The influence of the water with decreased content of heavy stable hydrogen and oxygen (18O) isotope on development of radiation injuries at gamma-radiation in low dose

The investigations of the water with decreased content of heavy stable hydrogen and of oxygen (18O) isotope received by the method of rectification were conducted on development of radiation injuries in organisms of the experimental animals. The objects of the investigation were male mice of Balb/c line. The animals were irradiated at the unit PX-gamma-30 with the gamma-radiation source 60Co in the dose of 25 sGy. It was observed that the water with decreased content of heavy stable hydrogen and of oxygen (18O) isotope influences positively the processes of restoration. It was manifested in decrease of the degree of gravity of radiation injuries in immune system organs (thymus and spleen), peripheral blood and marrow cells.     

Role of pelvimetry in active management of labour.

All cases referred for pelvimetry in 1970-1 and all breech presentations referred for pelvimetry in 1972-4 were reviewed. Indications for pelvimetry fell into four main categories: high head in the antenatal clinic (47-8%); high head in labour (13-9%); breech presentation (20-9%); and previous caesarean section (14-8%). In the first two categories pelvimetry rarely if ever influenced management, and it should not be performed routinely. In breech presentation and cases of caesarean section pelvimetry seemed to be of value, but in the latter group it should be performed puerperally to avoid the known radiation hazard to the fetus. A fairly close correlation between obstetric conjugate and pelvic capacity was shown, which suggested that a 3400-g baby might pass through a pelvis of obstetric conjugate of 10 cm as a cephalic trial of labour, but would need an obstetric conjugate of 11-7 cm for safe vaginal breech delivery.     

ATR signaling cooperates with ATM in the mechanism of low dose hypersensitivity induced by carbon ion beam

Little work has been done on the mechanism of low dose hyper-radiosensitivity (HRS) and later appeared radioresistance (termed induced radioresistance (IRR)) after irradiation with medium and high linear energy transfer (LET) particles. The aim of this study was to find out whether ATR pathway is involved in the mechanism of HRS induced by high LET radiation. GM0639 cells and two ATM deficient/mutant cells, AT5BIVA and AT2KY were irradiated by carbon ion beam. Thymidine block technique was developed to enrich the G2-phase population. Radiation induced early G2/M checkpoint was quantitatively assess with dual-parameter flow cytometry by detecting the cells positive for phospho-histone H3. The involvement of ATR pathway in HRS/IRR response was detected with pretreatment of specific inhibitors prior to carbon ion beam. The link between the early G2/M checkpoint and HRS/IRR under carbon ion beam was first confirmed in GM0639 cells, through the enrichment of cell population in G2-phase or with Aurora kinase inhibitor that attenuates the transition from G2 to M phase. Interestingly, the early G2/M arrest could still be observed in ATM deficient/mutant cells with an effect of ATR signaling, which was discovered to function in an LET-dependent manner, even as low as 0.2 Gy for carbon ion radiation. The involvement of ATR pathway in heavy particles induced HRS/IRR was determined with the specific ATR inhibitor in GM0639 cells, which affected the HRS/IRR occurrence similarly as ATM inhibitor. These data demonstrate that ATR pathway may cooperate with ATM in the mechanism of low dose hypersensitivity induced by carbon ion beam.     

The low dose and low dose rate cytogenetic effects induced by gamma-radiation in human blood lymphocytes in vitro. II. the results of cytogenetic study

The yield of chromosome aberrations induced by gamma-radiation of 60Co in human blood lymphocytes in vitro at low doses (30 divided by 600 mGy) and low dose rates (0.70, 5.05, 59.2 mGy/min) was investigated. It was found that the observed level of chromosomal aberrations induced by gamma-irradiation was unaffected by the value of the dose rate when using constant dose rate and obtaining different doses by altering the exposure time. However, a relatively enhanced level of chromatid aberrations was found at 5.05 and 59.2 mGy/min dose rates in the dose range less than 250 mGy. We have found that the observed level of the sum of chromosomal aberrations induced by gamma-irradiation at doses less than 250 mGy and a dose rate of 59.2 mGy/min was essentially larger compared with the level extrapolated from high doses (above 300 mGy) using a linear-quadratic dose curve. This complied with our previous finding in 1976, 1977 when the enhanced level of dicentrics was only found at a high dose rate approximately 500 mGy/min. Such a non-linear cytogenetic effect does not manifest itself statistically significantly at dose rates of 0.70 and 5.05 mGy/min for the sum of chromosomal aberrations and does not manifest itself at all for dicentrics at all the examined dose rates.     

