Microdosimetric considerations of effects of heavy ions on E. coli K-12 mutants.

The inactivation cross sections of E. coli K-12 recombination-deficient mutants, JC1553 (recA) and AB2470 (recB), for several MeV/u alpha-particles and N ions have been successfully analyzed by Katz's target theory in which radiosensitivity parameter E0 is assumed to be LET independent and equal to D37 for gamma-rays. For E. coli K-12 wild type, AB1157 (rec+, uvr+), however, it is impossible to interpret the inactivation cross section data by an LET-independent E0-value. In the latter case, as in the case of B. subtilis spore, it is necessary to assume that the radiosensitivity of the target for the core of a heavy ion is higher than that for delta-electrons. As well as Waligorski, Hamm and Katz's dose, the dose around the trajectory of an ion based on Tabata and Ito's energy deposition algorithm for electrons has been used in the course of analysis.     

Microdosimetric measurements of heavy ion tracks.

The biological effectiveness of radiations depends on the spatial pattern of ionizations and excitations produced by the charged particle tracks involved. Ionizations produced by both the primary ion and by energetic delta rays may contribute to the production of biologically relevant damage and to the concentration of damage which may effect the probability of repair. Although average energy concentration (dose) can be calculated using homogeneous track models, the energy is actually concentrated in small volumes containing segments of the ion and delta ray tracks. These local concentrations are studied experimentally using low pressure proportional counters, and theoretically, using Monte Carlo methods. Small volumes near an ion track may be traversed by a delta ray. If they are, the energy deposited will be similar to that produced by a single electron track in a low-energy x-ray irradiation. The probability of a delta ray interaction occurring decreases with the square of the radial distance from the track. The average energy deposited is the product of this probability and the energy deposited in an interaction. Average energy deposited calculated from measured interaction probability is in good agreement with the results of homogeneous track models.     

Quantitative interpretation of heavy ions effects: models for the biological effects of heavy ions.

Heavy ions are an important part of space radiation. Although they contribute only about 1 percent in number the fraction in terms of energy deposited is much higher. Also the quality of radiation is different from the other components since the LET is generally quite high. This poses the problem of Relative Biological Effectiveness (RBE). It is considerably more important in space than on earth because shielding measures are costly and sometimes not even feasible. Radiation hazards appear to be the limiting factor In long term space flights and their evaluation constitutes a major task. There is still no general agreement about RBE of earthbound radiation, and even less concerning the biological weighting of very heavy and very energetic ions in space. Because of the lack of experimental data--particularly for risk estimates in humans-- theoretical approaches may be very helpful in this respect and provide the only means to judge the radiation protection situation in outer space. In order to be useful careful checks of their consistency are necessary. This paper summarizes some of the more common approaches in a critical manner. The unhappy conclusion at the end will be that at present it is not possible to understand even heavy ion action on survival quantitatively with an acceptable precision.     

Mutagenic effects of heavy ions in bacteria.

Various mutagenic effects by heavy ions were studied in bacteria, irradiated at accelerators in Dubna, Prague, Berkeley or Darmstadt. Endpoints investigated are histidine reversion (B. subtilis, S. typhimurium), azide resistance (B. subtilis), mutation in the lactose operon (E. coli), SOS chromotest (E. coli) and lambda-prophage induction (E. coli). It was found that the cross sections of the different endpoints show a similar dependence on energy. For light ions (Z < or = 4) the cross section decreases with increasing energy. For ions of Z = 10, it is nearly independent of energy. For heavier ions (Z > or = 26) it increases with energy up to a maximum or saturation. The increment becomes steeper with increasing Z. This dependence on energy suggests a "mutagenic belt" inside the track that is restricted to an area where the density of departed energy is low enough not to kill the cell, but high enough to induce mutations.     

Mutagenic effects of heavy ions in bacteria.

