Protein Carbonyl Groups Research Articles

Dill tablet: A potential antioxidant and anti-diabetic medicine

ObjectiveTo evaluate the antiglycation and antioxidant properties of the dill tablet, an herbal product used in Iran as a hypolipidemic medicine. MethodsIn this descriptive study, the antioxidant and antiradical properties of dill tablet at different concentration (0.032, 0.065, 0.125, 0.25, 0.5 and 1 mg/mL) were measured. The total phenolic, flavonols and flavonoid, alkaloids, anthocyanin, tannin and saponin contents in dill tablet were determined. Furthermore, antiglycation properties of dill tablet were assayed. In the in vivo experiments, male rats were randomly divided into three groups (n = 6): Group 1: normal rats; Group 2: diabetic rats; Group 3: diabetic rats + 300 mg/kg dill tablet, and Group 4: diabetic rats + 100 mg/kg dill tablet. After 2 months, the blood glucose was measured enzymatically and advanced glycation end-products (AGEs) formation was determined using a fluorometric method. ResultsOur results illustrated that different concentrations of dill tablet had significant antioxidant activity. Dill tablet markedly declined AGEs formation and fructosamine levels (P < 0.001) compared with glycated sample. Oxidation of protein carbonyl and thiol group was significantly reduced by dill tablet in a dose dependent manner (P < 0.001). Formation of amyloid cross-β and fragmentation were markedly inhibited by dill tablet (P < 0.001) compared with glycated sample. After 2 months, fasting blood glucose levels (P < 0.001) and AGEs formation (P < 0.05) were significantly reduced by dill tablet in diabetic animals. ConclusionsDill tablet exhibited significant antiglycation and antioxidant activities. This study provides a scientific basis for using dill in treatment of diabetic patients.

Fluorimetric screening assay for protein carbonyl evaluation in biological samples

Many assays are available for the detection of protein carbonyls (PCs). Currently, the measurement of PC groups after their derivatization with 2,4-dinitrophenol hydrazine (DNPH) is widely used for measuring protein oxidation in biological samples. However, this method includes several washing steps. In this context, we have developed a rapid, sensitive, and accurate fluorimetric method adapted to 96-well microplates for the convenient assessment of protein carbonyl level in biological samples. The method reported here is based on the reaction of carbonyl content in proteins with 7-hydrazino-4-nitrobenzo-2,1,3-oxadiazole (NBDH) to form highly fluorescent derivatives via hydrazone formation. PCs were determined using the DNPH and NBDH assays in fully reduced bovine serum albumin (BSA) and plasma and liver homogenates obtained from healthy control rats up the addition of various amounts of HOCl-oxidized BSA (OxBSA). Using the NBDH assay, PC concentrations as low as 0.2nmol/mg were detected with precision as low as 5%. Matrix-assisted laser desorption/ionization time-of-flight (MALDI–TOF) mass spectroscopy was used to successfully identify the formation of the NBDH adducts after derivatization with standard oxidized peptides. Finally, the two methods were further used for PC determination in plasma and liver samples from diabetic and normal rats, showing that the NBDH assay can be reliably used in biological experiments.

Association between hyperleptinemia and oxidative stress in obese diabetic subjects.

