Arterial wall damage by X-rays and fast neutrons.

Radiation-induced atheromatosis has been studied for 200 kVp X-rays and 15 MeV neutrons. From the results of two earlier experiments, a r.b.e. less than 1 was expected for neutrons. (1) Irradiation of blood-vessels causes depolymerization of the mucopolysaccharides in the vessel wall, resulting in atheromatosis, and (2) in two other mucopolysaccharide systems a r.b.e. less than 1 is observed for fast neutrons. Irradiation of the carotid arteries of a total number of 120 hypercholesterolaemic rabbits, divided over several groups, with 500 and 1000 rad of X-rays or neutrons results, however, in atheromatous plaques which are more pronounced for neutrons than for X-rays at the 500 rad dose-level; for a dose of 1000 rad the effect of neutrons is less extensive than the effect of X-rays. These results lead to the assumption that the origin of the atheromatosis seems not only to be the mucopolysaccharide depolymerization, but that radiation induced damage at the cellular level must be taken into account.     

The effects of graded doses of 1 MeV fission neutrons or X rays on the murine hematopoietic stroma.

The acute radiosensitivity in vivo of the murine hematopoietic stroma for 1 MeV fission neutrons or 300 kVp X rays was determined. Two different assays were used: (1) an in vitro clonogenic assay for fibroblast precursor cells (CFU-F) and (2) subcutaneous grafting of femora or spleens. The number of stem cells (CFU-S) or precursor cells (CFU-C), which repopulated the subcutaneous implants, was used to measure the ability of the stroma to support hemopoiesis. The CFU-F were the most radiosensitive, and the survival curves after neutron and X irradiation were characterized by D0 values of 0.75 and 2.45 Gy, respectively. For regeneration of CFU-S and CFU-C in subcutaneously implanted femora, D0 values of 0.92 and 0.84 Gy after neutron irradiation and 2.78 and 2.61 Gy after X irradiation were found. The regeneration of CFU-S and CFU-C in subcutaneously implanted spleens was highly radioresistant as evidenced by D0 values of 2.29 and 1.49 Gy for survival curves obtained after neutron irradiation, and D0 values of 6.34 and 4.85 Gy after X irradiation. The fission-neutron RBE for all the cell populations was close to 3 and varied from 2.77 to 3.28. The higher RBE values observed for stromal cells, compared to the RBE of 2.1 reported previously for hemopoietic stem cells, indicate that stromal cells are relatively more sensitive than hemopoietic cells to neutron irradiation.     

Chromosomal rearrangements induced in mouse spermatogonia by 14.5-MeV neutrons

Inbred CBA male mice were irradiated with 14.5-MeV neutrons. Three acute doses, 75, 150 and 250 rad, and one chronic dose, 250 rad, were given. The percentages of affected spermatocytes as counted from reciprocal translocations which had been induced in spermatogonia were 0.7, 0.8 and 1.6 respectively for the acute series and 2.2 after chronic exposure. The data could be fitted to a linear or concave curvilinear regression line. There seemed to be a slight increase of damage with dose, even if the percentages were generally lower than those reported earlier for fast neutrons with energies around 1 MeV. The existence of dose-rate effects is discussed, and the conclusion drawn so far is that there seems to be no such effect either for 1-MeV fast neutrons or 14.5-MeV high energy neutrons. The term “reversed dose-rate effect”, as used earlier, relates to another phenomenon. The difference between the point estimates for the chronic and acute 250 rad series is not significant. The effectiveness of neutrons with energies around 14 MeV versus neutrons with energies around 1 MeV is discussed.     

Relative biological effectiveness of x-rays and fast neutrons in inducing translocations in mouse spermatogonia

The dose-response relationships for inducing translocations in the spermatogonia of mice were studied, and they were compared for 200 kVp X-rays and 2 MeV fast neutrons. The dose response for fast neutrons was markedly convex; more precisely, the response obtained was linear in the dose range from 24 to 94 rad with a regression coefficient of 11.36·10^?4, but decreased for a further increase in dose up to 267 rad. On the other hand, that for X-rays showed a linear dose-response relationship from 48 to 672 rad with a regression coefficient of 2.69·10^?4. The relative biological effectiveness for inducing translocations in the spermatogonia of mice was compared for the linear parts of the dose response in both types of radiation, and the relative biological effectiveness (RBE) value was 4.22.     