The peculiarities and mechanisms of the mutagenic action of gamma-rays and heavy ions on bacterial cells have been investigated. Direct mutations in the lac-operon of E. coli in wild type cells and repair deficient strains have been detected. Furthermore, the induction of revertants in Salmonella tester strains was measured. It was found that the mutation rate was a linear-quadratic function of dose in the case of both gamma-rays and heavy ions with LET up to 200 keV/micrometer. The relative biological effectiveness (RBE) increased with LET up to 20 keV/micrometer. Low mutation rates were observed in repair deficient mutants with a block of SOS-induction. The induction of SOS-repair by ionizing radiation has been investigated by means of the "SOS-chromotest" and lambda-prophage induction. It was shown that the intensity of the SOS-induction in E. coli increased with increasing LET up to 40-60 keV/micrometer.     

Biological effects of heavy ions in Arabidopsis seeds.

Irradiation of dry seeds of Arabidopsis with heavy ions (HZE-particles) produced by UNILAC-accelerator (GSI, Darmstadt) yielded aberrations in varied developmental endpoints such as survival rate and embryo vitality. The damage increased with particle density and charge. Cross sections in the range of 0.2-1.0 micrometer2 for Ne and Ar and 2.0-10.0 micrometers2 for Xe were estimated. Soaked seeds were more sensitive than dry seeds (cross-section 2.0-10.0 micrometers2 for Ar). The induced total damage in the irradiated seeds was estimated adding the different damages weighted by certain factors. These results will be used as base data for the interpretation and evaluation of spaceflight experiments on the biological effects of cosmic radiation.     

Biological effects of heavy ions from the standpoint of target theory.

The biological effect of heavy ions is best described through the action cross section, as a function of the end-point of interest and the charge and speed of the ion. In track theory this is called the "ion-kill" cross section, for it is the effect produced by a single heavy ion and its delta rays. As with nuclear emulsions the biological track structure passes from the grain count regime to the track width regime to the thindown region with an increase in LET. With biological cells, as with any detector capable of storing sublethal damage, with low LET irradiation the action cross section (in the ion-kill mode) is increasingly obscured by the effect of "gamma-kill", by the influence of overlapping delta rays from neighboring heavy ions. Thus at low LET response is dominated by the gamma-kill mode, so that the RBE approaches 1. The theory requires 4 radiosensitivity parameters for biological detectors, extracted from survival curves at several high LET bombardments passing through the grain count regime, and at high doses. Once these are known the systematic response of biological detectors to all high LET bombardments can be unfolded separating ion kill from gamma kill, predicting the response to a mixed radiation environment, and predicting low dose response even at the level of a single heavy ion. Cell killing parameters are now available for a variety of cell lines. Newly added is a set of parameters for cell transformation.     

Effect of heavy ions on neuro-endocrine regulations.

During American and Russian short and long-term space flights neuroimmune dysregulations have been observed in man and rats for up to three months after the return. During Extra-Vehicular Activity, radiation exposure risk is greater to elicit short and/or long-term deleterious effects on the functional capacity of the neuroimmune system. In order to assess the effects of high LET events on neuroimmune networks, our preliminary ground-based study was to investigate brain inflammatory responses in mouse after low dose radiation exposure with high LET particles (12C, 95MeV/u, 42 mGy). Plasma corticosterone levels were rapidly (6 hours) increased by two-fold, then decreased 24 hours post-irradiation. At 3 days plasma corticosterone and ACTH concentrations were also two- to three-fold increased. Plasma ACTH levels were still elevated up to seven days to two months. Furthermore immune functions are under current assessment. The results of this study should allow a greater understanding of the effects of high LET particles on neuroimmune system.     

Effect of heavy ions on bacterial spores.

Inactivation of B. subtilis spores has been studied using accelerated He, C, N, O and Ne ions. The energy dependence of the inactivation cross sections for heavy ions was very weak and the mean cross sections for carbon ions (0.6-4.7 MeV/amu), nitrogen ions (0.6-4.1 MeV/amu), oxygen ions (0.8-1.1 MeV/amu), and neon ions (2.2-3.7 MeV/amu) were found to be about 0.22, 0.23, 0.26, and 0.33 micrometer2 , respectively. Analysis was carried out along lines similar to Katz's target theory but the parameters were chosen so that they have an experimental basis.     

Effects of heavy ions on cycling stem cells.

