Different mutational function of low‐ and high‐linear energy transfer heavy‐ion irradiation demonstrated by whole‐genome resequencing of Arabidopsis mutants

Heavy‐ion irradiation is a powerful mutagen that possesses high linear energy transfer (LET). Several studies have indicated that the value of LET affects DNA lesion formation in several ways, including the efficiency and the density of double‐stranded break induction along the particle path. We assumed that the mutation type can be altered by selecting an appropriate LET value. Here, we quantitatively demonstrate differences in the mutation type induced by irradiation with two representative ions, Ar ions (LET: 290 keV μm^?1) and C ions (LET: 30.0 keV μm^?1), by whole‐genome resequencing of the Arabidopsis mutants produced by these irradiations. Ar ions caused chromosomal rearrangements or large deletions (≥100 bp) more frequently than C ions, with 10.2 and 2.3 per mutant genome under Ar‐ and C‐ion irradiation, respectively. Conversely, C ions induced more single‐base substitutions and small indels (<100 bp) than Ar ions, with 28.1 and 56.9 per mutant genome under Ar‐ and C‐ion irradiation, respectively. Moreover, the rearrangements induced by Ar‐ion irradiation were more complex than those induced by C‐ion irradiation, and tended to accompany single base substitutions or small indels located close by. In conjunction with the detection of causative genes through high‐throughput sequencing, selective irradiation by beams with different effects will be a powerful tool for forward genetics as well as studies on chromosomal rearrangements.     

DNA double-strand breaks in mammalian cells exposed to Á-rays and very heavy ions

The spatial distribution of DNA double-strand breaks (DSB) was assessed after treatment of mammalian cells (V79) with densely ionizing radiation. Cells were exposed to beams of heavy charged particles (calcium ions: 6.9 MeV/u, 2.1⋅10³ keV/μm; uranium ions: 9.0 MeV/u, 1.4⋅10⁴ keV/μm) at the linear accelerator UNILAC of GSI, Darmstadt. DNA was isolated in agarose plugs and subjected to pulsed-field gel electrophoresis under conditions that separated DNA fragments of size 50 kbp to 5 Mbp. The measured fragment distributions were compared to those obtained after γ-irradiation and were analyzed by means of a convolution and a deconvolution technique. In contrast to the finding for γ-radiation, the distributions produced by heavy ions do not correspond to the random breakage model. Their marked overdispersion and the observed excess of short fragments reflect spatial clustering of DSB that extends over large regions of the DNA, up to several mega base pairs (Mbp). At fluences of 0.75 and 1.5/μm², calcium ions produce nearly the same shape of fragment spectrum, merely with a difference in the amount of DNA entering the gel; this suggests that the DNA is fragmented by individual calcium ions. At a fluence of 0.8/μm² uranium ions produce a profile that is shifted to smaller fragment sizes in comparison to the profile obtained at a fluence of 0.4/μm²; this suggests cumulative action of two separate ions in the formation of fragments. These observations are not consistent with the expectation that the uranium ions, with their much larger LET, should be more likely to produce single particle action than the calcium ions. However, a consideration of the greater lateral extension of the tracks of the faster uranium ions explains the observed differences; it suggests that the DNA is closely coiled so that even DNA locations several Mbp apart are usually not separated by less than 0.1 or 0.2 μm. Received: 27 January 1998 / Accepted in revised form: 15 April 1998     

Characteristics of Myeloid Differentiation and Maturation Pathway Derived from Human Hematopoietic Stem Cells Exposed to Different Linear Energy Transfer Radiation Types