Measurement of the restricted dose mean LET outside the primary beam of a 60Co radiotherapy unit

A recently developed method for the direct measurement of the restricted dose mean LET (cutoff energy = 500 eV) of an unspecified photon or electron radiation field with the high-pressure ionization chamber has been utilized to investigate the variation of this radiobiologically important parameter outside the primary beam of a clinical 60Co unit. A small high-pressure tissue-equivalent ionization chamber was used, and its characteristics and experimental considerations for the present investigation are reported. Measurement of the restricted dose mean LET at the examined points outside the primary 60Co beam showed an increase of 50% with respect to the restricted dose mean LET of the uncollimated 60Co beam. No significant variation was noted with off-central axis distance, field size, wedge filter, or depth below Perspex slabs. Dose rates at the points of measurement outside the primary 60Co beam were 1-5% of the dose rate in the primary beam.     

Chromosome aberrations induced in human lymphocytes by neutron irradiation.

In vitro dose--response curves of unstable chromosome aberrations in human lymphocytes have been obtained for neutron spectra of mean energies 0-7, 0-9, 7-6 and 14-7 MeV. The aberration yields have been fitted to the quadratic function Y = alphaD + betaD2, which is consistent with the single-track and two-track model of aberration formation. However with high-LET radiation, the linear component of yield, corresponding to damage caused by single tracks, predominants, and this term becomes more dominant with increasing LET, so that for fission spectrum neutrons the relationship is linear, Y = alphaD. At low doses, such as those recieved by radiation workers, limiting r.b.e. values between 13 and 47 are obtained relative to 60Co gamma-radiation. At higher doses, as used in radiotherapy, the values are much lower; ranging from 2-7 to 8 at 200 rad of equivalent gamma-radiation. Both sets of r.b.e. values correlate well with track-averaged LET but not with dose-averaged LET. When the numbers of cells without aberrations are plotted against radiation dose, curves are obtained which are similar in shape to those for conventional cell-survival experiments with comparable neutron spectra. The Do values obtained in the present study are close to those from other cell system.     

Chromosome aberrations induced in human lymphocytes by radiation from 252Cf.

In vitro dose--response curves of unstable chromosome aberrations in human lymphocytes have been obtained for 252Cf neutron radiation. The aberration yields fitted best to the linear function Y=aD, which is consistent with the single-track model of aberration formation for high LET radiation. The curves have been compared with others previously produced in this laboratory for several energies of neutrons and for 60Co gamma radiation. The r.b.e. for 252Cf with respect to 60Co is 27 at very low doses, decreasing to 6 at an aberration yield equivalent to 400 rad of 18 rad/hour gamma radiation. A profile of chromosome-aberration induction with depth in a perspex phantom was obtained by placing blood samples at several distances over the range 0.65-2.0 cm from the californium source. This profile was compared with depth-damage calculations for a radium needle. The r.b.e. of 252Cf radiation relative to 226Ra gamma radiation increased with the distance from the source, implying that californium is more effective at greater distances in destroying the ability of cells to divide, which may be an advantage in the treatment of large tumours.     

Radiation-induced hemopoietic death in mice as a function of photon energy and dose rate

Radiation-induced hemopoietic death was measured in mice exposed to photons of four different energies: 250-kVp X rays, 60Co gamma rays (1.25 MeV), and 6- and 25-MV photons from a linear accelerator. For each radiation source, the lethal dose which killed 50% of the population in 30 days (LD50/30) associated with the hemopoietic syndrome was determined in groups of mice exposed to graded doses from 600 to 1150 cGy at dose rates of 20, 40, and 80 cGy/min. The calculated LD50/30 values for 25 and 6 MV were significantly different from each other at all exposure rates while no difference was observed between 6 MV and 60Co. Using 60Co gamma rays as the standard, the relative biologic effectiveness was as follows: 250 kVp greater than 25 MV greater than 6 MV = 60Co. The data suggest that there may be a greater damage to tissue within the marrow cavities following exposure to very high megavoltage radiation, a factor which must be considered with the increasing utilization of linear accelerators in the clinic and laboratory.     

Variation in U.V. primary fluorescence-intensity of vital cells depending on 60Co gamma-radiation dose.