Murine marrow stem cells assayed with the spleen colony assay have been shown to be largely in a noncycling state, Go. In the unirradiated animal where these spleen-colony forming units (CFUs) transit normally between a non-proliferative state and active proliferation, exposure to a sufficient dose of ionizing radiation increases the frequency (probability) of this transition. For low-LET irradiation, marrow stem cells are not induced into cycle until a threshold dose is achieved. This dose appears to be in the range 50 to 100 cGy, inducing proliferation in an all-or-nothing manner. For irradiation with heavy charged-particles, however, the threshold dose is dependent on mass and energy. Irradiation with particles of sufficient mass and energy stimulates active proliferation even at the smallest doses tested, 5 cGy. Further, this response does not appear to result from an all-or-nothing effect. Rather, individual animals with intermediate levels of stem cell cycling have been observed. These data support the notion that locally controlled hemopoiesis can be affected by local deposition of radiation damage.     

Fast heavy ions in the heliosphere

The study of heavy ions in space can provide a greater understanding of contemporary nucleosynthesis in our galaxy, as well as the acceleration and propagation of energetic particles in the interplanetary medium and the magnetosphere. We describe here a satellite experiment that uses plastic track detectors to study the age of cosmic-ray source material and the distribution of pathlengths over which the heaviest cosmic rays travel to earth. The experiment will also search for the singly charged particles, thought to make up the anomalous component, and for low-energy heavy ions deep in the magnetosphere.     

Mutation induction by heavy ions.

Mutation induction by heavy ions is compared in yeast and mammalian cells. Since mutants can only be recovered in survivors the influence of inactivation cross sections has to be taken into account. It is shown that both the size of the sensitive cellular site as well as track structure play an important role. Another parameter which influences the probability of mutation induction is repair: Contrary to naive assumptions primary radiation damage does not directly lead to mutations but requires modification to reconstitute the genetic machinery so that mutants can survive. The molecular structure of mutations was analyzed after exposure to deuterons by amplification with the aid of polymerase chain reaction. The results--although preliminary--demonstrate that even with densely ionizing particles a large fraction does not carry big deletions which suggests that point mutations may also be induced by heavy ions.     

Microscopic track structure of equal-LET heavy ions.

The spatial distributions of ionization and energy deposition produced by high-velocity heavy ions are crucial to an understanding of their radiation quality as exhibited eg., in track segment experiments of cell survival and chromosome aberrations of mammalian cells. The stopping power (or LET) of a high velocity ion is proportional to the ratio z2/v2, apart from a slowly varying logarithmic factor. The maximum delta-ray energy that an ion can produce is proportional to v2 (non-relativistically). Therefore, two HZE ions having the same LET, but in general differing z and v will have different maximum delta-ray energies and consequently will produce different spatial patterns of energy deposition along their paths. To begin to explore the implications of this fact for the microscopic dosimetry of heavy ions, we have calculated radial distributions in energy imparted and ionization for iron and neon ions of approximately equal LET in order to make a direct comparison of their delta-ray track structure. Monte Carlo techniques are used for the charged particle radiation transport simulation.     

Low energy ions in the heavy ions in space (HIIS) experiment on LDEF.

We present data from the Lexan top stacks in the Heavy Ions In Space (HIIS) experiment which was flown for six years (April 1984-Jan 1990) onboard the LDEF spacecraft in 28.5 degrees orbit at about 476 km altitude. HIIS was built of passive (i.e. no timing resolution) plastic track detectors which collected particles continuously over the entire mission. In this paper we present data on low energy heavy ions (10 < or = Z, 20MeV/nuc < E < 200 MeV/nuc). These ions are far below the geomagnetic cutoff for fully ionized ions in the LDEF orbit even after taking into account the severe cutoff suppression caused by occasional large geomagnetic storms during the LDEF mission. Our preliminary results indicate an unusual elemental composition of trapped particles in the inner magnetosphere during the LDEF mission, including both trapped anomalous cosmic ray species (Ne, Ar) and other elements (such as Mg and Fe) which are not found in the anomalous component of cosmic rays. The origin of the non-anomalous species is not understood, but they may be associated with the solar energetic particle events and geomagnetic disturbances of 1989.     

Microscopic track structure of heavy ions videoelectronically measured.

