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Journal Cover Redox Biology
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  This is an Open Access Journal Open Access journal
   ISSN (Online) 2213-2317
   Published by Elsevier Homepage  [2805 journals]
  • Role of nitrite, urate and pepsin in the gastroprotective effects of
           saliva

    • Abstract: Publication date: Available online 8 April 2016
      Source:Redox Biology
      Author(s): Bárbara S. Rocha, Jon O Lundberg, Rafael Radi, João Laranjinha
      Dietary nitrate is now recognized as an alternative substrate for nitric oxide (•NO) production in the gut. This novel pathway implies the sequential reduction of nitrate to nitrite, •NO and other bioactive nitrogen oxides but the physiological relevance of these oxidants has remained elusive. We have previously shown that dietary nitrite fuels an hitherto unrecognized nitrating pathway at acidic gastric pH, through which pepsinogen is nitrated in the gastric mucosa, yielding a less active form of pepsin in vitro. Here, we demonstrate that pepsin is nitrated in vivo and explore the functional impact of protein nitration by means of peptic ulcer development. Upon administration of pentagastrin and human nitrite-rich saliva or sodium nitrite to rats, nitrated pepsin was detected in the animal´s stomach by immunoprecipitation. •NO was measured in the gastric headspace before and after nitrite instillation by chemiluminescence. At the end of each procedure, the stomach´s lesions, ranging from gastric erosions to haemorrhagic ulcers, were scored. Nitrite increased gastric •NO by 200-fold (p<0.05) and nitrated pepsin was detected both in the gastric juice and the mucosa (p<0.05). Exogenous urate, a scavenger of nitrogen dioxide radical, blunted •NO detection and inhibited pepsin nitration, suggesting an underlining free radical-dependent mechanism for nitration. Functionally, pepsin nitration prevented the development of gastric ulcers, as the lesions were only apparent when pepsin nitration was inhibited by urate. In sum, this work unravels a novel dietary-dependent nitrating pathway in which pepsin is nitrated and inactivated in the stomach, preventing the progression of gastric ulcers.
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      PubDate: 2016-04-09T12:07:28Z
       
  • Tempol improves xanthine oxidoreductase-mediated vascular responses to
           nitrite in experimental renovascular hypertension

    • Abstract: Publication date: Available online 5 April 2016
      Source:Redox Biology
      Author(s): Gustavo H. Oliveira-Paula, Lucas C. Pinheiro, Danielle A. Guimaraes, Sandra O. Conde Tella, Ana L. Furlan Blanco, Celio D. Angelis, Alan N. Schechter, Jose E. Tanus-Santos
      Upregulation of xanthine oxidoreductase (XOR) increases vascular reactive oxygen species (ROS) levels and contributes to nitroso-redox imbalance. However, XOR can generate nitric oxide (NO) from nitrite, and increased superoxide could inactivate NO formed from nitrite. This study tested the hypothesis that XOR contributes to the cardiovascular effects of nitrite in renovascular hypertension, and that treatment with the antioxidant tempol (4-hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl) improves XOR-mediated effects of nitrite. Blood pressure was assessed weekly in two-kidney one-clip (2K1C) and control rats. After six weeks of hypertension, the relaxing responses to nitrite were assessed in aortic rings in the presence of the XOR inhibitor oxypurinol (or vehicle), either in the absence or in the presence of tempol. Moreover, in vivo hypotensive responses to nitrite were also examined in the presence of oxypurinol (or vehicle) and tempol (or vehicle). Aortic XOR activity and expression were evaluated by fluorescence and Western blot, respectively. Vascular ROS production was assessed by the dihydroethidium assay. 2K1C hypertensive rats showed increased aortic XOR activity and vascular ROS production compared with control rats. Oxypurinol shifted the nitrite concentration–response curve to the right in aortic rings from 2K1C rats (but not in controls). Oxypurinol also attenuated the hypotensive responses to nitrite in 2K1C rats (but not in controls). These functional findings agree with increased aortic and plasma XOR activity found in 2K1C rats. Tempol treatment enhanced oxypurinol-induced shift of the nitrite concentration–response curve to the right. However, antioxidant treatment did not affect XOR-mediated hypotensive effects of nitrite. Our results show that XOR is important to the cardiovascular responses to nitrite in 2K1C hypertension, and XOR inhibitors commonly used by patients may cancel this effect. This finding suggests that nitrite treatment may not be effective in patients being treated with XOR inhibitors. Moreover, while tempol may improve the vascular responses to nitrite, antihypertensive responses are not affected.
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      PubDate: 2016-04-06T11:54:55Z
       
  • CellNO trap: Novel device for quantitative, real-time, direct measurement
           of nitric oxide from cultured RAW 267.4 macrophages

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Weilue He, Megan C. Frost
      Nitric oxide (NO), is arguably one of the most important small signaling molecules in biological systems. It regulates various biological responses in both physiological and pathological conditions, often time producing seemingly contradictory results. The details of the effects of NO are highly dependent on the level of NO that cells experience and the temporal aspect of when and how long cells are exposed to NO. Herein, we present a novel measurement system (CellNO trap) that allows real-time NO measurement via chemiluminescence detection from general adhesive cultured cells using standard cell culture media and reagents that does not perturb the cells under investigation. Highly controlled light-initiated NO releasing polymer SNAP-PDMS was used to characterize and validate the quantitative data nature of the device. The NO generation profile from the macrophage cell-line RAW264.7 stimulated by 100ng/ml LPS and 10ng/ml IFN-γ was recorded. Measured maximum NO flux from RAW264.7 varied between around 2.5–9pmol/106 cell/s under 100ng/ml LPS and 10ng/ml IFN-γ stimulation, and 24h cumulative NO varied between 157 and 406 nmol/106cell depending on different culture conditions, indicating the conventional report of an average flux or maximum flux is not sufficient to represent the dynamic characters of NO. LPS and IFN-γ’s synergistic effect to RAW264.7 NO generation was also directly observed with the CellNO trap. The real-time effect on the NO generation from RAW264.7 following the addition of arginine, nor-NOHA and L-NAME to the cultured cells is presented. There is great potential to further our understanding of the role NO plays in normal and pathological conditions clearly understanding the dynamic production of NO in response to different stimuli and conditions; use of CellNO trap makes it possible to quantitatively determine the precise NO release profile generated from cells in a continuous and real-time manner with chemiluminescence detection.
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      PubDate: 2016-04-06T11:54:55Z
       
  • Glutathione S-transferase pi modulates NF-κB activation and
           pro-inflammatory responses in lung epithelial cells

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Jane T. Jones, Xi Qian, Jos L.J. van der Velden, Shi Biao Chia, David H. McMillan, Stevenson Flemer, Sidra M. Hoffman, Karolyn G. Lahue, Robert W. Schneider, James D. Nolin, Vikas Anathy, Albert van der Vliet, Danyelle M. Townsend, Kenneth D. Tew, Yvonne M.W. Janssen-Heininger
      Nuclear Factor kappa B (NF-κB) is a transcription factor family critical in the activation of pro- inflammatory responses. The NF-κB pathway is regulated by oxidant-induced post-translational modifications. Protein S-glutathionylation, or the conjugation of the antioxidant molecule, glutathione to reactive cysteines inhibits the activity of inhibitory kappa B kinase beta (IKKβ), among other NF-κB proteins. Glutathione S-transferase Pi (GSTP) is an enzyme that has been shown to catalyze protein S-glutathionylation (PSSG) under conditions of oxidative stress. The objective of the present study was to determine whether GSTP regulates NF-κB signaling, S-glutathionylation of IKK, and subsequent pro-inflammatory signaling. We demonstrated that, in unstimulated cells, GSTP associated with the inhibitor of NF-κB, IκBα. However, exposure to LPS resulted in a rapid loss of association between IκBα and GSTP, and instead led to a protracted association between IKKβ and GSTP. LPS exposure also led to increases in the S-glutathionylation of IKKβ. SiRNA-mediated knockdown of GSTP decreased IKKβ-SSG, and enhanced NF-κB nuclear translocation, transcriptional activity, and pro-inflammatory cytokine production in response to lipopolysaccharide (LPS). TLK117, an isotype-selective inhibitor of GSTP, also enhanced LPS-induced NF-κB transcriptional activity and pro-inflammatory cytokine production, suggesting that the catalytic activity of GSTP is important in repressing NF-κB activation. Expression of both wild-type and catalytically-inactive Y7F mutant GSTP significantly attenuated LPS- or IKKβ-induced production of GM-CSF. These studies indicate a complex role for GSTP in modulating NF-κB, which may involve S-glutathionylation of IKK proteins, and interaction with NF-κB family members. Our findings suggest that targeting GSTP is a potential avenue for regulating the activity of this prominent pro-inflammatory and immunomodulatory transcription factor.
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      PubDate: 2016-04-06T11:54:55Z
       
  • Oxidative instability of hemoglobin E (β26 Glu→Lys) is
           increased in the presence of free α subunits and reversed by
           α-hemoglobin stabilizing protein (AHSP): Relevance to
           HbE/β-thalassemia

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Michael Brad Strader, Tigist Kassa, Fantao Meng, Francine B. Wood, Rhoda Elison Hirsch, Joel M. Friedman, Abdu I. Alayash
      When adding peroxide (H2O2), β subunits of hemoglobin (Hb) bear the burden of oxidative changes due in part to the direct oxidation of its Cys93. The presence of unpaired α subunits within red cells and/or co-inheritance of another β subunit mutant, HbE (β26 Glu→Lys) have been implicated in the pathogenesis and severity of β thalassemia. We have found that although both HbA and HbE autoxidize at initially comparable rates, HbE loses heme at a rate almost 2 fold higher than HbA due to unfolding of the protein. Using mass spectrometry and the spin trap, DMPO, we were able to quantify irreversible oxidization of βCys93 to reflect oxidative instability of β subunits. In the presence of free α subunits and H2O2, both HbA and HbE showed βCys93 oxidation which increased with higher H2O2 concentrations. In the presence of Alpha-hemoglobin stabilizing protein (AHSP), which stabilizes the α-subunit in a redox inactive hexacoordinate conformation (thus unable to undergo the redox ferric/ferryl transition), Cys93 oxidation was substantially reduced in both proteins. These experiments establish two important features that may have relevance to the mechanistic understanding of these two inherited hemoglobinopathies, i.e. HbE/β thalassemia: First, a persistent ferryl/ferryl radical in HbE is more damaging to its own β subunit (i.e., βCys93) than HbA. Secondly, in the presence of excess free α-subunit and under the same oxidative conditions, these events are substantially increased for HbE compared to HbA, and may therefore create an oxidative milieu affecting the already unstable HbE.
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      PubDate: 2016-03-22T10:54:13Z
       
  • PARP-1 inhibition influences the oxidative stress response of the human
           lens

