Marker Of Peroxidation Research Articles

Increased non-protein bound iron in Down syndrome: contribution to lipid peroxidation and cognitive decline

Down syndrome (DS, trisomy 21) is the leading cause of chromosomal-related intellectual disability. At an early age, adults with DS develop with the neuropathological hallmarks of Alzheimer’s disease, associated with a chronic oxidative stress. To investigate if non-protein bound iron (NPBI) can contribute to building up a pro-oxidative microenvironment, we evaluated NPBI in both plasma and erythrocytes from DS and age-matched controls, together with in vivo markers of lipid peroxidation (F2-isoprostanes, F2-dihomo-isoprostanes, F4-neuroprostanes) and in vitro reactive oxygen species (ROS) formation in erythrocytes. The serum iron panel and uric acid were also measured. Second, we explored possible correlation between NPBI, lipid peroxidation and cognitive performance. Here, we report NPBI increase in DS, which correlates with increased serum ferritin and uric acid. High levels of lipid peroxidation markers and intraerythrocyte ROS formations were also reported. Furthermore, the scores of Raven’s Colored Progressive Matrices (RCPM) test, performed as a measure of current cognitive function, are inversely related to NPBI, serum uric acid, and ferritin. Likewise, ROS production, F2-isoprostanes, and F4-neuroprostanes were also inversely related to cognitive performance, whereas serum transferrin positively correlated to RCPM scores. Our data reveal that increased availability of free redox-active iron, associated with enhanced lipid peroxidation, may be involved in neurodegeneration and cognitive decline in DS. In this respect, we propose chelation therapy as a potential preventive/therapeutic tool in DS.

Acute cocoa flavanol intake affects antioxidative capacity and mediates the no- pathway in well trained athletes: Implications for exercise performance

Introduction: Evidence exists that cocoa flavanol (CF) can increase nitric oxide synthesis. This promotes vasodilation and several physiological processes, such as mitochondrial respiration and biogenesis, glucose uptake and SR-Ca handling, that may support exercise performance1. Moreover, CF intake is also known for its antioxidative capacities (in vitro and in vivo), resulting in reduced levels of oxidative stress2. During exhaustive exercise, free radical production increases dramatically, resulting in an altered redox state in the muscle, which possibly inhibits muscle contraction3. Acute intake of antioxidants may help neutralize free radicals and therefore directly prevent a decline in exercise performance4. Moreover, increased antioxidative capacity may repress the formation of peroxinitrite, which may elevate NO availability5. The aim of this study was to examine the effect of CF intake on 1) NO production in response to exhaustive exercise, 2) exercise-induced changes in antioxidative capacity and oxidative stress and 3) exercise performance in well-trained athletes. Materials and methods: Twelve well-trained men(PL36) (mean ± SD age, height, mass, VO2max: 30 ± 3 years, 177.9 ± 8.8 cm, 72.8 ± 7.8 kg , 63.0 ± 3.5 ml/kg/min) participated in this randomized, double-blind, cross-over study. Participants performed two 30-min time trials (TT) 1.5 and 3 hours after CF intake (900 mg) or placebo intake (PL 13 mg CF), interposed by passive rest. Lactate, glucose, heartrate, rate of perceived exertion (RPE) and power output were measured during the TTs. Blood samples were drawn at baseline, before and immediately after the two TTs and analysed for trolox equivalent antioxidative capacity (TEAC), Uric acid (UA), malonaldehyde (MDA), arginine and citrulline. TT performance was compared between PL and CF by paired t-test. Pacing strategy during TT and blood markers were analysed by repeated measured ANOVA. Results: Time to complete TT1 tended to be lower after CF intake compared to PL intake (29’13” ± 1’19” CF vs. 29’47” ± 1’58” PL, p=.09). After 25 minutes, a significant higher power output was observed in CF compared to PL. No differences in TT2 performance was observed between CF and PL. Glucose, lactate, heartrate and RPE increased significantly during both TTs, but were unaffected by CF intake. TEAC and UA significantly increased after both TTs. CF intake increased TEAC and UA, compared to PL intake (Figure 1A). MDA was increased by exercise, but remained unaffected by CF intake. Arginine was significantly lowered by exercise. Before TT1, CF intake tended to increase arginine, compared to PL intake. After TT1, arginine levels did not differ between CF and PL. Citrulline significantly increased by exercise, but was not altered by CF intake (Figure 1B). Conclusion: Acute CF intake tended to enhance performance on a TT 1.5 hours after intake. Increased power output was detected at the end of the TT. Exercise acutely increased antioxidative capacity, lipid peroxidation (marker of oxidative stress) and NO production (increased citrulline and decreased arginine). CF intake increased antioxidative capacity, but did not alter lipid peroxidation. CF intake tended to increased arginine levels, known to augment NO production and possibly explaining the improved exercise performance.

