Total Protein Carbonyl Research Articles

Protective Actions of Anserine Under Diabetic Conditions.

Background/Aims: In rodents, carnosine treatment improves diabetic nephropathy, whereas little is known about the role and function of anserine, the methylated form of carnosine. Methods: Antioxidant activity was measured by oxygen radical absorbance capacity and oxygen stress response in human renal tubular cells (HK-2) by RT-PCR and Western-Immunoblotting. In wildtype (WT) and diabetic mice (db/db), the effect of short-term anserine treatment on blood glucose, proteinuria and vascular permeability was measured. Results: Anserine has a higher antioxidant capacity compared to carnosine (p < 0.001). In tubular cells (HK-2) stressed with 25 mM glucose or 20–100 µM hydrogen peroxide, anserine but not carnosine, increased intracellular heat shock protein (Hsp70) mRNA and protein levels. In HK-2 cells stressed with glucose, co-incubation with anserine also increased hemeoxygenase (HO-1) protein and reduced total protein carbonylation, but had no effect on cellular sirtuin-1 and thioredoxin protein concentrations. Three intravenous anserine injections every 48 h in 12-week-old db/db mice, improved blood glucose by one fifth, vascular permeability by one third, and halved proteinuria (all p < 0.05). Conclusion: Anserine is a potent antioxidant and activates the intracellular Hsp70/HO-1 defense system under oxidative and glycative stress. Short-term anserine treatment in diabetic mice improves glucose homeostasis and nephropathy.

Free radical scavenging potential of Drosera indica L in presence of Dalton Ascites lymphoma (DAL) tumor bearing mice

The aim of the present study was to evaluate the antioxidant role of Drosera indica L. in Dalton Ascites lymphoma (DAL) bearing Swiss albino mice. The effect of ethanol and aqueous extracts of D.indica (EEDI and AEDI) were administered at a 250 and 500mg/kg once a day for 14 days, 24h after the inoculation of tumor cell line. After the treatment period, blood was collected from the animals and subsequently the animal were sacrificed for isolation of liver, brain, kidney and lungs for the observation of antioxidant status level. The parameters analyzed were catalase (CAT), superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), peroxidase (P), total protein (TP) and protein carbonyl content (PCC). Treatment with EEDI and AEDI significantly reduced the levels of MDA and increased the levels of GSH, SOD, CAT, P and TP in cancer induced animal and are similar as that of normal mice. The results suggest that the ethanol and aqueous extract of Drosera indica L. possess significant antioxidant effects in DAL bearing mice

Abstract 5842: Oxidative stress and protein carbonylation towards multi-drug resistance in cancer

Abstract Reactive oxygen species (ROS) are constantly produced under normal as well as stress conditions and play important roles in cell signaling and homeostasis. An elevated level of ROS has been reported in most cancers which may play a critical role in both cancer progression or cancer suppression. One proposed therapeutic strategy is to alter the oxidative stress from ROS-induced tumor promoting event to a ROS-induced apoptotic signaling. Most ROS inducing therapeutic agents initially induce ROS to kill cancer cells by apoptosis; however, prolonged exposure of drugs may result in reduced ROS level and an adaptive microenvironment that makes cancer cells resistant to the same ROS-inducing therapeutic drugs. Chronic exposure of ROS may also upregulate the multidrug resistance protein expression in cancer cells. We hypothesize that differences in oxidative stress and the antioxidant response between drug sensitive and drug resistance cells may play a key role in drug resistance to redox-active drugs. We used two pairs of cell lines, a mitoxantrone sensitive and resistance MCF7 human breast adenocarcinoma (MCF7/MCF7MX) and H460 non-small lung cancer cell lines (H460/H460MX) for our experiments. We employed western blot and ELISA assays to determine the total and specific protein oxidation by carbonylation. Both MCF7MX and H460MX showed slightly lower level of total protein carbonylation compared to their mitoxantrone sensitive counterparts. In western blot analysis, we observed two specific proteins that were differentially carbonylated in MCF7MX compared to and MCF7. Both cytoskeletal and multifunctional proteins were identified by mass spectrometry and confirmed by two-color western blot analysis. Investigation of the potential roles of these specific protein carbonylation in drug resistance will be presented. Citation Format: Baikuntha P. Aryal, Jean-Pierre Gillet, V Ashutosh Rao. Oxidative stress and protein carbonylation towards multi-drug resistance in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5842.

