Radiation quality dependence of DNA damage induction

Analysis of DNA fragmentation and repair in relation to radiation quality may give important information about the role of break complexity and correlated double strand breaks (DSBs). DNA fragment analysis was performed by pulsed-field gel electrophoresis after exposure to different radiation qualities. Normal human fibroblasts were irradiated with boron ions (40, 80 and 160 keV.micron-1), nitrogen ions (80, 125, 175 and 225 keV.micron-1) and neon ions (225 and 300 keV.micron-1). The amount of DNA less than 1.1 Mbp decreased with increasing linear energy transfer (LET) for all three ions. When theoretical random distributions were subtracted from the experimental data for 225 keV.micron-1 nitrogen ions in all size intervals (5-5700 kbp), there was a significant non-random distribution of DSBs for sizes up to 1-3 Mbp. This non-random distribution of breaks, probably produced by intra-track correlated DSBs, may constitute a substantial portion of the high-LET induced DSBs.     

Analysis of dose-LET distribution in the human body irradiated by high energy hadrons

For the purposes of radiological protection, it is important to analyse profiles of the particle field inside a human body irradiated by high energy hadrons, since they can produce a variety of secondary particles which play an important role in the energy deposition process, and characterise their radiation qualities. Therefore Monte Carlo calculations were performed to evaluate dose distributions in terms of the linear energy transfer of ionising particles (dose-LET distribution) using a newly developed particle transport code (Particle and Heavy Ion Transport code System, PHITS) for incidences of neutrons, protons and pions with energies from 100 MeV to 200 GeV. Based on these calculations, it was found that more than 80% and 90% of the total deposition energies are attributed to ionisation by particles with LET below 10 keV microm(-1) for the irradiations of neutrons and the charged particles, respectively.     

H2AX phosphorylation in response to DNA double-strand break formation during bystander signalling: effect of microRNA knockdown

Upon DNA double-strand break (DSB) formation, hundreds of H2AX molecules in the chromatin flanking the break site are phosphorylated on serine residue 139, termed gamma-H2AX, so that virtually every DSB site in a nucleus can be visualised within 10 min of its formation using an antibody to gamma-H2AX. One application of this sensitive assay is to examine the induction of DNA double-strand damage in subtle non-targeted cellular effects such as the bystander effect. Here whether microRNA (miRNA) serve as a primary signalling mechanism for bystander effect propagation by comparing matched human colon carcinoma cell lines with wild-type or depleted levels of mature miRNAs was investigated. No major differences were found in the levels of induced gamma-H2AX foci in the tested cell lines, indicating that though miRNAs play a role in bystander effect manifestation, they appear not to be the primary bystander signalling molecules in the formation of bystander effect-induced DSBs.     

Measurements of trapped protons and cosmic rays from recent Shuttle flights

Two new charged particle detectors have been flown in five recent Shuttle flights. The tissue-equivalent proportional counter measures the lineal energy spectrum of space radiation in the 0.26-300 keV micrometer-1 range. The charged particle spectrometer is a double dE/dx x E and dE/dx x Chrenekov detector system which provides a measurement of the differential energy spectrum of protons from 13 to 350 MeV and dose rate in silicon. In this paper the dose rate, equivalent dose rate, and radiation, quality factor for trapped protons and cosmic radiation are reported on separately. A comparison of the integral LET spectra with recent transport code calculations shows significant disagreement. Using the calculated dose rate from the omnidirectional AP8MAX model with IGRF reference magnetic field epoch 1970, and observed dose rate as a function of geographic latitude and longitude, the westward drift of the south Atlantic anomaly has been determined. The east-west effect has also been studied and a 'second' radiation belt observed. A comparison of the galactic cosmic radiation (GCR) lineal energy transfer spectra with model calculations shows disagreement comparable with those of the trapped protons.     

Responses of TLD Mg2SiO4:Tb and radiophotoluminescent glass to heavy charged particles and space radiation.

The LET dependences of thermoluminescence dosimeters of Mg2SiO4:Tb (TLMS) and radiophotoluminescent glass dosemeters (RPLG) were examined using high energy, heavy ion beams. TLMS kept its efficiency below 10 keV micrometer-1 and decreased almost linearly with the logarithm of LET for higher LET particles. The efficiency of RPLG decreased more gradually than TLMS although its reduction was observed at a lower LET region around 0.5 keV micrometer-1. Accordingly, the ratio of TLMS to RPLG valued showed a maximum peak around 20 keV micrometer-1 of LET. The results obtained with both dosemeters in the 40 day space mission in the Russian space station Mir showed that not only dose level but also radiation quality were varying considerable in the Mir Core Module.     

Passive spectrometry of linear energy transfer: development and use

A linear energy transfer (LET) spectrometer based on the evaluation of particle track parameters in a chemically etched polyallyldiglycolcarbonate (PADC) track detector has been developed at our laboratory. It permits us to determine LET spectra between 10 and 700 keV microm(-1) in tissues. The LET spectra obtained permit us to calculate total dose and dose equivalent corresponding to particles with etchable tracks also. We have recently been able to verify the calibration curves used by using C, Mg, Ne, Si and Fe ion beams with different energies. The calibration curves obtained are presented and compared with those originally used, and a good correlation is found. The LET spectrometer with new calibration was used to analyse the radiation quality of the radiotherapy proton beam at the Joint Institute for Nuclear Research (JINR). The radiation quality was studied along the proton's range, particular attention being devoted to the region of the Bragg peak. It was found that the biologically weighted effective dose (BWE) reaches a value of about 1.25 at the Bragg peak region. At the beam entrance this value increases to about 1.02 due to secondary particles created through primary proton nuclear reactions in tissues.     

