[Influence of indoinetophene on the antiradiation properties of indralin].

Radioprotective properties of indralin were studied at its combined administration with indometophene in the periods optimal for each preparation before acute radiation exposure. Animals were subjected to total radiation on the IGUR installation (137Cs): mice of the strain (CBA x C57B1) F1 at a dose of 9 Gy (LD100/30), purebred dogs--4 Gy (LD100/45). It was established in the experiments on mice that considerable radioprotective effect can be obtained by the use of indralin at a dose that is half the optimal radioprotective dose if it is applied against the background of indometophene administered at its optimal radioprotective dose four days before. The survival of mice increased on the average by 30-35% and provided the same effect of protection as a single indralin at the optimal radioprotective dose (100 mg/kg). The survivability of dogs after the combined application of the two radioprotectors makes up 43% against 14% after application of only indralin at a dose of 5 mg/kg (half the optimal radioprotective dose). Indometophene, along with strengthening the antiradiation activity of indralin at the ineffective (half the optimal) dose, allows the reduction of its undesirable postradiation effects in the hemopoietic tissue. The important role in the mechanism of the antiradiation activity of indometophene and indralin belongs to the increased ribonucleotide reductase activity and induction of the ribonucleotide synthesis that provides effective reparation of the damage to the DNA of the cells in radiosensitive tissues and organs as a result of administration of protective doses of radioprotectors at the optimal doses before radiation exposure.

[The use of EPR spectroscopy to control the changes of organism radiosensitivity/radioresistance. Experimental and clinical results].

The responses of deoxyribonucleotide (dNTP), DNA and protein synthesis systems in blood-forming organs of animals (dogs, mice) as well as changes in Fe(3+)-transferrin (Fe(3+)-TF) and Cu(2+)-ceruloplasmin (Cu(2+)-CP) pools in blood to gamma-irradiation and the administration of radioprotectors have been studied. It has been shown that changes in Fe(3+)-TF and Cu(2+)-CP pools in blood are indices of changes in the body radioresistance and are reliably controlled by the EPR technique. An increase in the Fe(3+)-TF pool promotes the activated synthesis of dNTP, DNA and Fe(3+)-containing proteins which are essential for the repair efficiency during the early post-irradiation time as well as for the development of compensatory and restorative reactions of cellular systems; i.e., they are responsible for the body resistance to DNA-damaging factors. It is important that the intensity of responses depends on the initial state of the organism. It has been shown, that changes in Fe(3+)-transferrin and Cu(2+)-ceruloplasmin pools, which are trust-worthy controlled by the EPR technique in whole blood, blood plasma, and serum, as well as the changes in the extracellular DNA content in blood plasma are the markers of the changes in the organism radioresistance. This has been proved during the medical examination of the Chernobyl accident recovery workers and civil population, including children, exposed to low-intensity radiation.

[The most important stages of the mechanism of action in vivo of carcinogen diethylnitrosoamine and its effect on the synthesis of RNA, proteins, DNA, and deoxyribonucleotides].

The mechanisms of nitric oxide (NO) generation from exogenous and endogenous sources, induced by the addition of the carcinogen diethylnitrosoamine (DENA) to rat organism have been studied. Within 15 h after the addition of DENA, the carcinogen itselt acts as an exogenous NO donor. The products of protein degradation (the process induced by DENA) act as endogenous donors of NO. It was shown that the generation of nitric oxide from diethylnitrosoamine leads to deep hemic and tissue hypoxia and induces the inactivation of oxygen-dependent enzymes, including ribonucleotide reductase, and the inhibition of ATP synthesis. Under these conditions, the protein synthesis and as a consequence the synthesis of deoxyribonucleotides and DNA are strongly suppressed; i.e., diethylnitrosoamine produces the effect similar to the action of the antibiotic cycloheximide, an inhibitor of translation. The administration of cycloheximide to the animal organism also led to the appearance of a considerable amount of nitric oxide in the blood. It is assumed that nitric oxide initiates (on the administration of the carcinogen) or at least enhances (on the administration of cycloheximide) the blockage of the synthesis of the protein, deoxyribonucleotides, and DNA. In response to the disturbance of protein synthesis, the complex of enzymes is activated that accomplish the utilization of the degradation products of proteins, including the inducible form of NO synthase.

