- Email: [email protected]
The activity of proline as singlet oxygen and superoxide scavenger
A. Stepanova et al. / Free Radical Biology and Medicine 120 (2018) S45–S166
S75
IDH2þ/þ mouse. These results indicate that IDH2 deficiency worsens cisplatin nephrotoxicity by increasing oxidative stress, suggesting that cisplatin-induced nephrotoxicity is associated with mitochondrial IDH2.
P-100
Effect of a healthy mixed diet on micronutrients intake and ROS regulation
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.248
Denise Zdzieblik, Carolin Brauchle, Christoph Centner, Albert Gollhofer, Daniel König Department of Sport and Sport Science, University of Freiburg, Freiburg, Germany
It is generally believed that several nutrients such as β-carotene, vitamin C, E and Zinc exert a protective effect against oxidative damage. However, there is little evidence if a healthy mixed diet affects antioxidant defense mechanisms through an increased intake of micronutrients. In an open-label study, we investigated the reactive oxygen species (ROS) production in 7 healthy subjects before and 2 weeks after the change from a rather unhealthy eating pattern (e.g. much fast food) to a healthy mixed diet according to the recommendations of the German Society of Nutrition (DGE). ROS production was determined by Electronic Spin Resonance technique (Bruker, Germany). Following an overnight fast, total ROS concentration in capillary blood was measured before and after the 2 weeks dietary intervention. The nutritional habits (dietary protocol) was analyzed by Nutriguides. The total ROS formation was significantly different following the dietary intervention (- 0.25 7 1.9 mM*min -1; p o 0.05) probably as a result of the significantly higher intake of antioxidants (vitamin C: þ136.42 787.42 mg, p o0.05; vitamin E: þ 6.247 1.24 mg, p o0.05 and β-carotene:þ9.46 74.78 mg; p o0.01). These results are indicative of an improved redox state by increasing the intake of antioxidant nutrients by a healthier eating pattern.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.247
P-101
IDH2 deficiency exacerbates cisplatin nephrotoxicity 1
2
Jee In Kim , Min Jung Kong , Kwon Moo Park
P-102
The involvement of the anti-oxidant protein DJ-1 in SOD1 maturation pathway: implications for neurobehavioral deficits in Drosophila Melanogaster Federica De Lazzari, Giovanni Frighetto, Matteo Stramazzo, Luigi Bubacco, Alexander Whitworth, Sandro Malacrida, Mauro Agostino Zordan, Aram Megighian, Federica Sandrelli, Marco Bisaglia University of Padova, Padova, Italy
The Parkinson's disease associated protein DJ-1 is as a multitasking protein with anti-oxidant activity. Indeed, DJ-1 has been shown to modulate signaling pathways involved in anti-oxidant defense. Nevertheless, no consensus has been found on its effective cellular function. Our group has demonstrated, in vitro, that DJ-1 can induce the activation of SOD1, the first line of defense against reactive oxygen species. SOD1 and DJ-1 are known to play an important role against oxidative stress. Although SOD1 activation normally relies on a dedicated copper chaperone, it has been shown to be partially activated also through an alternative pathway, to which DJ-1 may participate. To better clarify this mechanism in vivo we utilize Drosophila Melanogaster as model organism, which possesses homologue pathways for anti-oxidant responses. For this study, we are evaluating the behavioral traits of wild-type and mutant flies under normal conditions and under mild oxidative insults, assessing lifespan and complex locomotor features. Although DJ-1 loss does not affect fly lifespan under normal conditions, its absence compromises survival under paraquat exposure. Therefore, our work aims to elucidate the anti-oxidant role of DJ-1, focusing on its participation to SOD1 activation.