BackgroundObesity is a worldwide metabolic disorder affecting all types of people. The mechanism by which increased body fat mass that leads to insulin resistance and type 2 diabetes is not yet clearly known. There is a possible crosstalk between leptin, an adipokine and insulin signaling. Leptin mediates insulin sensitivity in hepatocytes; however, its concentration has found to be increased in obese and diabetic subjects. These subjects also have high incidence of oxidative stress status. Therefore, knowing the level of leptin present in obese diabetic subjects will be informative along with its relation to oxidative stress.MethodsA small population study was performed to explore the association between leptin concentration and oxidative stress status in control and obese type 2 diabetic subjects. Oxidative stress status parameters like malondialdehyde (MDA), superoxide dismutase activity (SOD), glutathione peroxidase activity (GSH-Px), and protein carbonyl (PCO) groups content was measured spectrophotometrically in serum of 43 subjects. Serum Leptin concentration was measured by quantikine sandwich ELISA assay.ResultsThe strong positive correlation between MDA (malondialdehyde) and leptin in obese diabetic patients (ρ = 0.787, P < 0.05) suggests close association between lipid peroxidation and hyperleptinemia. In addition, observed positive correlation between protein carbonyl groups and leptin level in obese diabetic subjects (ρ = 0.599, P = 0.001) suggest that hyperleptinemia might also be associated with increased protein oxidation. In multiple logistic regression analysis, leptin has shown a significant association with obese type 2 diabetes [odds ratio (OR): 1.161, 95% confidence interval (Cl): 1.027-1.312, P < 0.05], but the significance is lost after adjusting for Age, BMI, MDA and anti-oxidant parameters.ConclusionsIn the subjects with both obesity and diabetes, there is a significant degree of association between hyperleptinemia and oxidative stress. This association reinforces the existing understanding that obese subjects who also have diabetes are vulnerable to cardiovascular complications driven by increased oxidative stress and hyperleptinemia.

Oxidative state and oxidative metabolism in the brain of rats with adjuvant-induced arthritis

The purpose of the present study was to evaluate the oxidative status of the brain of arthritic rats, based mainly on the observation that arthritis induces a pronounced oxidative stress in the liver of arthritis rats and that morphological alterations have been reported to occur in patients with rheumatoid arthritis. Rats with adjuvant-induced arthritis were used. These animals presented higher levels of reactive oxygen species (ROS) in the total brain homogenate (25% higher) and in the mitochondria (+55%) when compared to healthy rats. The nitrite plus nitrate contents, nitric oxide (NO) markers, were also increased in both mitochondria (+27%) and cytosol (+14%). Arthritic rats also presented higher levels of protein carbonyl groups in the total homogenate (+43%), mitochondria (+69%) and cytosol (+145%). Arthritis caused a diminution of oxygen consumption in isolated brain mitochondria only when ascorbate was the electron donor. The disease diminished the mitochondrial cytochrome c oxidase activity by 55%, but increased the transmembrane potential by 16%. The pro-oxidant enzyme xanthine oxidase was 150%, 110% and 283% higher, respectively, in the brain homogenate, mitochondria and cytosol of arthritic animals. The same occurred with the calcium-independent NO-synthase activity that was higher in the brain homogenate (90%) and cytosol (122%) of arthritic rats. The catalase activity, on the other hand, was diminished by arthritis in all cellular fractions (between 30 and 40%). It is apparent that the brain of rats with adjuvant-induced arthritis presents a pronounced oxidative stress and a significant injury to lipids and proteins, a situation that possibly contributes to the brain symptoms of the arthritis disease.

Protective effect of Tetracera scandens L. leaf extract against CCl4-induced acute liver injury in rats

ABSTRACT Objective To investigate the protective potential of ethanolic extracts of Tetracera scandens L. (T. scandens) against CCl4 induced oxidative stress in liver tissues. Methods Dried leaf powder of T. scandens was extracted with ethanol and concentrated to yield a dry residue. Rats were administered with 100 mg/kg of ethanolic extracts orally once daily for one week. Animals were subsequently administered with a single dose of CCl4 (1 mL/kg body weight, intraperitoneal injection). Various assays, such as serum levels of alanine aminotransferase, aspartate aminotransferase, lipid peroxidation, protein oxidation (carbonyl protein group), tumor necrosis factor alpha, catalase, superoxide dismutase, and glutathione peroxidase, were used to assess damage caused by CCl4 and the protective effects of the ethanol extract on liver tissues. Results Hepatotoxicity induced by CCl4 was evidenced by a significant increase in serum aspartate aminotransferase and alanine aminotransferase level, lipid peroxidation, protein carbonyl group, and tumor necrosis factor alpha, as well as decreased activity of the hepatic antioxidant enzymes (catalase, superoxide dismutase, and glutathione peroxidase). Treatment with ethanolic T. scandens extracts prevented all of these typically observed changes in CCl4-treated rats. Conclusions Our findings indicate that T. scandens has a significant protective effect against CCl4 induced hepatotoxicity in rat, which may be due to its antioxidant properties.