Exposure of hematopoietic stem/progenitor cells (HSPCs) to ionizing radiation causes a marked suppression of mature functional blood cell production in a linear energy transfer (LET)- and/or dose-dependent manner. However, little information about LET effects on the proliferation and differentiation of HSPCs has been reported. With the aim of characterizing the effects of different types of LET radiations on human myeloid hematopoiesis, in vitro hematopoiesis in Human CD34⁺ cells exposed to carbon-ion beams or X-rays was compared. Highly purified CD34⁺ cells exposed to each form of radiation were plated onto semi-solid culture for a myeloid progenitor assay. The surviving fractions of total myeloid progenitors, colony-forming cells (CFC), exposed to carbon-ion beams were significantly lower than of those exposed to X-rays, indicating that CFCs are more sensitive to carbon-ion beams (D ₀ = 0.65) than to X-rays (D ₀ = 1.07). Similar sensitivities were observed in granulocyte-macrophage and erythroid progenitors, respectively. However, the sensitivities of mixed-type progenitors to both radiation types were similar.In liquid culture for 14 days, no significant difference in total numbers of mononuclear cells was observed between non-irradiated control culture and cells exposed to 0.5 Gy X-rays, whereas 0.5 Gy carbon-ion beams suppressed cell proliferation to 4.9% of the control, a level similar to that for cells exposed to 1.5 Gy X-rays. Cell surface antigens associated with terminal maturation, such as CD13, CD14, and CD15, on harvest from the culture of X-ray-exposed cells were almost the same as those from the non-irradiated control culture. X-rays increased the CD235a⁺ erythroid-related fraction, whereas carbon-ion beams increased the CD34⁺CD38⁻ primitive cell fraction and the CD13⁺CD14^+/−CD15⁻ fraction. These results suggest that carbon-ion beams inflict severe damage on the clonal growth of myeloid HSPCs, although the intensity of cell surface antigen expression by mature myeloid cells derived from HSPCs exposed to each type of radiation was similar to that by controls.     

Methodology in the radiolysis of biochemical compounds with cyclotron beams at low flux densities.

A modification of the standard beam-optics of the LBL 88-Inch Cyclotron now makes it practicable to use cyclotron radiations in the detailed study of the effects of linear energy transfer (LET) in the radiation chemistry of organic compounds in the solid state. Dosages and dose-rates are comparable to those employed in conventional γ-ray studies. The modification involves passing the focused beam through a pair of “beam-sweeping” electromagnets so that a circular target area up to 10 cm in diameter can be uniformly irradiated with beams in the nanoamp range. Yield data are given for the radiolysis of the Fricke dosimeter with beams of H⁺, H⁺², Be⁺⁴, C⁺⁶ and Ne⁺¹⁰ at ～10 MeV per nucleon. Preliminary data on the effects of LET in the radiolysis of solid glycine are reported.     

Induction and prevention of carcinogenesis in rat skin exposed to space radiation

Quantitative cancer incidence data exist for various laboratory animal models, but little of this information is usable for estimating human risks, primarily because of uncertainties about possible mechanistic differences among species. Acceptance and utilization of animal data for human risk assessment will require a much better understanding of the comparative underlying mechanisms than now exists. A dual-lesion, radiation-track model in rat skin has proven to be consistent with tumor induction data with respect to acute radiation doses ranging from 0.5 up to 10 Gy and higher, and average LETs ranging from 0.34 to 150 keV μm⁻¹ according to the form neoplastic risk (D,L) = CLD + BD². A recent result with the ⁵⁶Fe ion beam showed dose-response consistency for malignant (carcinomas) and benign (fibromas) tumor induction with earlier results utilizing argon and neon ion beams. A discrepancy between the model and experiment was found indicating that proportionality of cancer yield with LET did not occur at 150 versus 125 keV μm⁻¹, i.e. tumor yield did not increase in spite of a 20% increase of LET, which suggests that a LET response maximum exists at or within this dose range. Concordance between the model and tumor induction data in rat skin implies that potential intervening complexities of carcinogenic progression fail to obscure the basic radiobiological assumptions underpinning the model. Gene expression microarray analysis shows that vitamin A inhibits the expression of about 80% of the inflammation-related genes induced by the radiation and prevents about 46% of the neoplasms associated with ⁵⁶Fe ion radiation without appearing to interfere with the underlying dose and LET response patterns. Further validation is needed, but the model has the potential to provide quantitative estimates of cancer risk as a function of dose and LET for almost any type of radiation exposure and even for combinations of different radiations provided only three empirical parameters can be established for each type of radiation and organ system.     