[The use of EPR spectroscopy to control the changes in organism radioresistance. Clinical results].

It has been shown that changes in Fe(3+)-transferrin and Cu(2+)-ceruloplasmin pools, which are trust-worthy controlled by the EPR technique in whole blood, blood plasma, and serum, as well as changes in the extracellular DNA content in blood plasma are markers of changes in organism radioresistance. This has been proved during the medical examination of the Chernobyl accident recovery workers and civil population, including children, exposed to low-intensity radiation, as well as during clinical investigation of new radioprotectors.

[The use of EPR spectroscopy to control the changes in organism radioresistance. Experimental results].

The responses of deoxyribonucleotide (dNTP), DNA, and protein synthesis systems in blood-forming organs of animals (dogs, mice) as well as changes in Fe(3+)-transferrin (Fe(3+)-TF) and Cu(2+)-ceruloplasmin (Cu(2+)-CP) pools in blood to gamma-irradiation and the administration of radioprotectors have been studied. It has been shown that changes in Fe(3+)-TF and Cu(2+)-CP pools in blood are indices of changes of body radioresistance and are reliably controlled by the EPR technique. An increase in the Fe(3+)-TF pool promotes the activation of synthesis of dNTP, DNA, and Fe(3+)-containing proteins, which are essential for repair efficiency during early post-irradiation time as well as for the development of compensatory and restorative reactions of cellular systems; i.e., they are responsible for body resistance to DNA-damaging factors. It is important that the intensity of responses depends on the initial state of the organism. Thus, dogs with initial individual characteristics of blood typical for "suppressed" or "activated" states had abnormally high responses to irradiation by low doses of 0.25 and 0.5 Gy. This fact is important for the estimation of consequences of prolonged low-dose irradiation for human population. It has been shown that radioprotectors, efficient in survival test activate the synthesis of dNTP, DNA, and proteins in organs. The intensity of dNTP synthesis and the time when dNTP pools get maximum values determine the efficiency of protectors and the time of irradiation after their administration.

[The organization of medical aid to children of various cohorts of supervision, undergone to the influence of small dozes of radiation].

In clause the stages of organization of medical aid to children who undergone to the influence of the ecopathologic factors, including small dozes of ionizing radiation are submitted. The features of various stages of medical aid in nearest time after influence of the radiating agent are shown. The methodological complexities of diagnostic measures in the late times after the ecopathologic influence of small dozes of ionizing radiation are submitted, the role of the radiosensitivity in formation of pathological somatic diseases of condition at children is considered. The opportunity of the differential treating and preventive help rendering to the children various cohorts of supervision is shown.

[The mechanisms of nitric oxide production from exogenous and endogenous NO-donating compounds and its effect on deoxyribonucleotide and DNA synthesis].

The mechanisms of exogenous nitric oxide (NO) production through in vivo biotransformation of nitro-, nitroso- and amino-containing substances were discussed. In addition, the mechanisms of production and cellular sources of endogenous NO, appearing in the blood and tissues after the exposure to various DNA-damaging factors, have been considered. Considerable quantities of endogenous NO were detected in the body in the first hours after translation inhibition by cycloheximide or animal exposure to superlethal radiation doses, i.e., after the exposure to factors inducing destructive processes. The time and dose dependences of exogenous and endogenous NO production have been established. NO produced after a single or repeated administration of NO-donating compounds as well as endogenous NO proved to inhibit deoxyribonucleotide (dNTP) and DNA synthesis in animal tissues. Nonspecific compensatory responses to disturbed protein homeostasis included cyclic production of endogenous NO. The maximum levels of nitrosyl complexes were registered when the rate of protein synthesis decreased. The role of polyamines in the induction of macromolecule biosynthesis is discussed and NO production from these arginine-rich compounds is proposed. NO is released at the stage of polyamine inactivation. The inactivation mechanism includes the hydroxylation of aminogroups by NO synthase, the formation of nitroso intermediates, and their denitrosation with NO release.

The activation of ribonucleotide reductase in animal organs as the cellular response against the treatment with DNA-damaging factors and the influence of radioprotectors on this effect.

