Effects of 645 MeV and 9.2 GeV protons on Artemia eggs.

Developmental capacities of Artemia eggs have been studied after exposure to 645 MeV or 9.2 GeV protons. Effects of proton irradiation were studied in comparison with 60Co gamma ray irradiation, endpoints being emergence, hatching and 4-5 day old live nauplii percentages. Effectiveness of 645 MeV protons is greater than that of 9.2 GeV protons. R.b.e. values calculated for nauplius survival is 2.3 for 645 MeV protons and 1.5 for 9.2 GeV protons. These results can be taken into account in radiation hazard estimation during space flights.     

Response of human HTB140 melanoma cells to conventional radiation and hadrons

Conventional radiotherapy with X- and gamma-rays is one of the common and effective treatments of cancer. High energy hadrons, i.e., charged particles like protons and (12)C ions, due to their specific physics and radiobiological advantages are increasingly used. In this study, effectiveness of different radiation types is evaluated on the radio-resistant human HTB140 melanoma cells. The cells were irradiated with gamma-rays, the 62 MeV protons at the Bragg peak and in the middle of the spread-out Bragg peak (SOBP), as well as with the 62 MeV/u (12)C ions. The doses ranged from 2 to 24 Gy. Cell survival and proliferation were assessed 7 days after irradiation, whereas apoptosis was evaluated after 48 h. The acquired results confirmed the high radio-resistance of cells, showing better effectiveness of protons than gamma-rays. The best efficiency was obtained with (12)C ions due to higher linear energy transfer. All analyzed radiation qualities reduced cell proliferation. The highest proliferation was detected for (12)C ions because of their large killing capacity followed by small induction of reparable lesions. This enabled unharmed cells to preserve proliferative activity. Irradiations with protons and (12)C ions revealed similar moderate pro-apoptotic ability that is in agreement with the level of cellular radio-resistance.     

Modifications in repair and expression of potentially lethal damage (alpha-PLD) as measured by delayed plating or treatment with beta-araA in plateau-phase Ehrlich ascites tumor cells after exposure to charged particles of various specific energies

The ability of Ehrlich ascites tumor cells (EAT cells) to repair potentially lethal damage (alpha-PLD) as demonstrated by either an increase in survival after delayed plating or a decrease in survival after treatment with beta-arabinofuranosyladenine (beta-araA) was investigated after exposure to protons, deuterons, 3He, 4He, and heavy ions of various specific energies. A significant amount of repair or fixation was observed after delayed plating or treatment with beta-araA, respectively, in cells that were exposed to protons of 6-21 MeV energy, reflecting mainly variations in the survival curve shoulder width. Four-hour treatment with 80 microM/liter beta-araA resulted in an exponential survival curve for all proton energies tested. A decrease in particle energy increased killing and caused a reduction in Dq without a significant change in D0. The survival curve obtained after exposure of cells to 3.4 MeV protons had only a small shoulder and was only slightly modified by either delayed plating or treatment with beta-araA, suggesting a decrease in the induction rate of alpha-PLD. Similar results were also obtained after exposure to deuterons and 4He ions. The results are interpreted as indicating the importance of the specific particle energy and the delta-electron spectrum in the induction of alpha-PLD. When the results of delayed plating of cells exposed to protons, deuterons, or helium ions were pooled, an exponential relationship between Dq and penumbra radius was indicated. After exposure to 40Ar ions of 18 MeV specific energy, a shouldered survival curve was obtained, and beta-araA significantly enhanced killing by modifying Dq as well as D0, a result that also suggests induction of repairable damage by the delta particles produced and interaction of lesions induced within the core of the ion path with penumbra lesions. Based on these results a model is proposed assuming that alpha-PLD results from interaction, during the course of repair, of pairs of DNA lesions induced within a distance di. The model assumes the existence of a critical separation distance dic, with the property that pairs of lesions induced with separation distance shorter than dic (expressed as number of base pairs) will always be expressed as lethal, and the existence of a maximum separation distance dim, with the property that pairs of lesions induced with separation distance larger than dim will not interact.(ABSTRACT TRUNCATED AT 400 WORDS)     

Comparisons of LET distributions for protons with energies between 50 and 200 MeV determined using a spherical tissue-equivalent proportional counter (TEPC) and a position-sensitive silicon spectrometer (RRMD-III)

Experiments have been performed to measure the response of a spherical tissue-equivalent proportional counter (TEPC) and a silicon-based LET spectrometer (RRMD-III) to protons with energies ranging from 50-200 MeV. This represents a large portion of the energy distribution for trapped protons encountered by astronauts in low-Earth orbit. The beam energies were obtained using plastic polycarbonate degraders with a monoenergetic beam that was extracted from a proton synchrotron. The LET spectrometer provided excellent agreement with the expected LET distribution emerging from the energy degraders. The TEPC cannot measure the LET distribution directly. However, the frequency mean value of lineal energy, y(-)(f), provided a good approximation to LET. This is in contrast to previous results for high-energy heavy ions where y(-)(f) underestimated LET, whereas the dose-averaged lineal energy, y(-)(D), provided a good approximation to LET.     