Effect of high linear energy transfer radiation on biological membranes

Cellular membranes are vital elements, and their integrity is extremely essential for the viability of the cells. We studied the effects of high linear energy transfer (LET) radiation on the membranes. Rabbit erythrocytes (1×10⁷ cells/ml) and microsomes (0.6 mg protein/ml) prepared from liver of rats were irradiated with ⁷Li ions of energy 6.42 MeV/u and ¹⁶O ions of energy 4.25 MeV/u having maximum LET values of 354 keV/μm and 1130 keV/μm, respectively. ⁷Li- and ¹⁶O-induced microsomal lipid peroxidation was found to increase with fluence. The ¹⁶O ions were more effective than ⁷Li ions, which could be due to the denser energy distribution in the track and the yield of free radicals. These findings suggested that the biological membranes could be peroxidized on exposure to high-LET radiation. Inhibition of the lipid peroxidation was observed in the presence of a membrane-active drug, chlorpromazine (CPZ), which could be due to scavenging of free radicals (mainly HO⋅ and ROO⋅), electron donation, and hydrogen transfer reactions. The ⁷Li and ¹⁶O ions also induced hemolysis in erythrocytes. The extent of hemolysis was found to be a function of time and fluence, and showed a characteristic sigmoidal pattern. The ¹⁶O ions were more effective in the lower fluence range than ⁷Li ions. These results were compared with lipid peroxidation and hemolysis induced by gamma-radiation. Received: 10 March 1998 / Accepted in revised form: 6 July 1998     

重离子射线照射对家蚕的生物影响

为解明重离子射线的生物影响,调查了氖、碳及氦(20Ne8+,LET=300keV/μm;12C5+,LET=116keV/μm和4He2+,LET=16.2keV/μm)等重离子射线照射家蚕(Bombyxmori)后的存活率及形态变化。重离子射线照射不同发育时期的幼虫后所引起的生物影响不同,幼虫的发育时期越早,照射后引起的生物影响越大;对同一时期的幼虫,随着剂量的增加,照射的生物影响加大;以化蛹率和羽化率为指标的放射线感受性在供试的3种射线间具有相似的变化倾向,只是射线的射程越长,照射的生物影响越大;对熟蚕卵巢存在部位的局部照射也显示相似的结果。同一射线的不同LET轨迹位置对家蚕的卵巢及真皮细胞的生物影响不同,用Mylar薄膜覆盖调节碳离子射线的射程,卵巢及真皮细胞越是接近射线高LET的Bragg峰,照射个体的鳞毛及卵的形成被强烈抑制。因此,重离子射线对家蚕的生物影响与细胞及植物种子等小个体不同,对于全体照射,重离子射线的射程长短所造成的生物影响比射线的LET大小所引起的生物影响要大;而对于局部照射,目的器官越是接近射线的高LET轨迹,照射的生物影响越大。     