The track of an ionizing particle in a track detector, such as silver-chloride or nuclear emulsion, contains two measurable parameters, the "track width"--which corresponds to the integrated lateral energy deposition dE/dtau(x,r) over r along dx (dE/dx or LET)--and the lateral distribution of the track density along r, the radial structure of the track. A videoelectronic device consisting of a computer-controlled microscope, developed in Frankfurt, automatically reads out consecutive lateral profiles of the optical density of a preset track, following it through the depth of the detector. For a given track length (100 microns, 400 profiles for instance) the system evaluates the mean track width MTW and the variance sigma 2 . Both quantities are correlated so that a given MTW (LET) belongs to different values of sigma 2 depending on the charge of the particle. Videosystem and results are presented.     

Neoplastic transformation of hamster embryo cells by heavy ions

We have studied the induction of morphological transformation of Syrian hamster embryo cells by low doses of heavy ions with different linear energy transfer (LET), ranging from 13 to 400 keV/μm. Exponentially growing cells were irradiated with ¹²C or ²⁸Si ion beams generated by the Heavy Ion Medical Accelerator in Chiba (HIMAC), inoculated to culture dishes, and transformed colonies were identified when the cells were densely stacked and showed a crisscross pattern. Over the LET range examined, the frequency of transformation induced by the heavy ions increased sharply at very low doses no greater than 5 cGy. The relative biological effectiveness (RBE) of the heavy ions relative to 250 kVp X-rays showed an initial increase with LET, reaching a maximum value of about 7 at 100 keV/μm, and then decreased with the further increase in LET. Thus, we confirmed that high LET heavy ions are significantly more effective than X-rays for the induction of in vitro cell transformation.     

Direct radiation action of heavy ions on DNA as studied by ESR-spectroscopy.

The effects of heavy ions on the mechanisms of free radical formation in DNA-constituents as compared to low-LET irradiation are investigated by means of Electron Spin Resonance (ESR) spectroscopy. Dose-yield curves were measured at low (T < 100 K) and ambient temperatures in order to obtain the G-value, that is number of radicals formed per 100 eV absorbed energy. These G-values show a characteristic LET-dependence and are one to two orders of magnitude lower than for low-LET irradiation. Measurements on 2'Deoxycytidine at 300 K using combined heavy ion and X-ray irradiation methods suggested that this effect can be partially explained by a destruction of radicals during the irradiation.     

Mutation induction in yeast by very heavy ions.

Resistance to canavanine was studied in haploid yeast after exposure to heavy ions (argon to uranium) of energies between 1 and 10 MeV/u covering a LET-range up to about 10000 keV/micrometer. Mutations were found in all instances but the induction cross sections increased with ion energy. This is taken to mean that the contribution of penumbra electrons plays an important role. The probability to recover surviving mutants is highest if the cell is not directly hit by the particle. The experiments demonstrate that the geometrical dimensions of the target cell nucleus as well as its sensitivity in terms of survival have a critical influence on mutation induction with very heavy ions.     

On compositional variations of heavy ions during solar particle events

Intensity-time profiles of protons, alpha particles, and heavy ions (C, O, Fe) in the MeV/nucleon energy range have been analyzed for one solar particle event following the solar flare on September 23, 1978. The data have been obtained with the wide angle double dE/dx-E sensor of the Max-Planck-Institut/University of Maryland experiment onboard ISEE-3. We found time variations in the iron to helium ratio of up to 2 orders of magnitude and a significant variation of the O/He ratio during this event, whereas the C/O-ratio at the same energy/nucleon appears to be time independent. We investigated the influence of a rigidity dependent mean free path in interplanetary space and of rigidity dependent coronal propagation on heavy ion ratios during solar particle events. We found that both the magnitude and time scale of the ratio changes during the September 23 event cannot be explained by rigidity dependent interplanetary or coronal propagation alone. These ratio changes are probably caused by multiple injection at the sun.     

Genetic response of bacterial spores to very heavy ions.

Using spores of two Bacillus subtilis strains differing in repair capacity, we have studied repair and mutation induction in the spores after irradiation with very heavy ions up to uranium with specific particle energies up to 18.6 MeV/u. The results indicate that repair and mutation induction after heavy ion irradiation are closely related to each other and that both phenomena strongly depend on the atomic number and specific energy of the ions. The effects are discussed in comparison with results obtained after X-irradiation.