Chromosome aberrations induced by protons up to 31 MeV in cultured human cells

Summary Chromosome aberrations were induced in cultured human cells by proton beams of 31, 12, and 8 MeV. The frequencies of isocromatid breaks and dicentrics have been analysed as a function of proton energy and dose. Both effects are largely dependent on proton energy; isochromatid breaks increase linearly with the dose, whereas dicentrics show a definite parabolic behaviour. The experimental data were fitted to the analytic formY = KD ⁿ andY = D +D ² and the best fitted values of the parameters are reported and discussed. The values of RBE for the isochromatid breaks are in the ratio 1.7: 1.3: 1 for 8, 12, and 31 MeV respectively.In the case of the dicentrics the RBE values are dose-dependent function of the typeCD ^–n . The three distributions of dicentrics among the cells do not fit a Poisson distribution.Supported by a grant of CNR n^o 790067996 of the Finalized Project Tumour Growth Control201D; Consiglio Nazionale delle Ricerche, Italy     

Whole-cell imaging at nanometer resolutions using fast and slow focused helium ions

Observations of the interior structure of cells and subcellular organelles are important steps in unraveling organelle functions. Microscopy using helium ions can play a major role in both surface and subcellular imaging because it can provide subnanometer resolutions at the cell surface for slow helium ions, and fast helium ions can penetrate cells without a significant loss of resolution. Slow (e.g., 10–50 keV) helium ion beams can now be focused to subnanometer dimensions (∼0.25 nm), and keV helium ion microscopy can be used to image the surfaces of cells at high resolutions. Because of the ease of neutralizing the sample charge using a flood electron beam, surface charging effects are minimal and therefore cell surfaces can be imaged without the need for a conducting metallic coating. Fast (MeV) helium ions maintain a straight path as they pass through a cell. Along the ion trajectory, the helium ion undergoes multiple electron collisions, and for each collision a small amount of energy is lost to the scattered electron. By measuring the total energy loss of each MeV helium ion as it passes through the cell, we can construct an energy-loss image that is representative of the mass distribution of the cell. This work paves the way to use ions for whole-cell investigations at nanometer resolutions through structural, elemental (via nuclear elastic backscattering), and fluorescence (via ion induced fluorescence) imaging.     

Nonparametric method of determining the RBE coefficients of accelerated charged particles by the incidence of neoplasms in rats

The nonparametric method was used to determine RBE coefficients of accelerated charged particles (helium ions of 4 GeV/nucleon and 645 MeV protons) by the incidence of tumors localized in different rat organs or by the absence of tumors. The nonparametric method permitted to find the dose dependence of the RBE coefficients and to make statistical analysis of the results obtained with due regard for come features of developing damages which were not revealed by conventional methods of determining RBE coefficients.     

Relationship between linear energy transfer and behavioral toxicity in rats following exposure to protons and heavy particles.

Rats were exposed to protons (155 MeV) or to helium (165 MeV/amu), neon (522 MeV/amu) or argon (670 MeV/amu) particles to evaluate the behavioral toxicity of these types of radiations. Behavioral toxicity was assessed using the conditioned taste aversion paradigm. Exposure to all types of radiation produced dose-dependent increases in the intensity of the acquired taste aversion. However, the intensity of the aversions, measured as the dose that produced a 50% decrease in the intake of the sucrose-conditioned stimulus, did not show significant variation as a function of the linear energy transfer (LET) of the radiation. The results are discussed in terms of the relationship between LET and behavioral toxicity.     