Radiation-induced epigenetic alterations after low and high LET irradiations

Epigenetics, including DNA methylation and microRNA (miRNA) expression, could be the missing link in understanding radiation-induced genomic instability (RIGI). This study tests the hypothesis that irradiation induces epigenetic aberrations, which could eventually lead to RIGI, and that the epigenetic aberrations induced by low linear energy transfer (LET) irradiation are different than those induced by high LET irradiations. GM10115 cells were irradiated with low LET X-rays and high LET iron (Fe) ions and evaluated for DNA damage, cell survival and chromosomal instability. The cells were also evaluated for specific locus methylation of nuclear factor-kappa B (NFκB), tumor suppressor in lung cancer 1 (TSLC1) and cadherin 1 (CDH1) gene promoter regions, long interspersed nuclear element 1 (LINE-1) and Alu repeat element methylation, CpG and non-CpG global methylation and miRNA expression levels. Irradiated cells showed increased micronucleus induction and cell killing immediately following exposure, but were chromosomally stable at delayed times post-irradiation. At this same delayed time, alterations in repeat element and global DNA methylation and miRNA expression were observed. Analyses of DNA methylation predominantly showed hypomethylation, however hypermethylation was also observed. We demonstrate that miRNA expression levels can be altered after X-ray irradiation and that these miRNA are involved in chromatin remodeling and DNA methylation. A higher incidence of epigenetic changes was observed after exposure to X-rays than Fe ions even though Fe ions elicited more chromosomal damage and cell killing. This distinction is apparent at miRNA analyses at which only three miRNA involved in two major pathways were altered after high LET irradiations while six miRNA involved in five major pathways were altered after low LET irradiations. This study also shows that the irradiated cells acquire epigenetic changes suggesting that epigenetic aberrations may arise in the cell without initiating chromosomal instability.     

Heavy ion-induced DNA double-strand breaks with yeast as a model system

Cells of diploid yeast,Saccharomyces cerevisiae, were exposed to a variety of energetic heavy ions (provided by the UNILAC facility at the Gesellschaft für Schwerionenforschung, GSI),²⁴¹Am-particles and 80-keV x-rays after which they were assessed for DNA double-strand breaks (DSB) using either the neutral sedimentation or the pulsed-field gel electrophoresis (PFGE) technique. Both yielded comparable results. The DSB production cross-sections are compared with inactivation studies performed for the same cells under identical conditions. The measurements show that with lighter ions DSB induction cross-sections increase with linear energy transfer (LET), but the situation is less clear with the heavier ions. A close parallelism was found between DSB induction and cell inactivation in these yeast cells.Submitted paper presented at the International Symposium on Heavy Ion Research: Space, Radiation Protection and Therapy, Sophia-Antipolis, France, 21–24 March 1994     

LET dependence of the response of a PTW-60019 microDiamond detector in a 62 MeV proton beam

This study was initiated following conclusions from earlier experimental work, performed in a low-energy carbon ion beam, indicating a significant LET dependence of the response of a PTW-60019 microDiamond detector. The purpose of this paper is to present a comparison between the response of the same PTW-60019 microDiamond detector and an IBA Roos-type ionization chamber as a function of depth in a 62 MeV proton beam. Even though proton beams are considered as low linear energy transfer (LET) beams, the LET value increases slightly in the Bragg peak region. Contrary to the observations made in the carbon ion beam, in the 62 MeV proton beam good agreement is found between both detectors in both the plateau and the distal edge region. No significant LET dependent response of the PTW-60019 microDiamond detector is observed consistent with other findings for proton beams in the literature, despite this particular detector exhibiting a substantial LET dependence in a carbon ion beam.     

Cytologic and Autoradiographic Studies of the Micronucleus at Meiotic Prophase in Tetrahymena pyriformis

Micronuclear changes of variety 1 of Tetrahymena pyriformis during meiotic prophase have been observed by the light microscope. Morphologic changes in the micronucleus are divided into 6 stages. In stage I, chromatin begins to polarize; in stage II, the micronucleus becomes spindle shaped; and in stage III, one end of the micronucleus protrudes to form a “neck.” In stage IV, where the micronucleus elongates to maximal length, the whole micronucleus consists of 2 chromatin threads pairing longitudinally. One thread probably contains one genome. In stage V, the elongated thread becomes shorter and thicker. Finally, in stage VI, separate chromosomes appear and enter into metaphase. To discover the role of the elongation of the micronucleus, called crescent formation, autoradiographic analysis of RNA and DNA synthesis were undertaken using [³H]uridine and [³H]thymidine. Pulse label and chase experiments show that the crescent in stages II and III is actively synthesizing RNA. Though no remarkable DNA synthesis was observed during meiosis, a small amount of DNA synthesis occurred during the 1st and 2nd prezygotic divisions.     