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Andrew J.O. Smith, Simon S.R. Ball, Richard P. Bowater, I. Michael Wormstone
      Poly(ADP-ribose) polymerase-1 (PARP-1) is best characterised for its involvement in DNA repair. PARP-1 activity is also linked to cell fate, confounding its roles in maintaining genome integrity. The current study assessed the functional roles of PARP-1 within human lens cells in response to oxidative stress. The human lens epithelial cell line FHL124 and whole human lens cultures were used as experimental systems. Hydrogen peroxide (H2O2) was employed to induce oxidative stress and cell death was assessed by LDH release. The functional influence of PARP-1 was assessed using targeted siRNA and chemical inhibition (by AG14361). Immunocytochemistry and western blotting were used to assess PARP-1 expression and the alkaline comet assay determined the levels of DNA strand breaks. PARP-1 was generally observed in the cell nucleus in both the FHL124 cell line and whole human lenses. PARP-1 inhibition rendered FHL124 cells more susceptible to H2O2-induced DNA strand breaks. Interestingly, reduction of PARP-1 activity significantly inhibited H2O2-induced cell death relative to control cells. Inhibition of PARP-1 in whole human lenses resulted in a reduced level of lens opacity and cell death following exposure to H2O2 relative to matched pair controls. Thus, we show that PARP-1 could play a role in the fate of human lens cells, and these first observations in human lenses suggest that it could impact on lens opacity. Further studies are required to elucidate the regulatory processes that give rise to these effects.
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      PubDate: 2016-03-17T10:29:13Z
       
  • Nrf2 Silencing to Inhibit Proteolytic Defense Induced by Hyperthermia in
           HT22 Cells

    • Abstract: Publication date: Available online 3 March 2016
      Source:Redox Biology
      Author(s): Perinur Bozaykut, Nesrin Kartal Ozer, Betul Karademir
      Nrf2 pathway has been known to be protective against cancer progression however recent studies have revealed that the antioxidant activity of Nrf2 contributes to chemotherapy resistance. For many years, hyperthermia has been used as an additional therapy to increase the efficiancy of chemotherapy and radiotherapy. Besides the positive effects of hyperthermia during treatment procedure, thermotolerance has been found to develop against heat treatment. Although the involved molecular mechanisms have not been fully clarified, heat shock proteins (HSP) and proteasome activity are known to be involved in the acquisition of thermotolerance. The aim of this study was to investigate the potential beneficial effects of combining hyperthermia with Nrf2 silencing to inhibit molecular mechanisms leading to induction of defense mechanisms in transcription level. Following heat treatment of HT22 cells, HSP70 and the proteasome levels and as well as proteasome activity were found to be elevated in the nucleus. Our results demonstrated that Nrf2 silencing reduced defense mechanisms against heat treatment both in antioxidant and proteolytic manner and Nrf2 may be a potential target for therapeutic approach in order to improve the beneficial effects of hyperthermia in cancer therapy.
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      PubDate: 2016-03-08T09:45:14Z
       
  • Role of Heme Oxygenase-1 in Low Dose Radioadaptive Response

    • Abstract: Publication date: Available online 3 March 2016
      Source:Redox Biology
      Author(s): Lingzhi Bao, Jie Ma, Guodong Chen, Jue Hou, Tom K. Hei, K.N. Yu, Wei Han
      Radioadaptive response (RAR) is an important phenomenon induced by low dose radiation. However, the molecular mechanism of RAR is obscure. In this study, we focused on the possible role of heme oxygenase 1 (HO-1) in RAR. Consistent with previous studies, priming dose of X-ray radiation (1-10cGy) induced significant RAR in normal human skin fibroblasts (AG 1522 cells). Transcription and translation of HO-1 was up-regulated more than two fold by a priming dose of radiation (5cGy). Zinc protoporphyrin Ⅸ, a specific competitive inhibitor of HO-1, efficiently inhibited RAR whereas hemin, an inducer of HO-1, could mimic priming dose of X-rays to induce RAR. Knocking down of HO-1 by transfection of HO-1 siRNA significantly attenuated RAR. Furthermore, the expression of HO-1 gene was modulated by the nuclear factor (erythroid-derived 2)-like 2 (Nrf2), which translocated from cytoplasm to nucleus after priming dose radiation and enhance the antioxidant level of cells.
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      PubDate: 2016-03-08T09:45:14Z
       
  • The MLN4924 inhibitor exerts a neuroprotective effect against oxidative
           stress injury via Nrf2 protein accumulation

    • Abstract: Publication date: Available online 2 March 2016
      Source:Redox Biology
      Author(s): Ana Cristina Andérica-Romero, Jacqueline Hernández-Damián, Gustavo Ignacio Vázquez-Cervantes, Ismael Torres, José Pedraza-Chaverri
      It was explored the cytoprotective and antioxidant effect of MLN4924, a specific inhibitor of Nedd8-activating enzyme (NAE), against hydrogen peroxide (H2O2)-induced damage in cerebellar granule neurons (CGNs). Primary cultures of CGNs were exposed to H2O2 after preincubation with MLN4924. The compounds were removed, and CGNs were incubated in culture medium for 24h in order to determine cell viability by 3-[4,5-dimethylthiazol-2-yl)]-2,5-diphenyl-tetrazolium bromide (MTT) and fluorescein diacetate (FDA) assays. It was demonstrated that MLN4924 remarkably attenuated H2O2-induced cell damage. Meanwhile reactive oxygen species (ROS) production was evaluated with the fluorescent probe dihydroethidium (DHE). Interestingly H2O2-induced ROS production was inhibited by pretreatment with MLN4924. MLN4924 treatment in CGNs resulted in nuclear factor E2-related factor 2 (Nrf2) protein accumulation. Intriguingly this effect was observed in the cytosolic and nuclear compartments of the CGNs. The cytoprotective effect of MLN4924 was associated with its ability to diminish ROS production induced by H2O2 and the accumulation of Nrf2 protein levels in the cytoplasm and nucleus of the CGNs.
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      PubDate: 2016-03-02T08:33:30Z
       
  • Hemolytic and antimalarial effects of tight-binding glyoxalase 1
           inhibitors on the host-parasite unit of erythrocytes infected with
           plasmodium falciparum

    • Abstract: Publication date: Available online 2 March 2016
      Source:Redox Biology
      Author(s): Cletus A. Wezena, Miriam Urscher, Robert Vince, Swati S. More, Marcel Deponte
      Glyoxalases prevent the formation of advanced glycation end products by converting glycolysis-derived methylglyoxal to d-lactate with the help of glutathione. Vander Jagt and colleagues previously showed that erythrocytes release about thirty times more d-lactate after infection with the human malaria parasite Plasmodium falciparum. Functional glyoxalases in the host-parasite unit might therefore be crucial for parasite survival. Here, we determined the antimalarial and hemolytic activity of two tight-binding glyoxalase inhibitors using infected and uninfected erythrocytes. In addition, we synthesized and analyzed a set of diester derivates of both tight-binding inhibitors resulting in up to threefold lower IC50 values and an altered methemoglobin formation and hemolytic activity depending on the type of ester. Inhibitor treatments of uninfected erythrocytes revealed an extremely slow inactivation of the host cell glyoxalase, irrespective of inhibitor modifications, and a potential dispensability of the host cell enzyme for parasite survival. Our study highlights the benefits and drawbacks of different esterifications of glutathione-derived inhibitors and demonstrates the suitability of glyoxalase inhibitors as a tool for deciphering the relevance and mode of action of different glyoxalase systems in a host-parasite unit.
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      PubDate: 2016-03-02T08:33:30Z
       
  • TEMPOL increases NAD+ and improves redox imbalance in obese mice

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Mayumi Yamato, Kimika Kawano, Yuki Yamanaka, Misako Saiga, Ken-ichi Yamada
      Continuous energy conversion is controlled by reduction–oxidation (redox) processes. NAD+ and NADH represent an important redox couple in energy metabolism. 4-Hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl (TEMPOL) is a redox-cycling nitroxide that promotes the scavenging of several reactive oxygen species (ROS) and is reduced to hydroxylamine by NADH. TEMPOL is also involved in NAD+ production in the ascorbic acid–glutathione redox cycle. We utilized the chemical properties of TEMPOL to investigate the effects of antioxidants and NAD+/NADH modulators on the metabolic imbalance in obese mice. Increases in the NAD+/NADH ratio by TEMPOL ameliorated the metabolic imbalance when combined with a dietary intervention, changing from a high-fat diet to a normal diet. Plasma levels of the superoxide marker dihydroethidium were higher in mice receiving the dietary intervention compared with a control diet, but were normalized with TEMPOL consumption. These findings provide novel insights into redox regulation in obesity.
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      PubDate: 2016-03-02T08:33:30Z
       
  • Induction of mitochondrial reactive oxygen species production by GSH
           mediated S-glutathionylation of 2-oxoglutarate dehydrogenase

    • Abstract: Publication date: Available online 17 February 2016
      Source:Redox Biology
      Author(s): Ryan J. Mailloux, D. Craig Ayre, Sherri L. Christian
      2-oxoglutarate dehydrogenase (Ogdh) is an important mitochondria redox sensor that can undergo S-glutathionylation following an increase in H2O2 levels. Although S-glutathionylation is required to protect Ogdh from irreversible oxidation while simultaneously modulating its activity it remains unknown if glutathione can also modulate reactive oxygen species (ROS) production by the complex. We report that reduced (GSH) and oxidized (GSSG) glutathione control O2∙-/H2O2 formation by Ogdh through protein S-glutathionylation reactions. GSSG (1mM) induced a modest decrease in Ogdh activity which was associated with a significant decrease in O2∙-/H2O2 formation. GSH had the opposite effect, amplifying O2∙-/H2O2 formation by Ogdh. Incubation of purified Ogdh in 2.5mM GSH led to significant increase in O2∙-/H2O2 formation which also lowered NADH production. Inclusion of enzymatically active glutaredoxin-2 (Grx2) in reaction mixtures reversed the GSH-mediated amplification of O2∙-/H2O2 formation. Similarly pre-incubation of permeabilized liver mitochondria from mouse depleted of GSH showed an approximately ~3.5-fold increase in Ogdh-mediated O2∙-/H2O2 production that was matched by a significant decrease in NADH formation which could be reversed by Grx2. Taken together, our results demonstrate GSH and GSSG modulate ROS production by Ogdh through S-glutathionylation of different subunits. This is also the first demonstration that GSH can work in the opposite direction in mitochondria-amplifying ROS formation instead of quenching it. We propose that this regulatory mechanism is required to modulate ROS emission from Ogdh in response to variations in glutathione redox buffering capacity.
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      PubDate: 2016-02-20T07:32:53Z
       