Oxidative stress parameters as markers of the different trimesters in normal pregnancy

Pregnancy has been associated with changes in physiologic and metabolic functions accompanied by a high metabolic demand and elevated requirements for tissue oxygen with eventual increase in oxidative pressure on the antioxidant defence system of the body. Thus, it was based on this premise that some markers of pro-oxidant- antioxidant status of pregnancy compared with non-pregnant state were assessed including, marker of lipid peroxidation (malondialdehyde, MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidise (GPx). A total of 41 apparently healthy female volunteers were used for this study made up of 36 pregnant females at different trimester of pregnancy, categorised into three (3) groups of 12 each per trimester, and whose ages ranged between 19 years and 40 years. Standard biochemical assays were employed in the assessment of these markers. The level of lipid peroxidation (MDA) increased progressively for the pregnant subjects (p<0.05) at the third trimester (18.2mmole/ml) compared to the non-pregnant controls (2.3mmole/ml); an increase in SOD activity was also observed (p<0.05) at the third trimester (8.9U/ml) compared to the control (6.3U/ml) contrary to some other researches; while decrease in CAT (9.2U/ml) and GPx (7.8U/ml) activities at the third trimester compared to the non-pregnant control (p<0.05) (307.5U/ml and 9.4U/ml, respectively) were also observed. There was an observed increase in the body mass index, BMI, from first trimester, 26.4kg/m 2 and second trimester, 38.4 kg/m 2 to the third trimester, 42.7 kg/m 2 , compared to the control, 33.7 kg/m 2 supporting the claims that pathologic conditions associated with pregnancy become more prominent with increase in BMI. Thus, these biochemical markers of oxidative stress, viz., superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and malondialdehyde (MDA) can serve as markers to assess the stages of pregnancy and also can be used to differentiate pathologic and nonpathologic pregnant conditions. Keywords: Antioxidants; Pregnancy; Pro-oxidants; Trimesters; Free radicals; Body mass index

Protective role of vitamin C on sperm morphology and biochemical enzymes in lead nitrate treated albino rats

Lead is a well known heavy metal and is found in various components of earth environment and is a major industrial byproduct. Manipulation of lead from these sources causes toxicity in human beings by oxidative stress, which results in decrease fertility due to increase in the production of abnormal sperm population. Vitamin C is an antioxidant and acts by scavenging the free radicals, thus ameliorating the oxidative stress. The present study was carried out to evaluate the effect of lead nitrate and Vitamin C on testis. The study was conducted on 30 male albino rats of Wistar strain, weighing 150-250 gm. The rats were divided into 5 groups (6 rats in each group). Group 1 served as control and rest 4 groups were experimental. Except the control group, the animals of other 4 groups received lead nitrate and vitamin C orally for 6 days a week for 2 months. All the animals were sacrificed at the end of experiment and the result showed abnormal sperm count 5% in control group while it was 66% in group 4 and 17% in group 5. Sperm abnormalities observed were double head, banana head, amorphous head, defective head, headless, bent neck, bent tail, double tail, defective tail and looped tail. Malondialdehyde, a marker of lipid peroxidation was increased in experimental group while glutathione reductase (GR) enzyme was decreased in testicular tissue but this change in enzyme showed some improvement in vitamin C treated groups.

Protective effect of Naringin on experimental hindlimb ischemia/reperfusion injury in rats

This study was designed to investigate the antioxidant effects of Naringin, in ischemia/reperfusion (I/R)-induced skeletal muscle injury in rats. The rats were randomly allocated into three groups including control, I/R and I/R + Naringin groups. Muscle tissues of I/R groups revealed significantly higher antioxidant enzyme activities, and increased levels of malondialdehyde, as specific a marker of the lipid peroxidation and tissue damage, compared to the control group (p < 0.05). Levels of these parameters in muscle revealed significant reductions in the I/R + Naringin group compared to the I/R group (p < 0.05). Histopathological examination of ischemia muscles in the I/R group showed significant degeneration and inflammation compared to the control group, whereas ischemic muscles of Naringin-administered group showed significant reduction in degeneration and inflammation compared to the I/R group (p < 0.05). We suggest that the protective effect of Naringin may reduce the I/R injury in cases of extremity injuries with acute vascular complications, extremity surgery with prolonged tourniquet application.

