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Multiple cell hits by particle tracks in solid tissues.

Relative Biological Effectiveness (RBE) and Quality Factor (Q) at extreme values of Linear Energy Transfer (LET) have been determined on the basis of experiments with single-cell systems and specific tissue responses. In typical single cell systems, each heavy particle (Ar or Fe) passes through a single cell or no cell. In tissue end-point experiments each heavy particle passes through several cells, and the LET can exceed 200 keV/micrometer in every cell. In most laboratory animal tissue systems, however, only a small portion of the hit cells are capable of expressing the end-point of interest to the investigator, such as cell killing, mutation or carcinogenesis. The following question must therefore be addressed: Do RBE's and Q factors derived from single-cell experiments properly account for the increased probability of multiple-cell damage by HZE tracks? A model is offered in which measured radiation effects and known tissue properties are combined to estimate the value of a multiplier of damage effectiveness on the basis of number of cells at risk, p3n, per track containing a hit cell, where n is the number of cells per track, based on tissue and organ geometry, and P3 is the probability that a cell in the track is capable of expressing the experimental end-point.

Animals↗

The dependence of p53 on the radiation enhancement of thermosensitivity at different let.

PURPOSE: The aim of this study is to investigate the dependence of p53-gene status on the radiation enhancement of thermosensitivity at different levels of linear energy transfer (LET). METHODS AND MATERIALS: We used two kinds of human glioblastoma transfectants of A-172 cells bearing the wild-type p53 gene, A-172/neo cells with control vector containing the neo gene and A-172/mp53 cells with both the dominant negative mutated p53 gene and neo gene. We exposed these cells to X-rays and accelerated carbon-ion (C-) beams (13-200 KeV/microm) followed by heating at 44 degrees C. Cellular sensitivities were determined using clonogenic assay. RESULTS: The radiation enhancement of thermosensitivity was LET-dependent for the A-172/neo cells, but this was not clearly demonstrated in the A-172/mp53 cells. The supraadditive radiation enhancement of thermosensitivity was observed in A-172/neo cells at the LET range of 13 to 70 KeV/microm, though only an additive effect was observed at higher LET. In A-172/mp53 cells, only an additive effect was observed through all the LET examined. CONCLUSION: These results indicate that the radiation enhancement of thermosensitivity is p53- and LET-dependent. Our results suggest that the combined use of high-LET radiation and hyperthermia brings useful application for cancer therapeutic purposes.

Cell Survival↗

Chromatin loops are responsible for higher counts of small DNA fragments induced by high-LET radiation, while chromosomal domains do not affect the fragment sizes.

PURPOSE: To apply a polymer model of DNA damage induced by high-LET (linear energy transfer) radiation and determine the influence of chromosomal domains and loops on fragment length distribution. MATERIALS AND METHODS: The yields of DSB (double-strand breaks) induced by high-LET radiation were calculated using a track structure model along with a polymer model of DNA packed in the cell nucleus. The cell nucleus was constructed to include the chromosomal domains and chromatin loops. The latter were generated by the random walk method. RESULTS AND CONCLUSIONS: We present data for DSB yields per track per cell, DNA fragment sizes, the radial distribution of DSB with respect to the track center, and the distribution of 0, 1, 2, and more DSB from a single particle. Calculations were carried out for a range of particles including He (40 keV/microm), N (225 keV/microm), and Fe ions (150 keV/mum). Situations relevant to PFGE (pulsed-field gel electrophoresis) and microbeam experiments with direct irradiation of the cell nucleus were simulated to demonstrate the applicability of the model. Data show that chromosomal domains do not have a significant influence on fragment-size distribution, while the presence of DNA loops increases the frequencies of smaller fragments by nearly 30% for fragment sizes in the range from 2 kbp (bp = base pair) to 20 kbp.

Chromatin↗

Ex vivo determination of the effect of whole-body exposure to fast neutrons on murine spleen cell viability and apoptosis.

