Microwave Emissions and the Problem of Modern Viral Diseases.

The results of a study on the mechanisms of the influence of an increased level of microwave radiation on the growth of infectious, primarily viral, diseases in the environment are presented. This is the radiation of the earth's ionosphere, which reached its maximum in the late 1980s-early 2000s, following an increase in the level of solar activity since the 17th century. Over the past 30 years, the anthropogenic electromagnetic background has increased 100 times due to the development of cellular mobile communications and computerization. The predicted interaction of natural and anthropogenic sources of microwaves sharply increases their negative impact on the ecological situation. Of particular concern is the active spread in recent years of the new 5G communication standard; in the future, it is the development of the most dangerous millimeter range in our country. Energy from the environment in the microwave range can cause "unexpected behavior" in the DNA of viruses. Clarifications to the recommendations of experts on the protection of the population with the help of electromagnetic shielding, obtained in the framework of supramolecular physics of the environment, are proposed.

[Influence of silver and titanium dioxide nanoparticles on the expression of genes of biomarkers of inflammatory responses and apoptosis].

In order to evaluate the toxic effect of silver (AgNP) and titanium dioxide (TiO2) nanoparticles their influence on the expression of genes of biomarkers of inflammatory responses and apoptosis in human lymphocytes was studied. An increase in the IL-6, IL-8, TNF-α and p53 genes expression in the concentration range of silver and titanium dioxide nanoparticles of 10-40 µk g/ml was found. Increased expression of IL-6, IL-8, TNF-α and p53 genes under the nanoparticles action indicates the stimulation of the immune system and of apoptosis, respectively.

[Silver nanoparticles induce lipid peroxidation and morphological changes in human lymphocytes surface].

To evaluate toxicity of silver nanoparticles synthesized by using the reverse micelle formation method, the effects of nanoparticles on lipid peroxidation and morphological changes of cell membranes in human lymphocytes were studied. It was found that under the influence of nanoparticles a reduction in cell viability and formation of excessive levels of reactive oxygen species were observed. Silver nanoparticles at different concentrations activate the processes of lipid peroxidation and, as a consequence, led to morphological changes in human lymphocytes.

[ROS induction and structural modification in human lymphocyte membrane under the influence of carbon nanotubes].

To assess the potential risks of using the artificial nanostructures the structural state of the human lymphocyte membrane and lipid peroxidation under the influence of multi-walled carbon nanotubes with metal impurities was studied. The ability of carbon nanotubes to induce the formation of reactive oxygen species in cells was examined. A dose-dependent increase in reactive oxygen species formation in lymphocytes, which was not registered in cells pre-incubated with N-acetylcystein, after exposure to carbon nanotubes was shown. The addition of iron chelator deferoxamine to carbon nanotubes has also resulted in a decrease of reactive oxygen species. The mechanism of the activation of lipid peroxidation under the influence of carbon nanotubes and a structural modification of human lymphocyte membranes were discussed.

[Cloning and analysis of coding cDNA sequence of human lymphocyte Csk tyrosine kinase under normal conditions and in choroidal melanoma].

The main function of Csk tyrosine kinases is phosphorylation of the C-terminal part of Srk tyrosine kinases as a mechanism of their downregulation. A decrease in the expression of csk gene results in the enhancement of Srk tyrosine kinase activity. In this study, cDNA containing the full coding sequence of the human leukocyte Csk tyrosine kinase gene has been cloned. The protein encoded by a a 1624-bp cDNA fragment has 99% homology to human Csk tyrosine kinase. A comparative sequence analysis of full-length cDNAs for Csk tyrosine kinase of normal lymphocytes and lymphocytes of patients with choroidal melanoma revealed a nucleotide substitution in dexon 10 of the gene, which appears to be of diagnostic significance. It has been shown that the risk of choroidal melanoma correlated with the frequency of this allele.

[Obtaining a full-length encoding cDNA of Csk family tyrosine kinase from human lymphocytes].

