Cell hydration as a universal marker for detection of environmental pollution

At present, when the technological progress brings progressive increase in environmental pollutions by different chemical and physical (ionizing and non-ionizing radiations) factors, the detection of the safety of environmental medium from the point of public health is one of the fundamental problems of modern Life Sciences. This problem has especially disquieting character after the Chernobyl and Japan nuclear catastrophes, when the level of background ionizing radiation and chemical pollutions of environmental medium of the number of world’s regions are increased beyond safety doses. As the biological effect of weak environmental factors have nonlinear dose-dependent character, besides its thermodynamic characteristics it depends also on environmental composition and initial state of organism. Therefore, the current policy of World Health Organization and other international organizations whose mission is to establish safety standards for environmental pollutions by chemical and physical factors, based only on the their concentration or energy absorption rate by organism cannot be considered as adequate. It is suggested that the biological marker having universal sensitivity to different factors and determining the functional state of organisms could be used for estimation of the safety doses of environmental factors on organism. In present review are presented the data consisting of the hypothesis according to which the Na/K pump and Na/Ca-controlling cell hydration could serve as a universal and extra-sensitive cellular marker for detection of hazardous effect of environmental pollutions.     

High-affinity ouabain receptors: primary membrane sensors for ionizing radiation

Although the high sensitivity of the Na/K pump in cell membrane to ionizing radiation is well known in literature, the individual role of different isoforms of pump in determination of its radio-sensitivity is not clear yet. This is the subject of the present investigation. Using isotope, electro-physiological and enzymological methods, the effect of γ-ionizing radiation on cell membrane voltage-current characteristics, acetylcholine-induced membrane current, ²²Na⁺ and ⁴⁵Ca²⁺ exchange between cells and bathing solution, Na+K⁺-ATPase activity, dose-dependent ouabain binding with cell membrane, intracellular cAMP and membrane phosphorylation in snail neurons were studied. The changes in neurons as a result of 30-min γ-radiation exposure of snails to 5.16 Ci/kg at the end of the first 30 min of post-radiation period were as follows: the increase in membrane ionic conductance reversed the ouabain sensitivity of acetylcholine-induced currents, stimulation of ²²Na⁺ and ⁴⁵Ca²⁺ uptakes, inhibition of Na/K pump, activation Na/Ca exchange in reversed mode, increase in ouabain binding with high-affinity α₃ and decrease with α₂ middle-affinity receptors, decrease in intracellular cAMP content and membrane dephosphorylation. On the basis of the obtained data, it is suggested that both α₃ and α₂ catalytic subunits of Na⁺+K⁺-ATPase serve primary membrane sensors through the activation of which the biological effect of γ-radiation on neurons is realized. The IR has activation effects on α₃-dependent Na⁺/Ca²⁺ exchange in forward and its inactivation on α₂-dependent reverse modes.     

Age dependency of high-affinity ouabain receptors and their magnetosensitivity

Age dependency of [³H]-ouabain binding, ⁴⁵Ca²⁺ eflux and its magnetosensitivity in rats’ brain cortex and heart muscle tissues were studied. Curves of dose-dependent [³H]-ouabain binding consisted of three components with different affinities (10⁻⁷–10⁻⁴ M (α₁); 10⁻⁹–10⁻⁷ M (α₂); and 10⁻¹¹–10⁻⁹ M (α₃)). These curves were also characterized by different dose-dependent kinetics. [³H]-ouabain binding with α₃ receptors in brain cortex and heart muscle tissues of young and adult animals had a dose-dependent character, while that in old ones had a dose-independent character. A 0.2 T static magnetic field (SMF) exposure had modulation effect on ouabain binding with α₁, α₂ and α₃ receptors in young rats, while in adult ones, only α₃ receptors were magnetosensitive. In old animals, SMF exposure had no significant effect on ouabain binding with α₃ receptors in brain cortex, while in heart muscle, it had inhibitory effect on it. Age-dependent effect of ouabain impact on ⁴⁵Ca²⁺ efflux showed that all concentrations of ouabain lead to inhibitory effect in young animals’ brain cortex and heart muscle (with the exception of 10⁻¹⁰ and 10⁻⁶ M), while in old ones, it had activation effect as compared with data received without ouabain. SMF exposure in young animals had activation effect on ⁴⁵Ca²⁺ efflux from brain cortex and heart muscle in data without ouabain, and in old rats, ⁴⁵Ca²⁺ efflux from brain cortex was magnetic insensitive. In old animals, SMF increased ⁴⁵Ca²⁺ efflux even after extra low concentration of ouabain. It is suggested that α₃ receptors having a crucial role in the regulation of Na⁺/Ca²⁺ exchange serve as age-dependent magnetosensors of excitable cells.     