Anti-oxidant profiles and markers of oxidative stress in preterm neonates

Background: Preterm birth is associated with an increased oxidant burden which places these infants at a higher risk of injury. Aims: This prospective study aimed to assess levels of antioxidants and a marker of oxidative stress in preterm neonates. Objectives: (i) To compare levels of anti-oxidants [vitamin A, vitamin E, catalase, total anti-oxidant status (TAS)] as well as malondialdehyde level (MDA) (a marker of lipid peroxidation) between preterm and full-term neonates; (ii) to determine changes in the values of measured vitamins at birth and at discharge among preterm neonates; and (iii) to compare levels of anti-oxidants with MDA levels in relation to complications of prematurity and outcome. Methods: The study was undertaken in 100 preterm neonates and 100 full-term neonates as a control group. MDA was estimated by a thiobarbituric acid-reactive technique; TAS was determined using a Randox assay kit; catalase activity was measured spectrophotometrically and vitamin A and E levels were estimated by high performance liquid chromatography. Results: The plasma levels of vitamin A, vitamin E, TAS and catalase were significantly lower in the preterm than in the full-term group (P < 0.01), and the plasma level of MDA was significantly higher in preterm than full-term neonates (P < 0.01). Vitamin A and E levels in preterm neonates were significantly higher at discharge than at birth (P < 0.01). Vitamin A, vitamin E and catalase levels at birth were significantly lower in patients who developed necrotizing enterocolitis or bronchopulmonary dysplasia than in those who did not. Conclusion: Preterm neonates are exposed to increased oxidant stress at birth and are susceptible to anti-oxidant deficiencies. A higher dose of enteral vitamin A supplementation in preterm neonates might reduce morbidity and improve outcome. Further studies are warranted to evaluate the appropriate dose of oral vitamin E supplementation for preterm neonates.

Oxidative stress during spontaneous vaginal delivery: comparison between maternal and neonatal oxidative status

Introduction: Spontaneous vaginal delivery inflicts a change in the redox status of the mother and the neonate. Hypoxia followed by reperfusion during powerful labour contractions causes increased generation of free radicals affecting the maternal oxidative status. The difference that exists between the extra uterine and intra uterine environment in the partial pressures of oxygen can induce free radical formation which affects the antioxidant mechanism of the neonate during vaginal delivery. The present study was aimed at quantifying the changes with respect to oxidative stress that occurs in uncomplicated laborin both the mother and newborn. Method: 20 singlet on term pregnant women in labour without any complications, delivered by spontaneous vaginal delivery were included in our study. Estimation of plasma MDA (Malondialdehyde) as a marker of lipid peroxidation and antioxidants like SOD (Superoxide dismutase), Total reduced Glutathione (GSH), and Glutathione Peroxidase (GPX) in hemolysate were performed using spectrophotometric method from cord blood and women in labour respectively. Result: The results showed significantly elevated plasma MDA in cord blood compared to maternal plasma MDA, before (P<0.001) and after delivery (P<0.001). Statistically significant higher GSH levels in the cord blood was observed compared to maternal GSH levels both before (P <0.05) and after delivery (P< 0.05). Maternal GPX level was significantly elevated when compared to cord (P< 0.05). However, SOD levels did not show any significant change. Conclusion: Vaginal delivery predisposes the neonate to increased oxidative stress when compared to maternal system which is reflected as an alteration in their antioxidant levels.