Tyrosine kinases of Csk family play important role in the cell growth regulation and normal cell differentiation and also can participate in the process of cancer genesis as oncoproteins. The main function of these tyrosine kinases is the phosphorylation of the Src family tyrosine kinases at their carboxyl terminus, which is the basis of their activity negative regulation. The disturbance of the csk gene expression leads to the increase of the Src tyrosine kinase activity. We have cloned a full-length encoding cDNA of the tyrosine kinase csk gene of human lymphocytes. 1.6-kilobase cDNA encodes the protein, which consists of 12 exons with conserved SH2 and SH3 domains. The homology between this protein and human Csk tyrosine kinase is 99%. A full-length DNA-copy of human lymphocytes RNA can be used for the analysis of csk gene structure in normal and pathologically changed human cells.

[A role of protein kinase C in signal reactions in outer segments of the bovine retinal rods]. / Rol' proteinkinasy C v signal'nykh reaktsiiakh v naruzhnykh segmentakh palochek setchatki byka.

The effect of modulators of protein kinase C activity on Ca2+ translocation in dark-adapted and bleached retinal rod outer segments (ROS) was studied. The activators (1,2-diacyl glycerol and phorbol-12-myristate-13-acetate) and the inhibitor (chelerythrine chloride) of protein kinase C were shown to stimulate and inhibit the ATP-dependent Ca(2+)-uptake in dark-adapted retinal ROS, correspondingly. Apparently, this action is due to the influence of protein kinase C on Ca(2+)-ATPase activity in these vesicular structures. No involvement of modulators of protein kinase C activity on ATP-dependent Ca(2+)-uptake in bleached retinal ROS was found. The influence of protein kinase C on Ca(2+)-release from retinal ROS was observed. It was shown that the activators and inhibitors of protein kinase C increased the efficiency of this process both in dark-adapted and bleached retinal ROS. The mechanisms of action of the protein kinase C activity modulators on the Ca(2+)-uptake and Ca(2+)-release in retinal ROS are discussed.

Effect of modulators of protein kinase C activity on Ca2+ transport in retinal rod microsomes.

The effect of modulators of protein kinase C (PKC) activity on Ca2+ translocation in retinal rod microsomes was studied. It is shown that PKC activators (phorbol 12-myristate-13-acetate (PMA) and diacylglycerol (DAG)) and inhibitors (chelerythrine chloride, polymyxin B, and phloretin) stimulate and inhibit ATP-dependent Ca2+ uptake in retinal rod microsomes, respectively. This effect is apparently due to an influence of PKC on Ca-ATPase contained in these vesicular structures. It was found that PKC inhibitors (chelerythrine chloride, polymyxin B, and phloretin) and activators (PMA and DAG) potentiate Ca2+ release from Ca2+-loaded retinal rod microsomes. Specific and nonspecific mechanisms of Ca-release stimulation by the modulators of PKC activity are discussed.

[Isolation and identification of tyrosine kinase from the cytosolic fraction of bovine retinal rod outer segments]. / Vydelenie i identifikatsiia tirozinkinazy iz tsitozol'noi fraktsii naruzhnykh segmentov palocheksetchatki byka.

It was shown that the cytosol fraction of bovine retinal rod outer segments contains three forms of tyrosine kinase. One of them was purified 171-fold to attain a specific activity of 1.6 nmol/min per mg protein. The isolated protein had a molecular weight of about 54,000 in SDS electrophoresis. It was shown that this protein is a tyrosine-specific protein kinase, capable of autophosphorylation at the tyrosine residues and restoration of kinase activity upon denaturation-renaturation.

[Light inhibits tyrosine kinase in rod outer segments in vitro]. / Svet ingibiruet tirozinkinaznuiu aktivnost' v naruzhnykh segmentakh palochek setchatki in vitro.

It was shown that short-term (10 min) light exposure of dark-adapted retinal rod outer segments (ROS) leads to a threefold inhibition of the tyrosine kinase activity. Tyrosine kinase activity in the ROS from bleached retinas is by 30% lower than in the dark-adapted ROS. Prolonged illumination (60 min) of the dark-adapted ROS restores the tyrosine kinase activity to the level of ROS from the bleached retinas.