Chronic intermittent hypoxia increases rat sternohyoid muscle NADPH oxidase expression with attendant modest oxidative stress.

Chronic intermittent hypoxia (CIH) causes upper airway muscle dysfunction. We hypothesized that the superoxide generating NADPH oxidase (NOX) is upregulated in CIH-exposed muscle causing oxidative stress. Adult male Wistar rats were exposed to intermittent hypoxia (5% O2 at the nadir for 90 s followed by 210 s of normoxia), for 8 h per day for 14 days. The effect of CIH exposure on the expression of NOX subunits, total myosin and 4-hydroxynonenal (4-HNE) protein adducts in sternohyoid muscle was determined by western blotting and densitometry. Sternohyoid protein free thiol and carbonyl group contents were determined by 1D electrophoresis using specific fluorophore probes. Aconitase and glutathione reductase activities were measured as indices of oxidative stress. HIF-1α content and key oxidative and glycolytic enzyme activities were determined. Contractile properties of sternohyoid muscle were determined ex vivo in the absence and presence of apocynin (putative NOX inhibitor). We observed an increase in NOX 2 and p47 phox expression in CIH-exposed sternohyoid muscle with decreased aconitase and glutathione reductase activities. There was no evidence, however, of increased lipid peroxidation or protein oxidation in CIH-exposed muscle. CIH exposure did not affect sternohyoid HIF-1α content or aldolase, lactate dehydrogenase, or glyceraldehyde-3-phosphate dehydrogenase activities. Citrate synthase activity was also unaffected by CIH exposure. Apocynin significantly increased sternohyoid force and power. We conclude that CIH exposure upregulates NOX expression in rat sternohyoid muscle with concomitant modest oxidative stress but it does not result in a HIF-1α-dependent increase in glycolytic enzyme activity. Constitutive NOX activity decreases sternohyoid force and power. Our results implicate NOX-dependent reactive oxygen species in CIH-induced upper airway muscle dysfunction which likely relates to redox modulation of key regulatory proteins in excitation-contraction coupling.

Increased oxidative stress in patients with 3-hydroxy-3-methylglutaric aciduria.

3-hydroxy-3-methylglutaric aciduria (HMGA; OMIM 246450) is a rare autosomal recessive disorder, caused by the deficiency of 3-hydroxy-3-methylglutaryl-CoA lyase (4.1.3.4), which results in the accumulation of3-hydroxy-3-methylglutaric (HMG) and 3-methylglutaric (MGA) acids in tissues and biological fluids of affected individuals. Recent in vivo and in vitro animal studies have demonstrated that the accumulation of these metabolites can disturb the cellular redox homeostasis, which can contribute to the neurological manifestations presented by the patients. So, in the present work, we investigated oxidative stress parameters in plasma and urine samples from HMGA patients, obtained at the moment of diagnosis of this disorder and during therapy with low-protein diet and L-carnitine supplementation. It was verified that untreated HMGA patients presented higher levels of urinary di-tyrosine and plasma thiobarbituric acid-reactive substances (TBA-RS), which are markers of protein and lipid oxidative damage, respectively, as well as a reduction of the urinary antioxidant capacity. Treated HMGA patients also presented an increased protein oxidative damage, as demonstrated by their higher concentrations of plasma protein carbonyl groups and urinary di-tyrosine, as well as by the reduction of total sulfhydryl groups in plasma, in relation to controls. On the other hand, HMGA patients under therapy presented normal levels of TBA-RS and urinary antioxidant capacity, which can be related, at least in part, to the antioxidant and antiperoxidative effects exerted by L-carnitine. The results of this work are the first report showing that a redox imbalance occurs in patients with HMGA what reinforces the importance of the antioxidant therapy in this disorder.

