Markers Of Protein Oxidative Damage Research Articles

The kynurenine and serotonin pathway, neopterin and biopterin in depressed children and adolescents: an impact of omega-3 fatty acids, and association with markers related to depressive disorder. A randomized, blinded, prospective study.

https://www.isrctn.com/ISRCTN81655012, identifier ISRCTN81655012.

Oleuropein-Enriched Extract From Olive Mill Leaves by Homogenizer-Assisted Extraction and Its Antioxidant and Antiglycating Activities.

Olive oil consumption has increased in the last two decades and consequently, its wastes have increased, which generates a tremendous environmental impact. Among the by-products are the olive mill leaves, which are easier and inexpensive to treat than other olive by-products. However, little research has been done on their chemical composition and potential bioactivity. Hence, in this study, olive mill leaves were used to obtain Oleuropein-Enriched Extracts (OLEU-EE) using Conventional Extraction, Ultrasound-Assisted Extraction, and Homogenization-Assisted Extraction. These three techniques were evaluated using a Factorial Design to determine the parameters to obtain an OLEU-EE with high contents of Total Phenolic Compounds (TPC), Antioxidant Activity (AA), and Oleuropein concentration (OLEU). From the results, the Homogenizer-Assisted Extraction (HAE) technique was selected at 18,000 rpm, solid:liquid ratio 1:10, and 30 s of homogenization with 70% ethanol, due to its high TPC (5,196 mg GA/100 g), AA (57,867 μmol of TE/100 g), and OLEU (4,345 mg of OLEU/100 g). In addition, the antiglycating effect of OLEU-EE on the levels of (1) fluorescent Advanced Glycation End Products (AGEs) were IC50 of 0.1899 and 0.1697 mg/mL for 1λEXC 325/λEM 440 and 2λEXC 389/λEM 443, respectively; (2) protein oxidative damage markers such as dityrosine (DiTyr), N-formylkynurenine (N-formyl Kyn), and kynurenine (Kyn) were IC50 of 0.1852, 0.2044, and 0.1720 mg/mL, respectively. In conclusion, OLEU-EE from olive mill leaves has different capacities to inhibit AGEs evidenced by the IC50 of fluorescent AGEs and protein oxidation products, together with the scavenging free radical evidenced by the concentration of Trolox Equivalent. Therefore, OLEU-EE could be potential functional ingredients that prevent oxidative damage caused by free radicals and AGEs accumulation.

Oxidative damage and mitochondrial functionality in hearts from KO UCP3 mice housed at thermoneutrality.

The antioxidant role of mitochondrial uncoupling protein 3 (UCP3) is controversial. This work aimed to investigate the effects of UCP3 on the heart of mice housed at thermoneutral temperature, an experimental condition that avoids the effects of thermoregulation on mitochondrial activity and redox homeostasis, preventing the alterations related to these processes from confusing the results caused by the lack of UCP3. WT and KO UCP3 mice were acclimatized at 30°C for 4weeks and hearts were used to evaluate metabolic capacity and redox state. Tissue and mitochondrial respiration, the activities of the mitochondrial complexes, and the protein expression of mitochondrial complexes markers furnished information on mitochondrial functionality. The levels of lipid and protein oxidative damage markers, the activity of antioxidant enzymes, the reactive oxygen species levels, and the susceptibility to in vitro Fe-ascorbate-induced oxidative stress furnished information on redox state. UCP3 ablation reduced tissue and mitochondrial respiratory capacities, not affecting the mitochondrial content. In KO UCP3 mice, the mitochondrial complexes activities were lower than in WT without changes in their content. These effects were accompanied by an increase in the level of oxidative stress markers, ROS content, and in vitro susceptibility to oxidative stress, notwithstanding that the activities of antioxidant enzymes were not affected by UCP3 ablation. Such modifications are also associated with enhanced activation/phosphorylation of EIF2α, a marker of integrated stress response and endoplasmic reticulum stress (GRP778 BIP). The lack of UCP3 makes the heart more prone to oxidative insult by reducing oxygen consumption and increasing ROS. Our results demonstrate that UCP3 helps the cell to preserve mitochondrial function by mitigating oxidative stress.