The effects of high-linear energy transfer (LET) radiations on lymphoid tissues and lymphocytes are not well understood. As a first approach to delineate these effects, the present work was conducted to assess the effects of high-LET radiations on murine spleen cells ex vivo and in vitro. BALB/c mice were irradiated whole-body with 65 MeV neutrons or 15 MV X rays at doses ranging from 0.2 to 3 Gy. Spleens were removed 1 day postirradiation and weighed, and single cell suspensions were prepared and cultured for several days. Apoptosis occurring in vitro was determined at different times by flow cytometry analysis of cells labeled with propidium iodide. It was found that irradiation with fast neutrons reduced spleen weight and cellularity to a greater extent than photons. Considering the spleen cellularity as end point, the relative biological effectiveness (RBE) of fast neutrons was 2. However, for both modes of irradiation, apoptosis of recovered spleen cells in vitro increased as a function of dose and the duration of culture. The level of apoptosis occurring at various times postirradiation was found to be identical for high- and low-LET radiations. Taken together, these results suggest that external as well as cellular factors might differentially modulate the sensitivity of lymphocytes to fast neutrons and photons.

Animals↗

Effect of retinol on radiation- and estrogen-induced neoplastic transformation of human breast epithelial cells.

Clinical, epidemiological and experimental findings have provided evidence supporting a role of free radicals in the etiology of cancer. Free radical production is enhanced in many disease states, by carcinogen exposure, and under conditions of stress contributing widely to cancer development in humans. We have established an experimental breast cancer model to examine the effects of all-trans-retinol (retinol/vitamin A) on the production of free radicals in human breast epithelial cells induced by high linear energy transfer (LET)-radiation in the presence of 17beta estradiol. The following cell lines were used in these studies: the MCF-10F cell line, a spontaneously immortalized human breast epithelial cell line. Alpha 5 derived from MCF-10F cells irradiated with two separated doses of 60 cGy alpha particles in the presence of estrogens (60E/60E). Tumor 2, from a tumor formed in nude mice after injection with the cell line alpha 5. Tumor 3, from secondary tumor formed from injecting tumor 2 cells into nude mice. Each of the cell types examined had significantly elevated H(2)O(2) production levels compared to MCF-10F control cells (p<0.001). Retinol (1 microl/ml) significantly (p<0.05) decreased H(2)O(2) production in all cell types examined. Retinol significantly decreased (p<0.05) invasive capabilities of cells across matrigel coated invasion chambers and significantly reduced (p<0.05) PCNA, Fra-1, mutant p53 and increased Rb protein expression levels in comparison to non-retinol-treated ones when assayed using immunofluorescent staining coupled with confocal microscopy. The reduced H(2)O(2) production, decrease in cell invasive capabilities and alterations in protein expression levels suggest that retinol can be used as a chemopreventive agent in human breast cancer.

Animals↗

Unique biological aspects of radiation hazards--an overview.

Low orbit, geostationary, and deep-space flights differ from one another with respect to particle radiation environment, participating population size, mission duration, and biological risks other than radiation. It is proposed that all of these factors be considered in the setting of safety standards and, in particular, that the rem-dose concept is applicable only to radiations having low and intermediate linear energy transfer (electrons, protons, and helium ions), whereas the incidence of microlesions is a more meaningful indicator of the hazard due to higher-Z, high energy (HZE) particles. A microlesion is the biological injury inflicted in a specific tissue by a single HZE particle, and it is still in need of quantitative biological definition for specific mammalian tissues. If for example, a microlesion is taken as due to a HZE particle track 10 cell diameters long with LET > 200 KeV/micrometer in its core and > 25 rad dose in its penumbra at a distance of 10 micrometers, then the microlesion dose rate in geostationary orbit, for example, is about 9,000 microlesions per cm3 of tissue per month.

Aerospace Medicine↗

The U.S. National Research Council's views of the radiation hazards in space.

