Nowadays, there is growing interest in new compositions and drugs that can preserve and maintain human health. There are many studies on both known probiotics and new sources of biologically active substances which may potentially be new avenues for human longevity. Here, we describe the protective effect of cell lysates of the relict bacteria Bacillus sp. F (Bacilus Lyuba), isolated from the ancient permafrost of Central Yakutia, upon DNA damage, induced by hydrogen peroxide. The DNA damage was monitored by a comet assay in leukocytes isolated from the peripheral blood of experimental animals (BALB/c mice). Cell lysates were prepared using the French Press and ultrasonic treatment. The levels of DNA damage in the presence of bacterial lysates upon the stress test (20 μM hydrogen peroxide for 10 min at 37°C) were almost twice lower, compared with the buffer control. Of note, the maximal protective effect was observed in the temperature range of 50°C–60°C; with the increase of the temperature, this effect vanished. The obtained results do not give a clear answer on the mechanism of the protective properties of Bacillus sp. F. (Bacilus Lyuba) lysates. To answer this question, a detailed analysis of the protein composition of bacterial lysate at different temperatures should be performed.
The influence of different exposure regimes of low-intensity extremely high-frequency electromagnetic radiation on the growth rate of solid Ehrlich carcinoma in mice has been studied. It was shown that, at an optimum repetition factor of exposure (20 min daily for five consecutive days after the tumor inoculation), there is a clearly pronounced frequency dependence of the antitumor effect. The analysis of experimental data indicates that the mechanisms of antitumor effects of the radiation may be related to the modification of the immune status of the organism. The results obtained show that extremely high-frequency electromagnetic radiation at a proper selection of exposure regimes can result in distinct and stable antitumor effects.
Upon inducing acute local inflammation in the mouse hindpaw with zymosan, exposure to pulsed microwaves (35.27 GHz, peak power 20 kW, pulse duration 400–600 ns, repetition rate 5–500 s −1 ) could attenuate the exudative edema and local hyperthermia by 20%; in dynamics and scale these effects were similar to those of 3 μg/g Diclofenac. The anti-inflammatory effect increased roughly linearly with pulse duration at the same repetition rate, and showed a threshold dependence on average power flux density at fixed pulse duration. The dependence on the duration of exposure at fixed emission parameters was bell-shaped both in total irradiation and in local limb irradiation.
The capability of high peak-power pulsed electromagnetic radiation of extremely high frequency (35,27 GHz, pulse widths of 100 and 600 ns, peak power of 20 kW) to excite acoustic waves in model water-containing objects and muscular tissue of animals has been experimentally shown for the first time. The amplitude and duration of excited acoustic pulses are within the limits of accuracy of theoretical assessments and have a complex nonlinear dependence on the energy input of electromagnetic radiation supplied. The velocity of propagation of acoustic pulses in water-containing models and isolated muscular tissue of animals was close to the reference data. The excitation of acoustic waves in biological systems under the action of high peak-power pulsed electromagnetic radiation of extremely high frequency is the important phenomenon, which essentially contributes to the understanding of the mechanisms of biological effects of these electromagnetic fields.