Roux-en-Y Gastric Bypass and Sleeve Gastrectomy Differently Affect Oxidative Damage Markers and their Correlations with Body Parameters.

Bariatric surgery improves oxidative damage, but little is known about the differences between Roux-en-Y gastric bypass (RYGB) and sleeve gastrectomy (SG). This study compared changes in lipid and protein oxidative damage markers and their correlations with body parameters of patients before and after RYGB or SG. Body mass index (BMI), bioimpedance parameters, and biochemical parameters including lipid and protein oxidative damage markers were evaluated before and 6months after surgery. Data were analyzed by t test or Mann-Whitney rank sum test and Spearman's correlation coefficient between oxidative damage and other parameters. Twenty-five patients were submitted to RYGB and 14 to SG. There was a significant decrease of BMI, fat mass, fat-free mass, phase angle, serum total protein, transthyretin, and C-reactive protein in both groups (p< 0.05). Serum thiobarbituric acid reactive substances (TBARS), advanced oxidation protein products (AOPP), and serum lipids (p < 0.05) were significantly decreased in the RYGB group. TBARS levels were significantly correlated with serum total cholesterol (r = 0.468), LDL (r = 0.439), BMI (r = 0.424), and fat mass (r = 0.40) (p< 0.05). In the SG group, AOPP levels were significantly correlated with serum C-reactive protein (baseline: r = 0.53, 6months: r = 0.64) (p < 0.05). Alterations in these levels were negatively correlated with changes in BIA parameters [resistance (r = -0.574), reactance (r = -0.736), and phase angle (r = 0.549)] (p < 0.05). RYGB seems to be better in attenuating oxidative damage after 6months. The BMI reduction in the RYGB group suggests a concomitant decrease of lipid oxidative damage. In the SG group, changes in BIA parameters were inversely correlated with protein oxidative damage.

Nanomaterial categorization by surface reactivity: A case study comparing 35 materials with four different test methods

Nanomaterials (NMs) can be manufactured in plenty of variants differing in their physicochemical properties. Functional assays can be highly useful to cope with the enormous variability by supporting prioritization and categorization. Oxidative potential (OP) seems to be in particular important in this context and different assays are available. However, their reliability and predictivity are not well-characterized.This study compares four different test methods for measuring NM OP. Reactive oxygen species (ROS) generation was measured on a set of 35 different materials, all extensively characterized with respect to physicochemical properties and most of them with respect to toxicity. Different acellular assays were applied, namely electron spin resonance (ESR) spectroscopy using CPH spin probe and DMPO spin trap, and the ferric reduction ability of serum (FRAS) assay. In addition, protein carbonylation as a marker for oxidative protein damage was analyzed in NRK-52E cells. All assays were assessed individually for their predictivity compared to established toxicological endpoints. We also aimed to identify the optimal assay combination using multivariate logistic regression and other statistical measures.BET surface area-based doses were more suitable to relate surface reactivity to toxicity. In addition, normalization to the deposited dose was advantageous for cellular assays as it improved the predictivity for in vitro as well as in vivo toxicity. The carbonylation assay, potentially in combination with ESR (DMPO spin trap) or FRAS assay, led to the best predictive performance.In summary, we propose a testing strategy for NM OP and demonstrated the applicability in an extended case study on 35 materials. This work is an important contribution towards reliable grouping and testing strategies for NMs.

Behaviour of carbonyl groups in several clinical conditions: Analysis of our survey.