Relative effectiveness of neutrons and X-rays in induction of crossing over in drosophila males

Relative biological effectiveness of neutrons vs. X-rays in inducing crossing-over in males of D. melanogaster was investigated using 812 and 834 rad of neutrons and the same dose of X-rays. Crossing-over was induced in spermatocytes and spermatogonia of adults and pupae. Neutrons were 4 times more effective in spermatocytes of adults and their effectiveness in pupal spermatocytes was even more. Neutrons also induced more exchanges in spermatogonial cells including predefinitive spermatogonia. Higher effectiveness of neutrons can be attributed to their high linear energy transfer.     

Response of colony-forming units-spleen to heavy charged particles

Survival of colony-forming units-spleen (CFU-S) was measured after single doses of photons or heavy charged particles from the BEVALAC. The purposes were to define the radiosensitivity to heavy ions used medically and to evaluate relationships between relative biological effectiveness (RBE) and dose-averaged linear energy transfer (LET infinity). In in vitro irradiation experiments. CFU-S suspensions were exposed to 220 kVp X rays or to 20Ne (372 MeV/micron) or 40Ar (447 MeV/micron) particles in the plateau portion of the Bragg curve. In in vivo irradiation experiments, donor mice from which CFU-S were harvested were exposed to 12C (400 MeV/micron). 20Ne (400 or 670 MeV/micron), or 40Ar (570 MeV/micron) particles in Bragg peaks spread to 4 or 10 cm by spiral ridge filters. Based on RBE at 10 survival, the maximum RBE of 2.1 was observed for 40Ar particles characterized by an LET infinity of approximately 100 keV/micron. Lower RBEs were determined at lower or higher estimated values of LET infinity and ranged from 1.1 for low energy 40Ar particles to 1.5-1.6 for low energy 12C and 20Ne. The responses of CFU-S are compared with responses of other model systems to heavy charged particles and with the reported sensitivity of CFU-S to neutrons of various energies. The maximum RBE reported here, 2.1 for high energy 40Ar particles, is somewhat lower than values reported for fission-spectrum neutrons, and is appreciably lower than values for monoenergetic 0.43-1.8 MeV neutrons. Low energy 12C and 20Ne particles have RBEs in the range of values reported for 14.7 MeV neutrons.     

Induction of chromosome aberrations and cell killing in Syrian hamster fibroblasts by gamma-rays, X-rays and fast neutrons

Chromosome aberrations were scored in BHK21 C13 Syrian hamster fibroblasts, exposed to 60Co gamma-rays, 250 kV X-rays, 15 MeV neutrons or neutrons of mean energy 2.1 MeV produced from the 9Be(d,n)10B reaction. The cells were irradiated in stationary phase, where they are concentrated in the G1 phase of the cell cycle. Within experimental uncertainty there was no detectable difference between the responses to 60Co gamma-rays and to 250kV X-rays. The r.b.e. for the production of dicentrics, based on the 'one-hit' component of response, was (5 +/- 2) for the 15 MeV neutrons and (12 +/- 5) for the 2.1 MeV neutrons. For each radiation, a graph of the proportion of cells without a dicentric, centric ring or acentric fragment corresponded closely to the survival curve for stationary-phase cells obtained in the same experiment.     

Study of dose-rate effects of neutron radiation in a wild-type and a repair-deficient yeasts saccharomyces

We report here a comparative analysis of RBE for lethality of a single pulse (duration 65 micros) of fast neutron with ultra high dose rates (up to 6 x 10(6) Gy/s) and continuous neutron radiation (3.6 x 10(3) s) of the pulse reactor BARS-6. Three diploid strains, one haploid strain and three diploid repair-deficient strains (rad52-1/rad52-1; rad54/rad54; rad2/rad2) were used. The RBE values (D(0gamma)/1D(0n)) of a single pulse and continuous neutron irradiation were equal (1.7-1.8) with maximum RBE (4.1-3.1) in region of low doses (shoulder region). Haploid cells were found to be more (3 times) sensitive to both gamma-rays and neutrons than the wild type. There was no obvious decrease in the RBE of 1.9 in highly sensitive haploid cells as compared with highly resistant diploid cells. The repair-deficient strains (rad52-1/rad52-1; rad54/rad54) were more (up to 10 fold) sensitive to both neutrons and gamma-rays as compared with their parent line. The RBE values of 1.5-1.7 of neutrons for these mutants (independent by of the mode of irradiation) were found. The repair-deficient mutant rad2/rad2 had similar sensitivity as a wild type and a RBE value was 2.0. We have concluded that biological effectiveness of the neutrons of pulse reactor BARS-6 was independent of the dose-rate, differing up to 10(8) fold. The RBE didn't vary significantly with the capacity of cells to repair DNA damages.     