Protein carbonyl determination by a rhodamine B hydrazide-based fluorometric assay

A new fluorometric assay is presented for the ultrasensitive quantification of total protein carbonyls, and is based on their specific reaction with rhodamine B hydrazide (RBH), and the production of a protein carbonyl-RBH hydrazone the fluorescence of which (at ex/em 560/585 nm) is greatly enhanced by guanidine-HCl. Compared to the fluorescein-5-thiosemicarbazide (FTC)-based fluorometric assay, the RBH assay uses a 24-fold shorter reaction incubation time (1 h) and at least 1000-fold lower protein quantity (2.5 µg), and produces very reliable data that were verified by extensive standardization experiments. The protein carbonyl group detection sensitivity limit of the RBH assay, based on its standard curve, can be as low as 0.4 pmol, and even lower. Counting the very low protein limit of the RBH assay, its cumulative and functional sensitivity is 8500- and 800-fold higher than the corresponding ones for the FTC assay. Neither heme proteins hemoglobin and cytochrome c nor DNA interfere with the RBH assay.

Epigenetic modulation of Nrf2 and Ogg1 gene expression in testicular germ cells by methyl parathion exposure

Methyl parathion (Me-Pa) is an oxidizing organophosphate (OP) pesticide that generates reactive oxygen species (ROS) through its biotransformation. Some studies have also suggested that OP pesticides have the capacity to alkylate biomolecules, including DNA. In general, DNA methylation in gene promoters represses transcription. NRF2 is a key transcription factor that regulates the expression of antioxidant, metabolic and detoxifying genes through the antioxidant response element (ARE) situated in promoters of regulated genes. Furthermore, DNA repair genes, including 8-oxoguanine DNA glycosidase (OGG1), have been proposed as NRF2 target genes. Me-Pa exposure produces poor semen quality, genetic and oxidative damage in sperm cells, and reduced fertility. However, the Me-Pa effects on the methylation status and the expression of antioxidant (Nrf2) or DNA repair (Ogg1) genes in male germ cells have not been investigated. Therefore, mice were exposed to Me-Pa to evaluate the global (%5-mC) and specific methylation of Nrf2 and Ogg1 genes using pyrosequencing, gene expression, and total protein carbonylation in male germ cells. The results showed that Me-Pa significantly decreased the global DNA methylation pattern and significantly increased the methylation of two CpG sites within Ogg1 promoter and one CpG site within Nrf2 promoter. In addition, Ogg1 or Nrf2 expression did not change after Me-Pa exposure despite the oxidative damage produced. Altogether, our data suggest that Me-Pa toxicity alters Ogg1 and Nrf2 promoter methylation in male germ cells that may be modulating their gene expression.

The effects of resveratrol on markers of oxidative stress in patients with type 2 diabetes: a randomized, double-blind, placebo-controlled clinical trial.

Oxidative stress plays a pivotal role in the pathogenesis of type 2 diabetes (T2D). In vitro and animal studies have shown that resveratrol exerts an antioxidant effect, but clinical trials addressing this effect in patients with T2D are limited. The aim of this study was to determine whether resveratrol supplementation affects oxidative stress markers in a randomized, placebo-controlled, double-blind clinical trial. A total of 48 patients with T2D randomly were assigned to receive 800mg/day resveratrol or placebo for 2months. Plasma total antioxidant capacity, malondialdehyde concentration, protein carbonyl and total thiol contents, intracellular superoxide anion (O2-·) and hydrogen peroxide (H2O2) in PBMCs, the expression of genes involved in oxidative stress responses (Nrf2, SOD, Cat, HO-1, RAGE, NOS) in PBMCs, and metabolic and anthropometric parameters were measured at the baseline and at the trial end. Compared with the placebo group, resveratrol reduced plasma protein carbonyl content and PBMCs O2-· level and significantly increased plasma total antioxidant capacity and total thiol content. Furthermore, the expression of Nrf2 and SOD was significantly increased after resveratrol consumption. Resveratrol had no significant effects on the metabolic and anthropometric parameters except for a significant reduction in weight, BMI, and blood pressure levels. Resveratrol was well tolerated, and no serious adverse event was occurred. Our study demonstrated that 8weeks of supplementation with 800mg/day resveratrol has an antioxidant effect in the blood and PBMCs of patients with T2D. Clinical Trial Registry number and website IRCT registration number: IRCT2015072523336N1 and http://en.search.irct.ir/view/24752 .

