The 4Hz mechanical vibration-activated Na/Ca exchange as a quantum-sensitive novel target for pain therapy.

The activation of Na/40Ca exchange in reverse (R) mode leading to neuronal swelling and muscle contraction has been suggested as a mechanism for generation of pain signals. However, the activation of R Na/45Ca exchange, having higher rate than R Na/40Ca exchange, brings to depression of pain sensation, the mechanism of which is not clear. The previous data that the 4 Hz mechanical vibration (MV) has pain-relieving effects by activation of cGMP-dependent Na/Ca exchange in forward (F) mode, which leads to muscle hydration and neuronal dehydration, as in case of R Na/45Ca exchange activation, allow us to suggest that the comparative study of the effects of 4 Hz MV and R Na/45Ca exchange on thermal pain thresholds, tissue hydrations in different experimental conductions will make it possible to evaluate the mechanisms of R Na/45Ca exchange-induced inhibition of pain sensation. The obtained data show that the R Na/45Ca exchange-induced depression of pain sensation is due to high [Ca2+]i-inactivation of Na/K pump which brings to further increase of [Ca2+]i, while 4 Hz MV-activated F Na/Ca exchange-induced pain-relieving effect is due to Na/K pump activation by low [Ca2+]i. It is suggested that the R Na/45Ca exchange inhibits pain sensation, which is due to high [Ca2+]i-induced depression inhibition of muscle contractility, while the 4 Hz MV-induced pain-relieving effect is due to activation of Na/K pump by cGMP-dependent decrease of [Ca2+]i leading to muscle relaxation and neuronal dehydration. Therefore, the 4 Hz MV has been suggested as a novel quantum-mechanical sensitive target for pain therapy.

The Na/Ca Exchange as a Target for Antitumor Effect of 4Hz Pulsing Magnetic Field.

The correlation between cell hydration and proliferation is one of the controversial subjects in the present cancer research because the detailed mechanism(s) of correlation between these two cell parameters are not evaluated yet. The fact that magnetic fields have antitumor effects and our previous data that the effect of 4 Hz pulsing magnetic field (PMF) is realized by the activation of Cyclic guanosine monophosphate (cGMP)-dependent Na/Ca exchange allow us to suggest that through the study of PMF effects on cell hydration and proliferation in soft tissues of sarcoma-180 tumor-carrying mice compared with those of healthy ones will be possible to evaluate the character of correlation between cell hydration and proliferation in norm and pathology. The daily 30-min PMF (0.2 mT) exposed on tumor-carrying mice has lifetime prolongation, time-dependent depression of tumor growth, decrease of thermal threshold, and lactate concentration in blood. In the initial (6 days) period tumorigenesis PMF leads to dehydration in spleen cells, which is accompanied by the activation of cell proliferation, while in late periods of tumorigenesis it leads to cell hydration accompanied with proliferation. We are suggesting that PMF-induced overhydration and the increase of cell proliferation in cancer tissue as a consequence of high [Ca]i-induced activation of Ca-calmoduline-NO-cGMP pathway leads to the activation of FNa/Ca exchange, while its antitumor effect expressed in initial period of tumorigenesis is due to the activation of cGMP-dependent FNa/Ca, where because of low [Ca]i, it brings to cell dehydration as a result of reactivation of Na/K pump.

Cyclic AMP-dependent signaling system is a primary metabolic target for non-thermal effect of microwaves on heart muscle hydration.

Previously, we have suggested that cell hydration is a universal and extra-sensitive sensor for the structural changes of cell aqua medium caused by the impact of weak chemical and physical factors. The aim of present work is to elucidate the nature of the metabolic messenger through which physiological solution (PS) treated by non-thermal (NT) microwaves (MW) could modulate heart muscle hydration of rats. For this purpose, the effects of NT MW-treated PS on heart muscle hydration, [3H]-ouabain binding with cell membrane, 45Ca2+ uptake and intracellular cyclic nucleotides contents in vivo and in vitro experiments were studied. It is shown that intraperitoneal injections of both Sham-treated PS and NT MW-treated PS elevate heart muscle hydration. However, the effect of NT MW-treated PS on muscle hydration is more pronounced than the effect of Sham-treated PS. In vitro experiments NT MW-treated PS has dehydration effect on muscle, which is not changed by decreasing Na+ gradients on membrane. Intraperitoneal injection of Sham- and NT MW-treated PS containing 45Ca2+ have similar dehydration effect on muscle, while NT MW-treated PS has activation effect on Na+/Ca2+ exchange in reverse mode. The intraperitoneal injection of NT MW-treated PS depresses [3H]-ouabain binding with its high-affinity membrane receptors, elevates intracellular cAMP and decreases cGMP contents. Based on the obtained data, it is suggested that cAMP-dependent signaling system serves as a primary metabolic target for NT MW effect on heart muscle hydration.