Effects of Ramadan Fasting on Serum Amyloid A and Protein Carbonyl Group Levels in Patients With Cardiovascular Diseases.

Introduction: Serum amyloid-A (SAA) and protein carbonyl group are rigorously related with cardiovascular diseases (CVDs) as a sensitive marker of an acute inflammatory state and as an important index of oxidative stress, respectively. Moreover, diet is one of the main factors that can modify cardiovascular risks. Therefore, this study aimed to investigate the effects of Ramadan fasting on SAA and protein carbonyl group levels in patients with CVDs. Methods: Twenty-one patients (21 male; mean age 52±9 years old) with CVDs (coronary artery disease, cerebrovascular, or peripheral arterial diseases) were participated in this study. Biochemical parameters were measured in patients 2 days before and 2 days after Ramadan fasting. SAA levels were assessed using enzyme-linked immunosorbent assay and Cayman’s protein carbonyl colorimetric assay was provided for measuring protein carbonyl groups. Results: According to the findings of the study, post-Ramadan levels of inflammatory biomarker, SAA was decreased significantly in patients with CVDs in comparison with the baseline before-fasting values (16.84±8.20 vs. 24.40±6.72 μg/ml, P = 0.021). In addition, Ramadan fasting significantly reduced the levels of protein carbonyl group in patients as compared with those of baseline values (33.08±15.31 vs. 43.65±16.88 nmol/ml, P = 0.039). Conclusion: Ramadan fasting has impressive effects on modulating CVDs by decreasing inflammation and oxidative stress markers. However, to get a clear conclusion with more results, further investigation is warranted.

Protein carbonyl groups in trained subjects before and after a cardiopulmonary test

Physical exercise influences the body's oxidative status. The modifications can involve lipids, proteins and nucleic acids, and different effects seem to be induced by regular and acute exercise respectively. We examined protein oxidation, expressed as concentration of protein carbonyl groups (PC), in trained subjects before (time 0), 10 min (time 1) and 24 hours (time 2) after a cardiopulmonary test performed on a cycloergometer. We enrolled 38 trained subjects (26 men and 12 women), subdivided in two groups (A1 and B1) of 19 subjects each, according to the median value of VO2max, and in two groups (A2 and B2) of 19 subjects each, according to the median value of PC at baseline. PC concentration was measured by an enzyme-linked immunosorbent assay (ELISA). The groups A1 and B1 did not differ from each other as regards the basal PC level and groups A2 and B2 were not different as regards the VO2max. At time 1 PC showed a significant increase in comparison with baseline in trained subjects as a whole group, as well as in each subgroup. At time 2, PC were decreased in comparison with both times 0 and 1 in the whole group and in subgroups A1 and B2, whereas in subgroups A2 and B1 the PC value at time 2 was not different compared to time 0. The percentage increase of PC at time 1 vs time 0, as well as the percentage decrease at time 2 vs time 1 and time 0 respectively, were not different between subgroups A1 and B1. On the contrary, the percentage variations observed at each interval were significantly different between subgroups A2 and B2. The results suggest a reaction of antioxidant systems to acute exercise in trained subjects, influenced by basal PC levels more than by aerobic fitness.

Differential Measurements of Oxidatively Modified Proteins in Colorectal Adenopolyps.

