Involvement of ERK, Akt and JNK signalling in H2O2-induced cell injury and protection by hydroxytyrosol and its metabolite homovanillic alcohol

The olive oil polyphenol, hydroxytyrosol (HT), is believed to be capable of exerting protection against oxidative kidney injury. In this study we have investigated the ability of HT and its O-methylated metabolite, homovanillic alcohol (HVA) to protect renal cells against oxidative damage induced by hydrogen peroxide. We show that both compounds were capable of inhibiting hydrogen peroxide-induced kidney cell injury via an ability to interact with both MAP kinase and PI3 kinase signalling pathways, albeit at different concentrations. HT strongly inhibited death and prevented peroxide-induced increases in ERK1/2 and JNK1/2/3 phosphorylation at 0.3 microM, whilst HVA was effective at 10 microM. At similar concentrations, both compounds also prevented peroxide-induced reductions in Akt phosphorylation. We suggest that one potential protective effect exerted by olive oil polyphenols against oxidative kidney cell injury may be attributed to the interactions of HT and HVA with these important intracellular signalling pathways.

Synthesis and structure of the Mn II,II complex salt [Mn2(η1η1μ2-oda)(phen)4(H2O)2][Mn2][Mn2(η1η1μ2-oda)(phen)4(η1-oda)2].4H2O (odah2 = octanedioic acid): a catalyst for H2O2 disproportionation

The synthesis and X-ray crystal structure of the MnII,11 complex double salt [Mn2(η1η1µ2-oda)(phen)4(H2O)2][Mn2(η1η1µ2-oda(phen)4(η1-oda)2]·4H2O is reported, together with its catalytic activity towards the disproportionation of H2O2.

Regulation of Bcl-xL expression by H2O2 in cardiac myocytes.

Oxidative stress promotes cardiac myocyte apoptosis through the mitochondrial death pathway. Since Bcl-2 family proteins are key regulators of apoptosis, we examined the effects of H2O2 on the expression of principal Bcl-2 family proteins (Bcl-2, Bcl-xL, Bax, Bad) in neonatal rat cardiac myocytes. Protein expression was assessed by immunoblotting. Bcl-2, Bax, and Bad were all down-regulated in myocytes exposed to 0.2 mm H2O2, a concentration that induces apoptosis. In contrast, although Bcl-xL levels initially declined, the protein was re-expressed from 4-6 h. Bcl-xL mRNA was up-regulated from 2 to 4 h in neonatal rat or mouse cardiac myocytes exposed to H2O2, consistent with the re-expression of protein. Four different untranslated first exons have been identified for the Bcl-x gene (exons 1, 1B, 1C, and 1D, where exon 1 is the most proximal and exon 1D the most distal to the coding region). All were detected in mouse or rat neonatal cardiac myocytes, but exon 1D was not expressed in adult mouse hearts. In neonatal mouse or rat cardiac myocytes, H2O2 induced the expression of exons 1B, 1C, and 1D, but not exon 1. These data demonstrate that the Bcl-x gene is selectively responsive to oxidative stress, and the response is mediated through distal promoter regions.

Cardiac myocyte gene expression profiling during H2O2-induced apoptosis

High levels of oxidative stress promote cardiac myocyte death, though lower levels are potentially cytoprotective/anabolic. We examined the changes in gene expression in rat neonatal cardiac myocytes exposed to apoptotic (0.2 mM) or nontoxic (0.04 mM) concentrations of H2O2 (2, 4, or 24 h) using Affymetrix microarrays. Using U34B arrays, we identified a ubiquitously expressed, novel H2O2-responsive gene [putative peroxide-inducible transcript 1 (Perit1)], which generates two alternatively spliced transcripts. Using 230 2.0 arrays, H2O2 (0.04 mM) promoted significant changes in expression of only 32 genes, all of which were seen with 0.2 mM H2O2. We failed to detect any increase in the rate of protein synthesis in cardiac myocytes exposed to <0.1 mM H2O2, further suggesting that global, low concentrations of H2O2 are not anabolic in this system. H2O2 (0.2 mM) promoted significant (P < 0.05, >1.75-fold) changes in expression of 649 mRNAs and 187 RNAs corresponding to no established gene. Of the mRNAs, 114 encoded transcriptional regulators including Krüppel-like factors (Klfs). Quantitative PCR independently verified the changes in Klf expression. Thus, H2O2-induced cardiac myocyte apoptosis is associated with dynamic changes in gene expression. The expression of these genes and their protein products potentially influences the progression of the apoptotic response.

