The recovery parameters of DNA strand breaks of Ehrlich ascites cells after the combined action of ionizing radiation and hyperthermia

A mathematical model of DNA strand breaks postirradiation repair and the methodology allowing to differentiate the mechanism of inhibition of DNA strand breaks recovery after combined actions of ionizing radiation and hyperthermia have been described in this paper. Using this model and the results published by other authors for DNA strand breaks of Ehrlich ascites cells, there have been obtained the data showing that the portion of DNA-damages that the cell incapable to recover after consecutive thermoradiation action was risen with an increase in thermal load under insignificant change of repair constant. It means the mechanism of DNA strand breaks recovery inhibition is realized in a greater extent through the formation of irreversible damages but not through the damage of repair process itself.     

Qantitative description of mammalian cell recovery after combined exposure to ionizing radiation with chemical agents

The parameters of recovery of mammalian cells after exposure to ionizing radiation combined with chemical agents were calculated based on a mathematical model of post-radiation recovery. The data presented, in contrast to the previously published results, indicate that the inhibition of the recovery may be due either to the damage or disruption of the process of recovery or the increase in the part of irreversible damages from which the cells are incapable to recover; or both of these processes can be realized simultaneously. It is concluded that the combination of ionizing radiation with chemical agents that inhibit the recovery processes through the formation of irreversible damages or affect the probability of recovery can be a perspective way in terms of finding the most effective means to increase radiosensitivity.     

Revision of th distribution of chromosome aberrations induced by chemical mutagens using the BUDR label

Cell distribution was analysed with the help of the BrDU label for the number of chromosome aberrations and breaks induced by one-center (thiophosphamide and phosphamide) and two-center (dipine and fotrine) mutagens at the stage G0 in the Ist mitosis of human lymphocytes harvested at different times of culturing (from 56 to 96 h). The comparison was made between the type of aberration distribution in cells and the dependence of their frequency on the harvesting point for various mutagens. Poisson aberration distribution in cells for two-center mutagens was found to correspond to their constant frequency observed at different times of harvesting. On the other hand, for one-center mutagens, a geometrical distribution of chromosome breaks corresponded to an exponential decrease in their frequency in time. It is suggested that two-center chemical mutagens and ionizing radiation cause largely short-live damages which are realized into chromosome aberrations rather quickly (during one cell cycle). One-center mutagens, however, cause such damages that the probability of their transformation into chromosome aberrations is decreasing rather slowly in time, under the exponential law, and their realization into chromosome aberrations can occur in subsequent cell cycle.     

Recovery of yeast cells after exposure to ionizing radiation and hyperthermia

Quantitative regularities of recovery of wild-type diploid yeast cells irradiated with gamma-rays (60Co) simultaneously with exposure to high temperatures were studied. It was shown that in conditions of such a combined action the constant of recovery did not depend on the temperature at which the irradiation was carried out. However, with an increase of acting temperature an augmentation in the portion of irreversible component was registered. The analysis of cell inactivation revealed that the augmentation of the irreversible component was accompanied by a continuous increase of cell killing without any postirradiation division after which cells are incapable of recovery. The reproductive death was mainly exerted after ionizing radiation applied alone while in conditions of simultaneous thermoradiation action the interphase killing (cell death without division) predominated. It is concluded on this base that the mechanism of synergistic interaction of ionizing radiation and hyperthermia may be related with cardinal change in mechanisms of cell killing.     

Characterization of DNA repair in a mutant cell line derived from ICR 2A frog cells that is hypersensitive to non-dimer DNA damages induced by solar ultraviolet radiation

The level of excision repair and the inhibition and recovery of semiconservative DNA synthesis were examined following the induction of non-dimer DNA damages by solar ultraviolet radiation in a mutant cell line, DRP 36, derived from ICR 2A frog cells that is hypersensitive to these lesions. A relatively pure population of non-dimer photoproducts was produced by exposure of cells to the Mylar-filtered solar UV wavelengths produced by a fluorescent sunlamp followed by treatment with photoreactivating light (PRL) which removes most of the small yield of dimers induced by the irradiation. Using a modification of the bromodeoxyuridine (BrdUrd) photolysis assay, that enhances the sensitivity of this assay, it was found that DRP 36 cells perform a significantly lower level of excision repair following the induction of non-dimer DNA damages compared with the ICR 2A cells. In contrast, the level of excision repair of 254-nm-induced dimers was similar in the two cell lines. In addition, the induction of non-dimer damages caused a greater inhibition of DNA synthesis that persisted for a longer period of time in the mutant compared with the parental cells. Hence, these results indicate that the DRP 36 cells are deficient in the repair of at least one type of solar UV-induced non-dimer lesion.     