E-mail address: [email protected]
2
http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.249 1
Department of Molecular Medicine and ODR Medical Research Center, Keimyung University School of Medicine, Daegu, Republic of Korea 2 Department of Anatomy and BK 21 Plus2, School of Medicine, Kyungpook National University, Deagu, Republic of Korea
þ
Mitochondrial NADP -dependent isocitrate dehydrogenase (IDH2) is a major producer of NADPH in the mitochondria and plays a critical role in the redox balance. Cisplatin is an effective anticancer drug, however, its nephrotoxicity, by the breaking of redox balance, limits its use. Here, we investigated whether the cisplatin nephrotoxicity is associated with IDH2. IDH2 gene-deleted (IDH2-/-) and wild type (IDH2þ /þ ) mice were administrated cisplatin with or without Mito-TEMPO, a mitochondria-specific antioxidant. Cisplatin reduced IDH2 activity and expression and NADPH levels in the kidney together with mitochondrial oxidative injury. Mitochondrial damage after cisplatin injection was greater in the IDH2-/-than IDH2þ /þ kidneys. These changes were more pronounced in the IDH2-/than IDH2þ/þ kidneys, showing greater renal functional and morphological impairments in the IDH2-/-mouse. Mito-TEMPO reduced those cisplatin-induced kidney injuries both in IDH2-/- and IDH2þ /þ mouse. This reduction in the IDH2-/- mouse was more prominent than that in the
P-103
The activity of proline as singlet oxygen and superoxide scavenger Imre Vass, Ateeq ur Rehman Institute of Plant Biology, Biological Research Centre, Szeged, Hungary
Proline (Pro) is an important stress protecting molecule in plants and other cellular systems against oxidative damage. However, the actual sensitivity and selectivity of Pro against specific reactive oxygen species (ROS), especially singlet oxygen and superoxide, is debated. Here we present in vitro studies by using EPR and oxygen uptake detection of ROS showing that Pro can indeed scavenge both singlet oxygen and superoxide. The scavenging activity of Pro is compared to that of ascorbate (Asc), a known
S76
A. Stepanova et al. / Free Radical Biology and Medicine 120 (2018) S45–S166
quencher of singlet oxygen and superoxide. In contrast to Asc, which a chemical quencher, Pro eliminates singlet oxygen via a physical mechanism with ca. 1% efficiency that of Asc. The superoxide scavenging activity of Pro does not proceed via H2O2 production as in case of superoxide dismutase (SOD), but similar to that of Asc that includes electron transfer, and its efficiency is ca. 6% that of Asc on equal molar basis. Since Pro can accumulate up to 10-fold higher concentrations under stress conditions than Asc, the overall singlet oxygen and superoxide elimination activity of Pro can reach 10 and 60% of the activity of Asc, respectively. In stress exposed plants this could be physiologically relevant in cell compartments with low amount of more efficient scavengers.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.250
P-104
Cytochrome c - cardiolipin interaction leads to the cytochrome c modification and degradation via formation of cardiolipin hydroperoxides Uladzimir Barayeu 1,2, Mike Lange 1,2, Oleg Shadyro 3, Jürgen Arnhold 4, Jörg Flemmig 4, Maria Fedorova 1,2 1
Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, University of Leipzig, Germany 2 Center for Biotechnology and Biomedicine, University of Leipzig, Germany 3 Belarusian State University, Department of Chemistry, Minsk, Belarus 4 Institute of Medical Physics and Biophysics, Medical Faculty, University of Leipzig, Germany
Recently, the complex of cytochrome c (cyt c), a small heme mitochondrial protein, with mitochondria specific lipid cardiolipin (CL) was linked to the control of apoptotic cell death. Under apoptotic conditions cyt c - CL complex exhibits peroxidase activity, leading to the formation of CL- hydroperoxides. CL-hydroperoxides promote cyt c release to the cytoplasm, where cyt c participates in apoptosis pathway. However, not much known about the modifications of cyt c as a part of cyt c - CL complex. In the present work, the systematic study on the peroxidase activity of CL-cyt c complex in CL-containing liposomes was performed. CL-hydroperoxides stimulate cyt c peroxidase activity in the absence of hydrogen peroxide. Furthermore, in the presence of oxygen, CL-hydroperoxides are generated as the products of the CL - cyt c peroxidase reaction. During peroxidase cycle cyt c amino acid residues are modified by reactive unsaturated aldehydes including 4-hydroxy- and 4-oxo-nonenal, derived from CL- peroxidation. This leads to the consecutive cyt c inactivation and heme degradation accompanied by the release of free iron. The current study on CL-driven cyt c degradation and iron release probably plays an important role in the mitochondrial pathway of apoptosis induction.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.251
