Biomedical subjects
Publications and source records attributed to N K Chemeris.
At the early stage of trypsin treatment of mollusc neurons tetrodotoxin cannot block the Na+ current. In the course of further exposure of neurones to trypsin, tetrodotoxin-sensitivity is restored completely, so its temporal loss results from shielding rather than destruction of the tetrodotoxin-binding site. Pronase and papain do not affect the tetrodotoxin action on the Na+ current.
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The dose-inward current relationship for calcium channel blocking ions of Ni, Co, Mn and Mg was studied by means of intracellular dialysis and voltage clamp in isolated neurons from the mollusc Limnea stagnalis. For all the four ions the effect was described by the Langmuir isotherm with KMn = 0.36 +/- 0.05 m,, KNi = KCo = 0.74 +/- 0.07 mM, KMg = 18.2 +/- 5.00 mM. These bivalent cations compete with current-carrying ions for a common binding site in the channel's entrance.
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The dose-inward current relationships for current-carrying ions Ca2+ and Sr2+ were studied by means of intracellular dialysis and voltage clamp in isolated neurons of the mollusc Limnea stagnalis. For Ca2+ and Sr2+ ions these relationships had ceiling. The data obtained are explained in terms of activation theory using two-barrier model of calcium channel. Values of velocity constants vx for ion x2+ transition over the second energetic barrier and of constants Kx corresponding to effectiveness of binding in the channel's entrance were determined: vca:vsr = 1:2; KCa = 1400 M-1; KSr = 270 M-1.
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Frequency-dependent modifications of intracellular free calcium concentration ([Ca2+]i) in neutrophils exposures to modulated extremely high frequency electromagnetic radiation were analyzed using a special mathematical model for [Ca2+]i oscillations. The model took into account the activation of Ca2+ influx into the cell by cytosolic Ca2+ and Ca(2+)-induced Ca2+ release from intracellular stores. The calcium channels of plasma membrane were chosen as a target for the influence of harmonic signal and additive noise in the model. The model simulation showed that in response to modulating signal, the rise in [Ca2+]i, has frequency dependence and phase dependence in relation to the moment of chemical stimulation. The phase-frequency dependence of the effect was observed at a certain sequence of delivery of chemical stimulus and modulating signal to the cell. At intensities of modulating signals exceeding the threshold, a rise in [Ca2+]i, reaching a level of more than 50% of the initial level, was observed at a frequency of about 1 Hz and in the phase range of 0.3-2.5 radians. The effect was found only at high intensities of chemical stimulus. The additive noise introduced into the system modified qualitatively and quantitatively the phase-frequency characteristics of the cell response to the modulating signal. An increase in noise intensity resulted in a displacement of the average frequency of the band of rise in [Ca2+]i, and then the emergence of a set of bands with a greater Q-factors. The analysis of dynamics of the nonlinear system in terms of the stability theory showed that, as the intensity of chemical stimulus increases, the system transits by means of a series of bifurcations from regular driving to chaotic, and then to oscillations, induced by a modulating harmonic signal. The boundary of the transition of oscillations from chaotic to induced ones corresponds to a specific "threshold" of the intensity of chemical stimulus for the significant rise in [Ca2+]i in response to the modulating signal. The results of the model analysis are in good correspondence with the experimental data obtained earlier, namely, with the effects of modulated extremely high-frequency electromagnetic radiation on neutrophils, which were observed only in the presence of Ca2+ in extracellular medium and at high concentrations of calcium ionophore A23187. Thus, as the characteristic frequency of the quasi-periodic process of calcium signalling in the cell coincides with the frequency of external field, a narrow-band rise in [Ca2+]i is observed, which can result in a modification of the functional activity of the cell.
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.
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.
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.
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.
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.
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.
The anti-inflammatory effect of low-intensity extremely high-frequency electromagnetic radiation (EHF EMR, 42.0 GHz, 0.1 mW/cm2) was compared with the action of the known anti-inflammatory drug sodium diclofenac and the antihistamine clemastine on acute inflammatory reaction in NMRI mice. The local inflammatory reaction was induced by intraplantar injection of zymosan into the left hind paw. Sodium diclofenac in doses of 2, 3, 5, 10, and 20 mg/kg or clemastine in doses of 0.02, 0.1, 0.2, 0.4, and 0.6 mg/kg were injected intraperitoneally 30 min after the initiation of inflammation. The animals were whole-body exposed to EHF EMR for 20 min at 1 h after the initiation of inflammation. The inflammatory reaction was assessed over 3 - 8 h after the initiation by measuring the footpad edema and hyperthermia of the inflamed paw. Sodium diclofenac in doses of 5 - 20 mg/kg reduced the exudative edema on the average by 26% as compared to the control. Hyperthermia of the inflamed paw decreased to 60% as the dose of was increased diclofenac up to 20 mg/kg. EHF EMR reduced both the footpad edema and hyperthermia by about 20%, which was comparable with the effect of a single therapeutic dose of diclofenac (3 - 5 mg/kg). The combined action of diclofenac and the exposure to the EHF EMR caused a partial additive effect. Clemastine in doses of 0.02-0.4 mg/kg it did not cause any significant effects on the exudative edema, but in a dose of 0.6 mg/kg it reduced edema by 14 - 22% by 5 - 8 h after zymosan injection. Clemastine caused a dose-dependent increase in hyperthermia of inflamed paw at doses of 0.02-0.2 mg/kg and did not affect the hyperthermia at doses of 0.4 and 0.6 mg/kg. The combined action of clemastine and EHF EMR exposure caused a dose-dependent abolishment of the anti-inflammatory effect of EHF EMR. The results obtained suggest that both arachidonic acid metabolites and histamine are involved in the realization of anti-inflammatory effects of low-intensity
A short analytical review of Soviet and foreign studies devoted to metabolic regulation of neuron functioning is presented. It is shown that the electrical activity of neuron is accompanied by structural changes and an increase in RNA content. It is assumed that these changes are initiated by calcium ions, coming to the neuron during its excitation. The principles of metabolic regulation of functioning of electro- and chemoexcitable neuronal membranes are considered. It is shown that the metabolic reactions can be an intermediate between the activated chemoreceptor and the electrical response of the cell. Evidence is presented that the electroexcitable and chemosensitive structures in the membrane modulate each other. The role of intracellular messengers in regulating the activity of receptors and channels is considered. Possible mechanisms for their modifications under the influence of cellular factors are discussed. It is shown that the work of neuron is controlled by several metabolic regulatory systems simultaneously.
It has been shown that inactivation of cholinoreceptors observed in the region of neuron depolarization is conditioned by the supply of Ca ions into the cell through the electro--excitable Ca2+-channels. It is proved by three means: 1) variation of outer concentration of Ca2+(Ca2+h); 2) application of the blockers of electroexcitable Ca2+ channels; 3) variation of intracellular concentration Ca2+(Ca2+b).
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Dopamine has been found to decrease reversibly the action potential rising phase in identified L. stagnalis neurones. Using voltage-clamp and intracellular dialysis techniques it is shown that dopamine inhibits electroexcitable Ca-current, and changes in Ca-current determine those in the action potential shape. Serotonin and adrenaline influence the Ca-current in a similar way as dopamine does. Inhibition of Ca-conductance is found not to be related to possible effect of these neuromediators on intracellular level of free Ca ions or cyclic AMP.