Quantitative estimation of the recovery parameters of yeast cells irradiated in the presence of cysteamine

Study of the postirradiation recovery parameters of diploid yeast cells showed that the irreversible component of radiation damage was identical after the cell exposure in the presence and absence of cysteamine. On this basis, it is concluded that the radioprotector equally reduced the number of both irreversible and repairable primary radiation damages. Application of the mathematical model of recovery processes to the results obtained allowed us to draw a conclusion that the probability of cell recovery from the radiation damage per time unit was also identical after cell exposure in the presence and absence of cysteamine.     

Recovery of yeast cells following the combined action of hyperthermia and ionizing radiation

Consecutive action of elevated temperature (50 degrees C) and gamma-irradiation on yeast cells Saccharomyces cerevisiae was studied. It was shown that yeast cells can recover from lethal thermal and radiation lesions after the combined action of the two factors. The efficiency of recovery does not depend upon the sequence of treatments. Heating (50 degrees C) before or after gamma-irradiation increases the radiation response of yeast when plating the cells on a nutrient agar containing 1.5 M KCl. The synergistic effect decreases with yeast cells kept in water at 28 degrees C before plating. The influence of one factor on the effectiveness of recovery from damages induced by the other was estimated.     

Effects of a medium pH on the ability of Chinese hamster cells to repair radiation injuries]

In experiments with Chinese hamster cells at exponential and stationary growth phases, it has been shown that the postirradiation incubation of irradiated cells in a medium with low pH (up to 6.0) promotes the recovery of cells from potentially lethal damages; it has also been found that the recovery from sublethal radiation damages does not depend on the medium pH. The long-term incubation of nonirradiated cells with low pHc causes death of part of cells.     

Beta-Galactosidase messenger RNA made during recovery from inhibition of protein synthesis is not translated.

Bacteria that accumulate RNA in the course of inhibition of protein synthesis are impaired in their ability to synthesize beta-galactosidase during subsequent recovery. By contrast, constitutive enzyme synthesis in recovering cells is normal. Even though no beta-galactosidase is made during recovery from this inhibition, a substantial quantity of beta-galactosidase mRNA (as determined by DNA-RNA hybridization) is made. The beta-galactosidase mRNA made in vivo is functional in vitro. It is capable of directing the in vitro synthesis of a portion of the NH2-terminal beta-galactosidase molecule (in the alpha portion of the molecule). However, this protein is not made in vitro. It is concluded that the beta-galactosidase mRNA that is made during recovery from protein synthesis inhibition, although apparently at least partly normally transcribed in vivo and functional in vitro cannot be translated under these conditions in vivo.     

Dark recovery of diploid yeast cells after simultaneous exposure to UV-irradiation and hyperthermia

Quantitative regularities of dark recovery of wild-type diploid yeast cells of Saccharomyces cerevisiae simultaneously treated with UV-light (254 nm) and high temperatures (53-56 degrees C) were studied. Under this combined action, the constant of recovery, which defines the probability of elimination of the UV-radiation induced damage per unit of time, did not depend on the temperature of irradiation. It was shown that both the irreversible component of cell damage and the number of cells that died without division gradually increased as the temperature of exposure increased. It is concluded, on this basis, that the mechanism of synergistic interaction of UV-radiation and hyperthermia is related not to the inhibition of dark recovery itself, but to the increase in the shape of irreversibly damaged cells incapable of recovering from the induced damage.     

Premolecular period of the postradiation remodelling of cells

Recovery of yeast cells after exposure to ionizing radiation was found in 1957. During the first decade, i.e. in the "premolecular period" of studying the phenomenon, its basic features were revealed: dependence on ploidy of cells, on their energy exchange, on radiation LTE, and others. A mathematical model of recovery was proposed; the damages causing death of irradiated haploid and diploid cells were shown to be double strand breaks of DNA. The concepts of universal biological importance of the cell property to repair genetic damages were formulated.     

Repair of the radiation damage in hematopoietic stem cells from the mouse embryonic liver. Kinetic aspects]

The method of fractionated irradiation was used to study kinetic aspects of repair of sublethal radiation damages in precursor cells from mouse embryonal liver that form in vivo colonies on 8th and 11th days. It was shown that 11-day CFUs had a lesser ability to repair sublethal radiation damages than 8-day ones at different time-intervals between radiation fractions (from 2 to 6 h). These two CFUs sub-populations differed also in the repair kinetics.     