IntroductionAdenopolyps patients have a three-fold higher risk of colon cancer over the general population, which increases to six-fold if the polyps are multiple and with lower survival among African American population. Currently, 6% of CRC can be ascribed to mutations in particular genes. Moreover, the optimal management of patients with colorectal adenopolyps depends on the accuracy of appropriate staging strategies because patients with similar colorectal adenocarcinoma architecture display heterogeneity in the course and outcome of the disease. Oxidative stress, due to an imbalance between reactive oxygen species (ROS) and antioxidant capacities as well as a disruption of redox signaling, causes a wide range of damage to DNA, proteins, and lipids which promote tumor formation.Objective/MethodThis study applied spectrophotometric, dinitrophenylhydrazone (DNPH) assay, two-dimensional gel electrophoresis, and western blot analyses to assess the levels of oxidatively modified proteins in 41 pairs of primary colorectal tissues including normal/surrounding, adenopolyps (tubular, tubulovillous, villous, polypvillous) and carcinoma. Analysis of variance (ANOVA) and Student’s t-tests were utilized for the resulting data set.ResultsOur data showed that the levels of reactive protein carbonyl groups significantly increased as colorectal adenopolyps progresses to malignancy. No significant differences were found in the levels of carbonyl proteins between gender samples analyzed. For African American patients, there were, relative to Caucasians, 10% higher levels of reactive carbonyls in proteins of tubulovillous tissue samples (P < 0.05) and over 36% higher in levels in adenocarcinomas (P < 0.05). In normal tissues and tubular, there were no significant differences between the two groups in levels of protein carbonyls. Differences in the levels of protein carbonyl expression within individual patient samples with different number of tumor cells were notably evident.ConclusionResults suggested that oxidative stress could be involved in the modification of oxidatively carbonyl proteins in the precancer stages, leading to increased aggressiveness of colorectal polyps.

Endothelial dysfunction in subjects with chronic cadmium exposure.

Vascular endothelium is a target of cadmium (Cd) toxicity. Cd exposure has been reported to be associated with vascular disorders. In this study, we aimed to investigate the effects of Cd exposure on markers of endothelial function in human subjects chronically exposed to Cd. Based on blood Cd levels, seventy-five women were categorized into non-exposed, Cd-exposed and severely Cd-exposed groups. Nitrite, L-arginine, asymmetric dimethylarginine (ADMA), and soluble thrombomodulin levels in blood were measured. Nitrite levels were lower in Cd-exposed subjects than non-exposed subjects. Plasma L-arginine decreased while ADMA, an endogenous endothelial nitric oxide synthase (eNOS) inhibitor, increased in Cd-exposed subjects. Soluble thrombomodulin also increased in Cd-exposed subjects. In Cd-exposed subjects, plasma malondialdehyde and protein carbonyl groups increased while the erythrocytic glutathione decreased. Multiple linear regression analysis revealed a negative association between urinary Cd and nitrite levels in erythrocytes. Our research suggests that subjects with chronic Cd exposure have endothelial dysfunction.

Anti Oxidative Effect of Black Tea Theaflavin on Erythrocytes Subjected to Oxidative Stress

Tea (Camellia sinensis) is one of the most popularly consumed beverage worldwide. It is consumed mostly as green tea, oolong, or black tea. Black tea, good source of flavan-3-ols the theaflavins: mixture of theaflavin (TF), theaflavin-3-gallates, theaflavin-3′-gallates and theaflavin-3,3′-digallates, possess numerous biological and therapeutic properties. However, the exact mechanisms underlying these properties of TFs remain speculative. In the present study, we investigated the in vitro protective effect of theaflavin on membrane protein carbonyl group, sulfhydryl group (–SH) and erythrocyte hemolysis in rat. TFs (at micromolar concentration) showed significant antioxidant effect in protecting erythrocytes from oxidation-induced changes.

Piperine, a natural bioenhancer, nullifies the antidiabetic and antioxidant activities of curcumin in streptozotocin-diabetic rats.