Formula derived Maillard reaction products in post-weaning intrauterine growth-restricted piglets induce developmental programming of hepatic oxidative stress independently of microRNA-21 and microRNA-155.

We recently reported augmentation of lipid peroxidation products in the liver of intrauterine growth-restricted (IUGR) piglets fed a high load of Maillard reaction products (MRPs) during suckling period. The underlying mechanisms of MRPs effects remain unknown. Here, we studied the long-term impact of MRPs exposure on liver oxidative status of IUGR juvenile pigs. Livers of 54-day-old pigs suckled with formula containing either a high (HHF, n=8) or a low (LHF: n=8) load of MRPs were analyzed for protein carbonylation levels , activities and messenger RNA (mRNA) expression of glutathione (GSH) and main antioxidant regulators of redox homeostasis [Superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) were measured. In addition, mRNA levels of miRNA-21 and miRNA-155 were measured. The liver of HHF group exhibited a high level of lipid peroxidation with significantly increased expression and activity of SOD. Further in liver of HHF group, CAT activity was decreased as compared with LHF group, though with comparable total protein carbonyl contents, GSH contents, and expression of GPx and microRNAs (miRNA-21 and miRNA-155). Our findings suggest that the potential mechanism of MRPs-mediated oxidative stress programming in liver of IUGR piglets may occur via impairment of antioxidant defenses.

Impaired plasminogen binding in patients with venous thromboembolism: Association with protein carbonylation

IntroductionVenous thromboembolism (VTE) is associated with hypofibrinolysis. Its mechanisms are poorly understood. We investigated plasminogen-fibrin interaction and its association with fibrinolytic capacity and protein oxidation/carbonylation in VTE patients. Materials and methodsPlasma-purified plasminogen conversion to plasmin and surface plasmon resonance employed for plasminogen-fibrin interactions were individually evaluated in all healthy controls and non-anticoagulated patients following VTE, 10–23months since the event. We also assessed plasma fibrin clot permeability (Ks), clot lysis time (LT), activators and inhibitors of fibrinolysis together with oxidation/carbonylation markers. ResultsVTE patients had impaired plasminogen binding to fibrin (apparent Kd, +290%, p=0.002), reduced rate of plasmin generation (−4.7%, p=0.001), and longer LT (+18.6%, p<0.001) compared with controls. Fibrinogen and Ks were similar in both groups. Apparent Kd correlated with LT (r=0.43, p=0.037), tissue plasminogen activator-plasminogen activator inhibitor 1 (tPA-PAI-1) complexes (r=0.63, p=0.012), and active PAI-1 (r=0.49, p=0.03). Compared with controls, VTE patients had higher thiobarbituric acid reactive substances (TBARS), total protein carbonyl content (PC), and lower total antioxidant capacity (all p<0.001), that all were associated with LT (r=0.61, r=0.56, and r=−0.47, respectively, all p<0.05). Impaired plasminogen binding to fibrin reflected by apparent Kd positively correlated with TBARS (r=0.48, p=0.032) and PC (r=0.54, p=0.013) in the whole group. ConclusionsPlasminogen-fibrin interactions are altered in young and middle-aged VTE patients, without known thrombophilias, except increased factor VIII. The mechanisms underlying these phenomena remain to be established.

Assessment of the antioxidant and antimutagenic activity of extracts from goji berry of Greek cultivation.