Survival of diploid yeasts after gamma irradiation: genetic control of the maturation effect

A study was made of the influence of rad mutation, leading to radiosensitivity increase, on the effect of additional growth in diploid Saccharomyces cerevisiae yeast exposed to gamma-radiation. The most radiosensitive mutants of this series, rad52/rad52 and rad54/rad54, did not virtually vary from the wild type cells in the value of the additional growth effect. Some other mutants, for instance, rad53/rad53 and rad55/rad55, exhibited a significantly lesser effect of additional growth. It was shown that the effect of additional growth did not depend upon the rate of rapid and slow postirradiation recovery of the wild type cells. The results of the studies prompt the conclusion that the processes responsible for the additional growth effect and those responsible for recovery of cells from radiation damages are mutually independent.     

Comparison of the electrophysiological effect of amiodarone, lidocaine and quinidine on rat ventricular cells.

The effects of 20 microM each of amiodarone, lidocaine and quinidine on action potential and membrane currents were studied in rat ventricular cells. At a stimulation frequency of 0.1 Hz, quinidine prolonged the action potential duration (APD50) from 120 +/- 26 to 660 +/- 8 msec and increased the time to peak (Tp) amplitude from 7 +/- 1 msec to 32 +/- 6 msec. Lidocaine shortened APD50 from 123 +/- 15 to 83 +/- 6 msec without altering Tp. Amiodarone changed neither APD50 nor Tp. Voltage clamp study revealed that quinidine inhibited sodium inward current (INa) even when this current was elicited by depolarizing pulses at 0.1 Hz from a holding potential of -90 mV. For amiodarone and lidocaine, the inhibition was observed when INa was elicited from a holding potential of -70 mV. A frequency-dependent inhibition of INa by amiodarone and lidocaine was observed at frequencies higher than 1 Hz. Quinidine showed this inhibition even at 1 Hz. In correlation with the stronger frequency dependent inhibition of INa, a greater delay of the recovery and increase of the non-recovery fraction of INa was induced by quinidine. For lidocaine and amiodarone, only the recovery time constant was delayed. In cells treated with sea anemone toxin (ATX, 0.2 microM), APD50 was prolonged to 4-5 sec in 5 min. Quinidine, but not amiodarone, completely reversed the effect of ATX. Quinidine showed use-dependent inhibition of INa in these ATX-treated cells. Amiodarone, however, did not show this inhibition. It is likely that amiodarone suppresses INa by delaying the recovery of INa instead of blocking the open-state Na(+)-channels.(ABSTRACT TRUNCATED AT 250 WORDS)     

Trans effects in cell membrane transport

The efflux of a substrate from preloaded cells may be decelerated by an inhibitor in the external medium or accelerated by a compatible substrate in the external medium. The derivations of rate equations for the initial velocity of the zero-trans reaction, trans efflux inhibition, and accelerated exchange diffusion are described for steady state conditions. The rate constants making up the Michaelis constant for the trans inhibition reaction are the same as the corresponding parameters in the zero-trans reaction. The rate constants making up the Michaelis constant for the accelerated exchange reaction, however, are different from the corresponding parameters in the zero-trans reaction. The rate equation for trans inhibition shows that the velocity constant for recovery of the unloaded carrier may be determined with steady state experimental data. It is suggested that the observed recovery constant is independent of the substrates and trans inhibitors chosen for an assay of a particular carrier system. An experiment is briefly described to show a determination of a tentative value for the recovery constant of the unloaded nucleoside carrier in yeast cells and the apparent inhibition constant for a trans inhibitor.     

Effects of daunomycin and radiation on cell-survival and repair of DNA single-strand breaks.

The combined action of Daunomycin and irradiation was investigated using mouse L-929 cells in culture. Survival of cells was measured with the colony assay. Sedimentation in alkaline sucrose gradients was used to study repair of DNA single-strand breaks (SSB) in the presence of various concentrations of Daunomycin. A small increase in radio-sensitivity, as measured by decreasing Do, was obtained for doses of Daunomycin that are considerably toxic to the cells (0.1 microgram/ml). However, the Dq values remained constant even at high concentrations indicating that Daunomycin does not interfere with recovery processes. The rate of rejoining of SSB remained constant up to 1.0 microgram/ml, whereas concentrations of Daunomycin as high as 10 microgram/ml reduced the velocity of repair by a factor of 13. Our data show that concentrations of Daunomycin similar to those required for other DNA-binding drugs are required to inhibit SSB repair. For clinical purposes, no increase in tumour-killing efficiency may be expected from a combined treatment with Daunomycin and radiation.     