Using impulse-cytofluorophotometry in the ultra-violet spectral region the author has shown on vital, unstained Ehrlich ascites tumour cells that the primary fluorescence intensity of this tumour is on day 11 after transplantation 20 per cent higher than on day 8. Storage of the vital cells for 25 min at 20 degrees C has no effect on this result. When the cells are exposed to 60Co to gamma-radiation on day 6, a new stable fluorescence level is established afted 20 hours. Measurements of the primary fluorescence intensity depending on dose have shown a significant rise starting from 75 rad at 48 hours after irradiation. The fluorescence intensity rises by 42.5 per cent of the control value at 3000 rad, but only by 31.5 per cent on exposure to 4000 rad.     

Calculation of dose components in head phantom for boron neutron capture therapy.

Application of neutrons to cancer treatment has been a subject of considerable clinical and research interest since the discovery of the neutron by Chadwick in 1932 (3). Boron neutron capture therapy (BNCT) is a technique of radiation oncology which is used in treating brain cancer (glioblastoma multiform) or melanoma and that consists of preferentially loading a compound containing 10B into the tumor location, followed by the irradiation of the patient with a beam of neutron. Dose distribution for BNCT is mainly based on Monte Carlo simulations. In this work, the absorbed dose spatial distribution resultant from an idealized neutron beam incident upon ahead phantom is investigated using the Monte Carlo N-particles code, MCNP 4B. The phantom model used is based on the geometry of a circular cylinder on which sits an elliptical cylinder capped by half an ellipsoid representing the neck and head, both filled with tissue-equivalent material. The neutron flux and the contribution of individual absorbed dose components, as a function of depths and of radial distance from the beam axis (dose profiles) in phantom model, is presented and discussed. For the studied beam the maximum thermal neutron flux is at a depth of 2 cm and the maximum gamma dose at a depth of 4 cm.     

Radiation inactivation of enzymes at low dose rates: Identical molecular weights of rat liver cytosolic and lysosomal neuraminidases

A low-dose-rate gamma source (⁶⁰Co) was calibrated with enzymes of known radiation inactivation behaviors and used for molecular weight determination of rat liver neuraminidase (EC 3.2.1.18). The method allows direct comparison of radiation inactivation of standard and unknown enzymes under identical experimental conditions. The membrane-bound lysosomal neuraminidase had the same molecular weight (M_r = 56,000 ± 8500) as the soluble cytosolic neuraminidase (M_r = 56,000 ± 7000) although they differ in their kinetic properties and substrate specificity.     

Action of secondary radiation from 70-Gev protons on the cellular DNA of mammals

A study was made of the effect of secondary radiation of 70 GeV protons on DNA of Chinese hamster cells. With a reference to fibroblast DNA, lymphoid cell DNA, and the lethal effect of radiation on the survival of Chinese hamster cells the RBE was 1.6-7.6, 1.1-3.8 and 1.14-1.7, respectively. DNA breaks were repaired to an equal level after exposure to secondary radiation from the accelerator and gamma-radiation from 60Co in equally effective doses.     

Evaluation of the effect of 90Sr beta-radiation on human blood cells by chromosome aberration and single cell gel electrophoresis (comet assay) analysis

Among various environmental genotoxins, ionizing radiation has received special attention because of its mutagenic, carcinogenic and teratogenic potential. In this context and considering the scarcity of literature data, the objective of the present study was to evaluate the effect of 90Sr beta-radiation on human cells. Blood cells from five healthy donors were irradiated in vitro with doses of 0.2-5.0Gy from a 90Sr source (0.2Gy/min) and processed for chromosome aberration analysis and for comet assay. The cytogenetic results showed that the most frequently found aberration types were acentric fragments, double minutes and dicentrics. The alpha and beta coefficients of the linear-quadratic model, that best fitted the data obtained, showed that 90Sr beta-radiation was less efficient in inducing chromosome aberrations than other types of low linear energy transfer (LET) radiation such as 3H beta-particles, 60Co gamma-rays, 137Cs and 192Ir and X-rays. Apparently, 90Sr beta-radiation in the dose range investigated had no effect on the modal chromosome number of irradiated cells or on cell cycle kinetics. Concerning the comet assay, there was an increase in DNA migration as a function of radiation dose as evaluated by an image analysis system (tail moment) or by visual classification (DNA damage). The dose-response relation adequately fitted the non-linear regression model. In contrast to the cytogenetic data, 90Sr beta-radiation induced more DNA damage than 60Co gamma-radiation when the material was analyzed immediately after exposures. A possible influence of selective death of cells damaged by radiation was suggested.     