The aim of this study was to assess the antioxidant and antimutagenic activities of ultrasound assisted aqueous extracts from dry goji berry fruits cultivated in Greece. The extracts' free radical scavenging activity was assessed by the DPPH• and ABTS•+ assays. The results from both assays demonstrated that the extracts exhibited strong radical scavenging activity with IC50 values ranging from 1.29 to 3.00 mg/ml for DPPH• and from 0.39 to 1.10 mg/mL for ABTS•+ assay. The investigated extracts also inhibited free radical-induced DNA damage induced by peroxyl (ROO•) radicals with IC50 ranging from 0.69 to 6.90 mg/mL. Τhe antioxidant activity of the goji berry extract exhibited the highest potency in the above assays was also examined in muscle cells. In particular, muscle C2C12 cells were treated with the selected extract at non cytotoxic concentrations for 24 h and four oxidative stress markers were measured: total reactive oxygen species (ROS), glutathione (GSH), lipid peroxidation and protein carbonyl levels. The results showed that the extract at 25 and 100 μg/mL increased GSH levels up to 189.5% and decreased lipid peroxidation and protein carbonyls by 21.8 and 29.1% respectively. The present study was the first on the antioxidant effects of ultrasound assisted aqueous extracts from goji berry fruits in muscle cells.

Effects of l-Carnitine on the Follicle-Stimulating Hormone, Luteinizing Hormone, Testosterone, and Testicular Tissue Oxidative Stress Levels in Streptozotocin-Induced Diabetic Rats.

The present study was designed to investigate the antioxidant property of l-carnitine (LC) on serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (TH) and testis oxidative stress in streptozotocin (STZ)-induced diabetic rats. The rats were divided into the following groups: group I, control; group II, LC 100 mg/kg/d; group III, diabetic; and groups IV to VI, diabetic rats treated with 50, 100, and 200 mg/kg/d of LC, respectively. Daily injections were given intraperitoneally for 7 weeks. At the end of experimental period, after sacrificing the rats, FSH, LH, TH, total antioxidant capacity (TAC), malondialdehyde (MDA), glutathione (GSH), catalase (CAT), mitochondrial function (MTT), protein carbonyl (PC), and reactive oxygen species (ROS) levels were measured. STZ caused an elevation of MDA, ROS, and PC (P < .001) with reduction of GSH, CAT, TAC, and MTT (P < .001) in the serum levels. Group VI had significantly increased FSH, LH, and TH levels versus the untreated diabetic group (P < .001). Although groups V and VI significantly decreased MDA (P < .001), PC (P < .01), and ROS (P < .01) compared with the untreated diabetic group; only in group VI, the activity of GSH (P < .001), CAT (P < .01), TAC (P < .001), and MTT (P < .001) significantly increased. The results of the present study suggest that LC decreased diabetes-induced oxidative stress complications and also improved serum level of FSH, LH, and TH by reducing levels of lipid peroxidation and increasing antioxidant enzymes.

Oxidative Stress, Antioxidant Status and Neurodevelopmental Outcome in Neonates Born to Pre-eclamptic Mothers.

To measure the oxidative stress and antioxidant status in preeclamptic mother-newborn dyads and correlate them with neurodevelopmental outcome at one year of corrected age. This cohort study conducted in a tertiary care teaching hospital, south India included 71 preeclamptic and 72 normal mother-newborn dyads. Biochemical parameters including total antioxidant status (TAS), protein carbonyls and malondialdehyde levels (MDA) were measured in both maternal and cord blood. Infants in both the groups were followed up to one year of corrected age and neurodevelopmental assessment was done using Developmental Assessment Scale for Indian Infants (DASII). Correlation and multivariate regression analysis was done to evaluate the oxidative stress markers in relation to neurodevelopmental outcome. All oxidative stress markers were higher in maternal and cord blood of pre-ecclampsia group compared to the normal group. Maternal Total antioxidant status (M-TAS) was lower in pre-eclampsia group than normal group. More neonates in the pre-ecclampsia group were preterm and intrauterine growth restriction (IUGR) and had higher incidence of morbidities like respiratory distress syndrome (RDS) and early onset sepsis (EOS). Infants in the preeclampsia group had lower motor age, motor score and motor developmental quotient (MoDQ). On multivariate logistic regression analyses, lower M-TAS levels were strongly associated with poor neuro-motor outcomes at 1 y of corrected age. Maternal TAS with a cut-off value of 0.965mmol/L had a sensitivity of 77.8% and specificity of 55.3% in predicting MoDQ <70 at one year corrected age in infants born to preeclamptic mothers. Oxidative stress is increased in preeclamptic mother-newborn dyads. Low maternal TAS levels are associated with poor neuro-motor outcomes. Maternal TAS in preeclampsia is useful in predicting poor motor development at one year corrected age.