The author was the Chairman of a Task Group on the Biological Effects of Space Radiation formed as a result of discussions between NASA and the U.S. National Research Council's Committee on Space Biology and Medicine - a committee under the U.S. National Research Council's Space Studies Board. The Task Group was asked to review current knowledge on the effects of long-term exposure to radiation in space and to consider NASA radiation shielding requirements for orbital and interplanetary spacecraft. The group was charged with assessing the adequacy of NASA planning for the protection of humans from radiation in space and with making recommendations regarding needed research and/or new shielding requirements. This manuscript is a summary of the findings and recommendations of the Task Group. Beyond the protection of the Earth's atmosphere and its magnetosphere, the exposure to ionizing radiations far exceeds that on Earth. Of all the risks astronauts may face, this one is probably the most straightforward to control - by providing adequate shielding. However, because shielding adds weight, cost and complexity to space vehicles, it is important for designers to have a good quantitative understanding of the true risk and its degree of uncertainty so as not to under- or overshield spacecrafts. The extrapolations from our knowledge of ionizing radiation effects of low linear energy transfer (LET) to the risks from high-atomic-number high-energy energetic (HZE) cosmic rays are very uncertain because the necessary experiments on the effects of such particles have not been carried out and the extrapolation from low-LET to very high-LET has great uncertainties. These uncertainties were enumerated by the Task Group, and the types of experiments needed to minimize the uncertainties were described. The report found that, because of the small amounts of available time for biological research at HZE accelerators, it would take more than a decade of effort to obtain the answers to a narrow set of key questions that would facilitate reduction in risks and identification of the types of shielding needed.

Animals↗

Space radiation enhancement linked to geomagnetic disturbances.

Space radiation dosimetry measurements have been made on board the Space Shuttle. A newly developed active detector called "Real-time Radiation Monitoring Device (RRMD)" was used (Doke et al., 1995; Hayashi et al., 1995). The RRMD results indicate that low Linear Energy Transfer (LET) particles steadily penetrate around the South Atlantic Anomaly (SAA) without clear enhancement of dose equivalent and some daily periodic enhancements of dose equivalent due to high LET particles are seen at the lower geomagnetic cutoff regions (Doke et al., 1996). We also have been analyzing the space weather during the experiment, and found that the anomalous high-energy particle enhancement was linked to geomagnetic disturbance due to the high speed solar wind from a coronal hole. Additional analysis and other experiments are necessary for clarification of these phenomena. If a penetration of high-energy particles into the low altitude occurs by common geomagnetic disturbances, the prediction of geomagnetic activity becomes more important in the next Space Station's era.

Cosmic Radiation↗

Inverse radiation dose-rate effects on somatic and germ-line mutations and DNA damage rates.

The mutagenic effect of low linear energy transfer ionizing radiation is reduced for a given dose as the dose rate (DR) is reduced to a low level, a phenomenon known as the direct DR effect. Our reanalysis of published data shows that for both somatic and germ-line mutations there is an opposite, inverse DR effect, with reduction from low to very low DR, the overall dependence of induced mutations being parabolically related to DR, with a minimum in the range of 0.1 to 1.0 cGy/min (rule 1). This general pattern can be attributed to an optimal induction of error-free DNA repair in a DR region of minimal mutability (MMDR region). The diminished activation of repair at very low DRs may reflect a low ratio of induced ("signal") to spontaneous background DNA damage ("noise"). Because two common DNA lesions, 8-oxoguanine and thymine glycol, were already known to activate repair in irradiated mammalian cells, we estimated how their rates of production are altered upon radiation exposure in the MMDR region. For these and other abundant lesions (abasic sites and single-strand breaks), the DNA damage rate increment in the MMDR region is in the range of 10% to 100% (rule 2). These estimates suggest a genetically programmed optimatization of response to radiation in the MMDR region.

Animals↗

Radiation dosimetry measurements with real time radiation monitoring device (RRMD)-II in Space Shuttle STS-79.

