The influence of fractionation on cell survival and premature differentiation after carbon ion irradiation

Carbon ion radiotherapy/Fractionated irradiation/R-BE/Premature terminal differentiation. To investigate the influence of fractionation on cell survival and radiation induced premature differentiation as markers for early and late effects after X-rays and carbon irradiation. Normal human fibroblasts NHDF, AG1522B and WI-38 were irradiated With 250 kV X-rays, or 266 MeV/u, 195 MeV/u and I I MeV/u carbon ions. Cytotoxicity was measured by a clonogenic survival assay or by determination of the differentiation pattern. Experiments with high-energy carbon ions show that fractionation induced repair effects are similar to photon irradiation. The RBE10 values for clonogenic survival are 1.3 and 1.6 for irradiation in one or two fractions for NHDF cells and around 1.2 for AG1522B cells regardless of the fractionation scheme. The RBE for a doubling of post mitotic fibroblasts (PMF) in the population is I for both single and two fractionated irradiation of NHDF cells. Using I I MeV/u carbon ions, no repair effect can be seen in WI-38 cells. The RBE10 for clonogenic survival is 3.2 for single irradiation and 4.9 for two fractionated irradiations. The RBE for a doubling of PMF is 3.1 and 5.0 for single and two fractionated irradiations, respectively. For both cell lines the effects of high-energy carbon ions representing the irradiation of the skin and the normal tissue in the entrance channel are similar to the effects of X-rays. The fractionation effects are maintained. For the lower energy, which is representative for the irradiation of the tumor region. RBE is enhanced for clonogenic survival as well as for premature terminal differentiation. Fractionation effects are not detectable. Consequently, the therapeutic ratio is significantly enhanced by fractionated irradiation with carbon ions.

Induction of cytogenetic adaptive response in spermatogonia and spermatocytes by pre-exposure of mouse testis to low-dose C-12(6+) ions

To investigate the effects of pre-exposure of mouse testis to low-dose C-12(6+) ions on cytogenetics of spermatogonia and spermatocytes induced by subsequent high-dose irradiation. the testes of outbred Kun-Ming strain mice were irradiated with 0.05 Gy of C-12(6+) ions as the pre-exposure dose, and then irradiated with 2 Gy as challenging dose at 4 h after per-exposure. Poly(ADP-ribose) polymerase (PARPs) activity and PARP-1 protein expression were respectively measured by using the enzymatic and Western blot assays at 4 h after irradiation; chromosomal aberrations in spermatogonia and spermatocytes were analyzed by the air-drying method at 8 h after irradiation. The results showed that there was a significant increase in the frequency of chromosomal aberrations and significant reductions of PARP activity and PARP-1 expression level in the mouse testes irradiated with 2 Gy of C-12(6+) ions. However, pre-exposure of mouse testes to a low dose of C-12(6+) ions significantly increased PARPs activity and PARP-1 expression and alleviated the harmful effects induced by a subsequent high-dose irradiation. PARP activity inhibitor 3-aminobenzamide (3-AB) treatment blocked the effects of PARP-1 on cytogenetic adaptive response induced by low-dose C-12(6+) ion irradiation. The data suggest that pre-exposure of testes to a low dose of heavy ions can induce cytogenetic adaptive response to subsequent high-dose irradiation. The increase of PARP-1 protein induced by the low-dose ionizing irradiation may be involved in the mechanism of these observations. (C) 2008 Elsevier B.V. All rights reserved.

Vacuum window of intermediate energy heavy-ion microbeam irradiation facility

The beam must be extracted into the air through the vacuum window to irradiate the living cell. In the window design, the material and thickness must be chosen to compromise the beam spot size broadening and the window safety. The structure-static analysis on the window of different structures and materials is done with the finite element analysis method, and the deformation and the equivalent stress axe simulated. The safety of these candidates is investigated using the intensity theory. In addition, the small angle scattering and the transverse range of ions are simulated using SRIM code, including all the effects on the beam spot size broadening, such as the incident ion energy, the material and the thickness of the window and the air composing. At last, the appropriate vacuum windows are presented, including the structure, material and thickness.