Alterations in acetylcholinesterase and butyrylcholinesterase activities in chronic obstructive pulmonary disease: relationships with oxidative and inflammatory markers.

This work focused on finding a relationship between acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) activities and the development and severity of COPD. The possible link of these enzymes to oxidative and inflammatory processes was also investigated. The study included 229 healthy controls and 153 COPD patients. Erythrocyte AChE and plasma BChE activities were determined using spectrophotometric methods. Markers related to the oxidative status including thiobarbituric acid-reactive substances (TBARS), total protein carbonyls (PCs), advanced oxidation protein products (AOPP), reduced glutathione, nitric oxide, and peroxynitrite were measured. We also evaluated the activity of glutathione peroxidase, catalase, and superoxide dismutase in the plasma and erythrocytes. Serum levels of IL-6 and TNF-α were measured by the enzyme-linked immunosorbent assay. COPD patients showed increased AChE and BChE activities in comparison to healthy controls. Interestingly, AChE activity was higher in COPD smokers than in nonsmokers, while no difference was revealed for BChE. In addition, our results showed an inverse correlation between AChE activity and the levels of IL-6 in COPD smokers. Positive correlations were found, in COPD smokers, between plasma BChE activity and the levels of several biomarkers of protein oxidative damage including AOPP and PC. Our findings suggest that the alterations in AChE and BChE activities may be related to the oxidative and inflammatory processes in COPD patients rendering these enzymes as markers of COPD disease.

Uterine protein oxidative modifications may condition trophoblast function

Aim: Evaluate whether protein carbonylation resulting from uterine altered redox imbalance interferes with extravillous trophoblast viability. Introduction: Local redox homeostasis is believed to have a pivot role in uterine transformation necessary for blastocyst implantation and placenta development. By contrast, redox status imbalance plays a role in deficient placentation and the development of pregnancy-related complications (e.g. preeclampsia or gestational diabetes) with increased incidence in older women. Thus, it was hypothesized that at an older reproductive age, loss of redox homeostasis is a contributor to disruption of foetal/placental interactions and the development of such complications. Methods: Uterine human samples were collected at delivery by elective caesarean section. The protocol was approved by the ethical committee of “Centro Materno-Infantil do Porto”, volunteers gave written consent to be included in the study. Total protein carbonylation was detected by oxyblot and protein expression was quantified by western blotting. Specific protein carbonylation was verified by immunoprecipitation. Albumin was carbonylated using hydrogen peroxide (H2O2), followed by dialysis, and western blotting to confirm carbonylated albumin. HST-8SV neo extravillous trophoblasts were treated with carbonylated/non-carbonylated albumin, followed by cell viability assay. A P value less than 0.05 was assumed to denote significant difference. Results: At the placental site, carbonylated albumin normalized to total albumin expression showed a positive and significant association with maternal age. (r=0.6909, P=0.0021) In vitro, carbonylated albumin displayed a cytotoxic effect, at concentrations ranging from 10 to 100μg/ml. Lower concentrations did not affect trophoblast viability. Conclusion: Uterine aging is accompanied by selective albumin oxidative modifications, which appears to interfere with trophoblast ability to invade and transform the maternal placental site.

Cardamom supplementation improves inflammatory and oxidative stress biomarkers in hyperlipidemic, overweight, and obese pre-diabetic women: a randomized double-blind clinical trial.