Oxidative Stability and Physico-chemical Properties of Meat from Broilers Fed with Dietary Neem Leaf Powder, Spirulina and their Combination

This 6 week study was conducted to evaluate the antioxidant potential, maintaining quality and sensory properties of broiler meat from birds fed on dietary neem leaf powder (NLP). A total of 90 Vencobb broiler chicks were randomly allotted to 6 groups of 15 birds in each. Dietary treatments consisted of normal diet (control Group I), feed containing terramycin-200 (TM-200*) at the concentration of 0.05% (Group II), feed containing NLP of 0.2% (Group III), feed containing NLP of 0.2% and spirulina of 1% (Group IV), feed containing TM-200 at 0.05% and spirulina of 1% (Group V) and feed containing spirulina of 1% (Group VI). At the end of the experiment liver, kidney and muscle samples were collected to evaluate the tissue peroxidation (TBARS and protein carbonyls) and antioxidant markers (SOD). Physico-chemical quality determinants of both fresh and preserved meat viz. extract release volume (ERV), water holding capacity (WHC) and pH were also studied. TBARS protein carbonyls indicated a significant (P &lt; 0.05) decrease in all the treated groups when compared to control. Superoxide dismutase levels were found to be significantly increased in all the treated groups, in all the tissues collected. Compared to control group, favorable physico-chemical quality determinants were recorded in all the treated groups. The sensory attributes did not show significant (P &lt; 0.05) differences for color, flavor, juiciness, tenderness and overall acceptability. This study indicates enhanced stress tolerance levels, improved meat quality with unaffected consumer acceptance levels of the meat observed in the study, from broilers fed with neem and spirulina either alone or in combinationsș this points out that neem at 0.2% level can be used in poultry diets instead of antibiotic growth promoters (AGP).

Expression of glucokinase, glucose 6-phosphatase, and stress protein in streptozotocin-induced diabetic rats treated with natural honey

Diabetes mellitus is a metabolic disorder characterized by hyperglycemia and associated with oxidative stress. Based on literature, honey components could treat diabetes with unknown mechanism. The present study investigated the hypoglycemic effects and antioxidant activities of honey at the molecular level in STZ-induced diabetic rats. Using an animal model of diabetes, we investigated antidiabetic and antioxidant properties of natural honey. For this fasting blood glucose, malondialdehyde (MDA) level as a marker of lipid peroxidation and total antioxidant capacity were measured in diabetes-induced rats treated with natural honey. The transcript levels of stress proteins including heat shock protein 70 (HSP70), glucokinase (GK), and glucose 6-phosphatase (G6P) were determined using quantitative real-time PCR. Statistical analysis showed that honey significantly decreased MDA levels; in contrast, it increased total antioxidant capacity in diabetic rats (p < 0.05). Quantitative real-time PCR (QRT-PCR) analysis revealed that expressions of HSP70 and G6P decreased while the expression of GK increased in honey treatment groups in comparison with control group. These findings provide insight into the molecular mechanisms behind the hypoglycemic and antioxidative effects of honey that may be considered for further clinical studies in drug development.

Cardioprotective role of tadalafil against cisplatin-induced cardiovascular damage in rats

The present study investigated the possible cardioprotective effect of tadalafil (Tad) on cisplatin (CDDP)-induced cardiac and vascular damages in rats. A total number of seventy two healthy male albino rats initially weighting between 200 and 220g were used and randomly divided into four groups,18 rats in each. The control group received no treatment; CDDP group received a single dose of CDDP (4mg/kg) intraperitoneal (i.p.) per week for 4 weeks the duration of the experiment; Tad group received 0.4mg/kg BW Tad i.p. daily and Tad +CDDP group received 0.4mg/kg BW Tad i.p. +4mg/kg BW CDDP i.p. The results showed that Tad was able to decrease blood pressure, heart rate, levels of serum cardiac troponin (cTn-I), malondialdehyde (MDA) and increased levels of reduced glutathione (GSH) and nitric oxide (NO) in the heart homogenate sample from CDDP treated rats. Semi-quantitative analysis showed that Tad was able to decrease the histopathological scores of cardiac muscular hyalinzation and fibrosis in three sacrifices in CDDP treated rats. CDDP treated rats showed significantly increased thickening in wall of aorta with an irregular luminal layer of endothelial cell linings in three sacrifices when it was compared to other groups. Moreover, immunohistochemical labeling of α- smooth muscle actin (α-SMA) in aorta revealed significant lower scores in Tad +CDDP group when they were compared to CDDP group. In conclusion, Tad alone did not induce any harmful effects on blood pressure, selective antioxidant, peroxidation markers or cardiac histology, in addition, Tad has a cardio-protective role against CDDP.