Investigation of PL properties of C-doped SiO2/Si samples after high energy Pb ion irradiation

Amorphous SiO2 thin films with about 400-500 nm in thickness were thermally grown on single crystalline silicon. These SiO2/Si samples were firstly implanted at room temperature (RT) with 100 keV carbon ions to 2.0 x 10(17),5.0 X 10(17) or 1.2 x 10(18) ions/cm(2), then irradiated at RT by 853 MeV Pb ions to 5.0 x 10(11), 1.0 X.10(12) 2.0 x 10(12) or 5.0 x 10(12) ions/cm(2), respectively. The variation of photoluminescence (PL) properties of these samples was analyzed at RT using a fluorescent spectroscopy. The obtained results showed that Pb-ion irradiations led to significant changes of the PL properties of the carbon ion implanted SiO2 films. For examples, 5.0 x 10(12) Pb-ions/cm(2) irradiation produced huge blue and green light-emitters in 2.0 x 10(17) C-ions/cm(2) implanted samples, which resulted in the appearance of two intense PL peaks at about 2.64 and 2.19 eV. For 5.0 x 10(17) carbon-ions/cm(2) implanted samples, 2.0 x 10(12) Pb-ions/cm(2) irradiation could induce the formation of a strong and wide violet band at about 2.90 eV, whereas 5.0 x 10(12) Pb-ionS/cm(2) irradiation could,create double peaks of light emissions at about 2.23 and 2.83 eV. There is no observable PL peak in the 1.2 x 10(18) carbon-ions/cm(2) implanted samples whether it was irradiated with Pb ions or not. All these results implied that special light emitters could be achieved by using proper ion implantation and irradiation conditions, and it will be very useful for the synthesis of new type Of SiO2-based light-emission materials.

Development of an intermediate energy heavy-ion micro-beam irradiation system

The micro-beam irradiation system, which focuses the beam down to micron order and precisely delivers a predefined number of ions to a predefined spot of micron order, is a powerful tool for radio-biology, radio-biomedicine and micromachining. The Institute of Modern Physics of Chinese Academy of Sciences is developing a heavy-ion microbeam irradiation system up to intermediate energy. Based on the intermediate and low energy beam provided by Heavy Ion Research Facility of Lanzhou, the micro-beam system takes the form of the magnetic focusing. The heavy-ion beam is conducted to the basement by a symmetrical achromatic system consisting of two vertical bending magnets and a quadrupole in between. Then a beam spot of micron order is formed by a magnetic triplet quadrupole of very high gradient. The sample can be irradiated either in vacuum or in the air. This system will be the first opening platform capable of providing heavy ion micro-beam, ranging from low (10MeV/u) to intermediate energy (100MeV/u), for irradiation experiment with positioning and counting accuracy. Target material may be biology cell, tissue or other non-biological materials. It will be a help for unveiling the essence of heavy-ion interaction with matter and also a new means for exploring the application of heavy-ion irradiation.

Modification of Fe/Cu multilayers under 400 keV Xe20+ irradiation

Two kinds of Fe/Cu multilayers with different modulation wavelength were deposited on cleaved Si(100) substrates and then irradiated at room temperature using 400 keV Xe20+ in a wide range of irradiation fluences. As a comparison, thermal annealing at 300-900 degrees C was also carried out in vacuum. Then the samples were analyzed by XRD and the evolution of crystallite structures induced by irradiation was investigated. The obtained XRD patterns showed that, with increase of the irradiation fluence, the peaks of Fe became weaker, the peaks related to Cu-based fcc solid solution and Fe-based bcc solid solution phase became visible and the former became strong gradually. This implied that the intermixing at the Fe/Cu interface induced by ion irradiation resulted in the formation of the new phases which could not be achieved by thermal annealing. The possible intermixing mechanism of Fe/Cu multilayers induced by energetic ion irradiation was briefly discussed.