Interaction between checkpoint genes RAD9, RAD17, RAD24, and RAD53 involved in the determination of yeast Saccharomyces cerevisiae sensitivity to ionizing radiation

Mechanisms for genetic control of cell division cycle (checkpoint control) have been studied in most detail in yeast Saccharomyces cerevisiae. To clarify the role of checkpoint genes RAD9, RAD17, RAD24, and RAD53 in cell radioresistance, double mutants were analyzed for cell sensitivity to ionizing radiation. Double mutants carrying mutations in combination with mutation rad9Delta were shown to manifest the epistatic type of interaction. Our results suggest that checkpoint genes RAD9, RAD17, RAD24, and RAD53 belong to a single epistatic group designated RAD9 and govern the same pathway. Genes RAD9 and RAD53 have a positive effect on sensitivity to gamma-radiation, whereas RAD17 and RAD24 have a negative effect. Interactions between mutations may differ when considering their sensitivity to gamma-radiation and UV light; mutations rad9Delta and rad24Delta were shown to manifest the additive effect in the first case and epistatic effect in the second.     

Genetic Analysis of Gamma-Ray Mutagenesis in Yeast. I. Reversion in Radiation-Sensitive Strains

The frequency of revertants induced by 60Co gamma rays of the ochre allele, cyc1-9, has been measured in radiation-sensitive strains carrying one of 19 nonallelic mutations and in wild-type strains. The results indicate that ionizing radiation mutagenesis depends on the activity of the RAD6 group of genes and that the gene functions employed are very similar, but probably not identical, to those that mediate UV mutagenesis. Repair activities dependent on the functions of the RAD50 through RAD57 loci, the major pathway for the repair of damage caused by ionizing radiation, do not appear to play any part in mutagenesis. A comparison between the gamma-ray data and those obtained previously with UV (LAWRENCE and CHRISTENSEN 1976) and chemical mutagens (PRAKASH 1976) suggests that the RAD6 "mutagenic pathway" is in fact composed of a set of processes, some of which are concerned with error-prone, and some with error-free, recovery activities.     

Interaction between checkpoint genes RAD9, RAD17, RAD24, and RAD53 involved in the determination of yeast Saccharomyces cerevisiae sensitivity to ionizing radiation

Mechanisms for genetic control of cell division cycle (checkpoint control) have been studied in most detail in yeast Saccharomyces cerevisiae. To clarify the role of checkpoint genes RAD9, RAD17, RAD24, and RAD53 in cell radioresistance, double mutants were analyzed for cell sensitivity to ionizing radiation. Double mutants carrying mutations in combination with mutation rad9delta were shown to manifest the epistatic type of interaction. Our results suggest that checkpoint genes RAD9, RAD17, RAD24, and RAD53 belong to a single epistatic group designated RAD9 and govern the same pathway. Genes RAD9 and RAD53 have a positive effect on sensitivity to gamma-radiation, whereas RAD17 and RAD24 have a negative effect. Interactions between mutations may differ when considering their sensitivity to gamma-radiation and UV light; mutations rad9delta and rad24delta were shown to manifest the additive effect in the first case and epistatic effect in the second.     

Membrane structure alterations in rat blood lymphocytes under external low dose rate gamma-radiation exposure

The influence of a 0.72 cGy/day dose rate of gamma-radiation on plasma membranes of peripheral blood lymphocytes of rats exposed to the doses of 1.5, 15, 30, 60 and 100 cGy was studied. Parameters characterizing the viscosity and the polarity of lipid bilayer and also an external membrane surface properties were examined using fluorescent probes pyrene and 1-anilinonaphthalene-8-sulfonate (ANS). Was shown the membrane structural parameters alterations after animal exposure to the doses of 1.5, 15, 60 and 100 cGy, being of a nonmonotonous nature as the dose accumulated. After exposure to the doses lower then than 30 cGy spectral changes were revealed not in each particular experiment that was probably caused by the individual peculiarities of radiation response development. After exposure to the doses higher than 30 cGy the changes were of reproducible character. After a 1.5 cGy dose a slight lipid bilayer polarity decrease and ANS binding parameter multidirectional changes were observed. After exposure to 15, 60 and to 100 cGy was shown polarity elevation and repartition of polar groups within the bilayer, the increase of viscosity of more polar membrane regions and also ANS fluorescence reduction mostly at the expense of quantum yield decrease. After the exposure of 60 cGy was observed a viscosity decrease in hydrophobic regions along with viscosity increase in more polar regions and after a 100 cGy dose accumulation an essential surface charge shift was found. Revealed alterations indicate the reorganization of external membrane surface and of intensification of oxidative processes in lipid bilayer.     