A comparative study of the electrogeneration of hydrogen peroxide using Vulcan and Printex carbon supports

A comparative study of two different conductive carbon-black pigments, Vulcan XC-72 R and Printex L6, for the electrogeneration of hydrogen peroxide (H(2)O(2)) by reducing dissolved oxygen in an alkaline solution was performed. The materials were physically characterized by X-ray diffraction (XRD), Fourier transform infrared attenuated total reflection (ATR-FTIR) and X-ray photoelectron spectroscopy (XPS). XRD shows the presence of SO(2) and ATR-FTIR technique indicates a difference in NO and SO(2) functional groups between the two carbon pigments. XPS indicated presence of SO and NO and more oxygenated acid species on Printex L6. A rotating ring-disk electrode was used for electrochemical analysis of the oxygen reduction reaction (ORR). The results showed that the Printex L6 was better than Vulcan XC-72 R for H(2)O(2) production. Results also indicate that the number of electrons transferred in the ORR for Printex L6 and Vulcan XC-72 R were 2.2 and 2.9, respectively, while the percentages of H(2)O(2) formed were 88% and 51%. Scanning electrochemistry microscopy images confirmed the higher amount of H(2)O(2) formed in the Printex L6 pigment. Printex L6 was shown to be a more promising for H(2)O(2) production than Vulcan XC-72 R, while the latter was shown to have more potential for fuel cells. (C) 2011 Elsevier Ltd. All rights reserved.

TNF-alpha, H2O2 and NO response of peritoneal macrophages to Yersinia enterocolitica O : 3 derivatives

In this study, the effect of Yersinia derivatives on nitric oxide (NO), hydrogen peroxide (H2O2) and tumor necrosis factor-alpha (TNF-alpha) production by murine peritoneal macrophages was investigated. Addition of lipopolysaccharide (LPS) to the macrophage culture resulted in NO production that was dose dependent. on the other hand, bacterial cellular extract (CE) and Yersinia outer proteins (Yops) had no effect on NO production. The possible inhibitory effect of Yops on macrophage cultures stimulated with LPS was investigated. Yops partially inhibited NO production (67.4%) when compared with aminoguanidine. The effects of Yersinia derivatives on H2O2 production by macrophages were similar to those on NO production. LPS was the only derivative that stimulated H2O2 release in a dose-dependent manner. All Yersinia derivatives provoked the production of TNF-alpha, but LPS had the strongest effect, as observed for NO production. CE and Yops stimulated TNF-alpha production to a lesser extent than LPS. The results indicate the possibility that in vivo Yops may aid the evasion of the bacteria from the host defense mechanism by impairing the secretion of NO by macrophages. (C) 2003 Elsevier SAS. All rights reserved.

Transdentinal protective role of sodium ascorbate against the cytopathic effects of H2O2 released from bleaching agents

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Trans-enamel and trans-dentinal cytotoxic effects of a 35% H2O2 bleaching gel on cultured odontoblast cell lines after consecutive applications