Cellular requirements for deoxyribonucleotide (dNTP) pools during DNA synthesis are related to ensuring of the accuracy of DNA copying during replication and repair. This paper covers some problems on the reactions of dNTP synthesis system in organs of animals against the treatment with DNA-damaging agents. Ribonucleoside diphosphate reductase (NDPR) is the key enzyme for the synthesis of dNTP, since it catalyses the reductive conversion of ribonucleotides to deoxyribonucleotides. The results obtained show that the rapid and transient increase in NDPR activity in animal organs occurs as cellular response against the treatment with DNA-damaging agents (SOS-type activation). We have also found the intensive radioprotector-stimulated activation of deoxyribonucleotide synthesis as well as DNA and protein synthesis in mice organs within 3 days after the administration of two radioprotectors, indralin and indometaphen, that provide the high animal survival. Our studies suggest that these effects are the most important steps in the protective mechanism of the radioprotectors and are responsible for the high animal survival.

[Activation of deoxyribonucleotide synthesis by radioprotectants and antioxidants as a key stage in formation of body resistance to DNA-damaging factors].

The responses of the systems of synthesis of deoxyribonucleotides (dNTPs), DNA, and proteins in hematopoietic organs and liver of animals to gamma-radiation, administration of radioprotectants and antioxidants as well as the dependence of these responses on the doses of radiation and drugs were studied. Radioprotectants of acute (indralin) and durable effects (indomethaphen) as well as natural (alpha2-tocopherol) and synthetic anti-oxidants (ionol or 2,6-di-tert-butyl-4-methylphenol) efficient in survival test were used. Three stages could be recognized in the standard unspecific response of the studied systems to radiation: (1) immediate increase in ribonucleotide reductase activity in the tissues within the first 30 min as a part of the integrated SOS response to DNA damage, which activates dNTP synthesis; (2) inhibition of the synthesis of dNTPs, DNA, and and (3) restoring ribonucleotide reductase activity and integral increase in the production of dNTPs, DNA, and total protein, which is essential for the development of compensatory and restorative responses of the organism. The radioprotectants significantly increased ribonucleotide reductase activity, which increased intracellular concentrations of the four dNTP types in organs during radiation exposure and three following days. Within this period, ribonucleotide reductase activity was inhibited by 40-50% in animals not treated with radioprotectants as compared to control. Balanced high pools of dNTPs in the organs of radioprotectant-treated animals provided for high-performance repair of DNA damage. The radioprotectant-induced activation of dNTP synthesis during the development of compensatory and restorative responses provides for an earlier restoration of the cellular composition and functioning of the organs. Antioxidants stimulated the synthesis of dNTPs, DNA, and proteins in animal tissues in a strict dose interval. Their effect on the studied syntheses was dose-dependent: single or multiple long-term administration of high antioxidant doses inhibited synthesis of dNTPs, DNA, and proteins. Radioprotectants and antioxidants affected the pool of blood protein Fe3+-transferrin controlling the synthesis of iron-containing ribonucleotide reductase activity in hematopoietic organs, and hence, the iron-dependent stage in DNA synthesis--dNTP synthesis. Activation of protein synthesis in organs by the studied substances increased the pools of Fe3+-transferrin and Cu2+-ceruloplasmin in the blood, which activated dNTP and DNA synthesis. Activated synthesis of dNTP, DNA, and proteins in the organs and increased pools of studied plasma proteins underlay the formation of body resistance to DNA-damaging factors.

[Fungal infection of human organs by resistant melanin-synthesizing species is one of the pathogenic factors and one of the real consequences of the accident at Chernobyl power plant]. / Gribkovaia infektsiia organov cheloveka rezistentnymi melaninsinteziruiushchimi vidami iavliaetsia odnim iz patogennykh faktorov i real'nykh posledstvii avarii na ChAES.