The antiinflammatory effect of low-intensity extremely-high-frequency electromagnetic radiation (EHF EMR, 42.0 GHz, 0.1 mW/cm2) was studied in comparison to the effects of the antiinflammatory drug sodium diclofenac and the antihistamine clemastine in acute inflammatory reaction in mice of NMRI outbred stock. The local inflammatory reaction was induced by intraplantar injection of zymosan to the left hind paw. Intraperitoneal injections of 2, 3, 5, 10, and 20 mg/kg of sodium diclofenac or 0.02, 0.1, 0.2, 0.4, and 0.6 mg/kg of clemastine were made 30 min after the initiation of inflammation. An hour after the initiation of inflammation, animals were whole-body exposed to EHF EMR for 20 min. The inflammatory reaction was assessed 3–8 h after initiation by measuring the footpad edema and hyperthermia of the inflamed paw. Sodium diclofenac (5–20 mg/kg) reduced the exudative edema by ∼26% compared to the control. Hyperthermia of the inflamed paw decreased by 60% with an increase in the diclofenac dose to 20 mg/kg. EHF EMR reduced both the footpad edema and hyperthermia by ∼20%. This was comparable to the effect of a single therapeutic dose of diclofenac (3–5 mg/kg). The combination of diclofenac and exposure to EHF EMR produced a partial additive effect. Clemastine (0.02–0.4 mg/kg) did not affect the exudative edema, but at a dose of 0.6 mg/kg, edema was reduced by 14–22% five to eight hours after zymosan injection. Clemastine caused a dose-dependent increase in hyperthermia of inflamed paw at doses 0.02–0.2 mg/kg and did not affect the hyperthermia at doses 0.4 and 0.6 mg/kg. A combination of clemastine and EHF EMR exposure resulted in a dose-dependent abolishment of the antiinflammatory effect of EHF EMR. Our results suggest that both arachidonic acid metabolites and histamine are involved in the achievement of the antiinflammatory effects of low-intensity EHF EMR.
The radioprotective effects of guanosine and of inosine on bone marrow cells of mice exposed to acute X-rays (1.5 Gy) were studied by using the micronuclear test. The guanosine and inosine (riboxine) decrease the frequency of micronucleated polychromatic erythrocytes and significantly recover erythropoiesis. Also, radioprotective effects of the guanosine and of the inosine on the irradiated leucocytes of mice were tested by the alkaline comet assay. Was shown that purine ribonucleosides diminish quantity of DNA damage and activates repair processes in leucocytes under irradiation of blood and animals. The reactive oxygen species induced by ionizing radiation perform essential role in DNA damaging. Using a sensitive method of enhanced chemiluminescence in a peroxidase-luminol-p-iodophenol system for quantitative measurement of hydrogen peroxide and coumarin-3-carboxylic acid for quantitative measurement of hydroxyl radicals we have shown that guanosine and inosine essentially decrease the yield of hydrogen peroxide and hydroxyl radicals in X-ray-irradiated water. The results obtained indicate that radioprotective properties of guanosine and inosine (riboxine) in the blood cells are operative at the genome level.
Using the method of enhanced chemiluminescence in a peroxidase-luminol-p-iodophenol system, we found the formation of reactive oxygen species (in equivalent of hydrogen peroxide concentration) in 1 mM phosphate buffer under the exposure to high peak-power pulsed electromagnetic radiation of extremely high frequencies (37 GHz, peak power 20 kW, pulse width 400 ns, repetition rate 500 Hz). The results obtained show that the formation of hydrogen peroxide in aqueous solutions under the action of electromagnetic radiation is the result of the summary influence of heat and thermoacoustic waves excited in the solutions.
Low-intensity ultrahigh frequency electromagnetic radiation (42 GHz, 100 μW/cm2) reduces the severity of inflammation and inhibits production of active oxygen forms by inflammatory exudate neutrophils only in mice with inflammatory process. These data suggest that some therapeutic effects of electromagnetic radiation can be explained by its antiinflammatory effect which is realized via modulation of functional activity of neutrophils in the focus of inflammation.
Using a comet assay technique, it was shown for the first time that low-intensity extremely high-frequency electromagnetic radiation (EHF EMR) in vivo causes oppositely directed effects on spatial organization of chromatin in cells of lymphoid organs. In 3 hrs after single whole-body exposure of NMRI mice for 20 min at 42.0 GHz and 0.15 mW/cm2, an increase by 16% (p < 0.03 as compared with control) and a decrease by 16% (p < 0.001) in fluorescence intensity of nucleoids stained with ethidium bromide were found in thymocytes and splenocytes, respectively. The fluorescence intensity of stained nucleoids in peripheral blood leukocytes was not changed after the exposure. The exposure of cells of Raji hunan lymphoid line and peripheral blood leukocytes to the EHF EMR in vitro induced a decrease in fluorescence intensity by 23% (p < 0.001) and 18% (p < 0.05), respectively. These effects can be determined by changes in a number of physiological alkali-labile sites in DNA of exposed cells. We suggested that the effects of low-intensity EHF EMR on the immune system cells are realized with the participation of neuroendocrine and central nervous systems.
