O.I. Yablonskaya

A paradoxical effect of hydrated C60-fullerene at an ultralow concentration on the viability and aging of cultured Chinese hamster cells

The effect of an aqueous solution of hydrated C60-fullerene (HyFn) on the growth and “stationary phase aging” (accumulation of “age-related” changes in cultured cells during the slowing down of their proliferation within a single passage and the subsequent “aging” in the stationary phase of growth) of transformed B11-dii FAF28 Chinese hamster cells was studied. The final calculated concentration of HyFn in the growth medium was 10−19 M. A paradoxical result contrasting the available data on the absence of HyFn cytotoxicity at higher concentrations was obtained in our experiments: namely, HyFn decelerated cell proliferation (estimated by the growth of mass culture, as well as by the efficiency of colony formation) and accelerated the “stationary phase aging” of the cell culture. Moreover, repeated addition of an aqueous solution of HyFn (to the final calculated concentration of 10−19 M) to the cells that had already reached the stationary phase of growth caused a rapid (within no more than 24 h) death of a significant part of the cell population. The observed effect of HyFn at ultralow concentration is supposed to arise from the alterations in the properties of the water surrounding the fullerene molecule: namely, water becomes a donor and acceptor of electrons and regulates redox processes (especially those involving oxygen) in aqueous systems. This effect of HyFn at an ultralow concentration may be specific for transformed cells, and, therefore, experiments on normal fibroblasts with limited mitotic potential are planned as a continuation of the present study. It is also possible that the reported antiaging effect of HyFn in experimental animals is due to its anticancer, immunostimulatory, antiviral, and antibacterial properties manifested only at the whole-organism level.

The stable nonequilibrium state of bicarbonate aqueous systems

Data obtained by electron paramagnetic resonance (EPR) and chemiluminescence analysis indicate that in aqueous solutions of bicarbonates, superoxide radical and other reactive oxygen species (ROS) are constantly produced. The stationary level of the superoxide radical is found to increase when a solution is illuminated. Reactions involving ROS are shown to be accompanied by the generation of electron excitation energy, keeping bicarbonate solutions in a stable nonequilibrium state. The system can emit part of this energy. Variations in emitting activity are found to correlate with variations in the cosmophysical factors. The emitting activity of solutions is found to vary in the presence of low and ultralow concentrations of hydrated fullerenes. It is noted that the phenomenon of spontaneous charge separation in aqueous systems (G. H. Pollack) could play a role in maintaining a stable nonequilibrium state in bicarbonate systems where the reactions with ROS participation are catalyzed by forms of carbonate. It is concluded that the abovementioned properties of bicarbonate aqueous systems most likely keep living matter whose structural basis is formed by these systems in a stable excited state, thereby making it highly sensitive to the action of external factors with low and ultralow intensities.

Chemiluminescence of Cigarette Smoke: Salient Features of the Phenomenon.

The study disclosed herein provides for the first time a detailed experimental support for the general mechanism of the cigarette‐smoke‐derived chemiluminescence, as an example par excellence of the excited‐state generation in a chemically complex aerosol medium. The mechanism involves chemiexcitation in a unimolecular transformation of the smoke‐borne free radical species. However, the concentration of these radicals, [r∙] , obeys a bimolecular (second‐order) kinetics and depends on a particulate‐phase content (total particulate matter, TPM) of the cigarette smoke. The decrease in [r∙] with increasing the TPM amount manifests radical‐scavenging propensity of the smoke particulate phase. Astonishingly, no energy transfer takes place from the primary excited light‐emitting species to luminophoric molecules abundant in the smoke. The reported results build up fundamentals of a facile chemiluminescence assay for free radical properties of the smoke. The experimental approaches developed for this study are of general scope and may be used for mechanistic elucidation of the excited‐state generation in chemical systems and environments of an arbitrary complexity.

Stabilizing effects of hydrated fullerenes C60 in a wide range of concentrations on luciferase, alkaline phosphatase, and peroxidase in vitro

Abstract Hydrated fullerene (HyFnC60) is a highly hydrophilic supra-molecular complex consisting of unmodified С60 fullerene molecule enclosed into a hydrated shell. It has been shown in numerous experiments that aqueous solutions of HyFnC60 manifest a wide range of biological activities both in vivo and in vitro even at very low concentrations of HyFnC60. We used a spectrophotometric method and a method of biochemoluminescence to demonstrate that HyFnC60 in concentrations below 10−9 M down to 10−23 M stabilizes peroxidase, alkaline phosphatase, and bacterial luciferase against inactivation due to long-term incubation of the enzymes at room temperature and also against heat inactivation. In addition, HyFnC60 was able to “revive” heat inactivated enzymes. These effects cannot be explained by the direct action of the fullerene molecules upon the enzymes. We suggest that the effects of HyFnC60 on the enzymes are related to the ability of hydrated fullerene C60 molecules to organize thick aqueous shells around them. One of the specific properties of water phase in these shells is its ability to optimize redox reactions, which can support enzyme stability against factors deteriorating their structure.

Changes in chemiluminescence of whole blood of COPD patients treated with Hypoxen® and effects of C60 fullerenes on blood chemiluminescence

Summary Background Chronic obstructive pulmonary disease (COPD) is an inflammatory disease associated with reactive oxygen species (ROS) production. The aim of this study was to evaluate the effect of Hypoxen® treatment and the effect of HyFnC60 on ROS production in patients’ blood. Material/Methods ROS production in blood was estimated using chemiluminescence (CL) measurement with CL-amplifiers: luminol (LM), LM + zymosan (ZM) or lucigenin (LC) in the presence or absence of hydrated fullerenes (HyFnC60) added to blood in low concentrations. Results In all the patients with COPD in remission phase with Hypoxen® prescription, the LM-dependent CL (LM-CL) with ZM and LC-enhanced CL (LC-CL) decreased after the treatment. Parameters of CL and effects of HyFnC60 upon them depended on blood state. Addition of HyFnC60 to blood decreased data scattering and helped to improve discrimination between different groups of patients. Using the discriminator analysis, we found the most important time-points in the kinetic curves of CL for classification of patients into groups (eg, COPD patients before and after treatment with Hypoxen®; patients’ blood with different sensitivity to HyFnC60 concentration). Conclusions Monitoring of CL of non-diluted whole blood in COPD patients can be used for the estimation of the Hypoxen® efficiency in complex therapy. Addition of HyFnC60 to blood increases sensitivity of the method.

Chapter 11 - Exogenous and Endogenous Mediators of Oxygen Metabolism: Alternatives for Chemical and Biological Activity

Abstract The present chapter provides the survey of the contemporary current state of art in studying the salient mechanistic alternatives of involving the bioactive mediators of oxygen metabolism in oxidative developments and draws particular attention to unresolved issues, which merit in-depth investigation. The forthcoming critical overview encompasses both chemical and biological issues, which are discussed in terms of oxidative stress. However, as the performed analysis reveals, the very term “oxidative stress” is still “ill defined,” and for comprehension of the latter phenomenon, first of all, the existing terminology needs to be revisited to reach a general agreement on definitions. The problems reviewed and analyzed herein refer to understanding the role of their numerous endogenous and exogenous mediators as prooxidant and antioxidant reactants involved in oxidation processes in vivo, merging and interconverting their chemical activities, and overlapping their functions with the other physiologically active compounds (most prominently, exhibiting hormonal activity).