Protein carbonylation is a marker of oxidative protein damage, that is likely involved in the pathogenesis of several diseases. The aim of this study was to evaluate the protein carbonyl (PC) groups in different clinical conditions. It included different groups of subjects: 81 trained subjects; 23 subjects with mild essential hypertension; 31 middle-aged subjects with metabolic syndrome (MS); 106 subjects with MS not selected for age (subdivided into two subgroups, with and without diabetes mellitus); 91 obese adults subdivided in two subgroups (BMI 30-35 Kg/m2 and BMI > 35 kg/m2); 48 subjects with obstructive sleep apnea syndrome (OSAS) subdivided in accordance with the apnea/hypopnea index (AHI); 27 subjects with chronic kidney disease (CKD) on conservative therapy; 31 subjects with CKD on haemodialysis treatment; and 50 subjects with juvenile myocardial infarction. PC groups were reduced in trained subjects in comparison with sedentary controls, while no variation was observed in mild essential hypertension. PC groups were increased in MS subjects and in adult obese subjects. In MS subjects the PC groups were not influenced by the presence of diabetes mellitus and in adult obese subjects were not influenced by the obesity degree. In OSAS subjects only those with AHI > 30 showed an increase of PC groups. PC groups increased in CKD subjects undergoing conservative treatment and haemodialysis therapy. In dialyzed subjects, after a standard dialysis session, there was a marked increase in PC groups. In juvenile myocardial infarction PC groups were higher than in controls; there was no difference between STEMI and NSTEMI and their concentration was unaffected by the number of cardiovascular risk factors or stenosed coronary vessels.

Regional vulnerability to lipoxidative damage and inflammation in normal human brain aging.

Oxidative damage and inflammation coexist in healthy human brain aging. The present study analyzes levels of protein adduction by lipid peroxidation (LP) end-products neuroketal (NKT) and malondialdehyde (MDA), as markers of protein oxidative damage, cycloxygenase-2 (COX-2) levels, as a marker of inflammation, and cytochrome P450 2J2 (CYP2J2), responsible of generation of neuroprotective products, in twelve brain regions in normal middle-aged individuals (MA) and old-aged (OA) individuals. In addition, levels of these markers were evaluated as a function of age as a continuous variable and correction for multiple comparisons. Selection of regions was based on their different vulnerability to prevalent neurodegenerative diseases in aging. Our findings show region-dependent LP end-products, COX-2 and CYP2J2 changes in the aging human brain. However, no clear relationship can be established between NKT, MDA, COX-2 and CYP2J2 levels, and regional vulnerability to neurodegeneration in old age.

Relation of the protein glycation, oxidation and nitration to the osteocalcin level in obese subjects

Carboxylated osteocalcin (Gla-OC) contributes to the bone formation, whereas its undercarboxylated form (Glu-OC) takes part in the energy metabolism. In vitro studies had shown that treatment of osteoblast-like cells with advanced glycation end product-modified bovine serum resulted in reduced synthesis of collagen 1 and osteocalcin. The aim of this study was to find association between Gla-OC and markers of protein glycation, oxidation and nitration, as well as pro-inflammatory and antioxidant defense markers in obese subjects. Non-obese [(body mass index (BMI)<30 kg/m; n=34)] and obese subjects (30<BMI <40 kg/m2; n=98), both sexes, aged 25 to 65 years, were included in this study. Urinary glycation, oxidation and nitration free adduct concentrations were determined by stable isotopic dilution analysis liquid chromatography and mass spectrometry, and normalized to creatinine. Obese subjects had lower Gla-OC serum levels when compared to the non-obese controls. Obese subjects had increased serum concentrations of insulin, C reactive protein, interleukin 6, leptin and insulin resistance index (HOMA IR). Urinary early glycation and advanced glycation end product (AGE) free products, Nε-fructosyl-lysine and 3-deoxyglucosone-derived hydroimidazolone, respectively, and oxidative damage marker, N-formylkynurenine free adduct, were increased in the obese compared to the non-obese subjects. Serum Gla-OC was negatively correlated with urinary methylglyoxal-derived AGE, hydroimidazolone MG-H1, and N-formylkynurenine free adducts. The Gla-OC/Glu-OC index negatively correlated with the MG-H1 free adduct, and correlated positively with the antioxidant defense marker - the glutathione peroxidase activity. Our results suggest that increased AGEs and protein oxidative damage markers in the course of obesity may contribute to decreased Gla-OC level and, consequently, future risk of decreased bone formation.