Fast neutron r.b.e. for lethality and genotoxicity in a wild-type and a repair-deficient strain of yeast

The relative biological effectiveness (r.b.e.) of cyclotron-produced fast neutrons (11 MeV) in relation to 60Co gamma-rays, was studied in a wild-type and a DNA repair-deficient yeast strain for cell killing and genotoxicity. In the wild-type (D7) strain the r.b.e. varied from 2.7 to 4.1 for lethality, 2.8 to 7.1 for reverse mutation and 3.5 to 7.8 for mitotic gene conversion. At different survival levels, the repair deficient strain (D7 rad 52/rad 52) generally showed a lower r.b.e. for both cell killing and genotoxicity (25.2 to 37.2 per cent reduction for the cell death and 24.8 to 70.6 per cent for mutation and gene conversion) compared to the wild type. Except at very low dose levels, the r.b.e. values for cell killing and genotoxicity were similar within a given strain. At similar survival levels, neutrons were no more genotoxic than gamma-rays.     

Mortality of monkeys after exposure to fission neutrons and the effect of autologous bone marrow transplantation.

In order to assess the risk of exposure to ionizing radiation in man, and to evaluate the results of therapeutic measures, the mortality of rhesus monkeys irradiated with X-rays and fission neutrons and the effect of autologous bone marrow transplantation have been investigated. The LD50/30d values for X- and neutron-irradiated monkeys amount to 525 and 260 rad respectively, resulting in an r.b.e. of approximately 2 for the occurrence of the bone marrow syndrome. Protection of the animals by autologous bone marrow transplantation was observed up to doses of 860 rad of X-rays and 440 rad of fission neutrons. After both fission-neutron irradiation and X-irradiation in the lowest range of lethal doses, the bone marrow syndrome was found to occur without the concurrent incidence of the intestinal syndrome. The studies indicate that, for humans accidentally exposed to what would otherwise be lethal doses of fast neutrons, bone marrow transplantation may be beneficial.     

Somatic aberration induction in Tradescantia occidentalis by neutrons, x- and gamma-radiations. I. Dosimetry.

The dosimetry is described for an investigation of the induction of somatic aberrations in Tradescantia occidentalis by substantially mono-energetic neutrons in the energy range 100 keV to 15 MeV, by 200 keV X-rays and cobalt-60 gamma-radiation. Spectrometry was carried out for both neutrons and X-rays. Neutron fluence was measured by uranium fission chambers. Two types of ionization chamber were employed for dose measurement. One chamber was manufactured of CH-plastic and filled with acetylene and the other of graphite and filled with carbon dioxide. Dosimetry for X- and gamma-radiation was by means of lithium fluoride thermoluminescent dosemeters calibrated against a Victoreen ionization chamber.     

The mouse splenocyte assay, an in vivo/in vitro system for biological monitoring: studies with X-rays, fission neutrons and bleomycin.