The real-time measurement of radiation environment was made with an improved real-time radiation monitoring device (RRMD)-II onboard Space Shuttle STS-79 (S/MM#4: 4th Shuttle MIR Mission, at an inclination angle of 51.6 degrees and an altitude of 250-400km) for 199 h during 17-25 September, 1996. The observation of the detector covered the linear energy transfer (LET) range of 3.5-6000 keV/micrometer. The Shuttle orbital profile in this mission was equivalent to that of the currently planned Space Station, and provided an opportunity to investigate variations in count rate and dose equivalent rate depending on altitude, longitude, and latitude in detail. Particle count rate and dose equivalent rate were mapped geographically during the mission. Based on the map of count rate, an analysis was made by dividing whole region into three regions: South Atlantic Anomaly (SAA) region, high latitude region and other regions. The averaged absorbed dose rate during the mission was 39.3 microGy/day for a LET range of 3.5-6000 keV/micrometer. The corresponding average dose equivalent rates during the mission are estimated to be 293 microSv/day with quality factors from International Commission on Radiological Protection (ICRP)-Pub. 60 and 270 microSv/day with quality factors from ICRP-Pub. 26. The effective quality factors for ICRP-Pub. 60 and 26 are 7.45 and 6.88, respectively. From the present data for particles of LET > 3.5keV/micrometer, we conclude that the average dose equivalent rate is dominated by the contribution of galactic cosmic ray (GCR) particles. The dose-detector depth dependence was also investigated.

Aerospace Medicine↗

Space radiation enhancement linked to geomagnetic disturbances.

Space radiation dosimetry measurements have been made on board the Space Shuttle. A newly developed active detector called "Real-time Radiation Monitoring Device (RRMD)" was used (Doke et al., 1995; Hayashi et al., 1995). The RRMD results indicate that low Linear Energy Transfer (LET) particles steadily penetrate around the South Atlantic Anomaly (SAA) without clear enhancement of dose equivalent and some daily periodic enhancements of dose equivalent due to high LET particles are seen at the lower geomagnetic cutoff regions (Doke et al., 1996). We also have been analyzing the space weather during the experiment, and found that the anomalous high-energy particle enhancement was linked to geomagnetic disturbance due to the high speed solar wind from a coronal hole. Additional analysis and other experiments are necessary for clarification of these phenomena. If a penetration of high-energy particles into the low altitude occurs by common geomagnetic disturbances, the prediction of geomagnetic activity becomes more important in the next Space Station's era.

Atlantic Ocean↗

The F value for chromosome aberrations in atomic bomb survivors does not provide evidence for a primary contribution of neutrons to the dose in Hiroshima.

Brenner and Sachs (Radiat. Res. 140, 134-142, 1994) proposed that the ratio of interchromosomal to intrachromosomal exchanges, termed the F value, can be a cytogenetic fingerprint of exposure to radiations of different linear energy transfer (LET). Using published data, they suggested that F values are over 10 for low-LET radiations and approximately 6 for high-LET radiations. Subsequently, as F values for atomic bomb survivors were reported to be around 6, Brenner suggested that the biological effects of atomic bomb radiation in Hiroshima are due primarily to neutrons. However, the F values used for the survivors were means from individuals exposed to various doses. As the F-value hypothesis predicts a radiation fingerprint at low doses, we analyzed our own data for the survivors in relation to dose. G-banding data for the survivors showed F values varying from 5 to 8 at DS86 doses of 0.2 to 5 Gy in Hiroshima and around 6 in Nagasaki with no evidence of a difference between the two cities. The results are consistent with our in vitro data that the F values are invariably around 6 for X and gamma rays at doses of 0.5 to 2 Gy as well as two types of fission-spectrum neutrons at doses of about 0.2 to 1 Gy. Thus, apart from a possible effect at even lower doses, current data do not provide evidence to support the proposition that the biological effects of atomic bomb radiation in Hiroshima are caused mainly by neutrons.

Chromosome Aberrations↗

Power saturation of ESR signal in ammonium tartrate exposed to 60Co gamma-ray photons, electrons and protons.