  • Lung extracellular matrix and redox regulation

    • Abstract: Publication date: Available online 18 February 2016
      Source:Redox Biology
      Author(s): Walter H. Watson, Jeffrey D. Ritzenthaler, Jesse Roman
      Pulmonary fibrosis affects millions worldwide and, even though there has been a significant investment in understanding the processes involved in wound healing and maladaptive repair, a complete understanding of the mechanisms responsible for lung fibrogenesis eludes us, and interventions capable of reversing or halting disease progression are not available. Pulmonary fibrosis is characterized by the excessive expression and uncontrolled deposition of extracellular matrix (ECM) proteins resulting in erosion of the tissue structure. Initially considered an ‘end-stage’ process elicited after injury, these events are now considered pathogenic and are believed to contribute to the course of the disease. By interacting with integrins capable of signal transduction and by influencing tissue mechanics, ECM proteins modulate processes ranging from cell adhesion and migration to differentiation and growth factor expression. In doing so, ECM proteins help orchestrate complex developmental processes and maintain tissue homeostasis. However, poorly controlled deposition of ECM proteins promotes inflammation, fibroproliferation, and aberrant differentiation of cells, and has been implicated in the pathogenesis of pulmonary fibrosis, atherosclerosis and cancer. Considering their vital functions, ECM proteins are the target of investigation, and oxidation-reduction (redox) reactions have emerged as important regulators of the ECM. Oxidative stress invariably accompanies lung disease and promotes ECM expression directly or through the overproduction of pro-fibrotic growth factors, while affecting integrin binding and activation. In vitro and in vivo investigations point to redox reactions as targets for intervention in pulmonary fibrosis and related disorders, but studies in humans have been disappointing probably due to the narrow impact of the interventions tested, and our poor understanding of the factors that regulate these complex reactions. This review is not meant to provide a comprehensive review of this field, but rather to highlight what has been learned and to raise interest in this area in need of much attention.
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      PubDate: 2016-02-20T07:32:53Z
       
  • Direct spectrophotometric measurement of supra-physiological levels of
           ascorbate in plasma

    • Abstract: Publication date: Available online 13 February 2016
      Source:Redox Biology
      Author(s): Jordan R. Witmer, Bailey J. Wetherell, Brett A. Wagner, Juan Du, Joseph J. Cullen, Garry R. Buettner
      Background Supra-physiological concentrations of ascorbate, vitamin C, in blood, greater than 1mM, achieved through intravenous administration (IV), are being tested in clinical trials to treat human disease, e.g. cancer. These trials need information on the high levels of ascorbate achieved in blood upon IV administration of pharmacological ascorbate so appropriate clinical decisions can be made. Methods Here we demonstrate that in the complex matrix of human blood plasma supra-physiological levels of ascorbate can be quantified by direct UV spectroscopy with use of a microvolume UV-Vis spectrophotometer. Results Direct quantitation of ascorbate in plasma in the range of 2.9mM, lower limit of detection, up to at least 35mM can be achieved without any sample processing, other than centrifugation. Conclusions This approach is rapid, economical, and can be used to quantify supraphysiological blood levels of AscH- associated with the use of IV administration of pharmacological AscH- to treat disease.
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      PubDate: 2016-02-15T06:58:34Z
       
  • Reduction of PCN biosynthesis by NO in Pseudomonas aeruginosa

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Lei Gao, Yuying Zhang, Yan Wang, Xinhua Qiao, Jing Zi, Chang Chen, Yi Wan
      Pyocyanin (PCN), a virulence factor synthesized by Pseudomonas aeruginosa, plays an important role during clinical infections. There is no study of the effect of nitric oxide (NO) on PCN biosynthesis. Here, the effect of NO on PCN levels in Pseudomonas aeruginosa strain PAO1, a common reference strain, was tested. The results showed that the NO donor sodium nitroprusside (SNP) can significantly reduce PCN levels (82.5% reduction at 60μM SNP). Furthermore, the effect of endogenous NO on PCN was tested by constructing PAO1 nor (NO reductase gene) knockout mutants. Compared to the wild-type strain, the Δnor strain had a lower PCN (86% reduction in Δnor). To examine whether the results were universal with other P. aeruginosa strains, we collected 4 clinical strains from a hospital, tested their PCN levels after SNP treatment, and obtained similar results, i.e., PCN biosynthesis was inhibited by NO. These results suggest that NO treatment may be a new strategy to inhibit PCN biosynthesis and could provide novel insights into eliminating P. aeruginosa virulence as a clinical goal.
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      PubDate: 2016-02-15T06:58:34Z
       
  • Complex coordinated extracellular metabolism: Acid phosphatases activate
           diluted human leukocyte proteins to generate energy flow as NADPH from
           nucleotide ribose

    • Abstract: Publication date: Available online 2 February 2016
      Source:Redox Biology
      Author(s): John B. Hibbs, Zdenek Vavrin, James E. Cox
      Complex metabolism is thought to occur exclusively in the crowded intracellular environment. Here we report that diluted enzymes from lysed human leukocytes produce extracellular energy. Our findings involve two pathways: the purine nucleotide catabolic pathway and the pentose phosphate pathway, which function together to generate energy as NADPH. Glucose6P fuel for NADPH production is generated from structural ribose of purine ribonucleoside monophosphates, ADP, and ADP-ribose. NADPH drives glutathione reductase to reduce an oxidized glutathione disulfide-glutathione redox couple. Acid phosphatases initiate ribose5P salvage from purine ribonucleoside monophosphates, and transaldolase controls the direction of carbon chain flow through the nonoxidative branch of the pentose phosphate pathway. These metabolic control points are regulated by pH. Biologically, this energy conserving metabolism could function in perturbed extracellular spaces.
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      PubDate: 2016-02-10T06:42:37Z
       
  • Value of the 8-oxodG/dG ratio in chronic liver inflammation of patients
           with hepatocellular carcinoma

    • Abstract: Publication date: Available online 9 February 2016
      Source:Redox Biology
      Author(s): Pengcheng Li, Grant A Ramm, Graeme A Macdonald
      The aim of this study was to examine the role of oxidative DNA damage in chronic liver inflammation in the evolution of hepatocellular carcinoma. The accumulated data demonstrated that oxidative DNA damage and chronic liver inflammation are involved in the transformation of normal hepatocytes and their evolution towards hepatocellular carcinoma. However, the levels of 8-oxy−2′-deoxy-guanosine (8-oxodG), a biomarker of oxidative DNA damage, were overestimated and underestimated in previous reports due to various technical limitations. The current techniques are not suitable to analyze the 8-oxodG levels in the non-malignant liver tissues and tumors of hepatocellular carcinoma patients unless they are modified. Therefore, in this study, the protocols for extraction and hydrolysis of DNA were optimized using 54 samples from hepatocellular carcinoma patients with various risk factors, and the 8-oxodG and 2′-deoxyguanosine (dG) levels were measured. The patients enrolled in the study include 23 from The Princess Alexandra Hospital and The Royal Brisbane and Women’s Hospitals, Brisbane, Australia, and 31 from South Africa. This study revealed that the 8-oxodG/dG ratios tended to be higher in most non-malignant liver tissues compared to hepatocellular carcinoma tissue (p=0.2887). It also appeared that the ratio was higher in non-malignant liver tissue from Southern African patients (p=0.0479), but there was no difference in the 8-oxodG/dG ratios between non-malignant liver tissues and tumors of Australian hepatocellular carcinoma patients (p=0.7722). Additionally, this study also revealed a trend for a higher 8-oxodG/dG ratio in non-malignant liver tissues compared to tumoural tissues of patients with HBV. Significant differences were not observed in the 8-oxodG/dG ratios between non-cirrhotic and cirrhotic non-malignant liver tissues.
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      PubDate: 2016-02-10T06:42:37Z
       
  • In vivo evaluation of different alterations of redox status by studying
           pharmacokinetics of nitroxides using magnetic resonance techniques

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Goran Bačić, Aleksandra Pavićević, Fabienne Peyrot
      Free radicals, particularly reactive oxygen species (ROS), are involved in various pathologies, injuries related to radiation, ischemia-reperfusion or ageing. Unfortunately, it is virtually impossible to directly detect free radicals in vivo, but the redox status of the whole organism or particular organ can be studied in vivo by using magnetic resonance techniques (EPR and MRI) and paramagnetic stable free radicals – nitroxides. Here we review results obtained in vivo following the pharmacokinetics of nitroxides on experimental animals (and a few in humans) under various conditions. The focus was on conditions where the redox status has been altered by induced diseases or harmful agents, clearly demonstrating that various EPR/MRI/nitroxide combinations can reliably detect metabolically induced changes in the redox status of organs. These findings can improve our understanding of oxidative stress and provide a basis for studying the effectiveness of interventions aimed to modulate oxidative stress. Also, we anticipate that the in vivo EPR/MRI approach in studying the redox status can play a vital role in the clinical management of various pathologies in the years to come providing the development of adequate equipment and probes.
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      PubDate: 2016-01-30T21:09:54Z
       
  • Redox Homeostasis: The Golden Mean of Healthy Living

    • Abstract: Publication date: Available online 19 January 2016
      Source:Redox Biology
      Author(s): Fulvio Ursini, Matilde Maiorino, Henry Jay Forman
      The notion that electrophiles serve as messengers in cell signaling is now widely accepted. Nonetheless, major issues restrain acceptance of redox homeostasis and redox signaling as components of maintenance of a normal physiological steady state. The first is that redox signaling requires sudden switching on of oxidant production and bypassing of antioxidant mechanisms rather than a continuous process that, like other signaling mechanisms, can be smoothly turned up or down. The second is the misperception that reactions in redox signaling involve “reactive oxygen species” rather than reaction of specific electrophiles with specific protein thiolates. The third is that hormesis provides protection against oxidants by increasing cellular defense or repair mechanisms rather than by specifically addressing the offset of redox homeostasis. Instead, we propose that both oxidant and antioxidant signaling are main features of redox homeostasis. As the redox shift is rapidly reversed by feedback reactions, homeostasis is maintained by continuous signaling for production and elimination of electrophiles and nucleophiles. Redox homeostasis, which is the maintenance of nucleophilic tone, accounts for a healthy physiological steady state. Electrophiles and nucleophiles are not intrinsically harmful or protective, and redox homeostasis is an essential feature of both the response to challenges and subsequent feedback. While the balance between oxidants and nucleophiles is preserved in redox homeostasis, oxidative stress provokes the establishment of a new radically altered redox steady state. The popular belief that scavenging free radicals by antioxidants has a beneficial effect is wishful thinking. We propose, instead, that continuous feedback preserves nucleophilic tone and that this is supported by redox active nutritional phytochemicals. These nonessential compounds, by activating Nrf2, mimic the effect of endogenously produced electrophiles (parahormesis). In summary, while hormesis, although globally protective, results in setting up of a new phenotype, parahormesis contributes to health by favoring maintenance of homeostasis.
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      PubDate: 2016-01-23T20:21:28Z
       
  • Manipulation of environmental oxygen Modifies Reactive OXYGEN AND Nitrogen
           Species Generation DURING Myogenesis