Hypoxiс postconditioning is an effective method of protection from severe hypoxia induced lipid peroxidation and neuronal apoptosis in rats

Postconditioning (PostC) is an exposure of the damaged organism to extreme factors of the mild intensity to mobilize endogenous protective mechanisms. In our laboratory method of PostC using three daily trials of mild hypobaric hypoxia (MHH) was developed. It has been found that such method of the PostC effectively prevents degeneration of the hippocampal and neocortical neurons in rats, subjected to severe hypoxia (SH). Present study has been aimed at examination of the impact of oxidative processes in the development of the neuroprotection acquired in the course of hypoxic PostC during first three days of reoxygenation after SH in rats. The levels of thiobarbituric acid reactive substances (TBARS) and Schiff bases (SB) were used as markers of lipid peroxidation. In addition, the intensity of the apoptotic DNA fragmentation has been studied. During the three days after the SH a sustained increase of SB in the rat hippocampus was observed (700-1000% of the control value). After the first PostC episode the SB levels decreased to 150% of the baseline. Subsequently this parameter did not differ significantly from the control values. TBARS showed accumulation on the first day following the SH but afterwards its levels dropped to 40% of control and did not recover then to normal values. In the PostC animals, the levels of TBARS after each of three PostC episodes did not differ from the control values. These facts indicate that the PostC MHH balances the activity of pro- and antioxidant systems in vulnerable brain regions and promotes the effective utilization of components damaged by peroxidation. Fragments ladder typical for cells undergone apoptosis was obtained by the electrophoretic separation of the total DNA, extracted from a rat brain after one, two and three days after the SH. In the PostC group, the DNA fragmentation was revealed only after the first PostC episode, demonstrating antiapoptotic action PostC MHH.

Tert-butylhydroquinone (TBHQ) improves antioxidant status in rat tissues following chronic diazinon intoxication

Considering that the involvement of oxidative stress has been implicated in the toxicity of organophosphate insecticides, the objective of the present study was to investigate antioxidants and oxidative stress markers in the liver and kidney of Wistar rats treated with chronic doses of diazinon. In addition, the effect of Tert-butylhydroquinone (TBHQ), a food-additive antioxidant, on attenuation of diazinon-induced oxidative stress was evaluated. 28 rats were randomly assigned to 1 of 4 treatment groups: diazinon (10 mg/kg BW, once a day; n=7), TBHQ (0.028 g/kg of diet, once a day; n=7), TBHQ + diazinon (diazinon; 10 mg/kg BW, once a day + TBHQ; 0.028 g/kg of diet, once a day; n=7) and control (corn oil, as vehicle of diazinon and TBHQ, n=7). TBHQ, diazinon and corn oil were given to rats orally via gavage for 7 weeks. Total thiol groups, ferric reducing antioxidant power (FRAP) and malondialdehyde (MDA) levels in liver and kidney tissues were investigated at the end of 7th week. Treatment with diazinon significantly increased MDA level, as a marker of lipid peroxidation, in the kidney (p<0.05). Co-administration of TBHQ with diazinon significantly increased total thiol groups and FRAP levels in liver and kidney (p<0.05). However, TBHQ did not reduce lipid peroxidation in the tissues. The results of the present study showed that administration of TBHQ at a dose of 0.028 g/kg of diet, increased antioxidant capacity of tissues of rats treated with diazinon, though this increase was not sufficiently effective to reduce lipid peroxidation caused by diazinon. Higher amounts of TBHQ might be more effective in attenuation of diazinon-induced lipid peroxidation.

Oxidative Stress in Egyptian Children with Autism: Relation to Autoimmunity

Introduction Oxidative stress has been implicated in the pathogenesis of major psychiatric disorders, as the brain has comparatively greater vulnerability to oxidative damage Objectives and Aims We are the first to study the relationship between oxidative stress (by measuring plasma F2-isoprostane, as a marker of lipid peroxidation, and glutathione peroxidase, as an antioxidant enzyme) and autoimmunity (as indicated by serum antineuronal antibodies). Methods We studied these markers in a group of 44 Egyptian autistic children and 44 healthy matched-children. Results Our results showed that oxidative stress was found in 88.64% of autistic children. Oxidative stress, resulting from elevated plasma F2-isoprostane and/or reduced glutathione peroxidase, had significant risk for antineuronal positivity, which was found in 54.5% of autistic children, (odds ratio: 12.38 and 6.43, respectively, confidence interval: 1.37–112.10 and 1.21–34.19, respectively). Conclusions the strong association between oxidative stress and autoimmunity in autistic children may indicate the possible role of oxidative stress, through induction of autoimmunity, in some autistic patients. Therefore, studies considering the role of antioxidants and immunotherapy in amelioration of autistic manifestations are recommended.