Survival and recovery of yeast cells after combined treatment with ionizing radiation and heat

Cell survival, recovery kinetics and inactivation forms after successive and simultaneous treatments with gamma rays (60Co) and high temperatures were studied in diploid yeast cells capable of recovery. Both the extent and the rate of the recovery were shown to be greatly decreased with increase in the duration of heat treatment (60 degrees C) followed by radiation and with increase in exposure temperature after simultaneous treatment with heat and radiation. A quantitative approach describing the recovery process was used to estimate the probability of recovery per unit time and the irreversible component of damage after the combined treatment with heat and radiation. It was shown that the probability of recovery was independent of the conditions of the treatment with heat and radiation, while the irreversible component gradually increased as a function of the duration of heat treatment (60 degrees C) after sequential treatment with heat and radiation and as a function of the exposure temperature after simultaneous treatment with heat and radiation. The increase in the irreversible component was accompanied by an increase in cell death without postirradiation division. It is concluded on this basis that the synergistic interaction of ionizing radiation and hyperthermia in yeast cells is not related to the impairment of the recovery capacity itself and that it may be attributed to an increased yield of irreversible damage.     

Relation between radiation resistance and salt sensitivity of spores of five strains of Clostridium botulinum types A, B, and E.

The NaCl tolerance of different strains of Clostridium botulinum varies over a wide range, and the patterns of NaCl inhibition differ distinctly and characteristically from strain to strain. The more radiation-resistant strains, such as 33A, 62A, and 7272A, are more resistant to NaCl, whereas the more radiation-sensitive strains, such as 51B and 1304E, are more sensitive to NaCl. This rule appears to hold irrespective of whether the spores were unirradiated controls or whether they were radiation damaged prior to exposure to NaCl in the recovery media. The data seem to indicate that radiation doses in the shoulder portion of the radiation survival curves did not noticeably sensitive the spores to NaCl, whereas radiation doses in the exponential-decline portion of the survival curve invariably produced a distinct sensitization. Thus, strains 33A and 62A were not sensitized to NaCl by 0.3 to 0.4 Mrad, i.e., in the shoulder portion of the survival curve. Radiation-sensitive strain 51B, which shows no distinct shoulder in its survival curve, was sensitized to NaCl by 0.1 Mrad, the lowest radiation dose employed in this study. These observations seem to suggest a possible relationship between deoxyribonucleic acid repair capacity and salt tolerance.     

Morphofunctional characteristics of the pancreas in the progeny from females with experimental hypothyroidism]

Histophysiological pancreatic indices were studied in 1-, 21- and 30-day-old offsprings of white non-lineal female rats that were given antithyreoid drug mercasolil in the dose 3 mg/kg of body weight during pregnancy and lactation. Chemical inhibition of maternal thyreoid function results in decreasing weight index of the offspring's pancreas, in decreasing section area of the terminal secretory parts, in narrowing the homogenous zone of exocrine pancreocytes, in decreasing the portion occupied by the endocrine part on the section area, as well as in decreasing the index of B/A cells ratio of pancreatic islands and of nuclear volume of basophilic insulocytes. It is possible to conclude from the data obtained that maternal hypothyreosis produced by mercasolil unfavourably affects the process of structural and functional setting and the status of the pancreas in the offspring.     

Relation between radiation resistance and salt sensitivity of spores of five strains of Clostridium botulinum types A, B, and E.

The NaCl tolerance of different strains of Clostridium botulinum varies over a wide range, and the patterns of NaCl inhibition differ distinctly and characteristically from strain to strain. The more radiation-resistant strains, such as 33A, 62A, and 7272A, are more resistant to NaCl, whereas the more radiation-sensitive strains, such as 51B and 1304E, are more sensitive to NaCl. This rule appears to hold irrespective of whether the spores were unirradiated controls or whether they were radiation damaged prior to exposure to NaCl in the recovery media. The data seem to indicate that radiation doses in the shoulder portion of the radiation survival curves did not noticeably sensitive the spores to NaCl, whereas radiation doses in the exponential-decline portion of the survival curve invariably produced a distinct sensitization. Thus, strains 33A and 62A were not sensitized to NaCl by 0.3 to 0.4 Mrad, i.e., in the shoulder portion of the survival curve. Radiation-sensitive strain 51B, which shows no distinct shoulder in its survival curve, was sensitized to NaCl by 0.1 Mrad, the lowest radiation dose employed in this study. These observations seem to suggest a possible relationship between deoxyribonucleic acid repair capacity and salt tolerance.