The role of cell hydration in realization of biological effects of non-ionizing radiation (NIR).

The weak knowledge on the nature of cellular and molecular mechanisms of biological effects of NIR such as static magnetic field, infrasound frequency of mechanical vibration, extremely low frequency of electromagnetic fields and microwave serves as a main barrier for adequate dosimetry from the point of Public Health. The difficulty lies in the fact that the biological effects of NIR depend not only on their thermodynamic characteristics but also on their frequency and intensity "windows", chemical and physical composition of the surrounding medium, as well as on the initial metabolic state of the organism. Therefore, only biomarker can be used for adequate estimation of biological effect of NIR on organisms. Because of the absence of such biomarker(s), organizations having the mission to monitor hazardous effects of NIR traditionally base their instruction on thermodynamic characteristics of NIR. Based on the high sensitivity to NIR of both aqua medium structure and cell hydration, it is suggested that cell bathing medium is one of the primary targets and cell hydration is a biomarker for NIR effects on cells and organisms. The purpose of this article is to present a short review of literature and our own experimental data on the effects of NIR on plants' seeds germination, microbe growth and development, snail neurons and heart muscle, rat's brain and heart tissues.

Comparative study of time-dependent effects of 4 and 8 Hz mechanical vibration at infrasound frequency on E. coli K-12 cells proliferation.

The aim of the present work is to study the time-dependent effects of mechanical vibration (MV) at infrasound (IS) frequency at 4 and 8 Hz on E. coli K-12 growth by investigating the cell proliferation, using radioactive [(3)H]-thymidine assay. In our previous work it was suggested that the aqua medium can serve as a target through which the biological effect of MV on microbes could be realized. At the same time it was shown that microbes have mechanosensors on the surface of the cells and can sense small changes of the external environment. The obtained results were shown that the time-dependent effects of MV at 4 and 8 Hz frequency could either stimulate or inhibit the growth of microbes depending from exposure time. It more particularly, the invention relates to a method for controlling biological functions through the application of mechanical vibration, thus making it possible to artificially control the functions of bacterial cells, which will allow us to develop method that can be used in agriculture, industry, medicine, biotechnology to control microbial growth.

Cell hydration as a biomarker for estimation of biological effects of nonionizing radiation on cells and organisms.

"Changes in cell hydration" have been hypothesized as an input signal for intracellular metabolic cascade responsible for biological effects of nonionizing radiation (NIR). To test this hypothesis a comparative study on the impacts of different temperature and NIR (infrasound frequency mechanical vibration (MV), static magnetic field (SMF), extremely low frequency electromagnetic field (ELF EMF), and microwave (MW)) pretreated water on the hydration of barley seeds in its dormant and germination periods was performed. In dormant state temperature sensitivity (Q 10) of seed hydration in distilled water (DW) was less than 2, and it was nonsensitive to NIR treated DW, whereas during the germination period (48-72 hours) seeds hydration exhibited temperature sensitivity Q 10 > 2 and higher sensitivity to NIR treated DW. Obtained data allow us to suggest that the metabolic driving of intracellular water dynamics accompanied by hydrogen bonding and breaking is more sensitive to NIR-induced water structure changes in seed bathing aqua medium than the simple thermodynamic processes such as osmotic gradient driven water absorption by seeds in dormant state. Therefore, cell hydration is suggested to be a universal and extrasensitive biomarker for detection of biological effects of NIR on cells and organisms.

Age-dependent effect of static magnetic field on brain tissue hydration.

Age-dependent effect of Static Magnetic Field (SMF) on rats in a condition of active and inactive Na(+)/K(+) pump was studied for comparison of brain tissues hydration state changes and magnetic sensitivity. Influence of 15 min 0, 2 Tesla (T) SMF on brain tissue hydration of three aged groups of male albino rats was studied. Tyrode's physiological solution and 10(-4) M ouabain was used for intraperitoneal injections. For animal immobilization, the liquid nitrogen was used and the definition of tissue water content was performed by tissue drying method. Initial water content in brain tissues of young animals is significantly higher than in those of adult and aged ones. SMF exposure leads to decrease of water content in brain tissues of young animals and increase in brain tissues of adult and aged ones. In case of ouabain-poisoned animals, SMF gives reversal effects on brain tissue's hydration both in young and aged animals, while no significant effect on adults is observed. It is suggested that initial state of tissue hydration could play a crucial role in animal age-dependent magnetic sensitivity and the main reason for this could be age-dependent dysfunction of Na(+)/K(+) pump.

