- Email: [email protected]
Identification of novel carbonylated amino acids in proteins from human plasma
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D. Rossin et al. / Free Radical Biology and Medicine 108 (2017) S18–S107
irradiated with increasing UVR doses (0.5–2 MED) 3 times per week for 4 weeks. After the end of the irradiations, 5 formulations containing retinoic acid 0.05%, lactic acid 10%, salicylic acid 3%, epidermal growth factor 3.3% and the vehicle were applied on the back of the mice for 3 weeks. Hydration, transepidermal water loss (TEWL), elasticity and skin thickness, were frequently measured. After the end of applications, oxidative stress was evaluated in skin biopsies and strippings of stratum corneum using fluorescence spectroscopy, as well endogenous antioxidants using HPLC–EC detection. Epidermal thickening was evident in all mice after UV exposure. After treatment with lactic acid the thickness decreased significantly. Highest TEWL was observed in the retinoic acid group. The skin treated with salicylic acid demonstrated the lowest levels of ascorbic and uric acid. In strippings, lactic acid icreased the levels of antioxidants. Highest elasticity was measured in the retinoic acid group. In conclusion, retinoic and lactic acids seem to be the most effective treatments for the photodamaged skin. E-mail address: [email protected] (M. Karasmani) http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.231
P-147
In vitro evaluation of pine extracts antioxidant protection Crhysoula Zerva, Eleni Andrianna Kanari, Nikolaos Panagiotis Andreou, Vasilios Roussis, Emmanouil Vourakis, Maria Kyriazi, Georgios Papaioannou, Michail Rallis National Kapodistrian University of Athens, Greece
Oxidative stress is known to be responsible for a wide range of pathological conditions. Many skin diseases are initiated or/and promoted by oxidative stress. Oxidative stress could be caused by insufficient fuctioning of the antioxidant defence mechanisms or increased exposure to reactive oxygen species (ROS). The antioxidant and protective efficacy of aqueous extracts of various parts of the plants Pinus halepensis, Pinus brutia, Pinus nigra, and Eucalyptus globulus in comparison with aqueous extract of green tea dry leaves (Camelia sinensis, Theaceae), was evaluated. The plant selection was based on their content in phenolic antioxidant molecules. The comparison was made by carrying out in vitro assays of the plant extracts and observing their effects on cell culture of fibroblasts from mice fibroblasts, which were exposed to chemical oxidative stress. The resutls of this research showed the importance of the concentration of phenolic antioxidants contained into the extracts. The lower dose of 0,1 μg/ml of extract decrease oxidative stress and viability, while 1 μg/ml enhace oxidative stress and fibroblast viabilty. It seems that rates of mitosis is dependent of oxidative stress.
P-148
Identification of novel carbonylated amino acids in proteins from human plasma Adelina Rogowska-Wrzesinska University of Southern Denmark, Denmark Keywords: Human plasma; carbonylation; peroxyl and alkoxyl radicals
In order to fully understand the role of protein carbonylation in health and disease, identification of carbonylated protein(s), and their detailed characterisation is required. Mass spectrometry is a particularly suited for such studies due to its specificity and sensitivity. We have used biotin-hydrazide and mass spectrometry based approach for identification of up to 14 different types of carbonylated amino acids. In native human plasma we have observed 133 carbonylated sites in 36 proteins. The approach identified 10 hitherto undetected types of carbonylated amino acids in proteins: aldehyde and ketone modifications of leucine, valine, alanine, isoleucine, glutamine, lysine and glutamic acid (+14 Da), an oxidised form of methionine - aspartate semialdehyde (-32 Da) - and decarboxylated glutamic acid and aspartic acid (-30 Da). The carbonyl compounds reported are consistent with the chemistry of peroxyl and alkoxyl radicals generated on proteins. The consequence of the formation of these products has yet to be understood. However, it is important to note that some of these carbonyls can introduce changes to protein charge, give rise to Schiff base cross-links, and can lead to changes in residues that define protein structure (e.g. ring opening of P residues). These events may affect protein function (e.g. interaction with other) or protein conformation and activity. E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.233
P-149
Roles of branched-chain amino acids regulation in oxidative stress revealed by fibroblasts from classic Maple Syrup Urine Disease patients Paula Fernandez Guerra 1, Lei Cheng 2, Robert A. Fenton 2, Peter Bross 1, Pilar Rodriguez Pombo 3, Johan Palmfeldt 1 1
Research Unit for Molecular Medicine, Aarhus University and Aarhus University Hospital, Denmark 2 Department of Biomedicine, Aarhus University, Aarhus, Denmark 3 Centro de Diagnóstico de Enfermedades Moleculares (CEDEM), Dpto Biol. Mol., Centro de Biología Molecular-SO, UAM-CSIC, Centro de Investigación en Red de Enfermedades Raras, (CIBERER), IDIPAZ, Universidad Autónoma Madrid, Madrid, Spain
E-mail address: [email protected] (N. Panagiotis Andreou)
Keywords: Oxidative stress; mitochondrial superoxide; protein carbonylation; Maple Syrup Urine Disease; branched-chain amino acid catabolism
http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.232
Branched-chain amino acids (BCAAs) are essential amino acids commonly used in clinical procedures to improve patients outcome.However, their roles and cellular mechanisms are not clear. One effect of BCAAs supplementation is reduction of oxidative stress. BCAAs levels are regulated by their catabolism and the rate limiting enzyme is branched chain a-ketoacid dehydrogenase