P-105
Effect of a novel biscumarin derivative on human melanoma cells: Regulation of cell survival, proliferation, migration and kinases Qurat-ul-Ain 1,2, M. Iqbal Choudhary 1,2, Karin Scharffetter-Kochanek 2,3 1
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan 2 H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan 3 Department of Dermatology and Allergy, University of Ulm, Ulm, Germany
We have investigated the effects of a newly synthesized biscumarin derivative on cell survival, proliferation, migration and signal transduction pathways in the human metastatic melanoma cells A375. Among the modulated pathways of cell survival and tumor suppression we found highly significant effects of this novel synthetic derivative on TOR (Target of Rapamycin), Checkpoint Kinase 2 (Chk- 2), Tumor Suppressor Protein Kinase p53 ( P53), Additionally the compound was found to modulate a number of pathways of cell proliferation with varying degree of efficacy. We have shown that biscumarin derivative induced activation of Epidermal Growth Factor Receptor (EGFR), Focal Adhesion Kinase (FAK), cAMP response Element-Binding Protein (CREB), Glycogen Synthase Kinase (GSK3 α/f3) and f3 –catenin: this effect may be at thebases of observed motility inhibition of melanoma cell caused by the compound. We have shown that the biscumarin derivative diminish phosphorylation of CHK-2 protein, TOR and P53 an event that is likely be the basis of observed inhibition of melanoma cell survival and tumor growth. Inhibition of cell survival and growth possibly due to the deminish phosphorylation of the chk-2 kinase on Threonine: T68, of TOR on Serine: S2448 of P53 on Serine: S392.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.252
P-106
The redox-active thiosemicarbazone, Dp44mT promotes NDRG1 to down-regulate oncogenic signaling pathways in cancer Sharleen Menezes, Zaklina Kovacevic, Des Richardson The University of Sydney, Sydney, Australia
The redox-active thiosemicarbazone, Dp44mT has shown potent anticancer activity against various cancers. This occurs by its ability to sequester, bind iron, and also produce reactive oxygen species. The activity of this agent is in part mediated by its ability to upregulate the metastasis suppressor, N-myc Downstream Regulated Gene 1 (NDRG1). NDRG1 inhibits cell proliferation and migration by negatively regulating numerous oncogenic signaling pathways. However, the mechanisms by which it modulates these pathways remain to be elucidated. In this study we examined the effect of NDRG1 and Dp44mT on the ErbB-family of receptor tyrosine kinases namely epidermal growth factor receptor (EGFR), human epidermal growth factor
S75
IDH2þ/þ mouse. These results indicate that IDH2 deficiency worsens cisplatin nephrotoxicity by increasing oxidative stress, suggesting that cisplatin-induced nephrotoxicity is associated with mitochondrial IDH2.
P-100
Effect of a healthy mixed diet on micronutrients intake and ROS regulation
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.248
Denise Zdzieblik, Carolin Brauchle, Christoph Centner, Albert Gollhofer, Daniel König Department of Sport and Sport Science, University of Freiburg, Freiburg, Germany
It is generally believed that several nutrients such as β-carotene, vitamin C, E and Zinc exert a protective effect against oxidative damage. However, there is little evidence if a healthy mixed diet affects antioxidant defense mechanisms through an increased intake of micronutrients. In an open-label study, we investigated the reactive oxygen species (ROS) production in 7 healthy subjects before and 2 weeks after the change from a rather unhealthy eating pattern (e.g. much fast food) to a healthy mixed diet according to the recommendations of the German Society of Nutrition (DGE). ROS production was determined by Electronic Spin Resonance technique (Bruker, Germany). Following an overnight fast, total ROS concentration in capillary blood was measured before and after the 2 weeks dietary intervention. The nutritional habits (dietary protocol) was analyzed by Nutriguides. The total ROS formation was significantly different following the dietary intervention (- 0.25 7 1.9 mM*min -1; p o 0.05) probably as a result of the significantly higher intake of antioxidants (vitamin C: þ136.42 787.42 mg, p o0.05; vitamin E: þ 6.247 1.24 mg, p o0.05 and β-carotene:þ9.46 74.78 mg; p o0.01). These results are indicative of an improved redox state by increasing the intake of antioxidant nutrients by a healthier eating pattern.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.247
