Electrochemical sensing of hydrogen peroxide using a glassy carbon electrode modified with multiwalled carbon nanotubes and zein nanoparticle composites: application to HepG2 cancer cell detection.

A nanobiocomposite was prepared from multiwalled carbon nanotubes and zein nanoparticles. It was dispersed in water/ethanol and drop cast onto a glassy carbon electrode. The modified electrode can be used for electroreduction of H2O2 (typically at a working potential of -0.71 V vs. Ag/AgCl). The electrochemical properties of the electrode were investigated by cyclic voltammetry, linear sweep voltammetry, chronoamperometry and electrochemical impedance spectroscopy. Response to H2O2 is linear in the 0.049 to 22 µM concentration range, and the detection limit is 35 nM at pH 7.0. The sensor was successfully utilized for the measurement of H2O2 in a synthetic urine sample, and for monitoring the release of H2O2 from human dermal fibroblasts and human hepatocellular carcinoma cells. Graphical abstractSchematic representation of a novel metal- and enzyme-free electrochemical nanosensor. A glassy carbon electrode was modified with a nanocomposite prepared from multiwalled carbon nanotubes and zein nanoparticles. It was applied to the identification of liver cancer cells via sensing of H2O2 and has a very low detection limit.

Novel manganese(II)-based metal-organic gels: synthesis, characterization and application to chemiluminescent sensing of hydrogen peroxide and glucose.

A metal-organic gel (MOG) was synthesized that is composed of manganese(II) as the central ion and 1,10-phenanthroline-2,9-dicarboxylic acid as the ligand. The resulting MOG exhibits excellent activity for catalyzing the chemiluminescence (CL) of the luminol/hydrogen peroxide system. The CL system was characterized by CL spectra, UV-vis absorption spectra and by studying potential interferences by common radical scavengers. The CL reaction was exploited in a new scheme for the determination of hydrogen peroxide. CL intensity increases linearly in the 0.4 µM ~ 3 mM hydrogen peroxide concentration range, and the limit of detection (LOD) is 0.12 µM. The method was extended to an enzymatic assay for glucose by using glucose oxidase and by measurement of the enzymatically formed hydrogen peroxide. The assay works in the 0.2 µM ~ 3 mM glucose concentration range, and the LOD is 0.08 µM. Graphical abstract Schematic representation of the synthesized Mn-containing MOGs catalyzing luminol-hydrogen peroxide chemiluminescent reaction, which can be used to establish a new CL method for the detection of hydrogen peroxide and glucose.

Longer duration of obesity is associated with a reduction in urinary angiotensinogen in prepubertal children.

BACKGROUND: We aimed to study the impact of obesity on urinary excretion of angiotensinogen (U-AGT) in prepubertal children, focusing on the duration of obesity and gender. Also, we aimed to evaluate whether plasma angiotensinogen (P-AGT) and hydrogen peroxide (H2O2) play a role in the putative association. METHODS: Cross-sectional evaluation of 305 children aged 8-9 years (160 normal weight, 86 overweight, and 59 obese). Anthropometric measurements and 24-h ambulatory blood pressure monitoring were performed. Angiotensinogen (AGT) was determined by a commercial enzyme-linked immunosorbent assay (ELISA) kit and H2O2 by a microplate fluorometric assay. RESULTS: U-AGT and P-AGT levels were similar across body mass index (BMI) groups and between sexes. However, boys who were overweight/obese since the age of 4 years presented lower levels of U-AGT compared with those of normal weight at the same age. In children who were overweight/obese since the age of 4, urinary H2O2 decreased with P-AGT. CONCLUSIONS: A higher duration of obesity was associated with decreased U-AGT in boys, thus reflecting decreased intrarenal activity of the renin-angiotensin system. Also, children with a longer duration of obesity showed an inverse association between urinary H2O2 and P-AGT. Future studies should address whether these results reflect an early compensatory mechanism to limit obesity-triggered renal dysfunction.

Increase in oxidative stress levels following welding fume inhalation: a controlled human exposure study.