PADC nuclear track detector for ion spectroscopy in laser-plasma acceleration

The transparent polymer polyallyl-diglycol-carbonate (PADC), also known as CR-39, is widely used as detector for heavy charged particles at low fluence. It allows for detection of single protons and ions via formation of microscopic tracks after etching in NaOH or KOH solutions. PADC combines a high sensitivity and high specificity with inertness towards electromagnetic noise. Present fields of application include laser-ion acceleration, inertial confinement fusion, radiobiological studies with cell cultures, and dosimetry of nuclear fragments in particle therapy. These require precise knowledge of the energy-dependent response of PADC to different ion species. We present calibration data for a new type of detector material, Radosys RS39, to protons (0.2–3 MeV) and carbon ions (0.6–12 MeV). RS39 is less sensitive to protons than other types of PADC. Its response to carbon ions, however, is similar to other materials. Our data indicate that RS39 allows for measuring carbon ion energies up to 10 MeV only from the track diameters. In addition, it can be used for discrimination between protons and carbon ions in a single etching process.     

Effects of 1.9 MeV monoenergetic neutrons onVicia faba chromosomes: Microdosimetric considerations

Summary AeratedVicia faba root meristems were irradiated with 1.9 MeV monoenergetic neutrons. This source of neutrons optimally provides one class of particles (recoil protons) with ranges able to traverse cell nuclei at moderate to high-LET. The volumes of theVicia faba nuclei were log-normally distributed with a mean of 1100 µm³. The yield of chromatid-type aberrations was linear against absorbed dose and near-constant over 5 collection periods (2–12 h), after irradiation. Energy deposition events (recoil protons) determined by microdosimetry were related to cytological changes with the finding that 19% of incident recoil protons initiate visible changes inVicia faba chromosomes. It is probable that a substantial fraction of recoil proton track length and deposited energy is in insensitive (non-DNA containing) portions of the nuclear volume.     

A novel method for assessment of fragmentation and beam-material interactions in helium ion radiotherapy with a miniaturized setup

Radiotherapy with protons and carbon ions enables to deliver dose distributions of high conformation to the target. Treatment with helium ions has been suggested due to their physical and biological advantages. A reliable benchmarking of the employed physics models with experimental data is required for treatment planning. However, experimental data for helium interactions is limited, in part due to the complexity and large size of conventional experimental setups.We present a novel method for the investigation of helium interactions with matter using miniaturized instrumentation based on highly integrated pixel detectors. The versatile setup consisted of a monitoring detector in front of the PMMA phantom of varying thickness and a detector stack for investigation of outgoing particles. The ion type downstream from the phantom was determined by high-resolution pattern recognition analysis of the single particle signals in the pixelated detectors. The fractions of helium and hydrogen ions behind the used targets were determined. As expected for the stable helium nucleus, only a minor decrease of the primary ion fluence along the target depth was found. E.g. the detected fraction of hydrogen ions on axis of a 220 MeV/u ⁴He beam was below 6% behind 24.5 cm of PMMA. Monte-Carlo simulations using Geant4 reproduce the experimental data on helium attenuation and yield of helium fragments qualitatively, but significant deviations were found for some combinations of target thickness and beam energy.The presented method is promising to contribute to the reduction of the uncertainty of treatment planning for helium ion radiotherapy.     

Multinucleate cells and micronucleus formation in cultured human cells exposed to 12 MeV protons and gamma-rays

Cultured human cells of the EUE line were exposed to different doses of 12 MeV protons, plated and allowed to grow for 8 days; colonies were then scored for the presences of multinucleate cells and micronuclei. The frequency of both effects is an increasing function of the dose; the evaluated exponents of the dose-response equation (e = bDn) are n = 1.0 %/- 0.1 for multinucleate cells and n = 1.6 +/- 0.1 for micronuclei. By comparison with the results obtained with gamma irradiations, r.b.e. values were obtained for both effects. The correlation between the logarithm of the surviving fraction and the yield of the studied effects has been proved to be statiscally significant.     

Spatial distribution and yield of DNA double-strand breaks induced by 3-7 MeV helium ions in human fibroblasts