Free radical melanin centers have been detected in the cell concentrate of bronchoalveolar lavage (BAL) of liquidator of Chernobyl NPP accident. To identify the nature of these centers the EPR technique and the fluorescent technique were used to study BAL of liquidators with lung chronic pathology, their blood, blood components as well as model melanin- and lipofuscin-containing systems: synthetic DOPA-melanin, human melanosome, human lipofuscin, human melanolipofuscin. I Besides that we have investigated the samples of fungi, extracted from lung phlegm of liquidators (Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Penicillium Sp., Candida albicans) as well as the melanin, extracted from fungal conidium. It has been shown that the melanin centers found in BAL cells of liquidators is the melanin of melanin-synthesizing mutant fungi Aspergillus fumigatus and Aspergillus flavus. The prolonged gamma-irradiation at low dose rate and the effects of inhaled radioactive particles cause the adaptive mutation of micromycetes producing the chemo- and radioresistant population. We think that the radioactive dust and pathogenic mutant micromycetes were inhaled in lungs of liquidators during their work at the Chernobyl NPP. Thus, one of valid consequences of Chernobyl accident may be the wide fungous of human organs, in particularly, by Aspergillus mutant. The radiation-induced weakening of immune reactions of liquidators promotes the resistance of this fungus mutant infection.

[Reactions of deoxyribonucleotide synthesis system to irradiation and their modification by radioprotectors]. / Reaktsii sistemy sinteza dezoksiribonukleotidov na obluchenie i ikh modifikatsiia radioprotektorami.

The paper covers the problem on reactions of deoxyribonucleotide (dNTP) synthesis system in blood-forming organs of animals induced by irradiation. The synthesis of dNTP is a rate-limiting stage for DNA synthesis. Cellular requirements for dNTP pools during DNA synthesis are related with ensuring of the accuracy of DNA copying during replication and repair. It has been shown that organism defence mechanisms against irradiation include the following stages: 1. The prompt SOS-activation of dNTP synthesis 30 min later after irradiation, playing the important role in protecting of cell's genetic apparatus from damage. 2. The inhibition of dNTP synthesis within 3-24 h after irradiation resulting to the imbalance of four dNTP and the decrease of their pools. As result of that, the abnormal repair is observed due to depurinations, errors of base incorporations and "misrepair". 3. The restore of dNTP synthesis occurred 2 days later after irradiation. The increase of dNTP pools promotes the increase of DNA synthesis rate as well as proliferative activity of cells. Confirming the fact that the alterations in dNTP pools play essential role in the production of DNA lesions became an important step in understanding of the multistage process leading to radioprotection. To get high and balanced pools of dNTP needed for the increase in the volume of repair of DNA lesions the radioprotectors with high efficiency relative to the survival test were used in experiments. They induced the elevated dNTP synthesis in bone marrow and spleen during the time when the irradiation alone caused the essential prolonged suppression of dNTP synthesis as well as DNA and protein synthesis in organs of nonprotected animals. It has been shown that substances with antioxidant and antiradical activity induced the dNTP synthesis, too. In vivo regulatory factors of dNTP synthesis have been studied to elucidate the mechanisms of getting of high and balanced dNTP pools by using of different substances.

[Time- and dose-dependent post-irradiation changes of Fe3+-transferrin and Cu2+-ceruloplasmin pools in blood, their influence on ribonucleotide reductase activity in animal tissues and the effects of radioprotectors]. / Vremennye i dozozavisimye postradiatsionnye izmeneniia v soderzhanii Fe3+-transferrina i Cu2+ tseruloplazmina v krovi zhivotnykh i ikh vliianie na ribonukleotidreduktaznuiu aktivnost' tkanei.