Age-dependent magnetosensitivity of heart muscle ouabain receptors.

In our previous work we have shown that the age-dependent decrease in the magnetosensitivity of heart muscle hydration is accompanied by a dysfunction of the Na(+) /K(+) pump. The reciprocal relation between the Na(+/) K(+) pump and Na(+) /Ca(2+) exchange in development was suggested as a possible pathway for the age-dependent decrease in the magnetosensitivity of heart muscle hydration (water content). Because high and low affinity ouabain receptors in cell membranes are involved in Na(+) /Ca(2+) exchange and Na(+) /K(+) pump functions, respectively, the effect of a 0.2 T static magnetic field (SMF) on dose-dependent, ouabain-induced hydration and [(3) H]-ouabain binding with heart muscle tissues in young, adult and older rats was studied. Three populations of receptors in membranes with high (10(-11) -10(-9) M), middle (10(-9) -10(-7) M) and low (10(-7) -10(-4) M) affinity to [(3) H]-ouabain were distinguished, which had specific dose-dependent [(3) H]-ouabain binding kinetics and effects on muscle hydration. The magnetosensitivity of [(3) H]-ouabain binding kinetics with high affinity receptors was prominent in all the three age groups of animals, while with low affinity receptors it was more expressed only in the young group of animals. All three types of receptors that caused modulations of muscle hydration were age dependent and magnetosensitive. Based on the obtained data we came to the conclusion that heart muscle hydration in young animals is more magnetosensitive due to the intense expression of high affinity ouabain receptors, which declines with aging.

Bidirectional frequency-dependent effect of extremely low-frequency electromagnetic field on E. coli K-12.

In the present work, the frequency-dependent effects of extremely low-frequency electromagnetic field (ELF EMF) on Escherichia coli K-12 growth have been studied. The frequency-dependent effects of ELF EMF have shown that it can either stimulate or inhibit the growth of microbes. However, the mechanism by which the ELF EMF affects the bacterial cells is not clear yet. It was suggested that the aqua medium can serve as a target through which the biological effect of ELF EMF on microbes may be realized. To check this hypothesis, the frequency-dependent effects (2, 4, 6, 8, 10 Hz, B = 0.4 mT, 30 min) of ELF EMF on the bacterial growth were studied in both cases where the microbes were in the culture media during the exposure and where culture media was preliminarily exposed to the ELF EMF before the addition of bacteria. For investigating the cell proliferation, the radioactive [(3)H]-thymidine assay was carried out. It has been shown that EMF at 4 Hz exposure has pronounced stimulation while at 8 Hz it has inhibited cell proliferation.

The study of the effects of mechanical vibration at infrasound frequency on [(3)H]-thymidine incorporation into DNA of E. coli K-12.

The aim of the present work was to investigate the frequency-dependent effects of mechanical vibration at infrasound frequency (MV at IS frequency or MV) on E. coli K-12 growth by investigating the cell proliferation, using radioactive [(3)H]-thymidine assay. The frequency-dependent effects of MV were shown that it could either stimulate or inhibit the growth of microbes. However, the mechanism through which the MV effects affect the bacterial cells is not clear yet. It was suggested that the aqua medium can serve as a target through which the biological effect of MV on microbes could be realized. To check this hypothesis the frequency-dependent effect (2, 4, 6, 8, 10 Hz) of MV on the bacterial growth in cases of exposure the preliminary treated microbes-free medium and microbes containing medium were studied. It has been shown that MV at 4, 8, and 10 Hz frequency has inhibition effects, while at 2 and 6 Hz has stimulation effects on cell proliferation.

Age-dependent magnetosensitivity of heart muscle hydration.

The reason for hyper magnetosensitivity of young animals compared to older ones remains unclear. It has been suggested that age-induced tissue dehydration (decreased water content) could be a basis for the aging-related decrease in the organism's magnetosensitivity. To test this hypothesis, the effect of a 0.2 T static magnetic field (SMF) exposure on heart muscle hydration in three age groups of rats (young, adult, and older) was studied, with and without ouabain poisoning. The SMF exposure resulted in heart muscle dehydration of young (21%) and adult (6.2%) rats but had no effect on older animals. In young animals without ouabin poisoning, SMF exposure caused dehydration of the heart muscle while in the ouabain-poisoned animals it led to hydration (29.6%). These hydration effects were more pronounced in young animals than in adult and older animals. The increased hydration (5.7%) of heart muscles in older animals was evoked by providing distilled water for seven days, which elevated (by 12%) the SMF-induced heart muscle hydration effect. These results suggest that the hyper magnetosensitivity of the young heart muscle and the lower sensitivity of older animals are due to initial high (83.5%) and low (75.3%) tissue hydration levels, respectively. Therefore, the age-induced decrease in the magnetosensitivity of heart muscle is likely to be a result of Na(+)/K(+) pump dysfunction.