BACKGROUND: Tungsten inert gas (TIG) welding represents one of the most widely used metal joining processes in industry. It has been shown to generate a large majority of particles at the nanoscale and to have low mass emission rates when compared to other types of welding. Despite evidence that TIG fume particles may produce reactive oxygen species (ROS), limited data is available for the time course changes of particle-associated oxidative stress in exposed TIG welders. METHODS: Twenty non-smoking male welding apprentices were exposed to TIG welding fumes for 60 min under controlled, well-ventilated settings. Exhaled breathe condensate (EBC), blood and urine were collected before exposure, immediately after exposure, 1 h and 3 h post exposure. Volunteers participated in a control day to account for oxidative stress fluctuations due to circadian rhythm. Biological liquids were assessed for total reducing capacity, hydrogen peroxide (H2O2), malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG) concentrations at each time point. A linear mixed model was used to assess within day and between day differences. RESULTS: Significant increases in the measured biomarkers were found at 3 h post exposure. At 3 h post exposure, we found a 24 % increase in plasma-H2O2 concentrations ([95%CI: 4 % to 46 %], p = 0.01); a 91 % increase in urinary-H2O2 ([2 % to 258 %], p = 0.04); a 14 % increase in plasma-8-OHdG ([0 % to 31 %], p = 0.049); and a 45 % increase in urinary-8-OHdG ([3 % to 105 %], p = 0.03). Doubling particle number concentration (PNC) exposure was associated with a 22 % increase of plasma-8-OHdG at 3 h post exposure (p = 0.01). CONCLUSION: A 60-min exposure to TIG welding fume in a controlled, well-ventilated setting induced acute oxidative stress at 3 h post exposure in healthy, non-smoking apprentice welders not chronically exposed to welding fumes. As mass concentration of TIG welding fume particles is very low when compared to other types of welding, it is recommended that additional exposure metrics such as PNC are considered for occupational risk assessments. Our findings highlight the importance of increasing awareness of TIG welding fume toxicity, especially given the realities of welding workplaces that may lack ventilation; and beliefs among interviewed welders that TIG represents a cleaner and safer welding process.

Using inflammatory and oxidative biomarkers in urine to predict early acute kidney injury in patients undergoing liver transplantation.

OBJECTIVE: We examined the value of inflammatory and oxidative biomarkers in predicting acute kidney injury (AKI) following orthotopic liver transplantation (OLT). METHODS: Urinary excretion of tumour necrosis factor-α (TNF-α), interleukin-8 (IL-8), interleukin-10 (IL-10), superoxide dismutase (SOD), malondialdehyde (MDA), 6-keto prostaglandin F1α (6-keto-PGF1α), hydrogen peroxide (H2O2), and 8-keto prostaglandin F2α (8-iso-PGF2α), serum creatinine (SCr), blood urea nitrogen (BUN), urinary N-acetyl-beta-D-glucosaminidase (NAG), ß2-microglobulin (ß2-MG) and γ-glutamyl-transferase (γ-GT), were measured before surgery (baseline), at 2 h after graft reperfusion and 24 h after OLT in 28 liver transplantation patients. RESULTS: The levels of TNF-α, IL-8, IL-10, SOD, MDA, 6-keto-PGF1α, H2O2 and 8-iso-PGF2α in urine were all significantly higher in patients who had AKI than in those who did not at 2 h after graft reperfusion and 24 h after OLT (p < 0.01).

Paper-based electrochemical sensing platform with integral battery and electrochromic read-out.

We report a battery-powered, microelectrochemical sensing platform that reports its output using an electrochromic display. The platform is fabricated based on paper fluidics and uses a Prussian blue spot electrodeposited on an indium-doped tin oxide thin film as the electrochromic indicator. The integrated metal/air battery powers both the electrochemical sensor and the electrochromic read-out, which are in electrical contact via a paper reservoir. The sample activates the battery and the presence of analyte in the sample initiates the color change of the Prussian blue spot. The entire system is assembled on the lab bench, without the need for cleanroom facilities. The applicability of the device to point-of-care sensing is demonstrated by qualitative detection of 0.1 mM glucose and H(2)O(2) in artificial urine samples.