[Role of secondary phosphoinositide messengers in storage of Ca2+ ions in retinal rod microsomes]. / Rol' vtorichnykh mediatorov fosfoinozitidnoi prirody v deponirovanii ionov Ca2+ mikrosomami palochki setchatki.

The influence of guanosine-5'-triphosphate and secondary messengers forming in rod outer segment membranes during light-stimulated hydrolysis of phosphoinositides on the ATP-dependent Ca(2+)-uptake in microsomes of the retinal rod inner segment was studied. The water-soluble cytoplasmic components of the retinal rod outer segment were shown to be capable of stimulating the Ca(2+)-pump of endoplasmic reticulum after light illumination. This process is likely to proceed with the participation of 1,2-diacylglycerol localized in microsome membrane.

[The calcium-mobilizing effect of inositol-1,4,5-triphosphate in rod outer segments and disks]. / O Ca2+ mobilizuiushchem deistvii inozit-1,4,5-trifosfata v naruzhnykh segmentakh palochki i diskakh.

The effect of inositol-1,4,5-trisphosphate (IP3) on the release of calcium ions from retinal rod discs was studied. It was shown that the release of Ca2+ from discs is an electroneutral process. The intradiscal calcium concentration during the release of the ion from the organelle decreases by 1 mM. It was found that the IP3-dependent release of Ca2+ ions from discs is activated by guanosine triphosphate and beta gamma-transducin. The increase in calcium concentration in the medium also activates the IP3-dependent release of Ca2+ ions from discs, which probably is due to the stimulation of phospholipase C. It is suggested that the functional role of the release of ions in related not to phototransduction but to slow regulatory and adaptation processes in the photoreceptor cell.

[Inorganic phosphate is used for synthesis of phosphatidylinositol-4,5-diphosphate]. / Neorganicheskii fosfat ispol'zuetsia dlia sinteza fosfatidilinozitol- 4,5-difosfata.

It was shown that exogenous inorganic phosphate can be incorporated into newly synthesized phosphatidylinositol-4,5-bisphosphate without any participation of ATP.

[Phosphorylation of proteins and phosphoinositides in the axoneme of the bovine rod outer segments. The effect of structural factor]. / Fosforilirovanie belkov i fosfoinozitidov v aksoneme naruzhnogo segmenta palochki setchatki byka. Vliianie strukturnogo faktora.

The protein composition of microtubular structures of bovine retinal rod axoneme and phosphorylation of its proteins were studied. The axoneme was shown to consist of proteins of 25, 43, 51, 112, 145 and 240 kDa molecular mass. It was demonstrated that 32P is mainly incorporated into the 25, 43 and 51 kDa proteins. cGMP was found to enhance 32P. Incorporation into these proteins. In the presence of Ca(2+)-calmodulin the phosphorylation of the 43 and 51 kDa proteins was increased. The phosphoinositides were shown to be phosphorylated in axoneme. Phosphatidylinositol 4-monophosphate was found to be a principal substrate for phosphatidylinositol kinases to form phosphatidylinositol 4,5-bisphosphate. Ca2+, neomycin and colchicine inhibited 32P incorporation into phosphatidylinositol 4,5-bisphosphate indicating the dependence of phosphatidylinositol 4-monophosphate phosphorylation on structural state of axoneme. Phosphatidylinositol was found to diminish the axoneme ability for the polymerization of microtubules. Data was showed that kinases responsible for protein and phosphoinositides phosphorylation were tightly bound to axoneme structures.

betagamma-Transducin stimulates hydrolysis and synthesis of phosphatidylinositol 4,5-bisphosphate in bovine rod outer segment membranes.

T betagamma was shown to stimulate the hydrolysis and synthesis of PtdInsP2 in dark-adapted bovine retinal rod outer segments. In contrast, T alphaGDP blocked the effect of betagamma-transducin. It was also demonstrated that T betagamma was a stimulator of 32P incorporation into PtdInsP2 in ROS. These findings explain the modulating actions of GTP and light on PtdInsP2 hydrolysis and synthesis in ROS. The possible existence of cross-talk between the cGMP and phosphoinositide cascades in retinal rods was discussed.