    • Abstract: Publication date: Available online 21 January 2016
      Source:Redox Biology
      Author(s): Rachel McCormick, Timothy Pearson, Aphrodite Vasilaki
      Regulated changes in reactive oxygen and nitrogen species (RONS) activities are important in maintaining the normal sequence and development of myogenesis. Both excessive formation and reduction in RONS have been shown to affect muscle differentiation in a negative way. Cultured cells are typically grown in 20% O2 but this is not an appropriate physiological concentration for a number of cell types, including skeletal muscle. The aim was to examine the generation of RONS in cultured skeletal muscle cells under a physiological oxygen concentration condition (6% O2) and determine the effect on muscle myogenesis. Primary mouse satellite cells were grown in 20% or 6% O2 environments and RONS activity was measured at different stages of myogenesis by real-time fluorescent microscopy using fluorescent probes with different specificities i.e. dihydroethidium (DHE), 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM DA) and 5-(and-6)-chloromethyl-2′,7′ -dichlorodihydrofluorescein diacetate (CM-DCFH-DA). Data demonstrate that satellite cell proliferation increased when cells were grown in 6% O2 compared with 20% O2. Myoblasts grown in 20% O2 showed an increase in DCF fluorescence and DHE oxidation compared with myoblasts grown at 6% O2. Myotubes grown in 20% O2 also showed an increase in DCF and DAF-FM fluorescence and DHE oxidation compared with myotubes grown in 6% O2. The catalase and MnSOD contents were also increased in myoblasts and myotubes that were maintained in 20% O2 compared with myoblasts and myotubes grown in 6% O2. These data indicate that intracellular RONS activities in myoblasts and myotubes at rest are influenced by changes in environmental oxygen concentration and that the increased ROS may influence myogenesis in a negative manner.
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      PubDate: 2016-01-23T20:21:28Z
       
  • Low sulfide levels and a high degree of cystathionine β-synthase
           (CBS) activation by S-adenosylmethionine (SAM) in the long-lived naked
           mole-rat

    • Abstract: Publication date: Available online 14 January 2016
      Source:Redox Biology
      Author(s): Maja Dziegelewska, Susanne Holtze, Christiane Vole, Ulrich Wachter, Uwe Menzel, Michaela Morhart, Marco Groth, Karol Szafranski, Arne Sahm, Christoph Sponholz, Philip Dammann, Klaus Huse, Thomas Hildebrandt, Matthias Platzer
      Hydrogen sulfide (H2S) is a gaseous signalling molecule involved in many physiological and pathological processes. There is increasing evidence that H2S is implicated in aging and lifespan control in the diet-induced longevity models. However, blood sulfide concentration of naturally long-lived species is not known. Here we measured blood sulfide in the long-lived naked mole-rat and five other mammalian species considerably differing in lifespan and found a negative correlation between blood sulfide and maximum longevity residual. In addition, we show that the naked mole-rat cystathionine β-synthase (CBS), an enzyme whose activity in the liver significantly contributes to systemic sulfide levels, has lower activity in the liver and is activated to a higher degree by S-adenosylmethionine compared to other species. These results add complexity to the understanding of the role of H2S in aging and call for detailed research on naked mole-rat transsulfuration.
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      PubDate: 2016-01-18T19:54:04Z
       
  • Effects of High Fat Diets on Rodent Liver Bioenergetics and Oxidative
           Imbalance

    • Abstract: Publication date: Available online 14 January 2016
      Source:Redox Biology
      Author(s): Pâmela A. Kakimoto, Alicia J. Kowaltowski
      Human metabolic diseases can be mimicked in rodents by using dietary interventions such as high fat diets (HFD). Nonalcoholic fatty liver disease (NAFLD) develops as a result of HFD and the disease may progress in a manner involving increased production of oxidants. The main intracellular source of these oxidants are mitochondria, which are also responsible for lipid metabolism and thus widely recognized as important players in the pathology and progression of steatosis. Here, we review publications that study redox and bioenergetic effects of HFD in the liver. We find that dietary composition and protocol implementations vary widely, as do the results of these dietary interventions. Overall, all HFD promote steatosis, changes in β-oxidation, generation and consequences of oxidants, while effects on body weight, insulin signaling and other bioenergetic parameters are more variable with the experimental models adopted. Our review provides a broad analysis of the bioenergetic and redox changes promoted by HFD as well as suggestions for changes and specifications in methodologies that may help explain apparent disparities in the current literature.
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      PubDate: 2016-01-18T19:54:04Z
       
  • Molecular regulatory mechanisms of osteoclastogenesis through
           cytoprotective enzymes

    • Abstract: Publication date: Available online 11 January 2016
      Source:Redox Biology
      Author(s): Hiroyuki Kanzaki, Fumiaki Shinohara, Itohiya-Kasuya Kanako, Yuuki Yamaguchi, Sari Fukaya, Yutaka Miyamoto, Satoshi Wada, Yoshiki Nakamura
      It has been reported that reactive oxygen species (ROS), such as hydrogen peroxide and superoxide, take part in osteoclast differentiation as intra-cellular signaling molecules. The current assumed signaling cascade from RANK to ROS production is RANK, TRAF6, Rac1, and then Nox. The target molecules of ROS in RANKL signaling remain unclear; however, several reports support the theory that NF-κB signaling could be the crucial downstream signaling molecule of RANKL-mediated ROS signaling. Furthermore, ROS exert cytotoxic effects such as peroxidation of lipids and phospholipids and oxidative damage to proteins and DNA. Therefore, cells have several protective mechanisms against oxidative stressors that mainly induce cytoprotective enzymes and ROS scavenging. Three well-known mechanisms regulate cytoprotective enzymes including Nrf2-, FOXO-, and sirtuin-dependent mechanisms. Several reports have indicated a crosslink between FOXO- and sirtuin-dependent regulatory mechanisms. The agonists against the regulatory mechanisms are reported to induce these cytoprotective enzymes successfully. Some of them inhibit osteoclast differentiation and bone destruction via attenuation of intracellular ROS signaling. In this review article, we discuss the above topics and summarize the current information available on the relationship between cytoprotective enzymes and osteoclastogenesis.
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      PubDate: 2016-01-13T19:01:31Z
       
  • Prominent role of DPP III in the E2-mediated protection against hyperoxia
           in liver of female CBA/H mice

    • Abstract: Publication date: Available online 11 January 2016
      Source:Redox Biology
      Author(s): Sandra Sobočanec, Vedrana Filić, Mihaela Matovina, Dragomira Majhen, Željka Mačak Šafranko, Marijana Popović Hadžija, Željka Krsnik, Andrea Gudan Kurilj, Ana Šarić, Marija Abramić, Tihomir Balog
      A number of age-related diseases have a low incidence in females, which is attributed to a protective effect of sex hormones. For instance, the female sex hormone estrogen (E2) has a well established cytoprotective effect against oxidative stress, which strongly contributes to ageing. However, the mechanism by which E2 exerts its protective activity remains elusive. In this study we address the question whether the E2-induced protective effect against hyperoxia is mediated by the Nrf-2/Keap-1 signaling pathway. In particular, we investigate the E2-induced expression and cellular distribution of DPP III monozinc exopeptidase, a member of the Nrf-2/Keap-1 pathway, upon hyperoxia treatment. We find that DPP III accumulates in the nucleus in response to hyperoxia. Further, we show that combined induction of hyperoxia and E2 administration have an additive effect on the nuclear accumulation of DPP III. The level of nuclear accumulation of DPP III is comparable to nuclear accumulation of Nrf-2 in healthy female mice exposed to hyperoxia. In ovariectomized females exposed to hyperoxia, supplementation of E2 induced upregulation of DPP III, Ho-1, Sirt-1 and downregulation of Ppar-γ. While other cytoprotective mechanisms cannot be excluded, these findings demonstrate a prominent role of DPP III, along with Sirt-1, in the E2-mediated protection against hyperoxia.
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      PubDate: 2016-01-13T19:01:31Z
       
  • Lipidomics and H218O labeling techniques reveal increased remodeling of
           DHA-containing membrane phospholipids associated with abnormal locomotor
           responses in α-tocopherol deficient zebrafish (danio rerio) embryos

    • Abstract: Publication date: Available online 11 January 2016
      Source:Redox Biology
      Author(s): Melissa Q. McDougall, Jaewoo Choi, Jan F. Stevens, Lisa Truong, Robert L. Tanguay, Maret G. Traber
      We hypothesized that vitamin E (α-tocopherol) is required by the developing embryonic brain to prevent depletion of highly polyunsaturated fatty acids, especially docosahexaenoic acid (DHA, 22:6), the loss of which we predicted would underlie abnormal morphological and behavioral outcomes. Therefore, we fed adult 5D zebrafish (Danio rerio) defined diets without (E-) or with added α-tocopherol (E+, 500mg RRR-α-tocopheryl acetate/kg diet) for a minimum of 80 days, and then spawned them to obtain E- and E+ embryos. The E- compared with E+ embryos were 82% less responsive (p<0.01) to a light/dark stimulus at 96hours post-fertilization (hpf), demonstrating impaired locomotor behavior, even in the absence of gross morphological defects. Evaluation of phospholipid (PL) and lysophospholipid (lyso-PL) composition using untargeted lipidomics in E- compared with E+ embryos at 24, 48, 72, and 120 hpf showed that four PLs and three lyso-PLs containing docosahexaenoic acid (DHA), including lysophosphatidylcholine (LPC 22:6, required for transport of DHA into the brain, p<0.001), were at lower concentrations in E- at all time-points. Additionally, H2 18O labeling experiments revealed enhanced turnover of LPC 22:6 (p<0.001) and three other DHA-containing PLs in the E- compared with the E+ embryos, suggesting that increased membrane remodeling is a result of PL depletion. Together, these data indicate that α-tocopherol deficiency in the zebrafish embryo causes the specific depletion and increased turnover of DHA-containing PL and lyso-PLs, which may compromise DHA delivery to the brain and thereby contribute to the functional impairments observed in E- embryos.
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      PubDate: 2016-01-13T19:01:31Z
       
  • Cross-talk between two antioxidants, thioredoxin reductase and heme
           oxygenase-1, and therapeutic implications for multiple myeloma

    • Abstract: Publication date: Available online 11 January 2016
      Source:Redox Biology
      Author(s): Prahlad V. Raninga, Giovanna Di Trapani, Slavica Vuckovic, Kathryn F. Tonissen
      Multiple myeloma (MM) is characterized by an accumulation of abnormal clonal plasma cells in the bone marrow. Despite recent advancements in anti-myeloma therapies, MM remains an incurable disease. Antioxidant molecules are upregulated in many cancers, correlating with tumor proliferation, survival, and chemoresistance and therefore, have been suggested as potential therapeutic targets. This study investigated the cross-talk between two antioxidant molecules, thioredoxin reductase (TrxR) and heme oxygenase-1 (HO-1), and their therapeutic implications in MM. We found that although auranofin, a TrxR inhibitor, significantly inhibited TrxR activity by more than 50% at lower concentrations, myeloma cell proliferation was only inhibited at higher concentrations of auranofin. Inhibition of TrxR using lower auranofin concentrations induced HO-1 protein expression in myeloma cells. Using a sub-lethal concentration of auranofin to inhibit TrxR activity in conjunction with HO-1 inhibition significantly decreased myeloma cell growth and induced apoptosis. TrxR was shown to regulate HO-1 via the Nrf2 signaling pathway in a ROS-dependent manner. Increased HO-1 mRNA levels were observed in bortezomib-resistant myeloma cells compared to parent cells and HO-1 inhibition restored the sensitivity to bortezomib in bortezomib-resistant myeloma cells. These findings indicate that concurrent inhibition of HO-1 with either a TrxR inhibitor or with bortezomib would improve therapeutic outcomes in MM patients. Hence, our findings further support the need to target multiple antioxidant systems alone or in combination with other therapeutics to improve therapeutic outcomes in MM patients.
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      PubDate: 2016-01-13T19:01:31Z
       