Use of crude extract of lentil plant (Lens culinaris Medikus) in peroxidase-based analyses: fast kinetic determination of hydrogen peroxide and sarcosine in urine.

Peroxidase-catalysed reactions are used in a wide variety of analytical applications, most of them based on the final quantification of hydrogen peroxide. Clinical tests for glucose, cholesterol, creatine, creatinine or uric acid in blood or urine and enzyme-linked immunosorbent assays for pesticides, hepatitis or acquired immune deficiency syndrome are good examples of such applications. The most widely used and commercially available peroxidase for biotechnological processes and analytical applications is horseradish peroxidase followed, although in much lower proportion, by soybean peroxidase. The high commercial interest in peroxidases has led to the search for new sources of these enzymes. This work describes the analytical use of lentil plant peroxidase (LPP), which is a new peroxidase extracted from lentil plants (Lens culinaris Medikus); an abundant post-harvest agricultural waste in the area of Castilla y León (Spain). A procedure for the quantification of hydrogen peroxide in urine is first proposed using crude extract of lentil plant instead of the purified enzyme. This procedure is then applied to the determination of sarcosine; a natural amino acid that has attracted considerable interest in clinical diagnostics since urinary sarcosine was proposed and later questioned as a biomarker for prostate cancer. Under the action of sarcosine oxidase, sarcosine is oxidized by molecular oxygen to give glycine, formaldehyde and hydrogen peroxide that is quantified according to the previously proposed procedure. The limit of detection for both hydrogen peroxide and sarcosine is around 5 × 10(-7) M. In the determination of sarcosine, the high selectivity of the overall enzymatic reaction, the simple sample treatment and instrumentation, the high-sample throughput and the use of LPP in the plant extract instead of the purified enzyme provide a rapid and inexpensive procedure with characteristics very suitable for routine analysis in a clinical laboratory.

Proinflammatory gene expression and renal lipogenesis are modulated by dietary protein content in obese Zucker fa/fa rats.

Obesity is a risk factor for the development of chronic kidney disease (CKD) and end-stage renal disease. It is not clear whether the adoption of a high-protein diet in obese patients affects renal lipid metabolism or kidney function. Thus the aims of this study were to assess in obese Zuckerfa/fa rats the effects of different types and amounts of dietary protein on the expression of lipogenic and inflammatory genes, as well as renal lipid concentration and biochemical parameters of kidney function. Rats were fed different concentrations of soy protein or casein (20, 30, 45%) for 2 mo. Independent of the type of protein ingested, higher dietary protein intake led to higher serum triglycerides (TG) than rats fed adequate concentrations of protein. Additionally, the soy protein diet significantly increased serum TG compared with the casein diet. However, rats fed soy protein had significantly decreased serum cholesterol concentrations compared with those fed a casein diet. No significant differences in renal TG and cholesterol concentrations were observed between rats fed with either protein diets. Renal expression of sterol-regulatory element binding protein 2 (SREBP-2) and its target gene HMG-CoA reductase was significantly increased as the concentration of dietary protein increased. The highest protein diets were associated with greater expression of proinflammatory cytokines in the kidney, independent of the type of dietary protein. These results indicate that high soy or casein protein diets upregulate the expression of lipogenic and proinflammatory genes in the kidney.

Association between oxidative stress and masked hypertension in a multi-ethnic population of obese children and adolescents.

OBJECTIVE: To evaluate whether oxidative stress is correlated with adiposity, obesity-related metabolic abnormalities, and ambulatory blood pressure (ABP) in a multi-ethnic pediatric population. STUDY DESIGN: We conducted a prospective study enrolling 42 obese children (age, 12.8 ± 2.4 years) and 34 non-obese children (age, 11.8 ± 3.4 years). We measured urine 8-isoprostane and hydrogen peroxide (markers of oxidative stress) in both obese and non-obese groups. In the obese group, we measured the 24-hour ABP and obtained an oral glucose tolerance test, lipid panel, interleukin-6, and tumor necrosis factor-α. RESULTS: 8-isoprostane and hydrogen peroxide were correlated with body mass index standard deviation score and waist circumference. The mean 8-isoprostane and hydrogen peroxide levels of the obese group were higher than those of the non-obese group. In the subset of obese subjects who underwent ABP monitoring, 8-isoprostane was correlated with mean 24-hour systolic blood pressure: within the obese group, 8-isoprostane was higher in obese children with elevated mean 24-hour systolic blood pressure. CONCLUSIONS: Our findings provide evidence of a significant correlation between oxidative stress, adiposity, and blood pressure in children. Longitudinal studies in a larger population sample are needed to validate the association between elevated urine 8-isoprostane level and cardiovascular risk factors in an obese pediatric population.