Magnetically treated water at 4?Hz and 2.5?mT as a modulator of cisplatin effect on cell hydration and ouabain binding of sarcoma-180 tissue

There are many data about the extremely low-frequency electromagnetic fields (ELF-EMF) therapeutic use, especially in the field of oncology. Recent data suggest that 4 Hz EMF having dehydration effect on tissues has a pronounced antitumor activity on sarcoma-180 in mice. It was shown that 4 Hz EMF have pronounced effects on physicochemical properties of water and water solution. Therefore, the aim of the present work was the comparative study of the modulation effect of ELF-EMF on cisplatin-induced changes cell hydration and number of ouabain receptors in membrane of sarcoma-180 tumor tissues. Tissue hydration was measured as wet weight/dry weight and expressed as a water content of g/g in dry weight. The number of ³H-ouabain receptors in membrane was counted by isotope scintillation counter. In conclusion, ELF-EMF can be a possible tool for stimulation of cisPt antitumor effect.     

Comparative study of non-ionizing and ionizing radiation effect on hydration of winter wheat seeds in metabolic active and inactive states

Although the biological effect of non-ionizing (NIR) and ionizing radiation (IR) on plant seed potential is well documented, the mechanism of this impact is not clear yet. Since cell hydration, a fundamental parameter for determining cell function, is magneto-sensitive, in present work, a comparative study of extremely low frequencies (ELF) electromagnetic field (EMF), ELF EMF-modulated millimeter waves, and radiation exposure effects on winter wheat seeds water absorption and dry mass changes was performed. The seeds were incubated in physiological solutions (PS) for 2 and 72 h. In case of 2-h incubation, the water absorption determined by osmotic gradients between seeds and bathing medium, PS treated by all these factors leads to changes of water absorption. While in case of 72-h incubation (when germination processes are in active state) these impact had reversed character. The obtained data strongly suggest seeds bathing medium as a target through which the biological effects of NIR and IR are realized.     

Double-frequency impedance measurement method of determination of body hydration

The aim of the present study is to estimate the changes in cell hydration in different conditions using impedance measurements. Experiments were performed on adult and young male albino rats. Tissue hydration level was estimated as a difference between tissue wet and dry weight. Ouabain concentrations from 10⁻¹¹ to 10⁻⁴ M, physiological solution (PS), mannit and distillated water (DW) were used. The impedance measurements were taken with “Biophys-Expert-1”. The increase in water content in the brain cortex (through the injection) raises the active conductivity after intraperitoneally injections of PS, mannit and DW. The more pronounced increase was observed in case of DW injection. The values of reactive conductivity were higher when mannit injection was made. In case of different ouabain concentrations, the active and reactive components were higher in young animals than in adult ones. Considering obtained results, the impedance measurement can be provided as a method for the estimation of tissue hydration level.     

The frequency-dependent effect of extremely low-frequency electromagnetic field and mechanical vibration at infrasound frequency on the growth, division and motility of Escherichia coli K-12

The aim of this work was to investigate the frequency-dependent effects of extremely low-frequency electromagnetic field (ELF-EMF) and mechanical vibration at infrasound frequency (MV at IS frequency or MV) on growth and development of Escherichia coli K-12, by using classical microbiological (counting colony forming units), isotopic, spectrophotometric and electronmicroscopic methods. The frequency-dependent effects of MV and ELF-EMF were shown that they could either stimulate or inhibit the growth and the division of microbes depending on the periods following exposure. However, the mechanism through which the MV and ELF-EMF effects affect the bacteria cell is not clear yet. It was suggested that the aqua medium could serve a target through which the biological effect of MV and ELF-EMF on microbes could be realized. To check this hypothesis, the frequency-dependent effects (2, 4, 6, 8, 10 Hz) of both MV and ELF-EMF on the bacterial growth, division and their motility in cases of exposure, the preliminary treated microbes-free medium and microbes containing medium were studied. Both MV and ELF-EMF effect on microbes have frequency and post-exposure period duration-dependent characters. The [ ³ H]-thymidine involving experiments shown that EMF at 4 Hz exposure has pronounced stimulation effect on cell proliferation while 4 Hz MV has inhibition effect. But at 8–10 Hz, the both EMF and MV have inhibitory effects on cell proliferation. It is suggested that 4 and 8 Hz EMF have different biological effects on microbes.     