Cell bathing medium as a target for non thermal effect of millimeter waves.

Non thermal (NT) effect of direct radiation 4 Hz-modulated 90-160 GHz of Millimeter Waves (MMW) and preliminary MMW-treated physiological solution (PS) influence were studied on snail isolated neuron, rat's brain tissue hydration and skin penetration. It was shown that the 4 Hz-modulated low intensity 90-160 GHz MMW direct radiation and MMW-treated PS leads to on single neuron shrinkage, skin and brain tissue dehydration. On the basis of obtained data it was suggested that the cell bathing aqua medium serve as a target through which the NT effect of MMW on cell hydration is realized. The MMW-induced brain tissue dehydration can considering as consequence of MMW-induced skin water structural changes leading to unknown messenger formation able to modulate the brain cell hydration. The extrasensitivity of cell hydration to low intensity of MMW radiation allow to recommend cell hydration as a cellular marker for estimation of the NT biological effect of MMW on cells and organisms.

The density-dependency of dark- and low-background radiation effects on water and water solution properties.

The effects of dark -(Ev=0 lux) and low-background radiation (BGR), where R<1µRongen/h, on physicochemical properties (specific electrical conductivity, heat fusion, hydrogen peroxide (H2O2), and oxygen contents) of distilled water (DW) and physiological solution (PS) at 4°C and 18°C were studied. The incubation of DW and PS samples in dark and in low BGR (under dark) medium at 4°C and 18°C brings to changes of their physicochemical properties compared with DW and PS samples incubated in light and normal BGR condition (Ev=500-550 lux and R=17 µRoentgen/h). The observed changes of DW and PS properties depended on their initial temperature, density and ionic composition. It is suggested that water molecules dissociation and ions hydration are sensitive to illumination and BGR. Therefore, the cell-bathing medium can be considered as a messenger through which direct and non direct (by modulating of others factors-induced effects) influences of illumination and BGR on cell metabolism are realized.

The modulating impact of illumination and background radiation on 8 Hz-induced infrasound effect on physicochemical properties of physiolagical solution.

At present, when the level of background ionizing radiation is increasing in a number of world locations, the problem of the study of biological effect of high background radiation becomes one of the extremely important global problems in modern life sciences. The modern research in biophysics proved that water is a most essential target, through which the biological effects of ionizing and non-ionizing radiations are realized. Therefore, there is no doubt about the strong dependency of non-ionizing radiation-induced effect on the level of background radiation. Findings have shown that illumination and background radiation have a strong modulation effect on infrasound-induced impacts on water physicochemical properties, which could also have appropriate effect on living organisms.

The non thermal effect of weak intensity millimeter waves on physicochemical properties of water and water solutions.

The comparative study of the effects of 5.8 mW/cm(2) Millimeter Waves (MMW) and near Infrared (IR) irradiation on thermal properties, specific adsorption rate (SAR), specific electrical conductivity (SEC) and hydrogen peroxide (H(2)O(2)) content of distilled water (DW), and physiological solutions (PS) was performed. The thermal effect of MMW irradiation appeared only after the first minute of irradiation, while the IR heating started from the first minute of irradiation. The heat fusion of frozen MMW-treated DW and PS was significantly less than sham and IR-treated DW and PS. MMW irradiation had time-dependent elevation effect on water SEC and SAR, which was accompanied by the increase of H(2)O(2) formation in it. We suggest that the MMW-induced vibration of water dipole molecules caused the non thermal changes of physicochemical properties of DW and PS, which promote the formation of H(2)O(2) in water.

Exogenous hydrogen peroxide as a possible messenger for the stimulation effect of magnetized physiological solution on heart contractility.

The dual effect of magnetized physiological solution (MPS) on snail heart muscle contractility (muscle relaxation and stimulation of heartbeat) was shown previously. The MPS-induced relaxation of the heart muscle has been explained by activation of cGMP-dependent Ca(2+) effluxes from the muscle; however, the mechanism of the stimulating effect of MPS on heartbeat remains unclear. As in the presence of paramagnetic oxygen molecules, magnetic fields could generate the exogenous reactive oxygen species such as hydrogen peroxide (H(2)O(2)), we hypothesize that H(2)O(2) may play a role as the possible messengers through which the activation effect of MPS on heartbeat is realized. To test this hypothesis, the dose-dependent effects of exogenous H(2)O(2) on heart muscle contractility and (45)Ca uptake were studied. Here we compared the obtained data with the previous results of the effects of MPS on heart muscle contractility and (45)Ca uptake. We found that exogenous H(2)O(2) and MPS have similar effects on Na(+)-K(+) pump-induced transient inhibition of muscle contractility and (45)Ca uptake. The Na(+)-K(+) pump-induced depression of H(2)O(2)-sensitivity of muscle contractility is determined by activation of Ca(2+) efflux from the cell. On the basis of these data we suggest the exogenous H(2)O(2) as a possible messenger through which the stimulation effect of MPS on heart muscle is realized.