A non-enzymatic sensor for hydrogen peroxide based on polyaniline, multiwalled carbon nanotubes and gold nanoparticles modified Au electrode.

We describe the construction of a polyaniline (PANI), multiwalled carbon nanotubes (MWCNTs) and gold nanoparticles (AuNPs) modified Au electrode for determination of hydrogen peroxide without using peroxidase (HRP). The AuNPs/MWCNT/PANI composite film deposited on Au electrode was characterized by Scanning Electron Microscopy (SEM) and electrochemical methods. Cyclic voltammetric (CV) studies of the electrode at different stages of construction demonstrated that the modified electrode had enhanced electrochemical oxidation of H(2)O(2), which offers a number of attractive features to develop amperometric sensors based on split of H(2)O(2). The amperometric response to H(2)O(2) showed a linear relationship in the range from 3.0 µM to 600.0 µM with a detection limit of 0.3 µM (S/N = 3) and with high sensitivity of 3.3 mA µM(-1). The sensor gave accurate and satisfactory results, when employed for determination of H(2)O(2) in milk and urine.

Elevated levels of urinary hydrogen peroxide, advanced oxidative protein product (AOPP) and malondialdehyde in humans infected with intestinal parasites.

Oxidative stress has been implicated as an important pathogenic factor in the pathophysiology of various life-threatening diseases such as cancer, cardiovascular diseases and diabetes. It occurs when the production of free radicals (generated during aerobic metabolism, inflammation, and infections) overcome the antioxidant defences in the body. Although previous studies have implied that oxidative stress is present in serum of patients with parasitic infection there have been no studies confirming oxidative stress levels in the Malaysian population infected with intestinal parasites. Three biochemical assays namely hydrogen peroxide (H2O2), lipid peroxidation (LP) and advanced oxidative protein product (AOPP) assays were carried out to measure oxidative stress levels in the urine of human subjects whose stools were infected with parasites such as Blastocystis hominis, Ascaris, Trichuris, hookworm and microsporidia. The levels of H2O2, AOPP and LP were significantly higher (P<0.001, P<0.05 and P<0.05 respectively) in the parasite-infected subjects (n=75) compared to the controls (n=95). In conclusion, the study provides evidence that oxidative stress is elevated in humans infected by intestinal parasites. This study may influence future researchers to consider free radical-related pathways to be a target in the interventions of new drugs against parasitic infection and related diseases.

3D hierarchical hollow hydrangea-like Fe3+@ɛ-MnO2 microspheres with excellent electrochemical performance for dopamine and hydrogen peroxide.

Construction a sensor to accurately detect dopamine (DA) and hydrogen peroxide (H2O2) is meaningful due to their close relation to the health of organisms. In this work, one-step hydrothermal method was employed to synthesize hierarchical hollow hydrangea-like Fe3+@ɛ-MnO2 microspheres constructed by interconnected nanosheets, and the growth mechanism of the microspheres (nFe/nMn = 0.6) was also investigated in detail. The material was used to construct an electrochemical sensor for DA and H2O2 detection with the linear range of 0.02-78 µmoL-1 and 0.000133-5.19 mmoL-1, respectively. The detection limit and sensitivity for DA and H2O2 are 5 and 50 nmoL-1 (S/N = 3), 7034.1 and 242.6 µA m(mol·L-1)-1 cm-2, respectively. Furthermore, the sensor was successfully applied to the detection of DA and H2O2 in serum and urine samples, indicating a potential value of this work in the pharmaceutical and environmental fields.