Radioprotection by DMSO of mammalian cells exposed to X-rays and to heavy charged-particle beams

Populations of G1-phase Chinese hamster cells in stirred suspensions containing various concentrations of DMSO were irradiated with 250 kV X-rays or various heavy charged-particle beams. Chemical radioprotection of cell inactivation was observed for all LET values studied. When cell survival data were resolved into linear and quadratic components, the extent and concentration dependence of DMSO protection were found to be different for the two mechanisms. The chemical kinetics of radioprotection for single-events were similar for LET values up to those which gave the maximum RBE. DMSO protected to a lesser extent against energetic argon ions at an median LET of approximately 220 keV/micron. These data could indicate the contribution of indirect action by hydroxyl radicals and hydrogen atoms to cell inactivation by single-hit and double-hit mechanisms for various radiation qualities. The decrease in RBE observed at very high LET may result, in part, from reduced yields of water radicals at 10(-9)-10(-8) s resulting from radical recombination mechanisms within the charged particle tracks.     

LET and ion species dependence for cell killing in normal human skin fibroblasts

We studied the LET and ion species dependence of the RBE for cell killing to clarify the differences in the biological effects caused by the differences in the track structure that result from the different energy depositions for different ions. Normal human skin fibroblasts were irradiated with heavy-ion beams such as carbon, neon, silicon and iron ions that were generated by the Heavy Ion Medical Accelerator in Chiba (HIMAC) at the National Institute of Radiological Science (NIRS) in Japan. Cell killing was measured as reproductive cell death using a colony formation assay. The RBE-LET curves were different for carbon ions and for the other ions. The curve for carbon ions increased steeply up to around 98 keV/microm. The RBE of carbon ions at 98 keV/microm was 4.07. In contrast, the curves for neon, silicon and iron ions had maximum peaks around 180 keV/microm, and the RBEs at the peak position ranged from 3.03 to 3.39. When the RBEs were plotted as a function of Z*2/beta2 (where Z* is the effective charge and beta is the relative velocity of the ion) instead of LET, the discrepancies between the RBE-LET curves for the different ion beams were reduced, but branching of the RBE-Z*2/beta2 curves still remained. When the inactivation cross section was plotted as a function of either LET or Z*2/beta2, it increased with increasing LET. However, the inactivation cross section was always smaller than the geometrical cross section. These results suggest that the differences in the energy deposition track structures of the different ion sources have an effect on cell killing.     

Radioprotection of mouse colony forming units-spleen against heavy-charged particle damage by WR 2721

The effectiveness of S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR 2721) to protect against the heavy-charged particle beams with dose-averaged LET infinity's ranging from 26 to 260 keV/micron was studied using the marrow colony forming units-spleen as a model system. WR 2721 (400 mg/kg) was injected ip 30 min before whole-body irradiation in the plateau ionization region of the Bragg curve. Significant protection was observed at 26, 51, and 135 keV/micron LET values where the data were collected with 20Ne, 28Si, and 40Ar ions, respectively. The largest component of protection was the slope change, where at LET values of 26 and 51 keV/micron the DMFs (slope) were 2.1 and 2.3, respectively, which are very close to the gamma-ray value of 2.4 (gamma LET approximately equal to 0.2 keV/micron). Protection, however, decreased with increase in LET from 51 to 135 keV/micron to the DMF value of 1.2 and no significant protection was observed against 56Fe ions at 260 keV/micron. Significant increases in extrapolation number occurred with gamma rays and neon particles. The results are discussed in terms of charged particle track structure, radiation chemistry, and potential clinical applications.     

