AN EXPERIMENTAL AND THEORETICAL STUDY OF THE REACTIVITY OF SELECTED CATECHOLAMINES AND THEIR PRECURSORS TOWARDS ASCORBYL RADICAL

Ascorbyl radical is often used as a biomarker of oxidative stress in human organism. It is a product of the antioxidant activity of ascorbic acid and it is not expected to react further with biologically important molecules. For the first time, the reactivity of catecholamines and their precursors was investigated theoretically and experimentally towards ascorbyl radical and the main structural parameters governing activity were analyzed. It was shown that catechol moiety plays an important role, which classifies norepinephrine and 3,4-dihydroxyphenylacetic acid as the most reactive when compared to homovanillic acid, vanillylmandelic acid, and octopamine. DFT methods have been employed to determine the most probable mechanism of the reaction. Based on the change in reaction enthalpy it was concluded that Hydrogen Atom Transfer (HAT) is a preferred mechanism both in water and pentyl ethanoate. The stabilization interactions in ascorbic acid, its anion and radical are carefully analyzed in order to understand the preferability of the mentioned mechanism. The stability of the ascorbyl radical is explained in detail. The results prove that ascorbyl radical is not just a product of antioxidant activity, but also a potentially harmful species in an organism.

Electron Paramagnetic Resonance - A Powerful Tool of Medical Biochemistry in Discovering Mechanisms of Disease and Treatment Prospects

Electron Paramagnetic Resonance - A Powerful Tool of Medical Biochemistry in Discovering Mechanisms of Disease and Treatment ProspectsIn pathophysiological conditions related to oxidative stress, the application of selected antioxidants could have beneficial effects on human health. Electron paramagnetic resonance (EPR) spectroscopy is a technique that provides unique insight into the redox biochemistry, due to its ability to: (i) distinguish and quantify different reactive species, such as hydroxyl radical, superoxide, carbon centered radicals, hydrogen atom, nitric oxide, ascorbyl radical, melanin, and others; (ii) evaluate the antioxidative capacity of various compounds, extracts and foods; (iii) provide information on other important parameters of biological systems. A combination of EPR spectroscopy and traditional biochemical methods represents an efficient tool in the studies of disease mechanisms and antioxidative therapy prospects, providing a more complete view into the redox processes in the human organism.

Antioxidative Defense Enzymes in Placenta Protect Placenta and Fetus in Inherited Thrombophilia from Hydrogen Peroxide

Our aim was to investigate the activities of antioxidative defense enzymes in the placenta, fetal blood and amnion fluid in inherited thrombophilia. Thrombophilia was associated with nearly threefold increase of activity (p < 0.001) of the placental catalase (81.1 ± 20.6 U/mg of proteins in controls and 270.0 ± 69.9 U/mg in thrombophilic subjects), glutathione (GSH) peroxidase (C: 20.2 ± 10.1 U/mg; T: 60.0 ± 15.5 U/mg), and GSH reductase (C: 28.9 ± 5.6 U/mg; T: 72.7 ± 23.0 U/mg). The placental activities of superoxide dismutating enzymes—MnSOD and CuZnSOD, did not differ in controls and thrombophilia. Likewise, the activities of catalase and SOD in the fetal blood, and the level of ascorbyl radical which represents a marker of oxidative status of amniotic fluid, were similar in controls and thrombophilic subjects. From this we concluded that in thrombophilia, placental tissue is exposed to H2O2-mediated oxidative stress, which could be initiated by pro-thrombic conditions in maternal blood. Increased activity of placental H2O2-removing enzymes protects fetus and mother during pregnancy, but may increase the risk of postpartum thrombosis.