The effect of low-intensity extremely high-frequency electromagnetic radiation (EHF EMR, 42.0 GHz, 0.1 mW/cm(2), 20 min daily) on cell-mediated immunity and nonspecific inflammatory response in mice was studied. The intensity of cell-mediated immune response in the reaction of delayed-type hypersensitivity and nonspecific inflammation was estimated by a relative increase in the thickness of foot pad after immunization of animals by sheep red blood cells or zymosan. It was shown for the first time that the radiation reduces both immune and nonspecific inflammatory responses. It was shown with the use of models of acute inflammation and full-thickness skin wounds that EHF EMR suppresses the nonspecific inflammatory response but does not influence the duration of the pathological process. We suppose that the basis, of the effects revealed is the modification of functional activity of phagocytic cells under the influence of EHF EMR. The results suggest that some therapeutic effects of EHF EMR can be realized via the inhibition of inflammatory processes.
Low-intensity of electromagnetic radiation of extremely high frequencies (EHF EMR) is effectively used in medical practice for diagnostics, prevention and treatment of a broad spectrum of diseases of different etiology. However, in spite of existence of many hypotheses about mechanisms of EHF EMR effects on the molecular and cellular levels of organization of living systems, there is not conception that could explain all diversity of the EHF-therapy effects from unified approach. In our opinion, the problem of determination of mechanisms of EHF EMR effects on living organism is divided into two basic tasks: first, determining subcellular structures which can receive radiation, and, second, studying physiological reactions of the organism which are caused by radiation. It is obviously, that investigation of functions of single cells and subcellular elements can not entirely explain therapeutic effects and mechanisms of EHF EMR influence on multicellular organism on the whole. Plenty of functional relationships between organs and systems of organs should be taken into account. In the present review, a realization of the EHF-therapy effects due to the influence on immune system functions and start of system mechanisms of maintenance of the homeostasis on the organism level is hypothesized. Potential targets for EHF EMR acception on the level of different systems of the organism are analysed. The material is formed so that functional relations between immune system and other regulatory systems (nervous and endocrine systems) are traced.
It was shown by light and electron microscopy that local exposure of the projection of the MC-8 lao-gun acupuncture point in rat pad to low-intensity (0.05 mW/cm2) extremely high-frequency (42.0 GHz) electromagnetic radiation caused a degranulation of derma mast cells. It was suggested that the response of skin mast cells is an important amplifying mechanism in the chain of events leading to a systemic response of the organism to low-intensity electromagnetic radiation.
Using experimental and theoretical methods of dosimetry, the energy absorption of extremely high-frequency electromagnetic radiation (EHF EMR) in the skin of laboratory rats was analyzed. Specific absorption rate (SAR) in the skin was determined on the basis of both microthermometric measurements of initial rates of temperature rise in rat skin induced by the exposure and microcalorimetric measurements of specific heat of the skin. Theoretical calculations of SAR in the skin were performed with consideration for dielectric parameters of rat skin obtained from the measurements of the standing wave ratio upon reflection of electromagnetic waves from the skin surface and for the effective area of stationary overheating measured by infrared thermography. A numerical method was developed to determine electromagnetic wave energy reflected, absorbed, and transmitted in the model of flat layers. The algorithm of the method was realized in a computer program and used to calculate SAR in the skin on the basis of the complex dielectric constant of rat skin. The SAR values obtained from experimental measurements, theoretical calculations and numerical analysis are in good mutual correspondence and make about 220-280 W/kg at a frequency of 42.25 GHz and a power of 20 mW at the radiator output. The results obtained can be used for dosimetric supply of biomedical experiments on studying the physicochemical mechanisms of the biological effects of EHF EMR.