Abstract 39: Decreased Ferroxidase Activity in Cerebrospinal Fluid Following Subarachnoid Hemorrhage

Introduction: We previously reported on the relationship between unbound CSF iron and brain injury following SAH. The ferroxidase ceruloplasmin (Cp) regulates iron levels in the CNS and prevents free radical injury. Paradoxically, oxidative stress is known to cause structural modifications that result in decreased enzymatic activity of Cp. We sought to determine if CSF of patients with SAH exhibits impaired ferroxidase activity (FA). Methods: Samples of CSF were obtained on day 5 following aneurysm rupture and levels of malondialdehyde (MDA; a marker of lipid peroxidation) and ferroxidase activity (defined as the amount of enzyme that converts 1 μM of substrate into product per minute) were determined by means of absorbance and colorimetric assays. Ceruloplasmin concentration in CSF was measured with ELISA. Demographic and clinical data were recorded. Values are expressed as mean ± SEM. Comparisons were made using Wilcoxon rank sum test. Simple linear regression analysis was performed for comparison of continuous variables. Results: A total of 5 patients and 4 control subjects were included in this pilot study. Median Hunt-Hess and modified Fisher scores were 4 (IQR 1.5) and 4 (IQR 1), respectively. Levels of MDA in CSF (9.2 ± 0.9 vs. 0.09 ± 0.01 μM, p =0.02) were significantly higher in SAH patients, while FA was significantly reduced (64.8 ± 17.1 vs. 284.1 ± 64.9 U/L, p =0.01). Ceruloplasmin concentration was higher in SAH samples (49.4 ± 12 vs. 1.72 ± 0.7 mg/L, p =0.04), and patients who developed delayed cerebral ischemia had lower CSF FA (43.2 ± 6.3 vs. 97.2 ± 32.4 U/L, p =0.07), compared to those who did not. Conclusions: The ferroxidase activity of CSF following SAH is significantly impaired, in spite of higher ceruloplasmin concentrations. Evidence of increased CNS oxidative damage was also demonstrated in our cohort. We postulate that structural modifications in Cp mediated by an oxidative milieu occur in SAH and are associated with decreased enzymatic function. There is a possible inverse relationship between ferroxidase activity and risk of development of delayed cerebral ischemia. Further studies are needed to explore the significance of these findings.

Anti-inflammatory Effects of N-acetylcysteine against Carbosulfan-induced Hepatic Impairment in Male Rats

The present study is designed to investigate the anti-inflammatory potential of N-acetylcysteine (NAC) against carbosulfan (CB)-induced hepatic dysfunction in rats. Rats exposed to CB and NAC were examined for toxicity by assessing various biochemical alteration, inflammation (interferon gamma and leucocyte counts), and peroxidation marker, including in liver. Significant increases of blood alanine aminotransferase (ALT), alkaline phosphatase (ALP), and gamma glutamyltransferase (GGT) were detected in CB-treated rats. In addition, the level of pro-oxidative marker, such as malondialdehyde (MDA), was also increased. According to the results, rats exposed to CB showed a significant alteration in hepatic enzymes and an increase in the accumulation of lipid peroxidation index; also, CB induced an increase in pro-inflammatory cytokine in CB-treated rats when compared with their respective controls. Interestingly, administration of NAC to CB-treated rats attenuates the toxicity of CB, objectified by biochemical and oxidative improvement of liver. Thus, the present study reports for the first time that NAC could be a promising therapeutic agent against CB-induced liver impairment.