Defect and Additional Active Sites on the Basal Plane of Manganese-Doped Molybdenum Diselenide for Effective Enzyme Immobilization: In Vitro and in Vivo Real-Time Analyses of Hydrogen Peroxide Sensing.

The defect engineering makes the new concepts and designs to further enhance the electrocatalytic activity of layered structures. In this work, we demonstrated the synthesis of Mn-doped MoSe2 and reported the resultant defective sites. Subsequently, the MnMoSe2 was developed as a new type of electrocatalyst for electrochemical biosensors. The formation of defect/distortion and effective immobilization of myoglobin (Mb) were evidently confirmed by using the transmission electron microscopy and UV-vis spectroscopy analyses, respectively. The result of electrochemical impedance spectroscopy analysis reveals that the Mn doping not only helps  to enzyme immobilization but also enhances the electronic conductivity of layered material.  Owing to the multiple signal amplification strategies, the proposed Mb-immobilized MnMoSe2 (Mb@MnMoSe2) exhibited an ultralow detection limit (0.004 µM) and a higher sensitivity (222.78 µA µM-1 cm-2) of H2O2. In real-sample analysis, the Mb@MnMoSe2 showed a feasible recovery range of H2O2 detection in human serum (95.6-102.1%), urine (101.2-102.3%), and rain water (100.7-102.1%) samples. On the other hand, an in vivo study using HaCaT (7.1 × 105/mL) and RAW 264.7 (1 × 106/mL) living cells showed the feasible current responses of 0.096 and 0.085 µA, respectively. Finally, the Mn doping gives a new opportunity to fabricate a promising electrocatalyst for H2O2 biosensing.

Low-cost and environment-friendly synthesis of carbon nanorods assembled hierarchical meso-macroporous carbons networks aerogels from natural apples for the electrochemical determination of ascorbic acid and hydrogen peroxide.

In this work, the low-cost carbon nanorods assembled hierarchical meso-macroporous carbons networks aerogels (CNs-HMCNAs) was environment-friendly synthesized from a cheap and abundant biomass of apples (Malus pumila Mill) for the first time. The biomass of apples derived CNs-HMCNAs exhibited the unique hierarchical meso-macroporous structure with large specific surface area and high density of edge defective sites. At the CNs-HMCNAs modified GCE (CNs-HMCNAs/GCE), the electron transfer between the glassy carbon electrode (GCE) and the ascorbic acid (AA) (or hydrogen peroxide (H2O2)) was effectively enhanced, and thus induced a low overvoltage for AA electrooxidation (or H2O2 electroreduction). As an electrochemical AA (or H2O2) sensor, the CNs-HMCNAs/GCE exhibited wider linear range, lower detection limit, higher sensitivity and stability than GCE and the carbon nanotubes modified GCE (CNTs/GCE). In particular, the CNs-HMCNAs/GCE showed great potential feasibility in the practical determination of AA (in AA injection, Vitamin C tablet and kiwi juice) or H2O2 (in human urine, milk and beer).

Hydroxyhydroquinone interferes with the chlorogenic acid-induced restoration of endothelial function in spontaneously hypertensive rats.

BACKGROUND: Coffee is a rich source of antioxidative polyphenols, but epidemiological studies and interventional trials have failed to demonstrate any clear beneficial effects of coffee consumption on hypertension. The interaction between hydroxyhydroquinone (HHQ) and 5-caffeoylquinic acid (CQA) was examined, in an attempt to understand the controversial effects of coffee on hypertension. METHODS: Male Wistar Kyoto (WKY) rats or spontaneously hypertensive rats (SHRs, 14 weeks old) were divided into the following four groups; those on a control diet, 0.005% HHQ diet, 0.5% CQA diet, and HHQ plus CQA diet. The rats were fed the above diets for 8 weeks, and the tail arterial blood pressure was monitored in conscious rats at 2-week intervals. Urinary nitric oxide (NO) metabolites and hydrogen peroxide (H(2)O(2)) excretion were measured 8 weeks after the start of the experiment. Endothelium-dependent and -independent vasorelaxant responses and immunohistochemical staining for nitrotyrosine were examined in aortas. RESULTS: HHQ inhibited the CQA-induced improvement in hypertension, urinary NO metabolites or H(2)O(2) excretion, endothelial dysfunction, and nitrotyrosine deposits in aortas in SHR. However, the administration of HHQ alone had little effect on either strain. CONCLUSIONS: Based on the content ratio of HHQ and chlorogenic acids in coffee, HHQ interfered with the CQA-induced improvement in blood pressure and endothelial function in SHR. The results explain, at least in part, the conflicting action of coffee drinking on hypertension and vascular reactivity.