Knowing that curcumin has low bioavailability when administered orally, and that piperine has bioenhancer activity by inhibition of hepatic and intestinal biotransformation processes, the aim of this study was to investigate the antidiabetic and antioxidant activities of curcumin (90 mg/kg) and piperine (20 or 40 mg/kg), alone or co-administered, incorporated in yoghurt, in streptozotocin (STZ)-diabetic rats. The treatment for 45 days of STZ-diabetic rats with curcumin-enriched yoghurt improved all parameters altered in this experimental model of diabetes: the body weight was increased in association with the weight of skeletal muscles and white adipose tissues; the progressive increase in the glycemia levels was avoided, as well as in the glycosuria, urinary urea, dyslipidemia, and markers of liver (alanine and aspartate aminotransferases and alkaline phosphatase) and kidney (urinary protein) dysfunction; the hepatic oxidative stress was decreased, since the activities of the antioxidant enzymes superoxide dismutase, catalase and gluthatione peroxidase were increased, and the levels of malondialdehyde and protein carbonyl groups were reduced. The dose of 20 mg/kg piperine also showed antidiabetic and antioxidant activities. The treatment of STZ-diabetic rats with both curcumin and 20 mg/kg piperine in yoghurt did not change the antidiabetic and antioxidant activities of curcumin; notably, the treatment with both curcumin and 40 mg/kg piperine abrogated the beneficial effects of curcumin. In addition, the alanine aminotransferase levels were further increased in diabetic rats treated with curcumin and 40 mg/kg piperine in comparison with untreated diabetic rats. These findings support that the co-administration of curcumin with a bioenhancer did not bring any advantage to the curcumin effects, at least about the antidiabetic and antioxidant activities, which could be related to changes on its biotransformation.

Chemical-free inactivated whole influenza virus vaccine prepared by ultrashort pulsed laser treatment.

There is an urgent need for rapid methods to develop vaccines in response to emerging viral pathogens. Whole inactivated virus (WIV) vaccines represent an ideal strategy for this purpose; however, a universal method for producing safe and immunogenic inactivated vaccines is lacking. Conventional pathogen inactivation methods such as formalin, heat, ultraviolet light, and gamma rays cause structural alterations in vaccines that lead to reduced neutralizing antibody specificity, and in some cases, disastrous T helper type 2-mediated immune pathology. We have evaluated the potential of a visible ultrashort pulsed (USP) laser method to generate safe and immunogenic WIV vaccines without adjuvants. Specifically, we demonstrate that vaccination of mice with laser-inactivated H1N1 influenza virus at about a 10-fold lower dose than that required using conventional formalin-inactivated influenza vaccines results in protection against lethal H1N1 challenge in mice. The virus, inactivated by the USP laser irradiation, has been shown to retain its surface protein structure through hemagglutination assay. Unlike conventional inactivation methods, laser treatment did not generate carbonyl groups in protein, thereby reducing the risk of adverse vaccine-elicited T helper type 2 responses. Therefore, USP laser treatment is an attractive potential strategy to generate WIV vaccines with greater potency and safety than vaccines produced by current inactivation techniques.

Kinetics of glycoxidation of bovine serum albumin by glucose, fructose and ribose and its prevention by food components.

The aim of this study was to compare the kinetics of the glycoxidation of bovine serum albumin (BSA) as a model protein by three sugars: glucose, fructose and ribose, using fluorometric measurements of the content of advanced glycation end products (AGEs), protein-bound fructosamine, dityrosine, N'-formylkynurenine, kynurenine, tryptophan, the content of advanced oxidation protein products (AOPP), protein carbonyl groups, as well as thiol groups. Moreover, the levels of glycoalbumin and AGEs were determined by using an enzyme-linked immunosorbent assay. Based on the kinetic results, the optimal incubation time for studies of the modification of the glycoxidation rate by additives was chosen, and the effects of 25 compounds of natural origin on the glycoxidation of BSA induced by various sugars were examined. The same compounds were found to have different effects on glycoxidation induced by various sugars, which suggests caution in extrapolation from experiments based on one sugar to other sugars. From among the compounds tested, the most effective inhibitors of glycoxidation were: polyphenols, pyridoxine and 1-cyano-4-hydroxycinnamic acid.