Accelerated helium ions with mean energies at the target location of 3-7 MeV were used to simulate alpha-particle radiation from radon daughters. The experimental setup and calibration procedure allowed determination of the helium-ion energy distribution and dose in the nuclei of irradiated cells. Using this system, the induction of DNA double-strand breaks and their spatial distributions along DNA were studied in irradiated human fibroblasts. It was found that the apparent number of double-strand breaks as measured by a standard pulsed-field gel assay (FAR assay) decreased with increasing LET in the range 67-120 keV/microm (corresponding to the energy of 7-3 MeV). On the other hand, the generation of small and intermediate-size DNA fragments (0.1-100 kbp) increased with LET, indicating an increased intratrack long-range clustering of breaks. The fragment size distribution was measured in several size classes down to the smallest class of 0.1-2 kbp. When the clustering was taken into account, the actual number of DNA double-strand breaks (separated by at least 0.1 kbp) could be calculated and was found to be in the range 0.010-0.012 breaks/Mbp Gy(-1). This is two- to threefold higher than the apparent yield obtained by the FAR assay. The measured yield of double-strand breaks as a function of LET is compared with theoretical Monte Carlo calculations that simulate the track structure of energy depositions from helium ions as they interact with the 30-nm chromatin fiber. When the calculation is performed to include fragments larger than 0.1 kbp (to correspond to the experimental measurements), there is good agreement between experiment and theory.     

His+ reversions caused in Salmonella typhimurium by different types of ionizing radiation

The yield of his+ reversions in the Ames Salmonella tester strain TA2638 has been determined for 60Co gamma rays, 140 kV X rays, 5.4 keV characteristic X rays, 2.2 MeV protons, 3.1 MeV alpha particles, and 18 MeV/U Fe ions. Inactivation studies were performed with the same radiations. For both mutation and inactivation, the maximum effectiveness per unit absorbed dose was obtained for the characteristic X rays, which have a dose averaged linear energy transfer (LET) of roughly 10 keV/micron. The ratio of the effectiveness of this radiation to gamma rays was 2 for inactivation and about 1.4 for the his+ reversion. For both end points the effectiveness decreases substantially at high LET, i.e., for the alpha particles and the Fe ions. The composition of the bottom and the top agar was the one recommended by Maron and Ames [Mutat. Res. 113, 173-215 (1983)] for application in chemical mutagenicity tests. The experiments with the less penetrating radiations differed from the usual protocol by utilization of a technique of plating the bacteria on the surface of the top agar. As in an earlier study [Roos et al., Radiat. Res. 104, 102-108 (1985)] greatly enhanced yields of mutations, relative to the spontaneous reversion rate, were obtained in these experiments by performing the irradiations 6 h after plating, which differs from the conventional procedure to irradiate the bacteria shortly after plating.     

Induction of 8-azaguanine resistant mutants in human cultured cells exposed to 31 MeV protons

We report results on the induction of 8-azaguanine (8-AG)-resistant mutants in cultured human cells (EUE) exposed to 31 MeV protons. The spontaneous frequency of mutants was 5.6 +/- 0.7 x 10(-6) per viable cell. Gamma rays were taken as reference radiation. Expression times giving the highest frequency of mutants after 31 MeV protons and gamma irradiation were found to be about 10 days for both radiations. The dose-response relationship for mutant induction by protons, as determined at the optimal expression time, was compared to that obtained after gamma rays. The relative biological effectiveness (RBE) is 2.4 +/- 0.5, this value being higher than the RBE value determined for cell survival.     

Cellular and subcellular effects of very heavy ions

The biological effects of irradiation with ions of masses larger than 40 and energies up to 20 MeV per atomic mass unit are reviewed. The objects are viruses, bacterial spores, yeast and mammalian cells. Experimental parameters include loss of colony forming ability, induction of mutants, chromosomal aberrations, cell cycle progression, inhibition of biochemical activities and the formation of strand breaks. Some of the pertinent physical questions--e.g. track structure--are also discussed. It is shown that with very heavy ions the biological effectiveness is no longer unambiguously related to a single parameter like l.e.t. or Z*2/beta 2 but depends strongly on ion energy. This points to the importance of far-reaching delta-electrons. The analysis indicates also that even with very high l.e.t., cells are not killed by the passage of a single particle through their nucleus. Possible implications of the findings for fundamental radiation biology are outlined.     

Measuring the spectrum of mutation induced by nitrogen ions and protons in the human-hamster hybrid cell line A(L)C