Alterations of antioxidant status markers in dairy cows during lactation and in the dry period.

The purpose of the study reported in this Research Communication was to evaluate alterations in concentration of total antioxidant capacity (TAC) of plasma, plasma total thiols as markers of oxidative protein damage and malondialdehyde (as a final product of lipid peroxidation) in samples obtained at different stages of the lactation cycle and dry period of dairy cows. We found that TAC was significantly lower in the primiparous cows compared to multiparous cows. This study clearly demonstrates a need for monitoring primiparous cows during the production cycle, especially when they are faced with severe metabolic conditions. Furthermore, TAC may be a sensitive, reliable and useful indicator for measurement of cumulative effects of antioxidants as an addition to metabolic profile tests, which are currently used to analyse dairy cattle health.

Quantitation of plasma thiamine, related metabolites and plasma protein oxidative damage markers in children with autism spectrum disorder and healthy controls

Aims/hypothesis: To assess thiamine and related metabolite status by analysis of plasma and urine in autistic children and healthy controls, correlations to clinical characteristics and link to plasma protein markers of oxidative damage.Methods: 27 children with autism (21 males and 6 females) and 21 (15 males and 6 females) age-matched healthy control children were recruited. The concentration of thiamine and related phosphorylated metabolites in plasma and urine and plasma protein content of dityrosine, N-formylkynurenine and 3-nitrotyrosine was determined.Results: Plasma thiamine and thiamine monophosphate concentrations were similar in both study groups (median [lower–upper quartile]): autistic children – 6.60 nM (4.48–8.91) and 7.00 nM (5.51–8.55), and healthy controls – 6.82 nM (4.47–7.02) and 6.82 nM (5.84–8.91), respectively. Thiamine pyrophosphate (TPP) was decreased 24% in autistic children compared to healthy controls: 6.82 nM (5.81–8.52) versus 9.00 nM (8.41–10.71), p < .01. Urinary excretion of thiamine and fractional renal clearance of thiamine did not change between the groups. No correlation was observed between clinical markers and the plasma and urine thiamine concentration. Plasma protein dityrosine content was increased 88% in ASD. Other oxidative markers were unchanged.Conclusions/interpretation: Autistic children had normal plasma and urinary thiamine levels whereas plasma TPP concentration was decreased. The latter may be linked to abnormal tissue handling and/or absorption from gut microbiota of TPP which warrants further investigation. Increased plasma protein dityrosine may reflect increased dual oxidase activity in response to change in mucosal immunity and host–microbe homeostasis.

Association of GSTO1 and GSTO2 Polymorphism with Risk of End-Stage Renal Disease Development and Patient Survival.