The metabolic pathway of 4?Hz mechanical vibration-induced effect on snail heart muscle contractility

It was shown that the 4 Hz 30 dB mechanical vibration (MV) of physiological solution (PS) had modulation effect on snail heart contractility. However, the nature of metabolic pathway of MV-treated PS-induced effect is not clear yet. It was suggested that the MV-induced modulation of water molecules dissociation leads to the variation of reactive oxygen species’ level in cell bathing medium, which could serve as a messenger for switching on the metabolic pathway(s) responsible for modulation of muscle contractility. The aim of present work was to check this hypothesis and to elucidate the metabolic pathway through which the effect of MV-treated PS on heart contractility was realized. For this purpose, the effect of MV on heat fusion periods (HFP) and H₂O₂ content in PS, as well as on heart contractility, ⁴⁵Ca²⁺ efflux, intracellular levels of cGMP and cAMP, muscle hydration, and ouabain binding were studied. It was shown that MV treatment of PS increased the HFP-(21.33 ± 4%) and decreased the H₂O₂ content-(5 ± 0.9%). The intracardiac perfusion by MV-treated PS increased the amplitudes of heart contractility, which was accompanied by the increasing of ⁴⁵Ca²⁺ efflux (252.4 ± 16%), elevation of cGMP’s level (42.05 ± 7%), decreasing of cAMP’s level (82.67 ± 7%), increasing of the tissue hydration (18.64 ± 3%), and increasing of the number of ouabain binding sides (25 ± 4%). It was suggested that MV-induced increasing of heart muscle contraction amplitudes is due to the decreases of H₂O₂ content in the medium, which leads to the elevation of heart muscle contractility in result of activation of cGMP-dependent Na⁺/Ca²⁺ exchange in forward regime.     

The EMF-induced changes in aqua medium’s properties depend on background ionizing radiation, illumination and temperature

The effects of extremely low-frequency electromagnetic field (ELF EMF) and ELF-modulated millimeter waves (MMW) on physicochemical properties of physiological solution (PS) at different environmental medium were studied. The existence of frequency “windows” at 4 Hz and 8 Hz frequencies of ELF EMF and ELF-modulated MMW which have different effects on heat fusion period (HFP), hydrogen peroxide (H₂O₂) formation and oxygen (O₂) content of water solution and different dependency on temperature, background radiation (BGR) and illumination was shown. The obtained data allow us to suggest that cell bathing medium serving as a target through which the biological effects of ELF EMF and ELF-modulated MMW on cells are realized is extra sensitive to environmental factors. Therefore, the variability of experimental data on EMF biological effects, obtained in different laboratories can be explained by different environmental conditions of experiments, which very often are not considered adequately.     

The comparative study of the effects of 4 Hz Electromagnetic Fields-, Infrasound-treated and hydrogen peroxide containing physiological solutions on Na pump-induced inhibition of heart muscle contractility

The effects of 4 Hz extremely low frequency Electromagnetic Fields (ELF EMF), Infrasound (IS) and hydrogen peroxide (H₂O₂) on Na⁺–K⁺ pump-induced transient inhibition of heart muscle contractility was studied. It was documented that EMF-treated and H₂O₂-containing physiological solution had synergic depressing effect on Na⁺–K⁺ pump-induced inhibition of muscle contractility, while the IS had elevation effect on it. On the basis of the obtained data the H₂O₂ could be suggested as the messenger through which the stimulation effect of EMF on heart muscle is realised, while the relaxing effect of IS on heart contractility is due to the decrease of CO₂ solubility of muscle bathing aqueous medium.