Two dimensional α-MoO3-x nanoflakes as bare eye probe for hydrogen peroxide in biological fluids.

We report a rapid and facile method for detection of hydrogen peroxide (H2O2) in biological fluids using sub-stoichiometric two-dimensional (2D) molybdenum trioxide (α-MoO3-x) nanoflakes. The two-dimensional nanoflakes, initially blue in color, is oxidized after interaction with hydrogen peroxide thereby changing its oxidation state to form α-MoO3. The change in oxidation state of nanoflakes transforms from blue to a visually distinct hazy blue color with change in absorption spectrum. The phenomenal property is explored here in sensing up to 34 nM as limit of detection. The efficacy of the detection system was analyzed by "zone of inhibition" based agar diffusion assay with different concentrations of H2O2. The current approach is highly accurate, effective and reproducible for quantification of physiological concentration of H2O2 in biological fluid such as human urine.

A new 3D printed radial flow-cell for chemiluminescence detection: Application in ion chromatographic determination of hydrogen peroxide in urine and coffee extracts.

A new polymer flow-cell for chemiluminescence detection (CLD) has been designed and developed by diverging multiple linear channels from a common centre port in a radial arrangement. The fabrication of radial flow-cell by 3D PolyJet printing and fused deposition modeling (FDM) has been evaluated, and compared with a similarly prepared spiral flow-cell design commonly used in chemiluminescence detectors. The radial flow-cell required only 10 h of post-PolyJet print processing time as compared to ca. 360 h long post-PolyJet print processing time required for the spiral flow-cell. Using flow injection analysis, the PolyJet 3D printed radial flow-cell provided an increase in both the signal magnitude and duration, with an average increase in the peak height of 63% and 58%, peak area of 89% and 90%, and peak base width of 41% and 42%, as compared to a coiled-tubing spiral flow-cell and the PolyJet 3D printed spiral flow-cell, respectively. Computational fluid dynamic (CFD) simulations were applied to understand the origin of the higher CLD signal obtained with the radial flow-cell design, indicating higher spatial coverage near the inlet and lower linear velocities in the radial flow-cell. The developed PolyJet 3D printed radial flow-cell was applied in a new ion chromatography chemiluminescence based assay for the detection of H2O2 in urine and coffee extracts.

Changes in Urinary Hydrogen Peroxide and 8-Hydroxy-2'-Deoxyguanosine Levels after a Forest Walk: A Pilot Study.

Some studies have shown that exposure to forests has positive effects on human health, although the mechanisms underlying the health benefits of a forest environment have not been elucidated yet. The current study was aimed at examining how the levels of urinary hydrogen peroxide (H2O2) and 8-hydroxy-2'deoxyguanosine (8-OHdG) change after a forest or urban walk in healthy subjects. Twenty-eight volunteers (19 men and 9 women) participated in the study. The forest walks were carried out in a forest in Okayama Prefecture, Japan, and the urban walks (15 men and 7 women) were carried out in the downtown area of Okayama city, each for two hours. Spot urine samples were collected before the walk, the next day and one week after the forest or urban walk. Compared with pre-forest walk levels, urinary H2O2 (p < 0.1) and 8-OHdG (p < 0.1) concentrations significantly decreased in the participants the day after the forest walk; furthermore, urinary 8-OHdG remained at a low level even at one week after the forest walk (p < 0.05). However, there were no significant changes in the concentrations of these oxidative biomarkers after the urban walk. These findings suggest the possibility that exposure to forests may alleviate oxidative stress in the body.