To evaluate the trans-enamel and trans-dentinal cytotoxic effects of a 35% H2O2 bleaching gel on an odontoblast-like cell lines (MDPC-23) after consecutive applications.Fifteen enamel/dentine discs were obtained from bovine central incisor teeth and placed individually in artificial pulp chambers. Three groups (n = 5 discs) were formed according to the following enamel treatments: G1: 35% H2O2 bleaching gel (15 min); G2: 35% H2O2 bleaching gel (15 min) + halogen light (20 s); G3: control (no treatment). After repeating the treatments three consecutive times, the extracts (culture medium + gel components that had diffused through enamel/dentine discs) in contact with the dentine were collected and applied to previously cultured MDPC-23 cells (50 000 cells cm(-2)) for 24 h. Cell metabolism was evaluated by the MTT assay and data were analysed statistically (alpha = 5%; Kruskal-Wallis and Mann-Whitney U-test). Cell morphology was analysed by scanning electron microscopy.Cell metabolism decreased by 92.03% and 82.47% in G1 and G2 respectively. G1 and G2 differed significantly (P < 0.05) from G3. Regardless of halogen light activation, the application of the bleaching gel on the cultured odontoblast-like cells caused significantly more severe cytotoxic effects than those observed in the nontreated control group. In addition, significant morphological cell alterations were observed in G1 and G2.After three consecutive applications of a 35% H2O2 bleaching agent, the diffusion of the gel components through enamel and dentine caused severe toxic effects to cultured pulp cells.

TNF-alpha activates human monocytes for Paracoccidioides brasiliensis killing by an H2O2-dependent mechanism

Human monocytes activated by recombinant tumor necrosis factor alpha (TNF-alpha) exhibited significant fungicidal activity on the yeast cells of a highly virulent strain of Paracoccidioides brasiliensis. This process was significantly inhibited in the presence of catalase (CAT - a scavenger of H2O2), but not in the presence of superoxide-dismutase (SOD - a scavenger of superoxide anion) or N-G-monomethyl-L- arginine (N-G-MMLA - a nitric oxide inhibitor). Furthermore, there was a direct association between the intracellular killing of the fungus and the production of H2O2 by activated cells. These results strongly suggest a role for H2O2 in the killing of highly virulent strains of P. brasiliensis by TNF-alpha-activated human monocytes.

Production of reactive oxygen (H2O2) and nitrogen (NO) intermediates and TNF-α in mice genetically selected for high (H) and low (L) antibody response and experimentally infected with Leptospira serovar pomona

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Interleukin-10 but not Transforming Growth Factor beta inhibits murine activated macrophages Paracoccidioides brasiliensis killing: Effect on H2O2 and NO production

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Co-stressors chilling and high light increase photooxidative stress in diuron-treated red alga Kappaphycus alvarezii but with lower involvement of H2O2

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nb-containing hematites Fe2-xNbxO3: The role of Nb5+ on the reactivity in presence of the H2O2 or ultraviolet light

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Anti-inflammatory activity of Alchornea triplinervia ethyl acetate fraction: Inhibition of H2O2, NO and TNF-alpha

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Simple and fast spectrophotometric determination of H2O2 in photo-Fenton reactions using metavanadate

This work proposes a spectrophotometric method for the determination of hydrogen peroxide during photodegradation reactions. The method is based on the reaction of H2O2 with amonium metavanadate in acidic medium, which results in the formation of a red-orange color peroxovanadium cation, with maximum absorbance at 450 nm. The method was optimized using the multivariate analysis providing the minimum concentration of vanadate (6.2 mmol L-1) for the maximum absorbance signal. Under these conditions, the detection limit is 143 mu mol L-1. The reaction product showed to be very stable for samples of peroxide concentrations up to 3 mmol L-1 at room temperature during 180 h. For higher concentrations however, samples must be kept refrigerated (4 degrees C) or diluted. The method showed no interference of Cl- (0.2-1.3 mmol L-1), NO3- (0.3-1.0 mmol L-1), Fe3+, (0.2-1.2 mmol L-1) and 2,4-dichlorophenol (DCP) (0.2-1.0 mmol L-1). When compared to iodometric titration, the vanadate method showed a good agreament. The method was applied for the evaluation of peroxide consumption during photo-Fenton degradation of 2,4-dichlorophenol using blacklight irradiation. (c) 2004 Elsevier B.V. All rights reserved.