  • MELATONIN AND l-CARNITIN IMPROVES ENDOTHELIAL DISFUNCTION AND OXIDATIVE
           STRESS IN TYPE 2 DIABETİC RATS

    • Abstract: Publication date: Available online 13 January 2016
      Source:Redox Biology
      Author(s): Derya Selcen Salmanoglu, Tugba Gurpinar, Kamil Vural, Nuran Ekerbicer, Ertan Darıverenli, Ahmet Var
      Vascular dysfunction is thought to play a major role in the development of diabetic cardiovascular disease. The roles of endothelial dysfunction, oxidative stress, and dyslipidemia will be considered. Melatonin as well as L-carnitine were shown to possess strong antioxidant properties. Diabetes induced with high fat diet (for 8 weeks) and multipl low doses intraperitoneal injection of STZ (twice, 30mg/kg/d i.p). The diabetic animals were randomly assigned to one of the experimental groups as follows: Control group (C), high fat diet (HFD), STZ-induced diabetic group (HFD + STZ) , HFD + STZ diabetic group received melatonin (10mg/kg/d i.p), HFD + STZ diabetic group received L-carnitine (0.6g/kg/d i.p), and HFD +STZ diabetic group received glibenclamide (5mg/kg/d, oral). The serum fasting blood glucose, insulin, total cholesterol, HDL- cholesterol, LDL-cholesterol, triglyceride and malondialdehyde (MDA) levels were tested. Acetylcholine induced endothelium-dependent relaxation and sodium nitroprusside induced endothelium-independent relaxation were measured in aortas for estimating endothelial function. Also, glutathione peroxidase (GPx), superoxide dismutase (SOD) and nitric oxide (NO) levels activities were determined in rat liver. According to our results melatonin and L-carnitine treatment decreased fasting blood glucose, total cholesterol, and LDL levels. MDA levels significantly decreased with the melatonin treatment whereas SOD levels were not significantly changed between the groups. The results suggest that especially melatonin restores the vascular responses and endothelial dysfunction in diabetes.
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      PubDate: 2016-01-13T19:01:31Z
       
  • Plasma lipid oxidation induced by peroxynitrite, hypochlorite,
           lipoxygenase and peroxyl radicals and its inhibition by antioxidants as
           assessed by diphenyl-1-pyrenylphosphine

    • Abstract: Publication date: Available online 11 January 2016
      Source:Redox Biology
      Author(s): Mayuko Morita, Yuji Naito, Toshikazu Yoshikawa, Etsuo Niki
      Lipid oxidation has been implicated in the pathogenesis of many diseases. Lipids are oxidized in vivo by several different oxidants to give diverse products, in general lipid hydroperoxides as the major primary product. In the present study, the production of lipid hydroperoxides in the oxidation of mouse plasma induced by multiple oxidants was measured using diphenyl-1-pyrenylphosphine (DPPP) as a probe. DPPP itself is not fluorescent, but it reacts with lipid hydroperoxides stochiometrically to give highly fluorescent DPPP oxide and lipid hydroxides. The production of lipid hydroperoxides could be followed continuously in the oxidation of plasma induced by peroxynitrite, hypochlorite, 15-lipoxygenase, and peroxyl radicals with a microplate reader. A clear lag phase was observed in the plasma oxidation mediated by aqueous peroxyl radicals and peroxynitrite, but not in the oxidation induced by hypochlorite and lipoxygenase. The effects of several antioxidants against lipid oxidation induced by the above oxidants were assessed. The efficacy of antioxidants was dependent markedly on the type of oxidants. α-Tocopherol exerted potent antioxidant effects against peroxyl radical-mediated lipid peroxidation, but it did not inhibit lipid oxidation induced by peroxynitrite, hypochlorite, and 15-lipoxygenase efficiently, suggesting that multiple antioxidants with different selectivities are required for the inhibition of plasma lipid oxidation in vivo. This is a novel, simple and most high throughput method to follow plasma lipid oxidation induced by different oxidants and also to assess the antioxidant effects in biologically relevant settings.
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      PubDate: 2016-01-13T19:01:31Z
       
  • Mitochondrial thiol modification by a targeted electrophile inhibits
           metabolism in breast adenocarcinoma cells by inhibiting enzyme activity
           and protein levels

    • Abstract: Publication date: Available online 8 January 2016
      Source:Redox Biology
      Author(s): M. Ryan Smith, Praveen K. Vayalil, Fen Zhou, Gloria A. Benavides, Reena R. Beggs, Hafez Golzarian, Bhavitavya Nijampatnam, Patsy G. Oliver, Robin A.J. Smith, Michael P. Murphy, Sadanandan E. Velu, Aimee Landar
      Many cancer cells follow an aberrant metabolic program to maintain energy for rapid cell proliferation. Metabolic reprogramming often involves the upregulation of glutaminolysis to generate reducing equivalents for the electron transport chain and amino acids for protein synthesis. Critical enzymes involved in metabolism possess a reactive thiolate group, which can be modified by certain oxidants. In the current study, we show that modification of mitochondrial protein thiols by a model compound, iodobutyl triphenylphosphonium (IBTP), decreased mitochondrial metabolism and ATP in MDA-MB 231 (MB231) breast adenocarcinoma cells up to 6 days after an initial 24h treatment. Mitochondrial thiol modification also depressed oxygen consumption rates (OCR) in a dose-dependent manner to a greater extent than a non-thiol modifying analog, suggesting that thiol reactivity is an important factor in the inhibition of cancer cell metabolism. In non-tumorigenic MCF10A cells, IBTP also decreased OCR; however the extracellular acidification rate was significantly increased at all but the highest concentration (10µM) of IBTP indicating that thiol modification can have significantly different effects on bioenergetics in tumorigenic versus non-tumorigenic cells. ATP and other adenonucleotide levels were also decreased by thiol modification up to 6 days post-treatment, indicating a decreased overall energetic state in MB231 cells. Cellular proliferation of MB231 cells was also inhibited up to 6 days post-treatment with little change to cell viability. Targeted metabolomic analyses revealed that thiol modification caused depletion of both Krebs cycle and glutaminolysis intermediates. Further experiments revealed that the activity of the Krebs cycle enzyme, aconitase, was attenuated in response to thiol modification. Additionally, the inhibition of glutaminolysis corresponded to decreased glutaminase C (GAC) protein levels, although other protein levels were unaffected. This study demonstrates for the first time that mitochondrial thiol modification inhibits metabolism via inhibition of both aconitase and GAC in a breast cancer cell model.
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      PubDate: 2016-01-08T18:24:37Z
       
  • Effects of the isoflavone prunetin on gut health and stress response in
           male drosophila melanogaster

    • Abstract: Publication date: Available online 6 January 2016
      Source:Redox Biology
      Author(s): Stefanie Piegholdt, Gerald Rimbach, Anika Eva Wagner
      The traditional Asian diet is rich in fruits, vegetables and soy, the latter representing a significant source of dietary isoflavones. The isoflavone prunetin was recently identified to improve intestinal epithelial barrier function in vitro and to ameliorate general survival and overall health state in vivo in male Drosophila melanogaster. However, the prunetin-mediated health benefits in the fruit fly were ascertained under standard living conditions. As the loss of intestinal integrity is closely related to a reduction in Drosophila lifespan and barrier dysfunction increases with age, effects on prunetin-modulated gut health under oxidative or pathogenic stress provocation remain to be elucidated. In this study, male adult Drosophila melanogaster were administered either a prunetin or a control diet. Gut-derived junction protein expression and pathogen-induced antimicrobial peptide expressions as well as the stem cell proliferation in the gut were evaluated. Furthermore, survival following exposure to hydrogen peroxide was assessed. Prunetin ingestion did not attenuate bacterial infection and did not protect flies from oxidative stress. Intestinal mRNA expression levels of adherence and septate junction proteins as well as the stem cell proliferation were not altered by prunetin intake. Prunetin does not improve the resistance of flies against severe injuring, exogenous stress and therefore seems to function in a preventive rather than a therapeutic approach since the health-promoting benefits appear to be exclusively restricted to normal living circumstances.
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      PubDate: 2016-01-08T18:24:37Z
       
  • Cellular and subcellular oxidative stress parameters following severe
           spinal cord injury

    • Abstract: Publication date: Available online 30 December 2015
      Source:Redox Biology
      Author(s): Nishant P. Visavadiya, Samir P. Patel, Jenna L. VanRooyen, Patrick G. Sullivan, Alexander G. Rabchevsky
      The present study undertook a comprehensive assessment of the acute biochemical oxidative stress parameters in both cellular and, notably, mitochondrial isolates following severe upper lumbar contusion spinal cord injury (SCI) in adult female Sprague Dawley rats. At 24h post-injury, spinal cord tissue homogenate and mitochondrial fractions were isolated concurrently and assessed for glutathione (GSH) content and production of nitric oxide (NO•), in addition to the presence of oxidative stress markers 3-nitrotyrosine (3-NT), protein carbonyl (PC), 4-hydroxynonenal (4-HNE) and lipid peroxidation (LPO). Moreover, we assessed production of superoxide (O2 •¯) and hydrogen peroxide (H2O2) in mitochondrial fractions. Quantitative biochemical analyses showed that compared to sham, SCI significantly lowered GSH content accompanied by increased NO• production in both cellular and mitochondrial fractions. SCI also resulted in increased O2 •¯ and H2O2 levels in mitochondrial fractions. Western blot analysis further showed that reactive oxygen/nitrogen species (ROS/RNS) mediated PC and 3-NT production were significantly higher in both fractions after SCI. Conversely, neither 4-HNE levels nor LPO formation were increased at 24h after injury in either tissue homogenate or mitochondrial fractions. These results indicate that by 24h post-injury ROS-induced protein oxidation is more prominent compared to lipid oxidation, indicating a critical temporal distinction in secondary pathophysiology that is critical in designing therapeutic approaches to mitigate consequences of oxidative stress.
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      PubDate: 2016-01-03T17:47:57Z
       