SummaryBackgroundOxidative stress in patients with end-stage renal disease (ESRD) is associated with long-term cardiovascular complications. The cytosolic family of glutathione S-transferases (GSTs) is involved in the detoxication of various toxic compounds and antioxidant protection. GST omega class members, GSTO1 and GSTO2 possess, unlike other GSTs, dehydroascorbate reductase and deglutathionylation activities. The aim of this study was to clarify the role of genetic polymorphisms of GSTO1 (rs4925) and GSTO2 (rs156697) as risk determinants for ESRD development, as well as in the survival of these patients.MethodsA total of 199 patients and 199 healthy subjects were included in the study and genotyped for both GSTO1 and GSTO2 polymorphism. Protein thiol and carbonyl groups as markers of protein oxidative damage were determined spectrophotometrically. Cox proportional hazard model and Kaplan-Meier analysis were performed to investigate the role of GSTO1 and GSTO2 genetic polymorphism on mortality of ESRD patients during the follow-up period (36 month).ResultsIndividuals carrying the variant GSTO2 GG genotype were at 2.45-fold higher risk of ESRD development compared to the wild type GSTO2 AA genotype (OR=2.45; 95%CI=1.18–5.07; p=0.016). The results of GSTO1/ GSTO2 haplotype analysis showed that the haplotype combination of GSTO1 (*A)/GSTO2 (*A) (GSTO1 variant/GSTO2 wild type allele) was protective for ESRD (OR=0.23 95%CI=0.12-0.44, p=0.001). Patients carrying at least one GSTO1 reference allele have shorter mean overall (Log rank=2.844, p =0.241) and cardiovascular survival probability (Log rank=4.211, p=0.122).ConclusionsGSTO polymorphisms have been shown to act as significant markers in assessing the risk of ESRD development and patients’ survival.

Early and gender-specific differences in spinal cord mitochondrial function and oxidative stress markers in a mouse model of ALS.

IntroductionAmyotrophic lateral sclerosis (ALS) is a motor neuron disease with a gender bias towards major prevalence in male individuals. Several data suggest the involvement of oxidative stress and mitochondrial dysfunction in its pathogenesis, though differences between genders have not been evaluated. For this reason, we analysed features of mitochondrial oxidative metabolism, as well as mitochondrial chain complex enzyme activities and protein expression, lipid profile, and protein oxidative stress markers, in the Cu,Zn superoxide dismutase with the G93A mutation (hSOD1-G93A)- transgenic mice and Neuro2A(N2A) cells overexpressing hSOD1-G93A.Results and ConclusionsOur results show that overexpression of hSOD1-G93A in transgenic mice decreased efficiency of mitochondrial oxidative phosphorylation, located at complex I, revealing a temporal delay in females with respect to males associated with a parallel increase in selected markers of protein oxidative damage. Further, females exhibit a fatty acid profile with higher levels of docosahexaenoic acid at 30 days. Mechanistic studies showed that hSOD1-G93A overexpression in N2A cells reduced complex I function, a defect prevented by 17β-estradiol pretreatment. In conclusion, ALS-associated SOD1 mutation leads to delayed mitochondrial dysfunction in female mice in comparison with males, in part attributable to the higher oestrogen levels of the former. This study is important in the effort to further understanding of whether different degrees of spinal cord mitochondrial dysfunction could be disease modifiers in ALS.Electronic supplementary materialThe online version of this article (doi:10.1186/s40478-015-0271-6) contains supplementary material, which is available to authorized users.

Endothelial Cell Senescence with Age is Associated with Oxidative Stress‐Mediated Suppression of Endothelial Function in Healthy Adults