Using experimental and theoretical methods of dosimetry, the energy absorption of extremely high-frequency electromagnetic radiation (EHF EMR) in the skin of laboratory rats was analyzed. Specific absorption rate (SAR) in the skin was determined on the basis of both microthermometric measurements of initial rates of temperature rise in rat skin induced by the exposure and microcalorimetric measurements of specific heat of the skin. Theoretical calculations of SAR in the skin were performed with consideration for dielectric parameters of rat skin obtained from the measurements of the standing wave ratio upon reflection of electromagnetic waves from the skin surface and for the effective area of stationary overheating measured by infrared thermography. A numerical method was developed to determine electromagnetic wave energy reflected, absorbed, and transmitted in the model of flat layers. The algorithm of the method was realized in a computer program and used to calculate SAR in the skin on the basis of the complex dielectric constant of rat skin. The SAR values obtained from experimental measurements, theoretical calculations and numerical analysis are in good mutual correspondence and make about 220-280 W/kg at a frequency of 42.25 GHz and a power of 20 mW at the radiator output. The results obtained can be used for dosimetric supply of biomedical experiments on studying the physicochemical mechanisms of the biological effects of EHF EMR.
The dynamics of leukocyte number and functional activity of peripheral blood neutrophils under whole-body exposure of healthy mice to low-intensity extremely-high-frequency electromagnetic radiation (EHF EMR, 42.0 GHz, 0.15 mW/cm2, 20 min daily) was studied. It was shown that the phagocytic activity of peripheral blood neutrophils was suppressed by about 50% (p < 0.01 as compared with the sham-exposed control) in 2-3 h after the single exposure to EHF EMR. The effect persisted for 1 day after the exposure, and then the phagocytic activity of neutrophils returned to the norm within 3 days. A significant modification of the leukocyte blood profile in mice exposed to EHF EMR for 5 days was observed after the cessation of exposures: the number of leukocytes increased by 44% (p < 0.05 as compared with sham-exposed animals), mostly due to an increase in the lymphocyte content. The supposition was made that EHF EMR effects can be mediated via the metabolic systems of arachidonic acid and the stimulation of adenylate cyclase activity, with subsequent increase in the intracellular cAMP level. The results indicated that the whole-body exposure of healthy mice to low-intensity EHF EMR has a profound effect on the indices of nonspecific immunity.
The modification of indices of the humoral immune response to thymus-dependent antigen (sheep erythrocytes) after a whole-body exposure of healthy mice to low-intensity extremely-high-frequency electromagnetic radiation was studied. Male NMRI mice were exposed in the far-field zone of horn antenna at a frequency of 42.0 GHz and energy flux density of 0.15 mW/cm2 under different regimes: once for 20 min, for 20 min daily during 5 and 20 successive days before immunization, and for 20 min daily during 5 successive days after immunization throughout the development of the humoral immune response. The intensity of the humoral immune response was estimated on day 5 after immunization by the number of antibody-forming cells of the spleen and antibody titers. Changes in cellularity of the spleen, thymus and red bone marrow were also assessed. The indices of humoral immunity and cellularity of lymphoid organs changed insignificantly after acute exposure and series of 5 exposures before and after immunization of the animals. However, after repeated exposures for 20 days before immunization, a statistically significant reduction of thymic cellularity by 17.5% (p < 0.05) and a decrease in cellularity of the spleen by 14.5% (p < 0.05) were revealed. The results show that low-intensity extremely-high-frequency electromagnetic radiation with the frequency and energy flux density used does not influence the humoral immune response intensity in healthy mice but influences immunogenesis under multiple repeated exposures.
A theoretical analysis of the dependence of cell response to modulated electromagnetic fields on the parameters of modulating signals was performed on the basis of the model for calcium-dependent membrane-associated intracellular signaling systems. Expressed amplitude-frequency "windows" in the response of the nonlinear system to external influence were shown to depend on the effective time during which the amplitude of the influencing signal exceeded some threshold value, which is determined by the properties of the system. The presence of negative values in the influencing signal, i.e. the variations in the rate of some process around the inherent value, played an important role for the effect characteristics. The response of the nonlinear system to external influence strongly depended on the waveform of the modulating signal which is determined not only by frequency spectrum, but also by the initial phase of each frequency component. To ensure a regime for the optimal effect, the rates of increase and drop in the modulating signal amplitude should be synchronized with dynamic processes in the biological object being exposed.