  • Interplay between oxidant species and energy metabolism

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Celia Quijano, Madia Trujillo, Laura Castro, Andrés Trostchansky
      It has long been recognized that energy metabolism is linked to the production of reactive oxygen species (ROS) and critical enzymes allied to metabolic pathways can be affected by redox reactions. This interplay between energy metabolism and ROS becomes most apparent during the aging process and in the onset and progression of many age-related diseases (i.e. diabetes, metabolic syndrome, atherosclerosis, neurodegenerative diseases). As such, the capacity to identify metabolic pathways involved in ROS formation, as well as specific targets and oxidative modifications is crucial to our understanding of the molecular basis of age-related diseases and for the design of novel therapeutic strategies. Herein we review oxidant formation associated with the cell's energetic metabolism, key antioxidants involved in ROS detoxification, and the principal targets of oxidant species in metabolic routes and discuss their relevance in cell signaling and age-related diseases.
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      PubDate: 2016-01-03T17:47:57Z
       
  • Protein S-glutathionlyation links energy metabolism to redox signaling in
           mitochondria

    • Abstract: Publication date: Available online 31 December 2015
      Source:Redox Biology
      Author(s): Ryan J. Mailloux, Jason R. Treberg
      At its core mitochondrial function relies on redox reactions. Electrons stripped from nutrients are used to form NADH and NADPH, electron carriers that are similar in structure but support different functions. NADH supports ATP production but also generates reactive oxygen species (ROS) superoxide (O2 ●-) and hydrogen peroxide (H2O2). NADH-driven ROS production is counterbalanced by NADPH which maintains antioxidants in an active state. Mitochondria rely on a redox buffering network composed of reduced glutathione (GSH) and peroxiredoxins (Prx) to quench ROS generated by nutrient metabolism. As H2O2 is quenched, NADPH is expended to reactivate antioxidant networks and reset the redox environment. Thus, the mitochondrial redox environment is in a constant state of flux reflecting changes in nutrient and ROS metabolism. Changes in redox environment can modulate protein function through oxidation of protein cysteine thiols. Typically cysteine oxidation is considered to be mediated by H2O2 which oxidizes protein thiols (SH) forming sulfenic acid (SOH). However, problems begin to emerge when one critically evaluates the regulatory function of SOH. Indeed SOH formation is slow, non-specific, and once formed SOH reacts rapidly with a variety of molecules. By contrast, protein S-glutathionylation (PGlu) reactions involve the conjugation and removal of glutathione moieties from modifiable cysteine residues. PGlu reactions are driven by fluctuations in the availability of GSH and oxidized glutathione (GSSG) and thus should be exquisitely sensitive to changes ROS flux due to shifts in the glutathione pool in response to varying H2O2 availability. Here, we propose that energy metabolism-linked redox signals originating from mitochondria are mediated indirectly by H2O2 through the GSH redox buffering network in and outside mitochondria. This proposal is based on several observations that have shown that unlike other redox modifications PGlu reactions fulfill the requisite criteria to serve as an effective posttranslational modification that controls protein function.
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      PubDate: 2016-01-03T17:47:57Z
       
  • OXIDATIVE STRESS INFLUENCE ON RENAL DYSFUNCTION IN PATIENTS WITH
           OBSTRUCTIVE JAUNDICE: A CASE AND CONTROL PROSPECTIVE STUDY

    • Abstract: Publication date: Available online 29 December 2015
      Source:Redox Biology
      Author(s): David Martínez-Cecilia, María Reyes-Díaz, Juan Ruiz-Rabelo, Manuel Gomez-Alvarez, Carmen Muñoz Villanueva, José Álamo, Jordi Muntané, Francisco Javier Padillo Francisco
      Background: Obstructive Jaundice (OJ) is associated with a significant risk of developing acute renal failure (ARF). The involvement of oxidative stress in the development of cholestasis has been demonstrated in different experimental models. However, its role in the morbidity of human cholestasis is far to be elucidated. The aim of the study was the evaluation of oxidative stress markers in blood from patients with OJ and its relation to complications and benign/malignant evolution of cholestasis. Methods: A prospective cross-sectional study of 105 patients with OJ and 34 control subjects were included. Several markers of liver function and oxidative stress, such as lipoperoxides (LPO), as well as reduced glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities were assessed. Results: The patients with OJ showed a marked increase in plasma levels of LPO, SOD and GSH, while GSH-Px levels were decreased. The increase in lipid peroxidation products and the depletion of SOD activity in blood were also related to renal dysfunction. The highest level of LPO was associated with malignant etiology of the disease. The logistic regression analysis showed that the age of the patient and the levels of LPO in blood were predictors of renal dysfunction in OJ patients. Conclusions: This study demonstrates a correlation between oxidative stress and renal dysfunction patients with OJ.
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      PubDate: 2015-12-29T17:25:12Z
       
  • An unexplored role for Peroxiredoxin in exercise-induced redox
           signalling'

    • Abstract: Publication date: Available online 25 December 2015
      Source:Redox Biology
      Author(s): J. Wadley Alex, Sarah Aldred, Steven J. Coles
      Peroxiredoxin (PRDX) is a ubiquitous oxidoreductase protein with a conserved ionised thiol that permits catalysis of hydrogen peroxide (H2O2) up to a million times faster than any thiol-containing signalling protein. The increased production of H2O2 within active tissues during exercise is thought to oxidise conserved cysteine thiols, which may in turn facilitate a wide variety of physiological adaptations. The precise mechanisms linking H2O2 with the oxidation of signalling thiol proteins (phosphates, kinases and transcription factors) are unclear due to these proteins’ low reactivity with H2O2 relative to abundant thiol peroxidases such as PRDX. Recent work has shown that following exposure to H2O2 in vitro, the sulfenic acid of the PRDX cysteine can form mixed disulphides with transcription factors associated with cell survival. This implicates PRDX as an ‘active’ redox relay in transmitting the oxidising equivalent of H2O2 to downstream proteins. Furthermore, under oxidative stress, PRDX can form stable oxidised dimers that can be secreted into the extracellular space, potentially acting as an extracellular ‘stress’ signal. There is extensive literature assessing non-specific markers of oxidative stress in response to exercise, however the PRDX catalytic cycle may offer a more robust approach for measuring changes in redox balance following exercise. This review discusses studies assessing PRDX-mediated cellular signalling and integrates the recent advances in redox biology with investigations that have examined the role of PRDX during exercise in humans and animals. Future studies should explore the role of PRDX as a key regulator of peroxide mediated-signal transduction during exercise in humans.
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      PubDate: 2015-12-29T17:25:12Z
       
  • The thioredoxin system in breast cancer cell invasion and migration

    • Abstract: Publication date: Available online 19 December 2015
      Source:Redox Biology
      Author(s): Maneet Bhatia, Kelly L. McGrath, Giovanna Di Trapani, Pornpimol Charoentong, Fenil Shah, Mallory M. King, Frank M. Clarke, Kathryn F. Tonissen
      Metastasis is the most life threatening aspect of breast cancer. It is a multi-step process involving invasion and migration of primary tumor cells with a subsequent colonization of these cells at a secondary location. The aim of the present study was to investigate the role of thioredoxin (Trx1) in the invasion and migration of breast cancer cells and to assess the strength of the association between high levels of Trx1 and thioredoxin reductase (TrxR1) expression with breast cancer patient survival. Our results indicate that the expression of both Trx1 and TrxR1 are statistically significantly increased in breast cancer patient cells compared with paired normal breast tissue from the same patient. Over-expression of Trx1 in MDA-MB-231 breast cancer cell lines enhanced cell invasion in in vitro assays while expression of a redox inactive mutant form of Trx1 (designated 1SS) or the antisense mRNA inhibited cell invasion. Addition of exogenous Trx1 also enhanced cell invasion, while addition of a specific monoclonal antibody that inhibits Trx1 redox function decreased cell invasion. Over-expression of intracellular Trx1 did not increase cell migration but expression of intracellular 1SS inhibited migration. Addition of exogenous Trx1 enhanced cell migration while 1SS had no effect. Treatment with auranofin inhibited TrxR activity, cell migration and clonogenic activity of MDA-MB-231 cells, while increasing reactive oxygen species (ROS) levels. Analysis of 25 independent cohorts with 5910 patients showed that Trx1 and TrxR1 were both associated with a poor patient prognosis in terms of overall survival, distant metastasis free survival and disease free survival. Therefore, targeting the Trx system with auranofin or other specific inhibitors may provide improved breast cancer patient outcomes through inhibition of cancer invasion and migration.
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      PubDate: 2015-12-24T16:55:43Z
       
  • Photoprotection by dietary phenolics against melanogenesis induced by UVA
           through Nrf2-dependent antioxidant responses

    • Abstract: Publication date: Available online 19 December 2015
      Source:Redox Biology
      Author(s): Anyamanee Chaiprasongsuk, Tasanee Onkoksoong, Thanyawan Pluemsamran, Saowalak Limsaengurai, Uraiwan Panich
      Dietary phenolics may play a protective role in UV-mediated skin pigmentation through their antioxidant and UV-absorbing actions. In this study, we investigated whether genetic silencing of Nrf2, regulating the transcription of antioxidant genes, affected melanogenesis in primary human epidermal melanocytes (HEMn) and B16F10 melanoma cells subjected to UVA (8J/cm2) exposure. Then, we explored the antimelanogenic actions of phenolics; caffeic acid (CA) and ferulic acid (FA) providing partial UVA protection; quercetin (QU) and rutin (RU) providing strong UVA protection and; avobenzone (AV), an efficient UVA filter, in association with modulation of Nrf2-mediated antioxidant defenses in response to UVA insults in B16F10 cells. Upon oxidative insults, Nrf2 silencing promoted melanogenesis in both HEMn and B16F10 cells irradiated with UVA. Stimulation of melanogenesis by UVA correlated with increased ROS and oxidative DNA damage (8-OHdG), GSH depletion as well as a transient downregulation of Nrf2 nuclear translocation and of Nrf2-ARE signaling in B16F10 cells. All test compounds exerted anti-melanogenic effects with respect to their abilities to reverse UVA-mediated oxidative damage as well as downregulation of Nrf2 activity and its target antioxidants (GCLC, GST and NQO1) in B16F10 cells. In conclusion, defective Nrf2 may promote melanogenesis under UVA through oxidative stress mechanims. Compounds with antioxidant and/or UVA absorption properties could protect against UVA-induced melanogenesis through indirect regulatory effect on Nrf2-ARE pathway.
      Graphical abstract image

      PubDate: 2015-12-24T16:55:43Z
       
  • The Bioenergetic Health Index is a Sensitive Measure of Oxidative Stress
           in Human Monocytes