Early effects of carbon-ion irradiation on murine lymphocytes and thymocytes

To estimate the biological risks from space radiation encountered by cosmonauts in outer space, the effects from whole-body exposure to carbon ions or X-rays irradiations at 0, 0.39, 0.55 and 1 Gy at a dose rate of 0.2 Gy/min were investigated in BALB/c mice. The relative thymus and spleen weights were measured at 24 h after exposure, and the cell cycle distribution and percentage of apoptosis of thymocytes and spleen and peripheral blood lymphocytes were determined by flow cytometry. The data showed that exposure to carbon ions delayed cell progression of peripheral blood lymphocytes in S-phase, and delayed thymocytes and spleen lymphocytes in G(0)/G(1)-phase. Apoptosis of thymocytes and peripheral blood lymphocytes induced by carbon ions increased more rapidly with dose than was the case for X-rays. There were some differences between the relative weight loss of the thymus and the spleen with increasing dose of either carbon ions or X-rays. The results obtained provide evidence of dose- and organ-specific differences induced by carbon ions compared to X-rays, with increased apoptosis in peripheral blood lymphocytes and thymocytes, but not spleen lymphocytes. Our data may suggest that further work would be of interest to estimate risk of changes in immune function during particle radiation exposures in space travel. (c) 2007 COSPAR

The influence of carbon ion irradiation on sweet sorghum seeds

The aim of this study is to investigate the effects of different doses of 100 MeV/u carbon ions on sweet sorghum seeds in order to improve crop yields and their sugar content. After irradiation, seeds were germinated and grown to 30 days, and others were sown in the field. At the end of harvesting season all planted seeds were picked separately and M2 generations obtained. The differences among the treatments were examined using the RAPID procedure. In the study done by using 38 primers; according to the amplification results, the differences among the various doses treatment were shown.

Heavy-ion conformal irradiation in the shallow-seated tumor therapy terminal at HIRFL

Basic research related to heavy-ion cancer therapy has been done at the Institute of Modern Physics (IMP), Chinese Academy of Sciences since 1995. Now a plan of clinical trial with heavy ions has been launched at IMP. First, superficially placed tumor treatment with heavy ions is expected in the therapy terminal at the Heavy Ion Research Facility in Lanzhou (HIRFL), where carbon ion beams with energy up to 100 MeV/u can be supplied. The shallow-seated tumor therapy terminal at HIRFL is equipped with a passive beam delivery system including two orthogonal dipole magnets, which continuously scan pencil beams laterally and generate a broad and uniform irradiation field, a motor-driven energy degrader and a multi-leaf collimator. Two different types of range modulator, ripple filter and ridge filter with which Guassian-shaped physical dose and uniform biological effective dose Bragg peaks can be shaped for therapeutic ion beams respectively, have been designed and manufactured. Therefore, two-dimensional and three-dimensional conformal irradiations to tumors can be performed with the passive beam delivery system at the earlier therapy terminal. Both the conformal irradiation methods have been verified experimentally and carbon-ion conformal irradiations to patients with superficially placed tumors have been carried out at HIRFL since November 2006.

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 mu m(-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.

Pre-irradiation with low-dose C-12(6+) beam significantly enhances the efficacy of AdCMV-p53 gene therapy in human non-small lung cancer

The combination of ionizing radiation and gene therapy has been investigated. However, there are very few reports about the combination of heavy-ion irradiation and gene therapy. To determine if the pre-exposure to low-dose heavy ion beam enhances the suppression of AdCMV-p53 on non-small lung cancer (NSLC), the cells pre-irradiated or non-irradiated were infected with 20, 40 MOI of AdCMV-p53. Survival fraction and the relative biology effect (RBE) were determined by clonogenic assay. The results showed that the proportions of p53 positive cells in C-12(6+) beam induced AdCMV-p53 infected cells were more than 90%, which were significantly more than those in gamma-ray induced AdCMV-p53 infected cells. The pre-exposure to low-dose 12C6+ beam significantly prevented the G(0)/G(1) arrest and activated G(2)/M checkpoints. The pre-exposure to C-12(6+) beam significantly improved cell to apoptosis. RBEs for the C-12(6+)+ AdCMV-p53 infection groups were 30%-60%,20% -130% and 30%-70% more than those for the C-12(6+)_irradiated only, AdCMV-p53 infected only, and gamma-irradiation induced AdCMVp53 infected groups, respectively. The data suggested that the pre-exposure to low-dose C-12(6+) beam significantly promotes exogenous p53 expression in NSLC, and the suppression of AdCMV-p53 gene therapy on NSLC.