Micronucleus formation in lymphocytes of children from the vicinity of Chernobyl after <Superscript>131</Superscript>I therapy

After the Chernobyl accident a statistically significant increase in the number of children with thyroid tumours was observed. In this study 166 children with and 75 without thyroid tumours were analysed for micronucleus formation in peripheral blood lymphocytes using the cytochalasin B approach. The following factors did not significantly affect micronucleus formation: gender, age at the time of the first ¹³¹I treatment, tumour stage, tumour type, or metastases; a statistically significant increase in the number of micronuclei, however, was observed for the residents of Gomel compared to other locations, such as Brest, Grodno, and Minsk. The children with tumours received ¹³¹I treatment after surgical resection of the tumours. This gave us the opportunity to systematically follow the effect of ¹³¹I on micronucleus formation. A marked increase was observed 5 days after the ¹³¹I treatment followed by a decrease within a 4–7 months interval up to the next application, but the pre-treatment levels were not achieved. Up to 10 therapy cycles were followed each including an analysis of micronucleus formation before and 5 days after ¹³¹I application. The response of the children was characterised by clear individual differences and the increase/decrease pattern of micronucleus frequencies induced by iodine-131 was correlated with a decrease/increase pattern in the number of lymphocytes.     

Inactivation of aerobic and hypoxic cells from three different cell lines by accelerated (3)He-, (12)C- and (20)Ne-ion beams

The LET-RBE spectra for cell killing for cultured mammalian cells exposed to accelerated heavy ions were investigated to design a spread-out Bragg peak beam for cancer therapy at HIMAC, National Institute of Radiological Sciences, Chiba, prior to clinical trials. Cells that originated from a human salivary gland tumor (HSG cells) as well as V79 and T1 cells were exposed to (3)He-, (12)C- and (20)Ne-ion beams with an LET ranging from approximately 20-600 keV/micrometer under both aerobic and hypoxic conditions. Cell survival curves were fitted by equations from the linear-quadratic model and the target model to obtain survival parameters. RBE, OER, alpha and D(0) were analyzed as a function of LET. The RBE increased with LET, reaching a maximum at around 200 keV/micrometer, then decreased with a further increase in LET. Clear splits of the LET-RBE or -OER spectra were found among ion species and/or cell lines. At a given LET, the RBE value for (3)He ions was higher than that for the other ions. The position of the maximum RBE shifts to higher LET values for heavier ions. The OER value was 3 for X rays but started to decrease at an LET of around 50 keV/micrometer, passed below 2 at around 100 keV/micrometer, and then reached a minimum above 300 keV/micrometer, but the values remained greater than 1. The OER was significantly lower for (3)He ions than the others.     

Characterization of hydroxyl radical-induced damage after sparsely and densely ionizing irradiation

The extent of hydroxyl radical mediated cell inactivation was measured for a variety of particle beams ranging from 8.5 Me V/u neon ions to 570 Me V/u argon ions. In general, the fraction of the total radiosensitivity caused by OH decreases from close to 60 per cent at low ionization density or low linear energy transfer (low LET) to close to 25 per cent at high LET for aerobically irradiated mammalian cells. The extent of OH induced cell lethality can be explained in terms of LET infinity only for low energy or low atomic number particles where fragmentations and complicated track structures do not contaminate the characteristic particle LET. For example, at a calculated LET infinity of 100 ke V/micron, the OH mediated fraction of the total radiation damage is about 25 per cent for low energy carbon but close to 40 per cent for high energy carbon ions. For low energy charged nuclei of approximately the same energy, as the 5.4-13.4 MeV/u He, Li, C and Ne ions in this report, there is a predictable diminution of the OH mediated effect with increasing LET infinity; however, the biological effect cannot be predicted accurately from calculated LET infinity values for high energy particle irradiation, nor indeed from a variety of low energy charged particles of quite different energies (incident velocities). This illustrates the unsuitability of using LET as a unifying parameter, except under specific circumstances. As more is learned about the energy deposition for energized charged particles in terms of track structure (core and penumbra), it may be possible to characterize the radiobiological data with a better physical parameter than LET infinity.     