    • Abstract: Publication date: Available online 24 December 2015
      Source:Redox Biology
      Author(s): Balu K. Chacko, Degui Zhi, Victor M. Darley-Usmar, Tanecia Mitchell
      Metabolic and bioenergetic dysfunction are associated with oxidative stress and thought to be a common underlying mechanism of chronic diseases such as atherosclerosis, diabetes, and neurodegeneration. Recent findings support an emerging concept that circulating leukocytes and platelets can act as sensors or biomarkers of mitochondrial function in patients subjected to metabolic diseases. It is proposed that systemic stress-induced alterations in leukocyte bioenergetics are the consequence of several factors including reactive oxygen species. This suggests that oxidative stress mediated changes in leukocyte mitochondrial function could be used as an indicator of bioenergetic health in individuals. To test this concept, we investigated the effect of the redox cycling agent, 2,3 dimethoxynaphthoquinone (DMNQ) on the bioenergetic profiles of monocytes isolated from healthy human subjects using the extracellular flux analyzer. In addition, we tested the hypothesis that the bioenergetic health index (BHI), a single value that represents the bioenergetic health of individuals, is dynamically sensitive to oxidative stress in human monocytes. DMNQ decreased monocyte ATP-linked respiration, maximal respiration, and reserve capacity and caused an increase in proton leak and non-mitochondrial respiration compared to monocytes not treated with DMNQ. The BHI was a more sensitive indicator of the DMNQ-dependent changes in bioenergetics than any individual parameter. These data suggest that monocytes are susceptible to oxidative stress mediated by DMNQ and this can be accurately assessed by the BHI. Taken together, our findings suggest that the BHI has the potential to act as a functional biomarker of the impact of systemic oxidative stress in patients with metabolic disorders.
      Graphical abstract image

      PubDate: 2015-12-24T16:55:43Z
       
  • Detecting presence of cardiovascular disease through mitochondria
           respiration as depicted through biophotonic emission

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Nancy R. Rizzo, Nicole C. Hank, Jian Zhang
      Aims Increased production of reactive oxygen species (ROS) in mitochondria, play an important role in the cardiovascular system. Furthermore, oxidative metabolism of mitochondria comprised of biophoton emissions, are linked to ROS and oxidative stress. In this review we investigated the association between the ability of ClearViewTM system (ClearView) to indicate the presence or absence of cardiovascular disease through mitochondria respiration as depicted through biophotonic emission. Methods and results One hundred and ninety-five out of the three hundred and fifty-three human subjects enrolled in this prospective, single site study had at least one cardiovascular related diagnosis. Measurements with ClearView consisted of scanning all 10 fingers twice. Images were captured through the ClearView software and analyzed to produce a scale that indicates the presence or absence of cardiovascular disease. The association of ClearView's ability to indicate the presence or absence of cardiovascular disease with a physician's diagnosis was assessed using odds ratios (OR) and area under ROC curve (AUC). Adjusting for age, OR of ClearView measurements conducted with capacitive barrier was 3.44 (95%CI: 2.13, 5.55) and the OR without the capacitive barrier was 2.15 (95%CI: 1.42, 3.23). The OR in men were 5.91 (95%CI: 2.35, 14.85) and 2.88 (95%CI: 1.38, 6.01), adjusting for age and corresponding to with and without capacitive barrier. The OR in women were 3.50 (95%CI: 1.86, 6.59) and 2.09 (95%CI: 1.20, 3.64) with and without capacitive barrier. AUCs for measurements with capacitive barrier were >0.90. Conclusion ClearView detected the presence or absence of cardiovascular disease independent of other conditions.
      Graphical abstract image

      PubDate: 2015-12-24T16:55:43Z
       
  • Fatty acid nitroalkenes induce resistance to ischemic cardiac injury by
           modulating mitochondrial respiration at complex II

    • Abstract: Publication date: August 2016
      Source:Redox Biology, Volume 8
      Author(s): Jeffrey R. Koenitzer, Gustavo Bonacci, Steven R. Woodcock, Chen-Shan Chen, Nadiezhda Cantu-Medellin, Eric E. Kelley, Francisco J. Schopfer
      Nitro-fatty acids (NO2-FA) are metabolic and inflammatory-derived electrophiles that mediate pleiotropic signaling actions. It was hypothesized that NO2-FA would impact mitochondrial redox reactions to induce tissue-protective metabolic shifts in cells. Nitro-oleic acid (OA-NO2) reversibly inhibited complex II-linked respiration in isolated rat heart mitochondria in a pH-dependent manner and suppressed superoxide formation. Nitroalkylation of Fp subunit was determined by BME capture and the site of modification by OA-NO2 defined by mass spectrometric analysis. These effects translated into reduced basal and maximal respiration and favored glycolytic metabolism in H9C2 cardiomyoblasts as assessed by extracellular H+ and O2 flux analysis. The perfusion of NO2-FA induced acute cardioprotection in an isolated perfused heart ischemia/reperfusion (IR) model as evidenced by significantly higher rate-pressure products. Together these findings indicate that NO2-FA can promote cardioprotection by inducing a shift from respiration to glycolysis and suppressing reactive species formation in the post-ischemic interval.
      Graphical abstract image

      PubDate: 2015-12-24T16:55:43Z
       
  • The novel triterpenoid RTA 408 protects human retinal pigment epithelial
           cells against H2O2-induced cell injury via NF-E2-related factor 2 (Nrf2)
           activation

    • Abstract: Publication date: Available online 19 December 2015
      Source:Redox Biology
      Author(s): Xiaobin Liu, Keith Ward, Christy Xavier, Jamieson Jann, Abbot F. Clark, Iok-Hou Pang, Hongli Wu
      Oxidative stress-induced retinal pigment epithelial (RPE) cell damage is an important factor in the pathogenesis of age-related macular degeneration (AMD). Previous studies have shown that RTA 408, a synthetic triterpenoid compound, potently activates Nrf2. This study aimed to investigate the protective effects of RTA 408 in cultured RPE cells during oxidative stress and to determine the effects of RTA 408 on Nrf2 and its downstream target genes. Primary human RPE cells were pretreated with RTA 408 and then incubated in 200μM H2O2 for 6h. Cell viability was measured with the WST8 assay. Apoptosis was quantitatively measured by annexin V/propidium iodide (PI) double staining and Hoechst 33342 fluorescent staining. Reduced (GSH) and oxidized glutathione (GSSG) were measured using colorimetric assays. Nrf2 activation and its downstream effects on phase II enzymes were examined by Western blot. Treatment of RPE cells with nanomolar ranges (10 and 100nM) of RTA 408 markedly attenuated H2O2-induced viability loss and apoptosis. RTA 408 pretreatment significantly protected cells from oxidative stress-induced GSH loss, GSSG formation and decreased ROS production. RTA 408 activated Nrf2 and increased the expression of its downstream genes, such as HO-1, NQO1, SOD2, catalase, Grx1, and Trx1. Consequently, the enzyme activities of NQO1, Grx1, and Trx1 were fully protected by RTA 408 pretreatment under oxidative stress. Moreover, knockdown of Nrf2 by siRNA significantly reduced the cytoprotective effects of RTA 408. In conclusion, our data suggest that RTA 408 protect primary human RPE cells from oxidative stress-induced damage by activating Nrf2 and its downstream genes.


      PubDate: 2015-12-24T16:55:43Z
       
  • Impact of glutathione supplementation of parenteral nutrition on hepatic
           methionine adenosyltransferase activity

    • Abstract: Publication date: Available online 17 December 2015
      Source:Redox Biology
      Author(s): Wesam Elremaly, Ibrahim Mohamed, Thérèse Rouleau, Jean-Claude Lavoie
      Background & Aims The oxidation of the methionine adenosyltransferase (MAT) by the combined impact of peroxides contaminating parenteral nutrition (PN) and oxidized redox potential of glutathione is suspected to explain its inhibition observed in animals. A modification of MAT activity is suspected to be at origin of the PN-associated liver disease as observed in newborns. We hypothesized that the correction of redox potential of glutathione by adding glutathione in PN protects the MAT activity. Aim: to investigate whether the addition of glutathione to PN can reverse the inhibition of MAT observed in animal on PN. Methods Three days old guinea pigs received through a jugular vein catheter 2 series of solutions. First with methionine supplement, 1) Sham (no infusion); 2) PN: amino acids, dextrose, lipids and vitamins; 3) PN-GSSG: PN+10 μM GSSG. Second without nmethionine, 4) D: dextrose; 5) D+180μM ascorbylperoxide; 6) D+350μM H2O2. Four days later, liver was sampled for determination of redox potential of glutathione and MAT activity in presence or not of 1mM DTT. Data were compared by ANOVA, p<0.05. Results MAT activity was 45±4% lower in animal infused with PN and 23±7% with peroxides generated in PN. The inhibition by peroxides was associated with oxidized redox potential and was reversible by DTT. Correction of redox potential (PN+GSSG) or DTT was without effect on the inhibition of MAT by PN. The slope of the linear relation between MAT activity and redox potential was two fold lower in animal infused with PN than in others groups. Conclusion The present study suggests that prevention of peroxide generation in PN and/or correction of the redox potential by adding glutathione in PN are not sufficient, at least in newborn guinea pigs, to restore normal MAT activity.
      Graphical abstract image

      PubDate: 2015-12-19T16:20:04Z
       
  • Redox-Dependent Regulation Of Epidermal Growth Factor Receptor Signaling

    • Abstract: Publication date: Available online 8 December 2015
      Source:Redox Biology
      Author(s): David E. Heppner, Albert van der Vliet
      Tyrosine phosphorylation-dependent cell signaling represents a unique feature of multicellular organisms, and is important in regulation of cell differentiation and specialized cell functions. Multicellular organisms also contain a diverse family of NADPH oxidases (NOX) that have been closely linked with tyrosine kinase-based cell signaling and regulate tyrosine phosphorylation via reversible oxidation of cysteine residues that are highly conserved within many proteins involved in this signaling pathway. An example of redox-regulated tyrosine kinase signaling involves the epidermal growth factor receptor (EGFR), a widely studied receptor system with diverse functions in normal cell biology as well as pathologies associated with oxidative stress such as cancer. The purpose of this Graphical Redox Review is to highlight recently emerged concepts with respect to NOX-dependent regulation of this important signaling pathway.