P-101
IDH2 deficiency exacerbates cisplatin nephrotoxicity 1
2
Jee In Kim , Min Jung Kong , Kwon Moo Park
P-102
The involvement of the anti-oxidant protein DJ-1 in SOD1 maturation pathway: implications for neurobehavioral deficits in Drosophila Melanogaster Federica De Lazzari, Giovanni Frighetto, Matteo Stramazzo, Luigi Bubacco, Alexander Whitworth, Sandro Malacrida, Mauro Agostino Zordan, Aram Megighian, Federica Sandrelli, Marco Bisaglia University of Padova, Padova, Italy
The Parkinson's disease associated protein DJ-1 is as a multitasking protein with anti-oxidant activity. Indeed, DJ-1 has been shown to modulate signaling pathways involved in anti-oxidant defense. Nevertheless, no consensus has been found on its effective cellular function. Our group has demonstrated, in vitro, that DJ-1 can induce the activation of SOD1, the first line of defense against reactive oxygen species. SOD1 and DJ-1 are known to play an important role against oxidative stress. Although SOD1 activation normally relies on a dedicated copper chaperone, it has been shown to be partially activated also through an alternative pathway, to which DJ-1 may participate. To better clarify this mechanism in vivo we utilize Drosophila Melanogaster as model organism, which possesses homologue pathways for anti-oxidant responses. For this study, we are evaluating the behavioral traits of wild-type and mutant flies under normal conditions and under mild oxidative insults, assessing lifespan and complex locomotor features. Although DJ-1 loss does not affect fly lifespan under normal conditions, its absence compromises survival under paraquat exposure. Therefore, our work aims to elucidate the anti-oxidant role of DJ-1, focusing on its participation to SOD1 activation.
E-mail address: [email protected]
2
http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.249 1
Department of Molecular Medicine and ODR Medical Research Center, Keimyung University School of Medicine, Daegu, Republic of Korea 2 Department of Anatomy and BK 21 Plus2, School of Medicine, Kyungpook National University, Deagu, Republic of Korea
þ
Mitochondrial NADP -dependent isocitrate dehydrogenase (IDH2) is a major producer of NADPH in the mitochondria and plays a critical role in the redox balance. Cisplatin is an effective anticancer drug, however, its nephrotoxicity, by the breaking of redox balance, limits its use. Here, we investigated whether the cisplatin nephrotoxicity is associated with IDH2. IDH2 gene-deleted (IDH2-/-) and wild type (IDH2þ /þ ) mice were administrated cisplatin with or without Mito-TEMPO, a mitochondria-specific antioxidant. Cisplatin reduced IDH2 activity and expression and NADPH levels in the kidney together with mitochondrial oxidative injury. Mitochondrial damage after cisplatin injection was greater in the IDH2-/-than IDH2þ /þ kidneys. These changes were more pronounced in the IDH2-/than IDH2þ/þ kidneys, showing greater renal functional and morphological impairments in the IDH2-/-mouse. Mito-TEMPO reduced those cisplatin-induced kidney injuries both in IDH2-/- and IDH2þ /þ mouse. This reduction in the IDH2-/- mouse was more prominent than that in the
P-103
The activity of proline as singlet oxygen and superoxide scavenger Imre Vass, Ateeq ur Rehman Institute of Plant Biology, Biological Research Centre, Szeged, Hungary
Proline (Pro) is an important stress protecting molecule in plants and other cellular systems against oxidative damage. However, the actual sensitivity and selectivity of Pro against specific reactive oxygen species (ROS), especially singlet oxygen and superoxide, is debated. Here we present in vitro studies by using EPR and oxygen uptake detection of ROS showing that Pro can indeed scavenge both singlet oxygen and superoxide. The scavenging activity of Pro is compared to that of ascorbate (Asc), a known
S76
A. Stepanova et al. / Free Radical Biology and Medicine 120 (2018) S45–S166
quencher of singlet oxygen and superoxide. In contrast to Asc, which a chemical quencher, Pro eliminates singlet oxygen via a physical mechanism with ca. 1% efficiency that of Asc. The superoxide scavenging activity of Pro does not proceed via H2O2 production as in case of superoxide dismutase (SOD), but similar to that of Asc that includes electron transfer, and its efficiency is ca. 6% that of Asc on equal molar basis. Since Pro can accumulate up to 10-fold higher concentrations under stress conditions than Asc, the overall singlet oxygen and superoxide elimination activity of Pro can reach 10 and 60% of the activity of Asc, respectively. In stress exposed plants this could be physiologically relevant in cell compartments with low amount of more efficient scavengers.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.250