A modified mouse splenocyte culture system was standardized after testing different mitogens (i.e., phytohemagglutinin (PHA), concanavalin A (Con A)). The mitotic index was determined for comparison between different mitogens. Following selection of appropriate mitogen (PHA 16, Flow), a series of experiments were conducted to evaluate the application of a cytokinesis-block for scoring micronuclei and assays for chromosomal aberrations produced by treatment in G0 and G2 for the purposes of biological dosimetry following in vivo and/or in vitro exposure to X-rays, fission neutrons and bleomycin. In the X-irradiation studies, the frequencies of micronuclei and chromosomal aberrations (i.e., dicentrics and rings) increased in a dose-dependent manner. These data could be fitted to a linear-quadratic model. No difference was observed between irradiation in vivo and in vitro, suggesting that measurement of dicentrics and micronuclei in vitro after X-irradiation can be used as an in vivo dosimeter. Following in vivo irradiation with 1 MeV fission neutrons and in vitro culturing of mouse splenocytes, linear dose-response curves were obtained for induction of micronuclei and chromosomal aberrations. The lethal effects of neutrons were shown to be significantly greater than for a similar dose of X-rays. The relative biological effectiveness (RBE) was 6-8 in a dose range of 0.25-3 Gy for radiation-induced asymmetrical exchanges (dicentrics and rings), and about 8 for micronuclei in a dose range of 0.25-2 Gy. Furthermore, the induction of chromosomal aberrations by bleomycin was investigated in mouse G0 splenocytes (in vitro) and compared with X-ray data. Following bleomycin treatment (2 h) a similar pattern of dose-response curve was obtained as with X-rays. In this context a bleomycin rad equivalent of 20 micrograms/ml = 0.50 Gy was estimated.     

The response of mouse skin to hyperthermia combined with fast neutrons or X-rays

The effects of hyperthermia combined with fast neutrons (mean energy approximately 7.5 MeV) or X-rays (250 kVp) were studied in the skin of the mouse ear and foot. Hyperthermia was achieved by immersion in water at temperatures of 41.5-43.0 degrees C for 1 hour. The heat treatments used caused no observable tissue injury other than transient erythema but they enhanced the response to both neutrons and X-rays. The enhancement of neutron damage increased as the heating temperature was increased, as is well known for X-rays. When heat was given after irradiation the thermal enhancement ratio (t.e.r.) for neutrons was similar to that for X-rays. When heat was given before irradiation the neutron t.e.r. was less than that for X-rays. Consequently, the relative biological effectiveness of fast neutrons compared with X-rays was not altered by giving heat after irradiation but it was reduced by giving heat before irradiation.     

The relationship between o.e.r., r.b.e. and number of fractions for X-ray- or neutron-induced skin damage.

The skin reactions in aerated and hypoxic mouse tails after single or fractionated doses of 250 kV X-rays or fast neutrons (6 MeV deuterons on beryllium) have been measured. The o.e.r. for one to sixteen fractions of X-rays remains constant, while that for one to ten fractions of neutrons decreases with increasing neutron fractionation and decreasing neutron dose/fraction. The o.e.r. for X-rays was 1.7, for single-neutron doses 1.4, and for ten fractions of neutrons 1.25. It was anticipated that the o.e.r. for neutron-induced damage would decrease further as neutron fractionation is increased because the contribution to damage from the highest LET components of dose, the alpha and heavy recoil particles, would increase relative to the lowe LET components. The r.b.e. values obtained for skin damage were higher at all neutron doses/fraction examined in this study on tails than all those previously obtained in studies on skin at other sites on four species. This may be due to the influence of hypoxia on the r.b.e. measurements in the mouse tail.     

The relative biological effectiveness in V79 Chinese hamster cells of the neutron capture reactions in boron and nitrogen

V79 Chinese hamster cells were irradiated in the presence of different amounts of boric acid with thermal neutrons at the Medical Research Reactor at Brookhaven National Laboratory. From the linear dose-survival curves observed, a D0 value of 66 rad for the 10B(n, alpha) 7Li neutron capture reaction was obtained. No dependence of this value on the concentration of boric acid was found. Comparing this value to the D0 value of 150 rad obtained with 250 kVp X rays between 10 and 0.01% survival, an extrapolated RBE value of 2.3 was calculated. By irradiation of the same line of cells with cold neutrons at the Institut Laue - Langevin , a D0 value for the 14N(n,p)14C reaction of 77 rad was obtained, with a corresponding RBE value of 1.9. Comparison is made with previously published RBE values for the 10B(n, alpha) 7Li reaction.     

Survival and proliferative activity of L-cells after irradiation with neutrons of different energies

With L-cells exposed to neutrons and X-rays the RBE of fission spectrum neutrons (1.2 MeV) was 2.8, and that of high-energy neutrons (22 MeV), 1.3. X-Irradiation with small doses (0.25 to 0.50 Gy) exerted a stimulatory effect on the growth and division of cells.     

