Donika Ivanova

Magnetic resonance imaging of mitochondrial dysfunction and metabolic activity, accompanied by overproduction of superoxide.

This study shows that a mitochondria-penetrating nitroxide probe (mito-TEMPO) allows detection of superoxide and visualization of mitochondrial dysfunction in living cells due to the effect of T1 shortening in MRI. Mitochondrial dysfunction was induced by treatment of cells with rotenone and 2-methoxyestradiol (2-ME/Rot). The MRI measurements were performed on 7T MRI. The 2-ME/Rot-treated cells were characterized by overproduction of superoxide, which was confirmed by a conventional dihydroethidium test. In the presence of mito-TEMPO, the intensity of MRI signal in 2-ME/Rot-treated cells was ∼30-40% higher, in comparison with that in untreated cells or culture media. In model (cell-free) systems, we observed that superoxide, but not hydrogen peroxide, increased the intensity of T1-weighted MRI signal of mito-TEMPO. Moreover, the superoxide restores the T1-weighted MRI contrast of mito-TEMPOH, a noncontrast (diamagnetic) analogue of mito-TEMPO. This was also confirmed by using EPR spectroscopy. The results demonstrate that superoxide radical is involved in the enhancement of T1-weighted MRI contrast in living cells, in the absence and presence of mito-TEMPO. This report gives a direction for discovering new opportunities for functional MRI, for detection of metabolic activity, accompanied by overproduction of superoxide, as well as by disturbance of the balance between superoxide and hydrogen peroxide, a very important approach to clarify the fine molecular mechanisms in the regulation of many pathologies. The visualization of mitochondrial activity in real-time can be crucial to clarify the molecular mechanism of the functional MRI in its commonly accepted definition, as a method for detection of neurovascular coupling.

Vitamin K: Redox-modulation, prevention of mitochondrial dysfunction and anticancer effect

This review is directed to the redox-modulating properties and anticancer effect of vitamin K. The concept is focused on two aspects: (i) redox-cycle of vitamin K and its effect on the calcium homeostasis, “oncogenic” and “onco-suppressive” reactive oxygen species and the specific induction of oxidative stress in cancer; (ii) vitamin K plus C as a powerful redox-system, which forms a bypass between mitochondrial complexes II and III and thus prevents mitochondrial dysfunction, restores oxidative phosphorylation and aerobic glycolysis, modulates the redox-state of endogenous redox-pairs, eliminates the hypoxic environment of cancer cells and induces cell death. The analyzed data suggest that vitamin C&K can sensitize cancer cells to conventional chemotherapy, which allows achievement of a lower effective dose of the drug and minimizing the harmful side-effects. The review is intended for a wide audience of readers - from students to specialists in the field.

Effect of antigen stimulation on the oxidative stress parameters in sperm of outbred and inbred rabbits

ABSTRACT The presence of an increased oxidative stress during antigen stimulation was established in the sperm of inbred rabbits by evaluation of the level of real-time free radical formation (ROS and Asc•), final products of oxidation of lipids (MDA) and the activities of antioxidant defence enzymes superoxide dismutase (SOD) and catalase (CAT). There was not a significant difference between levels of ROS, Asc• and MDA in inbred and outbred rabbits before immunization. The immune response, represented by levels of ROS and Asc• of inbread and outbread rabbits were statistically significantly higher compared to those before immunization (p = 0.0001 and p = 0.00001) Moreover, the levels of ROS in the sperm of inbread rabbits were statistically significantly higher compared to outbread rabbits (p < 0.05). Oxidative stress was accompanied by an adaptive increase of SOD and CAT activities during the immune response, compared to those before immunization. Furthermore, the increased SOD and CAT activities appeared to be sufficient to inactivate the oxidative stress. We measured reduced levels of ROS, Asc• and MDA 30 days after immunization. When immune response reduced, the observed SOD and CAT activities tended to return to the values before immunization. That might have been connected with the decreased oxidative burden. However, CAT activities remained about 1.5 times higher than that before immunization. In conclusion, our results indicate that the administered antigen stimulation induces oxidative stress in both groups inbred and outbred rabbits.