Several preclinical studies have shown that spices may decrease the risk of chronic diseases. However, it has been suggested that more clinical trials be carried out to strengthen this preclinical evidence. The purpose of the present study was to evaluate the effects of cardamom (Elettaria cardamomum) supplementation on inflammation and oxidative stress in hyperlipidemic, overweight, and obese pre-diabetic women. This randomized, placebo-controlled, double-blind clinical trial was conducted on 80 pre-diabetic subjects. They randomly received the cardamom supplement (n = 40, 3 g d-1 ) or identical inert placebo (n = 40) for 8 weeks. Serum concentrations of high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), tumour necrosis factor α, total antioxidant capacity, malondialdehyde (MDA), protein carbonyl, and erythrocyte superoxide dismutase and glutathione reductase activity were analyzed at the baseline and after intervention. After the adjustment of some covariates, cardamom supplementation significantly decreased serum hs-CRP (P = 0.02), hs-CRP:IL-6 ratio (P = 0.008), and MDA (P = 0.009) compared with the placebo group. Cardamom could improve some parameters of inflammation and oxidative stress in pre-diabetic subjects. Thus it may be useful in reducing complications associated with inflammation and oxidative stress in these patients. Copyright © 2017 Society of Chemical Industry © 2017 Society of Chemical Industry.

Abstract 5486: Oxidative stress and selective protein carbonylation in human breast cancer tissue

Abstract Protein carbonylation is an irreversible modification to the side chain of amino acid residues induced by severe oxidative stress. Reactive oxygen species (ROS) are constantly produced under normal as well as stress-induced conditions, and play a role in both cancer progression and cancer suppression. Tumor tissue are known to have higher ROS levels compared to surrounding healthy tissue but ROS-induced specific protein carbonylation and its unique role in cancer progression/suppression are poorly understood. In this study we compared the relative total and specific protein carbonylation in flash frozen human breast cancer and matched adjacent normal tissue using ELISA, two-color western blot, mass spectrometry, and immunoprecipitation approaches. To understand antioxidant capacity in tumor tissue, we analyzed superoxide dismutase (SOD) for its antioxidant activity and protein level. Our results indicate that tumor tissue has greater total protein carbonylation, lower SOD activity, lower SOD protein levels, and elevated levels of autophagy compared to matched adjacent healthy tissue. We also identified three specific proteins that showed higher level of carbonylation selectively in tumor tissue compared to adjacent normal tissue. Our findings were further confirmed using the immortalized MDA-MB-231 breast cancer cell line and MCF-12A noncancerous normal human epithelial breast cell. Identification of selectively carbonylated proteins in cancer tissue, and understanding their specific role in cancer progression may promote the development of targeted therapeutic approaches to mitigate or enhance oxidative damage of such proteins selectively in tumor tissue. Citation Format: Baikuntha P. Aryal, Ashutosh Rao. Oxidative stress and selective protein carbonylation in human breast cancer tissue [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5486. doi:10.1158/1538-7445.AM2017-5486

P 36 Association of placental oxidative stress and pregnancy outcome in patients with hypertensive disorders of pregnancy – A pilot study in an Indian population

Objectives To assess the placental oxidative stress factors in various types of hypertensive disorders of pregnancy and to correlate with the pregnancy outcome. Methods Placental samples from Hypertensive disorders of pregnancy (HDP) patients and the control pregnant women (CPW) were collected. The levels of oxidative stress parameters MDA, Total antioxidant status (TAS) and protein carbonylation (PCO) were assessed in the placental lysates. Also the placental expression of antioxidant enzymes superoxide dismutase (SOD), Glutathione peroxidase (GPX1), Glutathione Reductase (GSH), catalase and stress sensitive serine kinase proteins ERK, p38-MAPK, JNK, IKBα and NOX-4 were found out by western blotting in HDP (gestational hypertension, Preeclampsia- eclampsia) and CPW. The Pregnancy outcomes like the gestational age at the time of delivery (GA) and birthweight (BW) and APGAR score of the baby also were measured. Results The GA, BW and APGAR score of the baby were found to be significantly lower in Preeclampsia and Eclampsia compared to CPW. The levels oxidative stress parameters MDA and PCO were elevated and the TAS were decreased in HDP compared to CPW. The placental expression of pERK, pNrf-2/Nrf-2, NOX-4 and IkBα were found to be elevated whereas the expression of CAT, JNK, pP38MAPK were found to be decreased in HDP when compared to CPW. Also a negative correlation was observed between the o0xidative stress markers and the pregnancy outcome in HDP. Conclusions An increased placental oxidative stress and a poor birth outcome were observed in hypertensive disorders of pregnancy. Download : Download high-res image (419KB) Download : Download full-size image Download : Download high-res image (231KB) Download : Download full-size image Download : Download high-res image (723KB) Download : Download full-size image