In this paper we present an investigation of the electron spin resonance (ESR) line shape of ammonium tartrate (AT) dosimeters exposed to radiation with different linear energy transfer (LET). We exposed our dosimeters to gamma-ray photons ((60)Co), 7 MeV and 14 MeV initial energy electrons, and 19.3 MeV initial energy protons. The differences in the power saturation behavior of ESR spectra of AT irradiated with photons, electrons and protons could be correlated to the effective LET of the radiation beams. We analyzed the behavior of peak-to-peak amplitude as a function of microwave power, and we developed a fitting procedure that permits us to obtain the dependence of the homogeneity parameter of the line shape on the LET of the radiation using the Castner saturation theory. This simple procedure allows us to distinguish the LET of the radiation beam.

Algorithms↗

High-LET radiolysis of liquid water with 1H+, 4He2+, 12C6+, and 20Ne9+ ions: effects of multiple ionization.

Monte Carlo simulations are used to investigate the effects of multiple ionization of water molecules on the yields of formation of free radical and molecular species, including molecular oxygen, in the radiolysis of pure, deaerated liquid water by using different types of radiation (1H+, 4He2+, 12C6+, and 20Ne9+ ions) up to approximately 900 keV/microm, at neutral pH and 25 degrees C. Taking into account the double, triple, and quadruple ionizations of water, the primary (or "escape") yields (at 10(-6) s) of the various radiolytic species (G(e(aq)-), G(H*), G(H2), G(*OH), G(HO2*/O2*-), and G(H2O2) are calculated as a function of the linear energy transfer (LET) of the radiation. Our results quantitatively reproduce the large increase observed in G(HO2*/O2*-) at high LET. Under the conditions of this study, the mechanisms of triple and quadruple ionizations contribute only weakly to the production of HO2*/O2*-. With the exception of protons, our calculations also simultaneously predict a maximum in G(H2O2) corresponding to the LET of approximately 4.5-MeV helium ions (approximately 100 keV/microm) and approximately 110-MeV carbon ions (approximately 180 keV/microm). This maximum occurs where G(HO2*/O2*-) begins to rise sharply, suggesting, in agreement with previous experimental data, that the yields of HO2*/O2*- and H2O2 are closely linked. Moreover, our results show a steep increase in the initial and primary yields of molecular oxygen with increasing LET, giving support to the "oxygen in heavy-ion tracks" hypothesis. By contrast, it is found that, in the whole LET range considered, the incorporation of multiple ionization in the simulations has only little effect on the variation of our computed G(e(aq)-), G(H*), G(H2), and G(*OH) values as a function of LET. As expected, G(e(aq)-) and G(*OH) decrease continuously with increasing LET. G(H*) at first increases and then decreases at high LET. Finally, G(H2) monotonically rises with increasing LET. Our calculated yield values compare generally very well with experiment.

Carbon↗

The relative biological effectiveness of accelerated carbon ions with different LET for inducing mitotic crossing over and intragenic reversion of the white-ivory allele in Drosophila larvae.

PURPOSE: To evaluate the relative biological effectiveness (RBE) of accelerated carbon ions generated with a synchrotron for inducing mutations as a function of linear energy transfer (LET), using the loss of heterozygosity for wing-hair mutations and the reversion of the mutant white-ivory eye-colour in Drosophila melanogaster. MATERIALS AND METHODS: The measurements were made using a combined mutation assay system so that induced mutant wing-hair clones as well as revertant eye-colour clones can be detected simultaneously in the same fly. Larvae were irradiated at the age of 72+/-6 h post-oviposition with X-rays or carbon ions with LET values of 13, 60 and 95 keV/microm. RESULTS: The RBE of carbon ions for producing wing-hair mosaic spots increased with increasing LET values. The RBE for the induction of eye-colour mutants did not change with LET. The estimated RBE values were found to be in the range 2 to 6.5 for the wing-hair and nearly unity for the eye-colour mosaic spot mutations. CONCLUSIONS: RBE-LET relationships were obtained for the induction of wing-hair and eye-colour mosaic spots. These relationships suggest that more complex types of DNA damage, such as nonrejoinable strand breaks that increase with LET, may be responsible for inducing the wing-hair mutation, while more simple forms of molecular damage induce reversion in the white-ivory allele.