[The inhibiting effect of 8-Cl-adenosine-3',5'-cyclophosphate on the growth of melanoma B-16 in mice]. / Tormoziashchii éffekt 8-VCl-adenozin-3',5'-tsiklofosfata na rost melanomy B-16 u myshei.

A site-selective analogue of the cyclic adenosine monophosphate 8-chloro-adenosine-3',5'-cyclophosphate was studied for its effects on the growth of transplanted murine melanoma B-16. When the agent was given to the mice, a substantial effect on the growth of the tumor was produced by a number of factors, which included the route of administration, concentration of the agent, the time and duration of therapy. Intraperitoneal injections of the agent in a dose of 20 mg/kg/day which were made during three consecutive days, beginning from day 5 after tumor transplantation caused a 58% decrease in tumor growth as compared to the controls. An examination of tumour biopsy specimen revealed that after a course of the injections there was a significant suppression of the activity of cAMP-dependent protein kinase, type I, and a drastic increase in that of cAMP-dependent protein kinase, type II.

[Interaction of rabbit skeletal muscle glycogen synthase I with substrate analogs--oxo-UDP hydrazones]. / Vzaimodeistvie glikogensintazy I skeletnykh myshts krolika s analogamisubstrata--gidrazonami okso-UDP.

Inhibition of rabbit skeletal muscle glycogen synthase I was studied by using several synthetic substrate analogs: dansylhydrazone of oxo-UDP, 3-hydroxy-2-naphthoylhydrazone of oxo-UDP, salicyloylhydrazone of oxo-UDP, 1-oxyl-2,2,5,5-tetramethylpyrrolidine-3-carbonylhydrazone of oxo-UDP, N'-(dansyl)hydrazinocarbonylhydrazone of oxo-UDP and N'-(fluorenylidene-9)-hydrazinocarbonylhydrazone of oxo-UDP. All these compounds (with the exception of the nitroxyl-containing hydrazone) were characterized by a nonlinear dependence of the reverse reaction rate on the analog concentration in Dixon coordinates. The parabolic type of inhibition was due to the fact that the analogs tested except for the nitroxyl-containing hydrazone were able to interact both with the active center of the enzyme and with the FMN-binding site. The inhibition constants for oxo-UDP hydrazones were calculated for the both centers; their comparison revealed that the affinity of the analogs for the FMN-binding site increased with an increase in the radical hydrophobicity. These data suggest that the site with a high binding affinity for FMN is hydrophobic in nature. Apparently, isoalloxasine-like compounds display the highest affinity for this site.

Specific cGMP binding by retinal rod axoneme and its modulation by calcium ions and calmodulin.

The high-affinity binding of cGMP to the retinal rod axoneme was identified. The axoneme was shown to contain two types of binding sites, with concentrations of 3.6 X 10(-10) and 5.8 X 10(-11) mol mg protein-1. The cGMP concentration for half-saturation of binding was 0.35 microM. The inhibition of cGMP binding by colchicine and vinblastine was 20% of Ca2+ and calmodulin control cGMP binding. The effect of calmodulin is explained by its interaction with specific binding sites which are possibly associated with Ca(2+)-induced depolymerization of axoneme microtubules.

[The study of nucleoside triphosphate-binding center of DNA-dependent RNA-polymerase of phage T7 using GTP analogs]. / Issledovanie nukleozidtrifosfatsviazyvaiushchego tsentra DNK- zavisimoi RNK-polimerazy faga T7 s pomoshch'iu analogov GTP.

The NTP binding site of bacteriophage T7 DNA-dependent RNA polymerase was studied using GTP analogs. For four analogs the irreversible inhibition was demonstrated. The kinetic parameters for competitive (Ki) and irreversible (KI and k3) inhibition were determined. One of the analogs, 5'[2-hydroxy(4-iodoacetamido)benzoyl]guanosine, was shown to inactivate the enzyme rapidly due to the modification of SH-groups. Some suggestions on the structure of the RNA polymerase active site have been made.