Astronauts can be exposed to charged particles, including protons, alpha particles and heavier ions, during space flights. Therefore, studying the biological effectiveness of these sparsely and densely ionizing radiations is important to understanding the potential health effects for astronauts. We evaluated the mutagenic effectiveness of sparsely ionizing 55 MeV protons and densely ionizing 32 MeV/nucleon nitrogen ions using cells of two human-hamster cell lines, A(L) and A(L)C. We have previously characterized a spectrum of mutations, including megabase deletions, in human chromosome 11, the sole human chromosome in the human-hamster hybrid cell lines A(L)C and A(L). CD59(-) mutants have lost expression of a human cell surface antigen encoded by the CD59 gene located at 11p13. Deletion of genes located on the tip of the short arm of 11 (11p15.5) is lethal to the A(L) hybrid, so that CD59 mutants that lose the entire chromosome 11 die and escape detection. In contrast, deletion of the 11p15.5 region is not lethal in the hybrid A(L)C, allowing for the detection of chromosome loss or other chromosomal mutations involving 11p15.5. The 55 MeV protons and 32 MeV/nucleon nitrogen ions were each about 10 times more mutagenic per unit dose at the CD59 locus in A(L)C cells than in A(L) cells. In the case of nitrogen ions, the mutations observed in A(L)C cells were predominantly due to chromosome loss events or 11p deletions, often containing a breakpoint in the pericentromeric region. The increase in the CD59(-) mutant fraction for A(L)C cells exposed to protons was associated with either translocation of portions of 11q onto a hamster chromosome, or discontinuous or "skipping" mutations. We demonstrate here that A(L)C cells are a powerful tool that will aid in the understanding of the mutagenic effects of different types of ionizing radiation.     

Interaction of ion tracks in spatial and temporal proximity

In the present work, a systematic analysis of the impact of spatial and temporal proximity of ion tracks on the yield of higher-order radiolytic species as well as of DNA damage patterns is presented. This potential impact may be of concern when laser-driven particle accelerators are used for ion radiation therapy. The biophysical Monte Carlo track structure code PARTRAC was used and, to this end, extended in two aspects: first, the temporal information about track evolution has been included in the track structure module and, second, the simulation code has been modified to enable parallel multiple track processing during simulation of subsequent modelling stages. Depending on the spatial and temporal separation between ion-track pairs, the yield of chemical species has been calculated for incident protons with start energies of 20 MeV, for He²⁺ ions with start energies of 1 and 20 MeV, and for 60 MeV C⁶⁺ ions. Provided the overlap of the considered ion tracks is sufficient in all four dimensions (space and time), the yield of hydroxyl radicals was found to be reduced compared to that of single tracks, for all considered ion types. The biological endpoints investigated were base damages, single-strand breaks, double-strand breaks, and clustered lesions for incident pairs of protons and He²⁺ ions, each with start energies of 20 MeV. The yield of clustered lesions produced by 20 MeV protons turned out to be influenced by the spatial separation of the proton pair; in contrast, no influence was found for different start times of the protons. The yield of single-strand breaks and base hits was found neither to depend on the spatial separation nor on the temporal separation between the incident protons. For incident 20 MeV He²⁺ ions, however, a dependence on the spatial and temporal separation of the ion pair was found for all considered biological endpoints. Nevertheless, spatial proximity conditions where such intertrack effects were obtained are not met in the case of tumour radiation therapy; thus, no impact on radiation effects due to short pulse duration of laser-driven accelerators can be expected from alterations during the chemical stage.     

不同浓度红景天苷对γ射线和质子辐照质粒DNA的影响

利用14．2MeV的质子和60↑Co产生的γ射线在200Gy的剂量下分别对添加了不同浓度红景天苷的pUC19质粒DNA样品进行了辐照,凝胶电泳分析结果表明,红景天苷在两种辐照过程中均对DNA具有一定的保护作用,并且随着浓度的增加保护作用增强。同时还发现质子辐照后DNA样品中出现了线性形态的,而γ射线辐照后则没有出现,再次证实质子对DNA的损伤作用强于γ射线,这应该和质子与物质相互作用复杂以及质子对物质的直接电离作用相关。     

Biological effectiveness of low energy protons. I. Survival of Chinese hamster cells

The biological effectiveness of monoenergetic protons was investigated with the track-segment method. Protons were accelerated by a Tandem Van de Graaff accelerator and their final energies were 3.0 and 7.4 MeV. The biological system used was Chinese hamster V-79 cells and their survival ability following proton irradiation was investigated. Cobalt-60 gamma-rays were used as reference radiation to assess proton relative biological effectiveness (RBE). Survival curves were obtained for the gamma-ray and proton irradiations, and the relation S = exp (-alpha D-beta D2) was fitted to the data and the parameters alpha and beta were determined. The RBE values, calculated on the basis of the mean inactivation dose D and other pertinent parameters, were found to be 1.7 +/- 0.1 and 2.8 +/- 0.2 for 7.4 and 3.0 MeV protons, respectively. Comparisons were made with the results published by other investigators and it was concluded that in this low energy range the biological effectiveness increases substantially with decreasing proton energy.