Oxidative stress in preeclamptic mother – newborn dyads and its correlation with early neonatal outcome – a case control study

Background: Preeclamptic mothers are likely to have increased oxidative stress during pregnancy which can adversely affect the outcome in their neonates. Objectives: To measure the oxidative stress in preeclamptic mother- newborn dyads and correlate it with the immediate neonatal outcome. Methods: This case control study conducted in a tertiary care teaching hospital, South India included 71 preeclamptic mothers – newborn dyads (cases) and 72 normal mothers – newborn dyads (controls). Biochemical parameters including total antioxidant status (TAS), protein carbonyls and malondialdehyde levels (MDA) were measured in both maternal and cord blood. Association between these oxidative stress parameters and early neonatal outcome was studied. Results: All oxidative stress markers were higher in the preeclampsia group compared to the controls. Cord blood protein carbonyl levels had significant correlation with maternal levels. Prematurity, low-birth weight, respiratory distress syndrome (RDS), early onset sepsis (EOS) and intra-uterine growth restriction (IUGR) were more among cases. Early neonatal outcomes like death, IUGR, EOS, and RDS had significant correlation with protein carbonyl levels among the cases. Conclusions: Oxidative stress is increased in preeclamptic mother – newborn dyads. Increased protein carbonyl levels in preeclampsia correlate with adverse early neonatal outcome.

Acute and chronic varied exercise intensity effects on total antioxidant capacity and protein carbonylation.

IntroductionExercise is known to have positive physiological adaptations to increase various aspects of health. However, during the course of exercise a significant amount of reactive oxygen species (ROS) are produced via increased metabolism. If not counteracted, ROS can oxidize proteins, lipids, and DNA, potentially leading to a disease state. Oxidized proteins may result in protein carbonyls, a marker of damage, particularly in plasma, and can be the result of increased ROS production during exercise metabolism. This could also initiate a cellular signal cascade up‐regulating the expression of antioxidants. Antioxidants are any substance that can delay or prevent oxidation of substrates and are classified as enzymatic (low molecular weight proteins that act by detoxifying free‐radicals in cells) or nonenzymatic (small molecules that directly scavenge ROS). Recently, total antioxidant capacity (TAC) has been utilized to illustrate general antioxidant defenses.MethodsEighteen of a 36 cohort of healthy, overweight (body mass: 87.64 kg; height: 1.68 m) middle‐aged women (age 43.89 years) randomized into two groups, exercised at HIGH (80% VO2peak) and LOW (40% VO2peak) aerobic intensity, along with identical resistance and flexibility training, were measured. Subjects exercised five days/week for 20 weeks with at least 85% adherence. Subjects were tested pre‐training, 10‐, and 20‐weeks in the laboratory where they underwent a maximal graded exercise (max GXT) test to assess fitness changes and other parameters. Venous blood samples were collected before and after the max GXT. TAC was assessed via spectrophotometry. Plasma samples were depleted of Albumin and IgG to remove the highly abundant proteins. Carbonlyated proteins were marked via 2,4‐Dinitrophenylhyrazine followed by one‐dimensional polyacrylamide gel electrophoresis and western blotting to determine the amount of carbonylation that occurred over the 20‐week period.ResultsTAC was significantly increased pre‐training following the max GXT in the LOW group and remained consistent over the 20‐week period, whereas the HIGH group was consistent but had a significant increase following the max GXT at 10‐weeks. Protein carbonylation had a significant decline from pre‐training to 10‐weeks in the HIGH group. The LOW group had significant increases in content between pre‐training and 20‐weeks and 10‐ to 20‐weeks.DiscussionThe highest record levels of TAC occurred (post‐exercise in the LOW group pre‐training), was where the lowest total protein carbonylation was seen. The highest TAC levels in the HIGH group occurred at 10‐weeks post‐exercise which is also where the lowest protein carbonyls in the HIGH group occurred. There was a steady increase in the protein carbonyls in the LOW group with significant differences seen between pre‐training and 20‐weeks and 10‐ to 20‐weeks. It is plausible that when TAC increased over what was a very similar response during the 20‐weeks of training, the consequence was a decrease in protein carbonyls at those times, indicating a protective effect.Support or Funding InformationResearch reported in this (publication, project, release) was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103449. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of NIGMS or NIH.