Biodosimetry estimate for high-LET irradiation

The purpose of this paper is to prepare for an easy and reliable biodosimeter protocol for radiation accidents involving high-linear energy transfer (LET) exposure. Human peripheral blood lymphocytes were irradiated using carbon ions (LET: 34.6 keV μm⁻¹), and the chromosome aberrations induced were analyzed using both a conventional colcemid block method and a calyculin A induced premature chromosome condensation (PCC) method. At a lower dose range (0–4 Gy), the measured dicentric (dics) and centric ring chromosomes (cRings) provided reasonable dose information. At higher doses (8 Gy), however, the frequency of dics and cRings was not suitable for dose estimation. Instead, we found that the number of Giemsa-stained drug-induced G2 prematurely condensed chromosomes (G2-PCC) can be used for dose estimation, since the total chromosome number (including fragments) was linearly correlated with radiation dose (r = 0.99). The ratio of the longest and the shortest chromosome length of the drug-induced G2-PCCs increased with radiation dose in a linear-quadratic manner (r = 0.96), which indicates that this ratio can also be used to estimate radiation doses. Obviously, it is easier to establish the dose response curve using the PCC technique than using the conventional metaphase chromosome method. It is assumed that combining the ratio of the longest and the shortest chromosome length with analysis of the total chromosome number might be a valuable tool for rapid and precise dose estimation for victims of radiation accidents.     

Relative biological effectiveness of high-energy iron ions for micronucleus formation at low doses

Dose-response curves for micronucleus (MN) formation were measured in Chinese hamster V79 and xrs6 (Ku80(-)) cells and in human mammary epithelial MCF10A cells in the dose range of 0.05-1 Gy. The Chinese hamster cells were exposed to 1 GeV/nucleon iron ions, 600 MeV/nucleon iron ions, and 300 MeV/nucleon iron ions (LETs of 151, 176 and 235 keV/microm, respectively) as well as with 320 kVp X rays as reference. Second-order polynomials were fitted to the induction curves, and the initial slopes (the alpha values) were used to calculate RBE. For the repair-proficient V79 cells, the RBE at these low doses increased with LET. The values obtained were 3.1 +/- 0.8 (LET = 151 keV/microm), 4.3 +/- 0.5 (LET = 176 keV/microm), and 5.7 +/- 0.6 (LET = 235 keV/microm), while the RBE was close to 1 for the repair-deficient xrs6 cells regardless of LET. For the MCF10A cells, the RBE was determined for 1 GeV/nucleon iron ions and was found to be 5.5 +/- 0.9, slightly higher than for V79 cells. To test the effect of shielding, the 1 GeV/nucleon iron-ion beam was intercepted by various thicknesses of high-density polyethylene plastic absorbers, which resulted in energy loss and fragmentation. It was found that the MN yield for V79 cells placed behind the absorbers decreased in proportion to the decrease in dose both before and after the iron-ion Bragg peak, indicating that RBE did not change significantly due to shielding except in the Bragg peak region. At the Bragg peak itself with an entrance dose of 0.5 Gy, where the LET is very high from stopping low-energy iron ions, the effectiveness for MN formation per unit dose was decreased compared to non-Bragg peak areas.     

Heavy ion effects on yeast: inhibition of ribosomal RNA synthesis

Diploid wild-type yeast cells were exposed to beams of heavy ions covering a wide range of linear energy transfer (LET) (43-13,700 keV/microns). Synthesis of ribosomal RNA (rRNA) was assessed as a functional measure of damage produced by particle radiation. An exponential decrease of relative rRNA synthesis with particle fluence was demonstrated in all cases. The inactivation cross sections derived were found to increase with LET over the entire range of LET studied. The corresponding values for relative biological effectiveness were slightly less than unity. Maximum cross sections measured were close to 1 micron 2, implying that some larger structure within the yeast nucleus (e.g., the nucleolus) might represent the target for an impairment of synthetic activity by very heavy ions rather than the genes coding for rRNA. Where tested, an oxygen effect for rRNA synthesis could not be demonstrated.