D. Rossin et al. / Free Radical Biology and Medicine 108 (2017) S18–S107
irradiated with increasing UVR doses (0.5–2 MED) 3 times per week for 4 weeks. After the end of the irradiations, 5 formulations containing retinoic acid 0.05%, lactic acid 10%, salicylic acid 3%, epidermal growth factor 3.3% and the vehicle were applied on the back of the mice for 3 weeks. Hydration, transepidermal water loss (TEWL), elasticity and skin thickness, were frequently measured. After the end of applications, oxidative stress was evaluated in skin biopsies and strippings of stratum corneum using fluorescence spectroscopy, as well endogenous antioxidants using HPLC–EC detection. Epidermal thickening was evident in all mice after UV exposure. After treatment with lactic acid the thickness decreased significantly. Highest TEWL was observed in the retinoic acid group. The skin treated with salicylic acid demonstrated the lowest levels of ascorbic and uric acid. In strippings, lactic acid icreased the levels of antioxidants. Highest elasticity was measured in the retinoic acid group. In conclusion, retinoic and lactic acids seem to be the most effective treatments for the photodamaged skin. E-mail address: [email protected] (M. Karasmani) http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.231
P-147
In vitro evaluation of pine extracts antioxidant protection Crhysoula Zerva, Eleni Andrianna Kanari, Nikolaos Panagiotis Andreou, Vasilios Roussis, Emmanouil Vourakis, Maria Kyriazi, Georgios Papaioannou, Michail Rallis National Kapodistrian University of Athens, Greece
Oxidative stress is known to be responsible for a wide range of pathological conditions. Many skin diseases are initiated or/and promoted by oxidative stress. Oxidative stress could be caused by insufficient fuctioning of the antioxidant defence mechanisms or increased exposure to reactive oxygen species (ROS). The antioxidant and protective efficacy of aqueous extracts of various parts of the plants Pinus halepensis, Pinus brutia, Pinus nigra, and Eucalyptus globulus in comparison with aqueous extract of green tea dry leaves (Camelia sinensis, Theaceae), was evaluated. The plant selection was based on their content in phenolic antioxidant molecules. The comparison was made by carrying out in vitro assays of the plant extracts and observing their effects on cell culture of fibroblasts from mice fibroblasts, which were exposed to chemical oxidative stress. The resutls of this research showed the importance of the concentration of phenolic antioxidants contained into the extracts. The lower dose of 0,1 μg/ml of extract decrease oxidative stress and viability, while 1 μg/ml enhace oxidative stress and fibroblast viabilty. It seems that rates of mitosis is dependent of oxidative stress.
P-148
Identification of novel carbonylated amino acids in proteins from human plasma Adelina Rogowska-Wrzesinska University of Southern Denmark, Denmark Keywords: Human plasma; carbonylation; peroxyl and alkoxyl radicals
In order to fully understand the role of protein carbonylation in health and disease, identification of carbonylated protein(s), and their detailed characterisation is required. Mass spectrometry is a particularly suited for such studies due to its specificity and sensitivity. We have used biotin-hydrazide and mass spectrometry based approach for identification of up to 14 different types of carbonylated amino acids. In native human plasma we have observed 133 carbonylated sites in 36 proteins. The approach identified 10 hitherto undetected types of carbonylated amino acids in proteins: aldehyde and ketone modifications of leucine, valine, alanine, isoleucine, glutamine, lysine and glutamic acid (+14 Da), an oxidised form of methionine - aspartate semialdehyde (-32 Da) - and decarboxylated glutamic acid and aspartic acid (-30 Da). The carbonyl compounds reported are consistent with the chemistry of peroxyl and alkoxyl radicals generated on proteins. The consequence of the formation of these products has yet to be understood. However, it is important to note that some of these carbonyls can introduce changes to protein charge, give rise to Schiff base cross-links, and can lead to changes in residues that define protein structure (e.g. ring opening of P residues). These events may affect protein function (e.g. interaction with other) or protein conformation and activity. E-mail address: [email protected] http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.233
P-149
Roles of branched-chain amino acids regulation in oxidative stress revealed by fibroblasts from classic Maple Syrup Urine Disease patients Paula Fernandez Guerra 1, Lei Cheng 2, Robert A. Fenton 2, Peter Bross 1, Pilar Rodriguez Pombo 3, Johan Palmfeldt 1 1
Research Unit for Molecular Medicine, Aarhus University and Aarhus University Hospital, Denmark 2 Department of Biomedicine, Aarhus University, Aarhus, Denmark 3 Centro de Diagnóstico de Enfermedades Moleculares (CEDEM), Dpto Biol. Mol., Centro de Biología Molecular-SO, UAM-CSIC, Centro de Investigación en Red de Enfermedades Raras, (CIBERER), IDIPAZ, Universidad Autónoma Madrid, Madrid, Spain
E-mail address: [email protected] (N. Panagiotis Andreou)
Keywords: Oxidative stress; mitochondrial superoxide; protein carbonylation; Maple Syrup Urine Disease; branched-chain amino acid catabolism
http://dx.doi.org/10.1016/j.freeradbiomed.2017.04.232
Branched-chain amino acids (BCAAs) are essential amino acids commonly used in clinical procedures to improve patients outcome.However, their roles and cellular mechanisms are not clear. One effect of BCAAs supplementation is reduction of oxidative stress. BCAAs levels are regulated by their catabolism and the rate limiting enzyme is branched chain a-ketoacid dehydrogenase