Estimation of malondialdehyde and vitamin-E levels in neonatal hyperbilirubinemia before and after phototherapy

Background: Hyperbilirubinemia is a common and benign problem in neonates worldwide. It is observed during the 1st week of life in approximately 60% of term neonates and 80% of preterm neonates. Phototherapy is most widely used as therapy for unconjugated hyperbilirubinemia. Phototherapy is related to oxidative stress and lipid peroxidation. The present study is undertaken to establish the relation between anti-oxidant status and the marker of lipid peroxidation in neonatal hyperbilirubinemia before and after phototherapy. Objectives: To estimate the levels of MDA (malondialdehyde) and vit-E levels in neonatal hyperbilirubinemia before and after phototherapy. Material and Methods: A total of 30 patients were eligible for the study who met the inclusion and exclusion criteria. Blood sample was collected from neonates of preterm/full term age 1 to 10 days with hyperbilirubinemia undergoing phototherapy. Total Bilirubin, Direct Bilirubin, MDA and vit-E levels were estimated in serum by thiobarbituric acid (TBA) method and Backer and Frank’s Method respectively. Results: The present study showed increase in Total bilirubin, Direct bilirubin, MDA and decrease in vit-E levels before phototherapy when compared to control group and same subjects after phototherapy showed decrease in Total bilirubin and Direct bilirubin, MDA and a further decrease in vit-E levels. Conclusion: From these results it is concluded though phototherapy had a beneficial effect in treatment of neonatal hyperbilirubinemia, supplementation of vit-E is necessary in addition to phototherapy.

Oxidative stress and anti oxidants status in pellagra

Background and objectives: Pellagra was vanished from most parts of the world where it was formerly present due to its dietary modification. However, it is still encountered among the jowar eating populations of India. The information about the role of oxidative stress in pellagra was not established. Therefore, in this study we assessed the oxidative stress status by using malondialdehyde (MDA), total anti oxidant status (TAOS) and redox ratio (RER) in clinically diagnosed pellagra patients. Materials and methods: Clinically diagnosed pellagra patients aged between 18 to 40 years, both male and females were recruited (n=78) from department of Dermatology. Age and gender matched controls (n=78) were recruited from the student and residents of the hospital. Malondialdehyde (MDA) is a marker of lipid peroxidation, Total Anti Oxidant Status (TAOS) and Redox Ratio (RER) markers were assessed by using commercially available kits. Results: There were no significant differences in the anthropometric parameters. However, the oxidative stress markers MDA (p<0.05), RER (p<0.001) were significantly high and TAOS was low (P<0.001) in pellagra patients in comparison with age and gender matched controls. Conclusion: The results of this study showed the increased MDA, RER levels and decreased TAOS levels. Estimation of these markers at early stage will help to take measures to prevent the progression of disease and develop antioxidant strategies.

Serum levels of high sensitivity C reactive protein and malondialdehyde in chronic kidney disease

Background: Chronic kidney disease cases are at increased risk for progression to end stage renal disease and accelerated atherosclerosis, with premature cardiovascular morbidity and mortality being the more frequent outcome. Aim: The study was taken up to find if there is any association between nontraditional cardiovascular risk markers like high sensitivity C reactive protein (marker of inflammation) and malondialdehyde (marker of lipid peroxidation) with the progression of chronic kidney disease. Methodology: The study included 44 pre dialysis chronic kidney disease cases and 44 healthy controls. Serum levels of creatinine, high sensitivity C reactive protein and malondialdehyde were estimated in both groups. The mean estimated glomerular filtration rate(eGFR) in chronic kidney disease patients was calculated by the MDRD formula. Results: The mean eGFR in cases was found to be 23.65 14.99 ml/min by MDRD formula. The serum hsCRP and malondialdehyde levels in cases was 11.8 7.24 mg/L and 3.02 1.24 nmol/ml respectively. Conclusion: There was a significant negative correlation (p<0.001) between high sensitivity C-reactive protein and malondialdehyde with eGFR. A highly significant positive correlation was found between serum hsCRP and malondialdehyde (p<0.001) in chronic kidney disease underlining the synergism between oxidative stress and inflammation, perpetuating to further deterioration of renal function and enhancing the predisposition to cardiovascular risk with the progression of chronic kidney disease.

Analysis of DNA damage, plasma lipid peroxidation, and antioxidant status in type 2 diabetes mellitus patients among a south Indian population

Oxidative stress is involved in the pathophysiology of diabetes mellitus, which in turn may also contribute to its complications. This case-control study was designed to find out the association between oxidative damage to lipids and DNA in type 2 diabetes mellitus. Sixty individuals with type 2 diabetes and age- and sex-matched 60 clinically healthy individuals without type 2 diabetes were included for the study. Oxidative stress was evaluated by malondialdehyde assay which is a marker of lipid peroxidation and plasma total antioxidant status was evaluated by the ferric reducing antioxidant power assay. Single-cell alkaline gel electrophoresis (comet assay) was employed to assess the level of DNA damage in peripheral lymphocytes. The extent of DNA damage was observed to be high in the diabetic patients when compared with the controls. The total antioxidant status was found to be significantly lower in the patients with diabetes mellitus when compared with the controls (p < 0.05). Lipid peroxidation status was significantly increased in diabetics (p < 0.05) when compared with the control group. The results of this study confirm that hyperglycemia contributes to increased generation of reactive oxygen species and decreased total antioxidant status which may be responsible for the oxidative DNA damage among south Indian population with diabetes mellitus.