The Correlation Between Urinary 8-Iso-Prostaglandin F2α and Hydrogen Peroxide Toward Renal Function in T2DM Patients Consuming Sulfonylurea and Combination of Metformin-Sulfonylurea.

BACKGROUND: Renal dysfunction is a common complication in type 2 diabetes mellitus patients associated with oxidative damage which could be characterized by 8-iso-prostaglandin F2α and hydrogen peroxide level as oxidative stress markers. OBJECTIVE: The aim of our study is to determine if there is a difference in 8-iso-prostaglandin F2α and hydrogen peroxide levels between sulfonylurea and combination of metformin-sulfonylurea in diabetic patients. We also wanted to determine if these oxidative stress markers correlate with the estimated Glomerular Filtration Rate (eGFR). SUBJECTS AND METHODS: We conducted a cross-sectional study with inclusion of 55 patients with type 2 diabetes mellitus in Dr. Sitanala Tangerang Hospital, Indonesia with purposive sampling. The value of eGFR was obtained by serum creatinine levels, while the level of 8-iso-prostaglandin F2α was measured by ELISA and urinary hydrogen peroxide using FOX-1 (Ferrous Ion Oxidation Xylenol Orange 1). RESULTS: There was no difference in 8-iso-prostaglandin F2α and hydrogen peroxide level between the two groups (p=0.088 and p=0.848). Moreover, there was no difference in eGFR values between the two groups, measured by Cockroft-Gault, MDRD, and CKD-EPI. 8-iso-prostaglandin F2α (n=55) was positively correlated with eGFR based on Cockroft-Gault (r=0.382; p=0.009), whereas urinary hydrogen peroxide (n=47) also generate significant positive correlation with eGFR based on the MDRD equation (r=0.326; p=0.021). Linear regression analysis showed that 8-iso-prostaglandin F2α is the most predictive factor and the only significant factor for eGFR in Cockroft-Gault, MDRD and also CKDEPI, even after controlled by gender, age, BMI, HbA1c, systole, and H2O2. CONCLUSION: The two treatments did not have any significant differences in antioxidant activity. However, an increase of urinary 8-iso-prostaglandin F2. and hydrogen peroxide which correlates with eGFR in the total sample may play a significant role in the pathophysiology of diabetic nephropathy.

Effect of oestrogen on reactive oxygen species production in the aortas of ovariectomized Dahl salt-sensitive rats.

OBJECTIVE: In the present study, we examined whether ovariectomy increases reactive oxygen species (ROS) and the expression of nicotinamide adenine dinucleotide phosphate, reduced form (NADPH) oxidase and modulates the scavenger enzymes for ROS in the aortas of Dahl salt-sensitive (DSS) rats fed a high salt diet. METHODS: DSS female rats were ovariectomized and fed a high salt diet (8% NaCl), or a high salt diet plus oestrogen supplement for 4 weeks. Urinary levels of hydrogen peroxide (H2O2) were measured by using 2',7'-dichlorofluorescein. The expression of an NADPH oxidase subunit p22phox, extracellular superoxide dismutase (ecSOD), glutathione peroxidase (GPx)1, GPx4 and monocyte chemoattractant protein 1 (MCP-1) messenger RNA was assessed by reverse transcription-polymerase chain reaction. The expression of MCP-1, and macrophage infiltration were examined by immunohistochemical analysis. RESULTS: Ovariectomy increased superoxide production and the expression of NADPH oxidase subunit p22phox mRNA and protein in the aortas of DSS rats fed a high salt diet. In contrast, ovariectomy reduced the expression of ecSOD mRNA and protein and the expression of GPx1 and GPx4 mRNA in the aorta. Ovariectomy increased MCP-1 mRNA and protein expression and ED1-positive cells in the aorta. CONCLUSIONS: Ovariectomy leads to an amplification of oxidative stress in DSS rats fed a high salt diet synergistically by an increase in the ROS-generating system and a decrease in the ROS-eliminating system, as shown in the increase in superoxide production and the urinary excretion of H2O2. Oestrogen supplementation counteracted these alterations, showing how oestrogen is antioxidative.