Senescence is emerging as a key mechanism of vascular aging, but evidence in healthy humans is lacking. We hypothesized that endothelial cells (EC) become senescent with age in healthy adults and that this is associated with endothelial dysfunction. In biopsied venous EC, expression of the cell cycle inhibitors p16 and p21 was greater in middle‐aged and older (MA/O, n=10, 60±1 yr) vs. young (Y, n=9, 22±1 yr) healthy adults (p16: 0.45±0.04 vs. 0.28±0.03 AU, P&lt;0.01; p21: 0.63±0.06 vs. 0.34±0.03 AU, P&lt;0.01; quantitative immunofluorescence) whereas p53 did not reach significance (P&gt;0.05). Similarly, EC expression of p16, p21, and p53 was positively related to age (p16: r=0.65, p&lt;0.01; p21: r=0.72, P&lt;0.01; p53: r=0.42, P&lt;0.01). Flow‐mediated dilation (FMD), a measure of endothelial function, was 47% lower in MA/O vs. Y (4.7±0.6 vs. 8.9±0.9%, P&lt;0.01) and was inversely related to EC p21 expression (r=‐0.43, P&lt;0.05). MA/O also had elevated EC abundance of nitrotyrosine (NT), a marker of oxidative protein damage, compared with Y (0.53±0.05 vs. 0.33±0.04 AU, P&lt;0.01) and greater expression of the pro‐oxidant enzyme NADPH oxidase (‐0.28±0.11 vs. ‐0.57±0.08 log(AU), P&lt;0.05). Oxidative stress‐mediated suppression of endothelial function, as indicated in the percent change in FMD following intravenous vitamin C (FMDvc), was greater in MA/O vs. Y (38±17 vs. ‐2±6 %Δ, P&lt;0.05). FMDvc was positively related to EC expression of p16, p21, and p53 (p16: r=0.63, P&lt;0.01; p21: r=0.46, P=0.05; p53: r=0.58, P&lt;0.05). These findings provide novel evidence that senescence occurs in human EC with age in healthy adults and is associated with oxidative stress‐mediated suppression of endothelial function.NIH AG044031 TR001082 AG013038

P87 - Mapping nitro-tyrosine modifications in fibrinogen by mass spectrometry as a biomarker for inflammatory disease

There is a growing awareness that inflammatory diseases have an oxidative pathology, which can result in specific oxidation of amino acids within proteins. It is known that patients with inflammatory disease have higher levels of plasma protein nitro-tyrosine than healthy controls. Fibrinogen is an abundant plasma protein, highly susceptible to such oxidative modifications, and is therefore a potential marker for oxidative protein damage. The aim of this study was to map tyrosine nitration in fibrinogen under oxidative conditions and identify susceptible residues. Fibrinogen was oxidised with 0.25mM and 1mM SIN-1, a peroxynitrite generator, and methionine was used to quench excess oxidant in the samples. The carbonyl assay was used to confirm oxidation in the samples. The carbonyl levels were 2.3, 8.72 and 11.5nmol/mg protein in 0, 0.25mM and 1mM SIN-1 samples respectively. The samples were run on a SDS-PAGE gel and tryptically digested before analysis by HPLC MS-MS. All 3 chains of fibrinogen were observed for all treatment conditions. The overall sequence coverage for fibrinogen determined by Mascot was between 60-75%. The oxidised samples showed increases in oxidative modifications in both alpha and beta chains, commonly methionine sulfoxide and tyrosine nitration, correlating with increasing SIN-1 treatment. Tyrosines that were most susceptible were Tyr135 (tryptic peptide YLQEIYNSNNQK) and Tyr277 (tryptic peptide GGSTSYGTGSETESPR), but several other nitrated tyrosines were also identified with high confidence. Identification of these susceptible peptides will allow design of sequences-specific biomarkers of oxidative and nitrative damage to plasma protein in inflammatory conditions.

Decreased melatonin levels and increased levels of advanced oxidation protein products in the seminal plasma are related to male infertility.

Melatonin, an indolamine secreted by the pineal gland, is known as a powerful free-radical scavenger and wide-spectrum antioxidant. Therefore, the aim of this study was to correlate markers of oxidative protein damage (advanced oxidation protein products, AOPPs) and the total antioxidant capacity (TAC) with melatonin levels in the seminal plasma of men with azoospermia (n=37), theratozoospermia (n=29) and fertile controls (normozoospermia, n=37). Melatonin concentration was measured by radioimmunoassay. The levels of AOPP as well as TAC efficiency (determined by the ferric reducing antioxidant power, FRAP) were estimated by spectrophotometric methods. The concentration of melatonin and AOPP significantly differed in azoospermic (P<0.0001) and theratozoospermic (P<0.0001) patients versus fertile men, and correlated negatively (r=-0.33, P=0.0016). The TAC levels were significantly higher in azoospermia than in theratozoospermia (P=0.0022) and the control group (P=0.00016). In azoospermia, the AOPP concentration was also significantly higher than that observed in theratozoospermia (P=0.00029). Decreased levels of melatonin together with elevated AOPP altered the oxidative-antioxidative balance in the ejaculate, thereby reducing fertility. Therefore, melatonin and AOPP levels may serve as additional diagnostic markers of semen quality and male reproductive potential.