Animals↗

High-LET ion radiolysis of water: visualization of the formation and evolution of ion tracks and relevance to the radiation-induced bystander effect.

Ionizing radiation-induced bystander effects, commonly observed in cell populations exposed to high-linear energy transfer (LET) radiations, are initiated by damage to a cellular molecule which then gives rise to a toxic signal exported to neighboring cells not directly hit by radiation. A major goal in studies of this phenomenon is the identification of this initial radiation-induced lesion. Liquid water being the main constituent of biological matter, reactive species produced by water radiolysis in the cellular environment are likely to be major contributors to the induction of this lesion. In this context, the radiation track structure is of crucial importance in specifying the precise location and identity of all the radiolytic species and their subsequent signaling or damaging effects. We report here Monte Carlo track structure simulations of the radiolysis of liquid water by four different impacting ions 1H+, 4He2+, 12C6+ and 20Ne10+, with the same LET ( approximately 70 keV/ microm). The initial radial distribution profiles of the various water decomposition products (eaq(-), *OH, H*, H2 and H2O2) for the different ions considered are presented and discussed briefly in the context of track structure theory. As an example, the formation and temporal evolution of simulated 24 MeV 4He2+ ion tracks (LET approximately 26 keV/microm) are reported for each radiolytic species from 1 ps to 10 micros. The calculations reveal that the ion track structure is completely lost by approximately 1 micros.

Bystander Effect↗

Radiosensitivity of V79 cells after alpha particle radiation at low doses.

Low dose hyper-radiosensitivity (HRS) of V79 cells was demonstrated after irradiation with gamma rays and 4He2+ ions of various linear energy transfer (LET) values (58.9, 79.3 and 101.7 keV.micron-1). In parallel, the cytogenetic analysis showed an LET dependence of aberrations at a dose of 1 Gy, while the observed chromatid fragments appeared to vary with the number of 4He2+ ions traversing the cell nucleus. The results of both studies are correlated so as to achieve a better understanding of the so-called induced radioresistance. The cell mechanism of radioresistance appears to be induced after a certain amount of energy is deposited in the cell nucleus. This amount depends both on the radiation quality as well as the number of particles traversing the cell, inducing chromosome alterations and chromatid damage.

Alpha Particles↗

RBE for carcinogenesis following exposure to high LET radiation.

Stochastic radiation effects following exposure to heavy ions and other high linear energy transfer (LET) radiation in space are a matter of concern when the long-term consequences of space flights are considered. This paper is an overview of the relevant literature, emphasizing uncertainties entailed from estimates of relative biological effectiveness (RBE) for different experiment endpoints, making the choice of a single weighting factor for the prediction of cancer risk in man extremely difficult. Life-span-shortening studies in mice exposed to heavy ions and ongoing large-scale experiments in monkeys exposed to protons suggest that RBEs for all cancers are lower than 5. This does not exclude a much higher RBE for rare tumors such as brain tumors in monkeys or promoted Harderian gland tumours in mice at LET >80 keV/mu m. Skin cancer studies in rats exposed to neon or argon resulted in similar RBE. Exposure to fission neutrons led to high RBE in all species, not excluding values much higher than 20 for specific cancers such as lung tumors in mice and all cancers in rats. The estimate of maximal RBE is, however, extremely dependent on the hypothesis made on the shape of the dose-response curves in the lower range of doses. These results suggest that neutrons may be the most hazardous component of high-LET radiation. There is only limited evidence from cancer experiments that LET >150 keV/mu m results in highly decreased efficiency, but this has been found for bone cancer induction following exposure to fission fragments.

Animals↗