      PubDate: 2015-12-09T14:59:59Z
       
  • NAC selectively inhibit cancer telomerase activity: A higher redox
           homeostasis threshold exists in cancer cells

    • Abstract: Publication date: Available online 8 December 2015
      Source:Redox Biology
      Author(s): Pengying Li, Feizhou Xie, Jing Wang, Yilun Sui, Michael Halim, Shanlin Liu, Dongyun Shi
      Telomerase activity controls telomere length, and this plays an important role in stem cells, aging and tumors. Antioxidant was shown to protect telomerase activity in normal cells but inhibit that in cancer cells, but the underlying mechanism is elusive. Here we found that 7721 hepatoma cells held a higher redox homeostasis threshold than L02 normal liver cells which caused 7721 cells to have a higher demand for ROS; MnSOD over-expression in 7721 decreased endogenous reactive oxygen species (ROS) and inhibited telomerase activity; Akt phosphorylation inhibitor and NAC both inhibited 7721 telomerase activity. The over-elimination of ROS by NAC resulted in the inhibition of Akt pathway. Our results suggest that ROS is involved in the regulation of cancer telomerase activity through Akt pathway. The different intracellular redox homeostasis and antioxidant system in normal cells and tumor cells may be the cause of the opposite effect on telomerase activity in response to NAC treatment. Our results provide a theoretical base of using antioxidants selectively inhibit cancer telomerase activity. Findings of the present study may provide insights into novel approaches for cancer treatment.
      Graphical abstract image

      PubDate: 2015-12-09T14:59:59Z
       
  • Formation of 2-nitrophenol from salicylaldehyde as a suitable test for low
           peroxynitrite fluxes

    • Abstract: Publication date: April 2016
      Source:Redox Biology, Volume 7
      Author(s): Yuliya Mikhed, Kai Bruns, Stefan Schildknecht, Michael Jörg, Mobin Dib, Matthias Oelze, Karl J. Lackner, Thomas Münzel, Volker Ullrich, Andreas Daiber
      There has been some dispute regarding reaction products formed at physiological peroxynitrite fluxes in the nanomolar range with phenolic molecules, when used to predict the behavior of protein-bound aromatic amino acids like tyrosine. Previous data showed that at nanomolar fluxes of peroxynitrite, nitration of these phenolic compounds was outcompeted by dimerization (e.g. biphenols or dityrosine). Using 3-morpholino sydnonimine (Sin-1), we created low fluxes of peroxynitrite in our reaction set-up to demonstrate that salicylaldehyde displays unique features in the detection of physiological fluxes of peroxynitrite, yielding detectable nitration but only minor dimerization products. By means of HPLC analysis and detection at 380nm we could identify the expected nitration products 3- and 5-nitrosalicylaldehyde, but also novel nitrated products. Using mass spectrometry, we also identified 2-nitrophenol and a not fully characterized nitrated dimerization product. The formation of 2-nitrophenol could proceed either by primary generation of a phenoxy radical, followed by addition of the NO2-radical to the various resonance structures, or by addition of the peroxynitrite anion to the polarized carbonyl group with subsequent fragmentation of the adduct (as seen with carbon dioxide). Interestingly, we observed almost no 3- and 5-nitrosalicylic acid products and only minor dimerization reaction. Our results disagree with the previous general assumption that nitration of low molecular weight phenolic compounds is always outcompeted by dimerization at nanomolar peroxynitrite fluxes and highlight unique features of salicylaldehyde as a probe for physiological concentrations of peroxynitrite.
      Graphical abstract image

      PubDate: 2015-11-29T13:20:49Z
       
  • Breast cancer 1 (BRCA1)-deficient embryos develop normally but are more
           susceptible to ethanol-initiated DNA damage and embryopathies

    • Abstract: Publication date: April 2016
      Source:Redox Biology, Volume 7
      Author(s): Aaron M. Shapiro, Lutfiya Miller-Pinsler, Peter G. Wells
      The breast cancer 1 (brca1) gene is associated with breast and ovarian cancers, and heterozygous (+/−) brca1 knockout progeny develop normally, suggesting a negligible developmental impact. However, our results show BRCA1 plays a broader biological role in protecting the embryo from oxidative stress. Sox2-promoted Cre-expressing hemizygous males were mated with floxed brca1 females, and gestational day 8 +/− brca1 conditional knockout embryos with a 28% reduction in protein expression were exposed in culture to the reactive oxygen species (ROS)-initiating drug ethanol (EtOH). Untreated +/− brca1-deficient embryos developed normally, but when exposed to EtOH exhibited increased levels of oxidatively damaged DNA, measured as 8-oxo-2'-deoxyguanosine, γH2AX, which is a marker of DNA double strand breaks that can result from 8-oxo-2'-deoxyguanosine, formation, and embryopathies at EtOH concentrations that did not affect their brca1-normal littermates. These results reveal that even modest BRCA1 deficiencies render the embryo more susceptible to drug-enhanced ROS formation, and corroborate a role for DNA oxidation in the mechanism of EtOH teratogenesis.
      Graphical abstract image

      PubDate: 2015-11-29T13:20:49Z
       
  • The antimalarial drug primaquine targets Fe–S cluster proteins and
           yeast respiratory growth

    • Abstract: Publication date: April 2016
      Source:Redox Biology, Volume 7
      Author(s): Anaïs Lalève, Cindy Vallières, Marie-Pierre Golinelli-Cohen, Cécile Bouton, Zehua Song, Grzegorz Pawlik, Sarah M. Tindall, Simon V. Avery, Jérôme Clain, Brigitte Meunier
      Malaria is a major health burden in tropical and subtropical countries. The antimalarial drug primaquine is extremely useful for killing the transmissible gametocyte forms of Plasmodium falciparum and the hepatic quiescent forms of P. vivax. Yet its mechanism of action is still poorly understood. In this study, we used the yeast Saccharomyces cerevisiae model to help uncover the mode of action of primaquine. We found that the growth inhibitory effect of primaquine was restricted to cells that relied on respiratory function to proliferate and that deletion of SOD2 encoding the mitochondrial superoxide dismutase severely increased its effect, which can be countered by the overexpression of AIM32 and MCR1 encoding mitochondrial enzymes involved in the response to oxidative stress. This indicated that ROS produced by respiratory activity had a key role in primaquine-induced growth defect. We observed that Δsod2 cells treated with primaquine displayed a severely decreased activity of aconitase that contains a Fe–S cluster notoriously sensitive to oxidative damage. We also showed that in vitro exposure to primaquine impaired the activity of purified aconitase and accelerated the turnover of the Fe–S cluster of the essential protein Rli1. It is suggested that ROS-labile Fe–S groups are the primary targets of primaquine. Aconitase activity is known to be essential at certain life-cycle stages of the malaria parasite. Thus primaquine-induced damage of its labile Fe–S cluster – and of other ROS-sensitive enzymes – could inhibit parasite development.
      Graphical abstract image

      PubDate: 2015-11-29T13:20:49Z
       
  • Cysteines 208 and 241 in Ero1α are required for maximal catalytic
           turnover

    • Abstract: Publication date: April 2016
      Source:Redox Biology, Volume 7
      Author(s): Thomas Ramming, Shingo Kanemura, Masaki Okumura, Kenji Inaba, Christian Appenzeller-Herzog
      Endoplasmic reticulum (ER) oxidoreductin 1α (Ero1α) is a disulfide producer in the ER of mammalian cells. Besides four catalytic cysteines (Cys94, Cys99, Cys394, Cys397), Ero1α harbors four regulatory cysteines (Cys104, Cys131, Cys208, Cys241). These cysteines mediate the formation of inhibitory intramolecular disulfide bonds, which adapt the activation state of the enzyme to the redox environment in the ER through feedback signaling. Accordingly, disulfide production by Ero1α is accelerated by reducing conditions, which minimize the formation of inhibitory disulfides, or by mutations of regulatory cysteines. Here we report that reductive stimulation enhances Ero1α activity more potently than the mutation of cysteines. Specifically, mutation of Cys208/Cys241 does not mechanistically mimic reductive stimulation, as it lowers the turnover rate of Ero1α in presence of a reducing agent. The Cys208/Cys241 pair therefore fulfills a function during catalysis that reaches beyond negative regulation. In agreement, we identify a reciprocal crosstalk between the stabilities of the Cys208–Cys241 disulfide and the inhibitory disulfide bonds involving Cys104 and Cys131, which also controls the recruitment of the H2O2 scavenger GPx8 to Ero1α. Two possible mechanisms by which thiol–disulfide exchange at the Cys208/Cys241 pair stimulates the catalytic turnover under reducing conditions are discussed.
      Graphical abstract image

      PubDate: 2015-11-24T12:59:47Z
       
  • l-dehydroascorbic acid can substitute l-ascorbic acid as dietary vitamin C
           source in guinea pigs

    • Abstract: Publication date: April 2016
      Source:Redox Biology, Volume 7
      Author(s): Henriette Frikke-Schmidt, Pernille Tveden-Nyborg, Jens Lykkesfeldt
      Vitamin C deficiency globally affects several hundred million people and has been associated with increased morbidity and mortality in numerous studies. In this study, bioavailability of the oxidized form of vitamin C (l-dehydroascorbic acid or DHA)—commonly found in vitamin C containing food products prone to oxidation—was studied. Our aim was to compare tissue accumulation of vitamin C in guinea pigs receiving different oral doses of either ascorbate or DHA. In all tissues tested (plasma, liver, spleen, lung, adrenal glands, kidney, muscle, heart, and brain), only sporadic differences in vitamin C accumulation from ascorbate or DHA were observed except for the lowest dose of DHA (0.25mg/ml in the drinking water), where approximately half of the tissues had slightly yet significantly less vitamin C accumulation than from the ascorbate source. As these results contradicted data from rats, we continued to explore the ability to recycle DHA in blood, liver and intestine in guinea pigs, rats and mice. These investigations revealed that guinea pigs have similar recycling capacity in red blood cells as observed in humans, while rats and mice do not have near the same ability to reduce DHA in erythrocytes. In liver and intestinal homogenates, guinea pigs also showed a significantly higher ability to recycle DHA compared to rats and mice. These data demonstrate that DHA in guinea pigs—as in humans—is almost as effective as ascorbate as vitamin C source when it comes to taking up and storing vitamin C and further suggest that the guinea pig is superior to other rodents in modeling human vitamin C homeostasis.
      Graphical abstract image

      PubDate: 2015-11-24T12:59:47Z
       
  • Curcumin inhibits activation of TRPM2 channels in rat hepatocytes

    • Abstract: Publication date: April 2016
      Source:Redox Biology, Volume 7
      Author(s): E. Kheradpezhouh, G.J. Barritt, G.Y. Rychkov
      Oxidative stress is a hallmark of many liver diseases including viral and drug-induced hepatitis, ischemia-reperfusion injury, and non-alcoholic steatohepatitis. One of the consequences of oxidative stress in the liver is deregulation of Ca2+ homeostasis, resulting in a sustained elevation of the free cytosolic Ca2+ concentration ([Ca2+]c) in hepatocytes, which leads to irreversible cellular damage. Recently it has been shown that liver damage induced by paracetamol and subsequent oxidative stress is, in large part, mediated by Ca2+ entry through Transient Receptor Potential Melastatin 2 (TRPM2) channels. Involvement of TRPM2 channels in hepatocellular damage induced by oxidative stress makes TRPM2 a potential therapeutic target for treatment of a range of oxidative stress-related liver diseases. We report here the identification of curcumin ((1E,6E)-1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione), a natural plant-derived polyphenol in turmeric spice, as a novel inhibitor of TRPM2 channel. Presence of 5µM curcumin in the incubation medium prevented the H2O2- and paracetamol-induced [Ca2+]c rise in rat hepatocytes. Furthermore, in patch clamping experiments incubation of hepatocytes with curcumin inhibited activation of TRPM2 current by intracellular ADPR with IC50 of approximately 50nM. These findings enhance understanding of the actions of curcumin and suggest that the known hepatoprotective properties of curcumin are, at least in part, mediated through inhibition of TRPM2 channels.
      Graphical abstract image

      PubDate: 2015-11-24T12:59:47Z
       
 
 
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