Oxidative modification of serum proteins in multiple sclerosis

Multiple sclerosis (MS) has been demonstrated to involve oxidative stress and augmented glycoxidation. In this study, several markers of protein oxidative damage and glycoxidation have been compared in 14 relapsing remittent in MS (RRMS) patients without immunomodifying treatment, 10 patients in clinical relapse, and clinically stable patient groups treated with interferon β 1a (18) , β 1b (19) and glatiramer acetate (GA; 6) in relation to healthy subjects (12). The glycophore content was increased in RRSM patients without treatment and in patients treated with GA. The level of advanced protein oxidation products (AOPP) was increased in RRSM patients without treatment and in patients with clinical relapse. The level of protein carbonyls was elevated in RRSM patients without treatment and in patients treated with interferon β 1b. The levels of dityrosine level and N′-formylkynureine were elevated in RRSM patients without treatment while serum protein thiol groups were decreased in RRSM patients in clinical relapse as well as RRMS patients treated with interferon β 1a. Several markers of protein modification showed correlation with the C-reactive protein level and white blood cell count, suggesting that oxidative protein modifications are linked to the inflammatory processes in MS. Results of this study confirm the occurrence of protein oxidative and glycoxidative damage in MS and show that spectrophotometric and fluorimetric markers of this damage, especially the AOPP level, may be useful in monitoring oxidative stress in the course of therapy of MS.

Assessing of plasma protein denaturation induced by exposure to cadmium, electromagnetic fields and their combined actions on rat

In our environment, we have numerous chances to be exposed to not only electromagnetic fields (EMFs) but also many chemicals containing mutagens. Therefore, the aim of this study was to estimate whether rat’s exposure to cadmium and/or EMFs could cause oxidative damage to molecular structure of proteins and whether and to what extent the effects of co-exposure differ from those observed under the treatment with each exposure alone. Thirty-two rats were divided into four groups. Group 1 was termed as control, group 2 was treated with cadmium (3.0 mg/Kg), group 3 was exposed to EMF (10 mT/h/day) and group 4 was treated with cadmium and exposed to EMF. Protein carbonyls (PCO) in the plasma as a marker of oxidative protein damage and total oxidant status (TOS), as well as electrical conductivity and SDS electrophoresis to estimate changes in molecular structure of protein, were determined. The exposure to Cd and/or EMF led to oxidative protein damage (increased PCO and TOS) accomplished by increased stress of electrical charges on the surface of the protein molecule (increased electrical conductivity) and changes in the molecular structure of protein. The effects were more pronounced after treatment with both Cd and EMF than at the treatment with each exposure alone. The serious damage to proteins at the co-exposure to Cd and EMF seems to be due to the interference of the EMF with the toxic activity of cadmium. This work concluded that combined exposure to Cd and EMFs might increase the risk of plasma damage via enhancing free radical generation and protein oxidation.

Two different markers of oxidative protein damage in plasma of alloxan-induced hyperglycemic rabbits: effects of pioglitazone and repaglinide. Oxidative protein damage and two antidiabetics