The biological effect of extremely low frequency electromagnetic fields and vibrations on barley seed hydration and germination.

The changes of wet and dry weights and germination of barley seed in different periods of its swelling in nontreated (control), extremely low frequency electromagnetic fields (ELF EMF) )-treated, and extremely low frequency vibrations (ELFV)-treated cold (4 degrees C) and warm (20 degrees C) distilled water (DW) were studied. The metabolic-dependent seed hydration, dry weight dissolving, germination, and water binding in seed were modulated by preliminary EMF- and ELFV-treated DW. Frequency "windows" for the effect of EMF and ELFV on seed hydration, solubility, water binding in seed, and germination were discovered. These "windows" were different for EMF and ELFV, as well as in various phases of seed swelling. It is suggested that EMF-induced water structure modification has a different biological effect on the process of seed hydration, solubility, water binding in seed, and germination compared to ELFV.

On the modulation effect of pulsing and static magnetic fields and mechanical vibrations on barley seed hydration.

The changes of wet and dry weights of barley seed in different periods of swelling were studied in seeds treated with Extremely Low Frequency Electromagnetic Fields (ELF EMF), Static Magnetic Fields (SMF) and Mechanical Vibrations (MV) in cold (4 degrees C) and warm (20 degrees C) distilled water as well as in seeds non-treated (control). The metabolic dependent seed hydration, dry weight loss and water binding in seed were modulated by preliminary EMF, SMF and MV-induced treatment of distilled water. The specific electrical conductivity (SEC) of control and treated distilled water was measured before the seed incubation. Frequency and intensity "windows" (i.e. range of frequency or intensity) for the effect of EMF, MV and SMF (correspondingly) on seed hydration, solubility and water binding in seed were studied. These "windows" were different in various phases of seed swelling. It is suggested that water structure modification is the result of valence angle changes (SMF and EMF) and dipole molecules vibration (EMF and MV) has different effects on the process of hydration, solubility and water binding in seed. These results are important from the point of understanding the mechanisms of the biological effect of EMF, as well as from the point of agriculture.

Effect of hydrogen peroxide on neuron sensitivity to acetylcholine.

The dose-dependent effect of hydrogen peroxide on snail neuromembrane chemosensitivity was studied by means of standard voltage-clamp method. Short-term exposure (7 min) of neurons to H(2)O(2) (10(-11)-10(-4) M) caused dose-dependent depression of Acetylcholine (Ach)-induced ionic currents in the membrane. The H(2)O(2)-induced depression of Ach-sensitivity of membrane was more pronounced in K(+)-free solution than in normal physiological solution and it disappeared in cold medium (5 degrees C). The H(2)O(2) (10(-11)-10(-4) M) decreased membrane electrical conductivity and cell volume. The dose-dependent decrease in Ach-sensitivity of the snail neuromembrane by H(2)O(2) may be due to a decrease in the number of functionally active membrane receptors caused by a decrease in membrane active surface. H(2)O(2)-induced decrease in Ach-sensitivity has a metabolic but Na(+)-K(+) pump independent character, the nature of which is the subject for current investigation.

Metabolic pathway of magnetized fluid-induced relaxation effects on heart muscle.

The effect of magnetized physiological solution (MPS) on isolated, perfused snail heart muscle contractility, (45)Ca uptake and intracellular level of cAMP, and cGMP was studied. The existence of the relaxing effect of MPS on heart muscle at room temperature (22 degrees C) and its absence in cold medium (4 degrees C) was shown. The MPS had a depressing effect on (45)Ca uptake by muscles and intracellular cAMP content and an elevating effect on intracellular cGMP level. It is suggested that the relaxing effect of MPS on heart muscle is due to the decrease of intracellular Ca ions as the result of activation of cGMP-dependent Ca efflux. The MPS induced decrease of intracellular cAMP content can be considered as a consequence of intracellular Ca loss, leading to the Na + K-ATPase reactivation, and causing the decrease of the intracellular level of ATP, serving as a substrate and positive modulator of cyclase activity.