The time- and dose-dependent changes of Fe(3+)-transferrin (Fe(3+)-TF) and Cu(2+)-ceruloplasmin (Cu(2+)-CP) pools, of superoxide dismutase activity and the inhibitory activity of alpha 2-macroglobulin in blood as well as changes in synthesis rates of deoxyribonucleotides (dNTP), DNA and proteins in organs (spleen, liver, bone marrow, thymus) of mice and dogs given total body irradiation have been studied using of ESR spectroscopy, radioisotope techniques and biochemical determination of enzymatic activity. The experimental data have allowed us to reveal the sequence of organism's response reactions against irradiation and their modifications by radioprotectors. Changes in blood Fe(3+)-TF pool is one of the most informative, highly radiosensitive and rapidly reactive marker against irradiation and drug administrations. This irontransport protein controls a rate-limiting iron-dependent stage for DNA synthesis--the synthesis of dNTP, catalyzed by iron-containing ribonucleotide reductase (Fe(3+)-RR). It has been shown that time-dependent post-irradiation changes of Fe(3+)-TP pool in blood are characterized by three distinct stages: 1) the prompt increase of pool (SOS-type response) playing the important role in protecting of cell's genetic apparatus from damage; 2) the decrease of its pool within 3-18 h after irradiation resulting in the loss of Fe(3+)-RR activity in tissues of blood-forming organs that make more stronger radiation-induced damage; 3) the following phase-dependent increase in Fe(3+)-TF pool at the 2-nd, 6th, 10-17th days after irradiation due to an increase in transferrin synthesis. This increase may be considered as compensatory reaction of blood-forming organs directed at restoring blood and organ's cells. The time-dependent courses of the reactions are independent from radiation doses indicating to the universal and nonspecific response of organism against irradiation. But, the intensity of this compensatory-adaptive response at 2-nd and 6th days grows with increasing radiation dose up to lethal that, and organism's response becomes abnormal and physiologically hypertrophic. The prolonged "stressful syndrome of biochemical tense state" should be attributed to negative effects for organism, since it may result in the failure of compensatory adaptive organism's reactions and animal killing. The radioprotectors ward off the appearance of this dangerous state. Dogs with initial individual characteristics of blood which were typical for "suppressed" or "activated" states had abnormal response against irradiation by low doses 0.25 or 0.5 Gy. In these cases the intensity of response reactions of organism was essentially increased and markedly deviated from linear dose dependence. The phase-dependent increase of Fe(3+)-TF pool in blood in post-irradiation time resulted to the increase of Fe(3+)-RR activity in blood-forming organs. The key event ensuring the development of compensatory adaptive reactions is the increase of capacity of protein-synthesizing apparatus, the activation of biosynthesis of dNTP and DNA against the treatment with damaging factors.

[Identification of electron spin resonance signals of ribonucleotide reductase in animal tissues with high proliferative activity]. / Identifikatsiia signala EPR ribonukleotidreduktazy v zhivotnykh tkaniakh s vysokoi proliferativnoi aktivnost'iu.

A new ESR signal (doublet with hyperfine splitting of 20 gauss and g = 2.005) was found for animal tissues having high level of proliferative activity. This signal is shown to be due to radical enzyme ribonucleotide reductase. The results reported for spleen development suggest that there is a correlation between changes in the enzyme activity during the first week of neonatal development and intensity of ESR doublet signal.

[Effect of ionol on oxygenase level in proliferating animal tissues]. / Vliianie ionola na soderzhanie oksigenaz v proliferiruiushchikh tkaniakh zhivotnykh.

It has been shown by low temperature ESR-spectroscopy that BHT (butylated hydroxytoluene) decreases the content of Fe2+ . . . O2 complexes being active centres of oxygenase type enzymes in loose connective tissue, regenerating liver and also in tumor and liver of tumor bearing mice. This effect is closely related to BHT presence in animal tissues.

[Electron paramagnetic resonance study of photosensitized transport and localization of electrons in enzyme-substrate complexes. Lysozyme and its inhibitor]. / Izuchenie fotosensibilizirovannogi perenosa i lokalizatsii élektronov v ferment-substratnykh kompleksakh pri pomoshchi élektronnogo paramagnitnogo rezonansa. Lizotsim i ego ingibitory

Photosensitized electron transfer are studied in three systems: lysozyme, its inhibitors (oligosaccharides) and their enzyme-inhibitor complexes. Electron donors were either tryptophane amino acid residues of lysozyme or tryptophane. It is shown that N-acetyl group of inhibitor molecules is a single electron--acceptor group in the inhibitor molecule as well as in the lysozyme-inhibitor complex. It is stated that the localization of unpaired electrons in the mixtures of lysozyme modification products and substrate--inhibitors depends on the state of enzyme molecule.