Estrogen plus progestin increase superoxide dismutase and total antioxidant capacity in postmenopausal women

Objective This cross-sectional study aimed to evaluate the behavior of blood antioxidant enzymes (superoxide dismutase (SOD), catalase and glutathione peroxidase), plasma total antioxidant capacity and oxidative damage (lipid oxidation and protein carbonyl levels) and their relationship with the serum levels of steroid hormones in premenopausal and postmenopausal women without and with estrogen alone (ET) or estrogen plus progestin therapy (EPT).Methods Blood was collected from four groups of subjects: premenopausal women (n = 24), postmenopausal women without hormone therapy (n = 31), postmenopausal women with ET (n = 12) and postmenopausal women with EPT (n = 16).Results The activities of the different SOD isoforms (CuZnSOD and MnSOD) and the plasma total antioxidant power were significantly higher in the postmenopausal women under EPT than in the postmenopausal women without hormone replacement therapy (HRT). Only CuZnSOD activity was increased in women receiving ET compared to the postmenopausal women without HRT. However, no differences were observed in the levels of lipid or protein oxidation or in the non-enzymatic plasma antioxidants (uric acid and albumin) among the groups. The duration of HRT and serum estrogen levels were positively correlated to the blood CuZnSOD activity and to plasma total antioxidant power, whereas the serum progesterone levels were positively correlated to CuZnSOD activity and negatively correlated to protein carbonyl groups. Interestingly, the total antioxidant power of plasma was positively correlated to CuZnSOD and glutathione peroxidase activities.Conclusion We conclude that EPT increases blood MnSOD and CuZnSOD activity in postmenopausal women, leading to an increased plasma total antioxidant capacity. This finding may be relevant to the prevention of oxidative stress-related disorders in postmenopausal women.

Abstract C05: Analysis of oxidative modified proteins in serous subtype of ovarian carcinoma

Abstract Serous subtype of ovarian cancer accounts for approximately 60% to 80% of ovarian cancer cases and is by far the most aggressive histology. Fewer than 25% of the serous ovarian cancer cases are detected at an early stage (stages I and II), a statistic which reflects grimly on the low survival rates, especially among African-American women. Elevated levels of reactive oxygen species (ROS) are one of many of known and suspected risk factors associated with ovarian cancer. Oxidative stress, which results from an imbalance between ROS and antioxidant capacities, can cause a wide range of direct or indirect DNA damage. There are extensive DNA repair systems that can correct DNA damage caused by ROS before cell replication and mutation fixation. Given that serous cancer subtype is considered to have originated from the fallopian epithelium mucosa that was exposed to immense physiological changes, driven by cyclically altering hormone levels as a result of ovulation. The influence of ovulation is a cause of increased oxidative stress. In this study, we applied spectrophotometric DNPH assay, two dimension gel electrophoresis (2-D SDS-PAGE), and Oxidized protein western blot analysis to identify and evaluate the level of oxidative modified protein, using derivatisation of protein carbonyl group with 2,4-dinitrophenylhyrdozine(DNPH) assay in the obtained 155 snap-freeze primary serous epithelial ovarian tissues including, normal, normal surrounding, cystoadenoma, borderline and carcinoma with age range of 65±12, (55% Caucasian 45% Africa American). Our results showed that the index of carbonyl protein /total protein increased by onset and progression of the serous ovarian malignancy. The elevation of oxidized protein among African American was 40 % higher than Caucasian in advanced stages of disease. SDS-PAGE profiles and western blot analysis of oxidized proteins showed a distinct protein pattern among cancerous and non-cancerous tissues. Our study suggests that level of oxidative stress, may involve the selective modification of some enzymes and structural proteins and play a role in etiological differences that may exist between stages of benign and invasive epithelial ovarian tumors. This study was funded in part as a result of the grants from NIH-CHDRD CA150317 and NCI CA150039. Citation Format: Sharifeh Mehrabi, Shakeria Cohen, Edward E. Partridge, Felix O. Aikhionbare. Analysis of oxidative modified proteins in serous subtype of ovarian carcinoma. [abstract]. In: Proceedings of the Sixth AACR Conference: The Science of Cancer Health Disparities; Dec 6–9, 2013; Atlanta, GA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2014;23(11 Suppl):Abstract nr C05. doi:10.1158/1538-7755.DISP13-C05

Radical scavenging and antioxidant effects of Matricaria chamomilla polyphenolic–polysaccharide conjugates