Reactive oxygen and nitrogen species are known to contribute to different pathological alterations in hyperglycemic state. Especially proteins are major targets resulting from their rapid reactions with many oxidants. Therefore, the present study was undertaken to examine some common markers of oxidative/nitrosative stress in plasma of alloxan-induced hyperglycemic rabbits with special regard to protein oxidation. The second goal was to investigate which of two antidiabetics, pioglitazone from peroxisome proliferator-activated receptor gamma agonists, and repaglinide from non-sulfonylurea KATP channel blockers, could be more effective in ameliorating these oxidative/nitrosative changes. In our hyperglycemic animals glutathione decreased by 41% as compared to control group. Simultaneously, lipid peroxidation products increased by 62%, nitrites by 59%, protein carbonyl groups by 71% and advanced oxidized protein products by 53%. In hyperglycemic-treated animals pioglitazone increased glutathione above control values. In turn, lipid peroxidation was reduced by 29%, nitrites by 22%, advanced oxidized protein products by 37% and protein carbonyl groups by 42% in relation to hyperglycemic-non treated rabbits. After repaglinide treatment, lipid peroxidation decreased by 17% but the levels of glutathione, nitrites and both markers of oxidized proteins, were not affected. Our results confirmed that the action of pioglitazone is more comprehensive than that of repaglinide, independently of their action on hyperglycemia.

Assessment of nitric oxide, advanced oxidation protein products, malondialdehyde, and thiol levels in patients with restless legs syndrome

We aimed to determine the importance of oxidative stress in the pathogenesis of restless legs syndrome (RLS) by quantification of advanced oxidation protein products and total thiol levels (as markers of oxidative protein damage), nitric oxide levels (as an antioxidant and endothelial function), and malondialdehyde levels (as a marker of lipid peroxidation) in patients with RLS. A total of 22 patients with primary RLS were enrolled in the study and 20 age-and-gender-matched healthy subjects were enrolled as a control group. Serum nitric oxide, malondialdehyde, thiol levels, and plasma advanced oxidation protein products levels were determined by spectrophotometric methods. Serum nitric oxide and thiol levels were lower in the patient group than in controls (p = 0.007 and p = 0.017, respectively). Plasma advanced oxidation protein products levels and serum malondialdehyde levels were found to be higher in patients with RLS than in controls (p = 0.017 and p = 0.008, respectively). Serum malondialdehyde level was found to be positively correlated with plasma advanced oxidation protein products levels (p = 0.039). Serum thiol level was found to be negatively correlated with plasma advanced oxidation protein products levels (p = 0.030). Increased advanced oxidation protein products, malondialdehyde levels, and decreased thiol and nitric oxide levels, may suggest that patients with RLS are under oxidative stress. Although both lipid peroxidation and protein oxidation may have a role in atherosclerosis in RLS, those factors may be related to the pathogenesis of RLS.

Ozone oxidative post-conditioning reduces oxidative protein damage in patients with disc hernia

Introduction/objectives: Although inflammation in disc hernia (DH) has been recognized and it is a well-known process mediated by loss of the cellular redox balance, only a few studies about the impact of chronic oxidative stress on this neurological disorder have been made. Ozone therapy has been widely used with clinical efficacy in DH. This work aimed at characterizing the systemic redox status of patients with low back pain and neck pain as well as studying if ozone oxidative post-conditioning modified the pathological oxidative stress and protected against oxidative protein damage and if there is any relationship between oxidative changes and pain in both DH.Methods: Redox status of 33 patients with diagnosis of DH by computerized axial tomography, nuclear magnetic resonance, and clinical evaluations was studied. Ozone was administered by paravertebral way. After ozone treatment, plasmatic levels of antioxidant/pro-oxidant markers, pain, and life quality disability parameters were evaluated.Results: One hundred percent of patients showed a severe oxidative stress. Major changes in superoxide dismutase activity, total hydroperoxides, advanced oxidation protein products, fructolysine content, and malondialdehyde were observed. After ozone oxidative post-conditioning, there was a re-establishment of patients’ cellular redox balance as well as a decrease in pain in both DH. A relationship between indicators of oxidative protein damage and pain was demonstrated.Conclusions: Ozone therapy protected against oxidation of proteins and reduced the pain. Relationship between markers of oxidative protein damage, disability parameters, and pain suggests the role of oxidative stress in the pathological processes involved in DH.