The Effects of Hyperbaric Oxygen Treatment on Total Antioxidant Capacity and Prolidase Activity after Bile Duct Ligation in Rats

ABSTRACTBackground: Hyperbaric oxygen (HBO) therapy may improve cholestasis, increase hepatic regeneration, and decrease oxidative stress in liver. In this study, we aimed to investigate the effects of HBO therapy on hepatic oxidative stress parameters, such as total thiol groups (T-SH), protein carbonyl (PCO), and total antioxidant capacity (TAC) as well as the predictive value of the noninvasive biochemical marker, sialic acid (SA), and prolidase activity in bile duct ligation (BDL)-induced oxidative damage and fibrosis in rats. Methods: We divided 32 adult male Sprague Dawley rats into four groups: sham, sham + HBO, BDL, and BDL + HBO; each group contained eight animals. We placed the sham + HBO and BDL + HBO groups in an experimental hyperbaric chamber, in which we administered pure oxygen at 2.5 atmospheres for 90 min on 14 consecutive days. Results: The application of BDL significantly increased PCO levels and prolidase activity, and decreased T-SH and TAC levels. HBO significantly decreased PCO levels and prolidase activity and increased T-SH and TAC levels in the liver tissues. There was no significant difference in sialic acid levels between any groups. Conclusions: These results indicate that HBO therapy has hepatoprotective effects on BDL-induced injury by decreasing PCO and prolidase activity and increasing antioxidant activities. We therefore suggest that HBO therapy may be useful after BDL-induced injury.

Protective effect of Allium sativum (garlic) aqueous extract against lead-induced oxidative stress in the rat brain, liver, and kidney.

The present investigation was undertaken to evaluate the ameliorative activity of Allium sativum against lead-induced oxidative stress in the brain, liver, and kidney of male rats. Four groups of male Wistar strain rats (100-120g) were taken: group 1 received 1000mg/L sodium acetate and group 2 was given 1000mg/L lead acetate through drinking water for 2weeks. Group 3 and 4 were treated with 250mg/kg body weight/day of A. sativum and 500mg/kg body weight/day of A. sativum, respectively, by oral intubation for a period of 2weeks along with lead acetate. The rats were sacrificed after treatment and the brain, liver, and kidney were isolated on ice. In the brain, four important regions namely the hippocampus, cerebellum, cerebral cortex, and brain stem were separated and used for the present investigation. Blood was also drawn by cardiac puncture and preserved in heparinized vials at 4°C for estimation of delta-aminolevulinic acid dehydratase (ALAD) activity. The results showed a significant (p<0.05) increase in reactive oxygen species (ROS), lipid peroxidation products (LPP), total protein carbonyl content (TPCC), and lead in the selected brain regions, liver, and kidney of lead-exposed group compared with their respective controls. Blood delta-ALAD activity showed a significant (p<0.05) decrease in the lead-exposed rats. However, the concomitant administration of A. sativum resulted in tissue-specific recovery of oxidative stress parameters namely ROS, LPP, and TPCC. A. sativum treatment also restored the blood delta-ALAD activity back to control. Overall, our results indicate that A. sativum administration could be an effective antioxidant treatment strategy for lead-induced oxidative insult.