Abstract 3139: Nurr1, a novel target of 1,1-bis(3’-indolyl)-1-(p-chlorophenyl) methane for inhibition of the initiation and progression of skin cancer tumorigenesis

Abstract The objective of this study was to demonstrate the chemopreventive potential of 1,1-bis(3′-indolyl)-1-(p-chlorophenyl methane) (DIM-D) in skin cancer using an in vitro and in vivo models. In vitro cell cytotoxicity and viability assays were carried out in A431 human epidermoid carcinoma cell line (A431) and normal human epidermal keratinocytes (NHEK) respectively. Apoptosis induction and accumulation of ROS following UVB exposure in DIM-D pretreated NHEK cells (2 hr prior) was also evaluated. Immunocytochemistry and western blot analysis were performed to determine proapoptotic and proinflammatory markers expressions in DIM-D treated A431 cells and in UVB irradiated NHEK cells. For in vivo experiments, DIM-D was encapsulated within nanocarriers by hot melt homogenization using Nano DeBEE. Nanocarriers were surface modified with oleic acid and incorporated into a gel using established procedures (G-NOD) and characterized for particle size, zeta potential, entrapment efficiency, rheology, skin permeation and drug release. The chemo-preventive efficiency of G-NOD was evaluated using UVB-induced skin cancer model in SKH hairless mice and skin samples were collected and investigated for lipid peroxidation, protein carbonyls, and molecular markers by western blot. The IC50 values of DIM-D were 24.5±2.6, 17.2±3.6and 4.1±1.1 whilst for EGCG were 192.1±3.8, 85.3±2.4 and 26.0 ±1.4 for 24, 48 and 72 hr treatments respectively. DIM-D induced higher expression in A431 cells compared to EGCG of cleaved caspase 3 (3.0-fold vs. 2.4-fold changes), Nurr1 (2.7-fold vs. 1.7-fold changes) and NFκB (1.3-fold vs. 1.1-fold changes). Nanocarriers containing DIM-D were 188.00±8.00 nm in size with polydispersity of 0.59±0.01. The zeta potential and entrapment efficiency were 37.17±0.90mV and 89±0.50%, respectively. There was significant (p&amp;lt;0.05) reduction in tumor size/number for G-NOD pretreated group (2 tumors/mouse, average diameter, 2±0.50 mm) compared to DIM-PG (4 tumors/mouse, average diameter, 4±0.45mm) and EGCG PEG-gel (EGCG-PG; 6 tumors/mouse, average diameter, 5±0.25mm). IHC and western blot results indicated significant increase (p&amp;lt;0.05) in expression of Nurr1 in G-NOD pretreated group compared to EGCG-PG. There was however pronounced reduction in expression of STAT3, NFkB, cleaved Caspase-3, BCl2 and 8-OHdG for G-NOD pretreated group compared to DIM-PG and EGCG-PG. Lipid peroxidation assay revealed decrease in amount of Malondialehyde (MDA) produced in G-NOD pretreated group prior to UV exposure by 2 fold compared to DIM-PG and 1.3 fold compared to EGCG-PG treatments. Therefore, findings in both in vitro and in vivo studies suggest that the enhanced percutaneous delivery of DIM-D reduces UVB-induced damage to skin lipids and protein and inhibits initiation and progression of skin photocarcinogenesis in SKH mice via the transactivation of Nurr1. Citation Format: Ravi Doddapaneni, Cedar Boakye, Punit Shah, Apurva R. Patel, Chandraiah Godugu, Stephen Safe, Santosh Katiyar, Mandip Sachdeva. Nurr1, a novel target of 1,1-bis(3’-indolyl)-1-(p-chlorophenyl) methane for inhibition of the initiation and progression of skin cancer tumorigenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3139. doi:10.1158/1538-7445.AM2014-3139