Chromatid-type aberrations following irradiation in G0 lymphocytes with heavy ions

We describe a low level of chromatid-type aberrations which included the relatively rare isochromatid/chromatid triradial in peripheral blood lymphocytes that were irradiated, ostensibly in GO, with accelerated heavy C-12 ions. These were produced only at the energies of 69 MeV/n (34.6 keV/ mu m), almost absent at the energy of either 58.6 McV/n (46.07 keV/mu m) or 19.3 MeV/n (97 keV/mu m), nor were they found after low-LET X-rays. Mechanisms potentially responsible for their formation are discussed.

Effects of annealing on the photoluminescence of He ion implanted sapphire after 230-MeV Pb ion irradiation

Single crystal sapphire (Al2O3) samples implanted with 110 keV He and irradiated at 320 K by Pb-208(27), ions with energy of 1.1 MeV/u to the fluences ranging from 1 X 10(12) to 5 X 10(14) ion/cm(2) and subsequently annealed at 600, 900 and 1100 K. The obtained PL spectra showed that emission peaks centred at 375, 390, 413, and 450 nm appeared in irradiated samples. The peak of 390 ran became very intense after 600 K annealing. The peak of 390 nm weakened and 510 nm peak started to build up at 900 K annealing, the peak of 390 nm vanished and 510 nm peak increased with the annealing temperature rising to 1100 K. Infrared spectra showed a broadening of the absorption band between 460 cm(-1), and 510 cm(-1) indicating strongly damaged regions being formed in the Al2O3 samples and position shift of the absorption band at 1000-1300 cm(-1) towards higher wavenumber after Pb irradiation.

Adaptive hormetic response of pre-exposure of mouse brain with low-dose C-12(6+) ion or Co-60 gamma-ray on growth hormone (GH) and body weight induced by subsequent high-dose irradiation

The brain of the Kun-Ming strain mice were irradiated with 0.05 Gy of C-12(6+) ion or Co-60 gamma-ray as the pre-exposure dose, and were then irradiated with 2 Gy of 12C6+ ion or Co-60 gamma-ray as challenging irradiation dose at 4 h after per-exposure. Body weight and serum growth hormone (GH) concentration were measured at 35th day after irradiation. The results showed that irradiation of mouse brain with 2 Gy of C-12(6+) ion or Co-60 gamma-ray significantly diminished mouse body weight and level of serum GH. The relative biological effectiveness values of a 2 Gy dose of C-12(6+) ion calculated with respect to Co-60 gamma-ray were 1.47 and 1.34 for body weight and serum GH concentration, respectively. Pre-exposure with a low-dose (0.05 Gy) of C-12(6+) ion or Co-60 gamma-ray significantly alleviated reductions of mouse body weight and level of serum GH induced by a subsequent high-dose (2 Gy) irradiation. The data suggested that low-dose ionizing irradiation can induce adaptive hormetic responses to the harmful effects of pituitary by subsequent high-dose exposure.

Conformal irradiation to moving targets in heavy ion radiotherapy with raster-scanning beam delivery system: (II) feasibility experiments

Within the framework of the pilot heavy-ion therapy facility at GSI equipped with an active beam delivery system of advanced raster scanning technique, a feasibility study on actively conformal heavy-ion irradiation to moving tumors has been experimentally conducted. Laterally, real-time corrections to the beam scanning parameters by the raster scanner, leading to an active beam tracing, compensate for the lateral motion of a target volume. Longitudinally, a mechanically driven wedge energy degrader (called depth scanner) is applied to adjust the beam energy so as to locate the high-dose Bragg peak of heavy ion beam to the slice under treatment for the moving target volume. It has been experimentally shown that compensations for lateral target motion by the raster scanner and longitudinal target shift by the depth scanner are feasible.