveniles. Vitamin C content was also higher in the heart of high-altitude frogs compared to low-altitude individuals. In summary, the high-altitude population of N. parkeri showed more robust hematological parameters, less oxidative damage, and stronger antioxidant defenses than the low-altitude population, all contributing to increased protection for survival in high-altitude environments.