Responses of the beagle to protracted irradiation. I. Effect of total dose and dose rate

The effects of protracted exposure to 60Co gamma rays on survival and tumor induction in the beagle were investigated. Total accumulated doses of 450, 1050, 1500, and 3000 cGy were given at rates of 3.8, 7.5, 12.8, and 26.3 cGy/day. Hazard models were used to identify trends in mortality associated with radiation exposure. The probability of an acute death (related to hematopoietic aplasia) was positively associated with the total dose received and the rate at which the dose was delivered. Once an animal survived the initial hematopoietic effects of radiation exposure, the risk of death from causes other than cancer, while elevated, was far less responsive than the neoplastic end points. No relationship between tumor or chronic nontumor deaths and dose rate could be identified. However, survival curves for tumor mortality did separate into a pattern clearly dependent on the accumulated dose.     

Ionizing radiation can overcome resistance to TRAIL in TRAIL-resistant cancer cells

Although the majority of cancer cells are killed by TRAIL (tumor necrosis factor-related apoptosis-inducing ligand treatment), certain types show resistance to it. Ionizing radiation also induces cell death in cancer cells and may share common intracellular pathways with TRAIL leading to apoptosis. In the present study, we examined whether ionizing radiation could overcome TRAIL resistance in the variant Jurkat clones. We first selected TRAIL-resistant or -sensitive Jurkat clones and examined cross-responsiveness of the clones between TRAIL and radiation. Treatment with gamma-radiation induced significant apoptosis in all the clones, indicating that there seemed to be no cross-resistance between TRAIL and radiation. Combined treatment of radiation with TRAIL synergistically enhanced killing of TRAIL-resistant cells, compared to TRAIL or radiation alone. Apoptosis induced by combined treatment of TRAIL and radiation in TRAIL-resistant cells was associated with cleavage of caspase-8 and the proapoptotic Bid protein, resulting in the activation of caspase-9 and caspase-3. No changes in the expressions of TRAIL receptors (DR4 and DR5) and Bcl-2 or Bax were found after treatment. The caspase inhibitor z-VAD-fmk completely counteracted the synergistic cell killing induced by combined treatment of TRAIL and gamma-radiation. These results demonstrated that ionizing radiation in combination with TRAIL could overcome resistance to TRAIL in TRAIL-resistant cells through TRAIL receptor-independent synergistic activation of the cascades of the caspase-8 pathway, suggesting a potential clinical application of combination treatment of TRAIL and ionizing radiation to TRAIL-resistant cancer cells.     

Low doses of radiation decrease the level of spontaneous and gamma-induced chromosomal mutagenesis in bone marrow cells of mice in vivo

Low doses of ionizing radiation are known to induce adaptive response (AR), which is characterized in most cases by temporary nature, though the possibility of long-term persistence of AR is not ruled out. In this investigation we studied the effect of low doses of gamma-radiation on both high-dose radiation-induced and spontaneous level of cytogenetic damage throughout the life of mice. SHK male mice 2 months old were used. Priming doses of 0.1 and 0.2 Gy (0.125 Gy/min, gamma-radiation from 60Co) were used. A challenging dose of 1.5 Gy (1 Gy/min) was used in the experiments using a routine AR experimental design. The frequency of micronucleated polychromatic erythrocytes in bone marrow cells of primed, primed and challenged, and control groups was assessed at various times of animal life span. It was shown that: a) single low-dose gamma-irradiation induces a cytogenetic AR which can be revealed at 1, 3, 6, 9, 12 months after priming; b) single low-dose gamma-irradiation decreases the cytogenetic damage to a level below the spontaneous rate at the end of lifetime (20 months) of animals; c) ability to induce adaptive response does not depend on the age of animals at the moment of priming irradiation. In conclusion, the mechanisms underlying AR not only protect from chromosome damage induced by high-dose irradiation but also may play a role in spontaneous mutagenesis during aging of animals.