Lethal and mutagenic effect of fast neutrons of different energies on the spores of Streptomyces griseus

A study was made of lethal and mutagenic effects of fast neutrons of different energy on spores of prototrophic and auxotrophic strains of Streptomyces griseus. Relative biological effectiveness of fast neutrons is higher than that of gamma-rays and depends on beam energy. Neutrons of 22-50 MeV induce Streptomyces griseus mutations more frequently (by one order of magnitude) than neutrons of 1.4-1.6 MeV do. The obtained mutants can be used in studying Streptomyces griseus genetics.     

Radioresistance of mongolian gerbils. Fast neutrons.

Seventy-six 8 week old Mongolian gerbils were exposed to acute, whole-body fast neutrons produced by The University of Michigan 83-in. cyclotron. Groups of seven or eigth gerbils were given doses between 485 and 881 rad at 25 rad per minute. The LD 50/30 determined by probit analysis was 750 rad, with 95 per cent fiducial limits of 733 and 776. For the 50 per cent mortality level, an r.b.e. of fast neutrons compared with cobalt-60 of 1-45 was determined. For the same end-point, the r.b.e. for fast neutrons compared with X-rays is 1-33. Mortality data, body-weight and microhaematocrit changes are discussed.     

Cytogenetic effects induced by neutrons in human peripheral blood lymphocytes in vitro. I. Dose-effect relationship for different types of chromosome aberrations when exposed to neutrons with different energies]

Human lymphocytes were irradiated in vitro during G0 stage by graded doses of thermal neutrons and of neutrons with mean energy of 0.04; 0.09; 0.35; 0.85 and 14.7 MeV as well as by 60Co gamma-rays. The data were fitted to the linear and linear-quadratic relations. The neutrons of low and intermediate energies showed the linear dependence on the dose, 14.7 MeV neutrons and gamma-rays--a linear-quadratic one, whereas the data obtained with 0.85 MeV neutrons fitted well the both models. Terminal and interstitial deletions produced by both gamma-rays and neutrons showed different dependencies upon the dose. Some qualitative pecularities of aberration spectra were found in the experiments with neutrons as compared with the data on gamma-irradiation: the ratio of exchanges to fragments was greater, and aberrations of chromatid type were produced. The specially designed experiments and calculations showed that the last effect was not connected with induced radioactivity.     

The response of spermatogonia and spermatocytes of the Northern vole Microtus oeconomus to the induction of sex-chromosome nondisjunction, diploidy and chromosome breakage by X-rays and fast fission neutrons

Microtus males were exposed to different doses of 250 kV X-rays or fast fission neutrons of 1 MeV mean energy. Early (= round) spermatids were analyzed for the presence of extra sex chromosomes, diploidy and micronuclei at different time intervals corresponding with treated differentiating spermatogonia and spermatocytes. Induction of nondisjunction of sex chromosomes could not be detected. In contrast, induction of diploids by both types of radiation was statistically significant at all sampling times. Dose-effect relationships for most of the sampling times were linear and sometimes linear-quadratic concave upward or downward. There were pronounced stage-specific differences in sensitivity as reflected by differences in doubling doses that ranged from 4 to 22 cGy for X-rays and from 0.4 to 4 cGy for neutrons. Spermatocytes at pachytene were the most sensitive cells and proliferating spermatogonia the least sensitive ones. The relative biological effectiveness (RBE) of neutrons depended on the cell stage treated and fluctuated between 1.4 and 9.2. Evidence for radiation-induced chromosomal breakage events was obtained via detection of micronuclei. Induction of micronuclei by X-rays or neutrons was statistically significant at all spermatocyte stages tested. There was no effect in spermatogonia. With a few exceptions dose-effect relationships were linear. Differences in stage sensitivity were clearly present as evidenced by doubling dose which ranged from 5 to 29 cGy for X-rays and from 1 to 3 cGy for neutrons. RBE values varied from 5.2 to 12.7. Maximum sensitivity was detected in spermatocytes at diakinesis, MI and MII. Resting primary spermatocytes (G1 and S phase) were somewhat less sensitive and actively proliferating spermatogonia were the least sensitive cells. The pattern of stage sensitivity for induction of diploids was distinctly different from that for induction of chromosomal breakage.