[EPR and low temperature spectrophotometric study of the phototransformation products of bacteriorhodopsin]. / Izuchenie produktov fotoprevrashcheniia bakteriorodopsina metodami EPR i nizkotemperaturnoi spektrofotometrii

It is shown that BR and intermediate products of its phototransformation P600, P550 and P415 (the maximum at -196 degrees C at 419 nm) are not paramagnetic. Illumination of samples containing P415 (P419) at -- 196 degrees C with light in the region of 360-480 nm results in the formation of paramagnetic centres with a sunglet spectrum deltaH=18 Oe and g=2.002 (R1). In parallel formation of a new photoproduct P421 in the absorption spectrum is observed. During subsequent heating at -140 degrees C formation of an asymmetric signal with deltaH=45 Oe and g=2.006 and g=2.03 was observed. In the absorption spectra a dark transition. P421-P565 was observed under the same conditions. P565 differs from initial BR P570 as to its photochemical properties. R1 is identified as retinal radical, R2 as a peroxide radical of the BR-complex lipids. Paramagnetic, spectral, and photochemical properties of some products of BR transformation are compared. A scheme of oxidative-phosphorylation processes with participation of Mn ions in BR phototransformation.

[Localization of unpaired electrons in molecules of lysozyme substrate-inhibitors. I. N-acetylglucosamine]. / Issledovanie lokalizatsii nesparennykh élektronov v molekulakh substrat-ingibitorov lizotsima. I. N-atsetilgliukozamin

Electron structure of N-acetylglucoseamine molecule and its ion-radicals has been studied by quantum-chemical methods CNDO2 and JNDO, and by ESR as well. N-acetyl group is shown to be the only electron acceptor group of AGA molecule. Phototransformations of paramagnetic centres are studied at different pH values.

[Effect of inhibitors of free radical processes on the "dark" paramagnetic centers of the melanoprotein granules of the pigment epithelium of the eye]. / Vliianie ingibitorov svobodnoradikal'nykh protsessov na "temnovye" paramagnitnye tsentry melanoproteinovykh granul pigmentnogo épiteliia glaza.

Effect of three inhibitors of free radical processes (IFRP) differing in antiradical activity (2-tret. butyl-6-methyl-3-oxipyridin, 2-ethyl-6-methyl-3-oxipyridin and 3-oxipyridin) on paramagnetic centres of melanoprotein granules of pigment epithelium was studied by ESR method at room temperature. It was shown that IFRP decreased the dark signal and that the reaction of paramagnetic particles dissociation proceeded according to the equation of the first order reaction. The rate constants of these reactions were calculated. It was found that the relative amount of reacting and non-reacting paramagnetic particles with the inhibitor depended on the nature and concentration of IFRP.

[Localization of unpaired electrons in molecules of the substrate inhibitors of lysozyme. II. Oligosaccharides]. / Issledovanie lokalizatsii nesparennykh élektronov v molekulakh substrat-ingibitorov lizotsima. II. Oligosakharidy.

It has been shown by the method of electron photosensitized transfer that N-acetyl group is the main electron-acceptor group in oligomeres of N-acetyl glucose amine (AGA). In n-AGA molecules (n-3.5) interacting with an electron radical products of the breakage of glycoside bond are observed, their concentration rising with an increase of the chain length. The ionic strength does not affect the photosensitized transfer of the electron in chitin oligomeres. Formation of paramagnetic centres in the molecule penta-AGA depends on the medium pH.

[Study of the responses of tumor and normal tissues to exposure to ionizing radiation by the EPR method. I. Comparison of the low temperature EPR spectra of irradiated tumor and normal tissues]. / Izuchenie reaktsii opukholevykh i normal'nykh tkanei na deistvie ioniziruiushchego izlucheniia metodom EPR. I. Sravnenie nizkotemperaturnykh spektrov EPR obluchennykh poukholei i normal'nykh tkanei.

ESR spectra of liver samples of normal mice and liver tumour samples of mice (line C3HA) injected hepatomo 22-a irradiated in the dose of 1 Mrad at --196 degrees C were studied. The ESR spectra of normal and tumour tissues irradiated at --196 degrees C are shown to differ. Only the ESR spectra of tumour and liver of the tumour host is characterized by an asymmetric signal with deltaH=6 Oe and g=2.0005 ("tumour" signal). The intensity of the "tumour" signal depends on the developmental stage of hepatoma. It is changed after the animals are injected with ionol.