P-104
Cytochrome c - cardiolipin interaction leads to the cytochrome c modification and degradation via formation of cardiolipin hydroperoxides Uladzimir Barayeu 1,2, Mike Lange 1,2, Oleg Shadyro 3, Jürgen Arnhold 4, Jörg Flemmig 4, Maria Fedorova 1,2 1
Institute of Bioanalytical Chemistry, Faculty of Chemistry and Mineralogy, University of Leipzig, Germany 2 Center for Biotechnology and Biomedicine, University of Leipzig, Germany 3 Belarusian State University, Department of Chemistry, Minsk, Belarus 4 Institute of Medical Physics and Biophysics, Medical Faculty, University of Leipzig, Germany
Recently, the complex of cytochrome c (cyt c), a small heme mitochondrial protein, with mitochondria specific lipid cardiolipin (CL) was linked to the control of apoptotic cell death. Under apoptotic conditions cyt c - CL complex exhibits peroxidase activity, leading to the formation of CL- hydroperoxides. CL-hydroperoxides promote cyt c release to the cytoplasm, where cyt c participates in apoptosis pathway. However, not much known about the modifications of cyt c as a part of cyt c - CL complex. In the present work, the systematic study on the peroxidase activity of CL-cyt c complex in CL-containing liposomes was performed. CL-hydroperoxides stimulate cyt c peroxidase activity in the absence of hydrogen peroxide. Furthermore, in the presence of oxygen, CL-hydroperoxides are generated as the products of the CL - cyt c peroxidase reaction. During peroxidase cycle cyt c amino acid residues are modified by reactive unsaturated aldehydes including 4-hydroxy- and 4-oxo-nonenal, derived from CL- peroxidation. This leads to the consecutive cyt c inactivation and heme degradation accompanied by the release of free iron. The current study on CL-driven cyt c degradation and iron release probably plays an important role in the mitochondrial pathway of apoptosis induction.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.251
P-105
Effect of a novel biscumarin derivative on human melanoma cells: Regulation of cell survival, proliferation, migration and kinases Qurat-ul-Ain 1,2, M. Iqbal Choudhary 1,2, Karin Scharffetter-Kochanek 2,3 1
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan 2 H. E. J. Research Institute of Chemistry, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, Pakistan 3 Department of Dermatology and Allergy, University of Ulm, Ulm, Germany
We have investigated the effects of a newly synthesized biscumarin derivative on cell survival, proliferation, migration and signal transduction pathways in the human metastatic melanoma cells A375. Among the modulated pathways of cell survival and tumor suppression we found highly significant effects of this novel synthetic derivative on TOR (Target of Rapamycin), Checkpoint Kinase 2 (Chk- 2), Tumor Suppressor Protein Kinase p53 ( P53), Additionally the compound was found to modulate a number of pathways of cell proliferation with varying degree of efficacy. We have shown that biscumarin derivative induced activation of Epidermal Growth Factor Receptor (EGFR), Focal Adhesion Kinase (FAK), cAMP response Element-Binding Protein (CREB), Glycogen Synthase Kinase (GSK3 α/f3) and f3 –catenin: this effect may be at thebases of observed motility inhibition of melanoma cell caused by the compound. We have shown that the biscumarin derivative diminish phosphorylation of CHK-2 protein, TOR and P53 an event that is likely be the basis of observed inhibition of melanoma cell survival and tumor growth. Inhibition of cell survival and growth possibly due to the deminish phosphorylation of the chk-2 kinase on Threonine: T68, of TOR on Serine: S2448 of P53 on Serine: S392.
E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2018.04.252
P-106
The redox-active thiosemicarbazone, Dp44mT promotes NDRG1 to down-regulate oncogenic signaling pathways in cancer Sharleen Menezes, Zaklina Kovacevic, Des Richardson The University of Sydney, Sydney, Australia
The redox-active thiosemicarbazone, Dp44mT has shown potent anticancer activity against various cancers. This occurs by its ability to sequester, bind iron, and also produce reactive oxygen species. The activity of this agent is in part mediated by its ability to upregulate the metastasis suppressor, N-myc Downstream Regulated Gene 1 (NDRG1). NDRG1 inhibits cell proliferation and migration by negatively regulating numerous oncogenic signaling pathways. However, the mechanisms by which it modulates these pathways remain to be elucidated. In this study we examined the effect of NDRG1 and Dp44mT on the ErbB-family of receptor tyrosine kinases namely epidermal growth factor receptor (EGFR), human epidermal growth factor