Matricaria chamomilla L. (MC), a member of the Asteraceae family, is one of the oldest medicinal plants, widely used worldwide for a variety of healing applications. Its recommendations, derived from both traditional and modern medicine, include numerous disorders such as inflammation, ulcers, wounds, gastrointestinal disorders, stomach ache, pharyngitis, rheumatic pain, as well as the other ailments.This work is focused on another aspect of the biological activity of chamomile polyphenolic–polysaccharide conjugates – their antioxidant properties in the protection of blood plasma components against in vitro oxidative stress.Measurements of DPPH and ABTS radical scavenging indicated considerable anti-free radical action of MC. Pre-incubation of blood plasma with MC considerably diminished the extent of ONOO−-induced oxidative modifications such as protein carbonyl groups, SH groups, 3-nitrotyrosine, as well as the formation of lipid hydroperoxides. The analysis of the FRAP assay result shows a considerable increase of ferric reducing ability of blood plasma in the presence of MC.The results obtained in this study indicate that polyphenolic–polysaccharide conjugates isolated from M. chamomilla substances possess antioxidant properties. The M. chamomilla macromolecular glycoconjugates may be useful in the creation of new natural-based medications or dietary supplements, helpful in the prevention and treatment of oxidative stress-mediated disorders.

P7 - Oxi-DIGE: A novel proteomic approach for detecting and quantifying carbonylated proteins

Proteins are involved in key cellular functions and our health and wellness depends on their quality. Accumulation of oxidatively damaged proteins is a hallmark of deleterious processes such increased oxidative stress, chronic inflammation, ageing and age-related diseases. Thus, quantifying and identifying oxidized proteins is a biomarker of choice for monitoring biological ageing and/or the efficiency of anti-oxidant, ant-inflammatory and anti-ageing therapies. However, the absence of reliable tools for analyses has inhibited its establishment as the gold standard for measuring the efficacy of anti-ageing and age related diseases interventions. Herein, we present a novel proteomics technology, named Oxi-DIGE?, which provides a significant improvement in terms of specificity, reproducibility and statistical support for proteomic analysis of carbonylated proteins. In Oxi-DIGE, protein carbonyls are labelled with fluorescent hydrazide probes that bind specifically to carbonyl groups in proteins. Experimental groups (e.g. control and experimental samples) are labelled with different flurophore-binded hydrazides that fluoresce light at different wavelengths, producing different colour fluorescence. Thus samples from different experimental groups are co-resolved on a single 2D gel. Increased accuracy is provided due to: (i) reduced false positives by using an exogenous synthetic fluorescent tag; (ii) multiplexing, that is the possibility to run multiple samples on the same gel, (iii) the use of an internal standard on each gel which eliminates inter-gel variations and provides an increased statistical confidence. In addition, the resolution of the carbonyl groups is improved, forming distinct spots that can be identified by mass spectrometry.?Patent Application (M. Baraibar, R. Ladouce., B. Friguet, A method for detecting and/or quantifying carbonylated proteins (WO/2012/175519) filed by UPMC and referring to the technology described in this abstract.

EFFECT OF DIFFERENT SALINITY LEVELS ON VEGETATIVE GROWTH AND CHEMICAL CONTENTS OF JOJOBA (Simmondsia chinensis) Seedlings

Two pots experiments had done at Faculty of Agriculture's Nursery in MansouraUniversity , Mansoura city , Egypt . In two summer seasons (2011 and 2012) to study the effect of Sodium chloride salinity levels on (vegetative growth) and (chemical contents) of Jojoba ( Simmondsia chinensis ) seedlings . Five treatments were arranged in complete block randomize with five salinity levels the control ( Tap water ) , 2000 , 5000 , 7000 and 10000 PPM . The most important finding could be summarized as follows: - The increasing of the salinity from 2000 to 10000 PPM decreased all plant vegetative growth characters and the total chlorophyll content compared to control . - A marked increasing in the ion leakage , the protein carbonyl group and the lipid peroxidation number in both seasons with the salinity increasing from 2000 to 10000 PPM compared to control .