A database of frequency distributions of energy depositions in small-size targets by electrons and ions

Linear energy transfer (LET) is an average quantity, which cannot display the stochastics of the interactions of radiation tracks in the target volume. For this reason, microdosimetry distributions have been defined to overcome the LET shortcomings. In this paper, model calculations of frequency distributions for energy depositions in nanometre size targets, diameters 1-100 nm, and for a 1 μm diameter wall-less TEPC, for electrons, protons, alpha particles and carbon ions are reported. Frequency distributions for energy depositions in small-size targets with dimensions similar to those of biological molecules are useful for modelling and calculations of DNA damage. Monte Carlo track structure codes KURBUC and PITS99 were used to generate tracks of primary electrons 10 eV to 1 MeV, and ions 1 keV μm(-1) to 300 MeV μm(-1) energies. Distribution of absolute frequencies of energy depositions in volumes with diameters of 1-100 nm randomly positioned in unit density water irradiated with 1 Gy of the given radiation was obtained. Data are presented for frequency of energy depositions and microdosimetry quantities including mean lineal energy, dose mean lineal energy, frequency mean specific energy and dose mean specific energy. The modelling and calculations presented in this work are useful for characterisation of the quality of radiation beam in biophysical studies and in radiation therapy.     

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.     

Efficiency-corrected dose verification with thermoluminescence dosemeters in heavy-ion beams

One of the most essential difficulties in heavy-ion dosimetry by means of thermoluminescence dosemeters (TLDs)--often seen as a serious disadvantage of TLD utilisation--regards the changing TL-efficiency with increasing linear energy transfer (LET) of the particle. This behaviour leads to a falsification of absorbed dose that can be significant for many applications, e.g. in space or radiotherapeutic dosimetry. The high-temperature TL emission of LiF:Mg,Ti TL detectors can be exploited to obtain information about the LET of the heavy-ion radiation field under study. The high-temperature ratio (HTR) is used as a parameter to determine average LET. To correct the absorbed dose according to the TL-efficiency, the detailed dependence of HTR- and TL-efficiency on LET was recorded. These investigations were accomplished at the Heavy Ion Medical Accelerator (HIMAC) in Chiba, Japan, with a variety of high-energy ion beams (helium, carbon, neon, silicon and iron) ranging in LET from 2.2 to 393 keV microm(-1). The obtained relationships HTR vs. LET and TL-efficiency vs. LET were combined into a TL efficieny vs. HTR relationship. This enables correction of the absorbed dose (HTR-B method). The methodology is demonstrated by means of TLD 700 ((7)LiF:Mg,Ti) measurements in carbon beams of 290 and 400 MeV n(-1) available from HIMAC.     

Spectra of linear energy transfer and other dosimetry characteristics as measured in C290 MeV/n MONO and SOBP ion beams at HIMAC-BIO (NIRS, Japan) with different detectors

Active mobile dosimetry unit (Liulin), passive plastic nuclear track detectors (PNTD) and thermoluminescent detectors (TLD) were exposed in a C290 MeV/n beam at HIMAC-BIO (NIRS, Japan). Two different types of beam configuration were used--monoenergetic beam (MONO) and spread-out Bragg peak (SOBP); the detectors were placed at several depths from the entrance up to the depths behind the Bragg peak. Relative response of TLDs in beams has been studied as a function of the depth, and it was re-proved that it can depend on the linear energy transfer (LET). Liulin measures energy deposition in Si; the spectra of energy deposited in Si can be transformed to the spectra of lineal energy or LET. PNTDs are able to determine the LET of registered particles directly. The limitation of both methods is in the range in which they can determine the LET-Liulin is able to measure perpendicularly incident charged particles up to ～35 keV/μm (in water), PNTD can measure from ～7 to 400 keV/μm, independently of the registration angle. The results from both methods are compared and combined for both beams' configuration, and a good agreement is observed.     

The efficiency of various thermoluminescence dosemeter types to heavy ions

Dose verification in heavy-ion beams using passive dosemeter systems, e.g. thermoluminescence dosemeters (TLDs), is crucial due to the changing efficiency of the dosemeters for different ion species and linear energy transfer (LET) values. This behaviour leads to a falsification of absorbed dose that can be significant for many applications, e.g. in space or radiotherapeutic dosimetry. TLDs can only be established as a 'reference' system in heavy-ion beams or other radiation fields if the efficiency functions for all contributing ion species and LET values are provided. In the framework of a research project of the Atominstitute of the Austrian Universities irradiations with various ions were performed in the years 2001-2003 at the Heavy Ion Medical Accelerator (HIMAC) of the National Institute for Radiological Sciences (NIRS) in Chiba, Japan. Efficiency values were recorded in dependence on ion species and LET in a range from 2 to 400 keV microm(-1). The efficiencies of five different commercially available TLD materials namely TLD 600, TLD 700, TLD 700H, TLD 300 and TLD 200 were investigated.     

Significance of fractionated irradiation for the biological therapeutic gain of carbon ions

It is well established that the RBE (relative biological effectiveness) for cell killing depends on LET (linear energy transfer), and that a maximum RBE is observed at approximately 150 keV.micron-1. However, the therapeutic gain depends on the ratio of the RBEs for the effects on the cancer cell population and the effects on normal tissues. The RBE of a given radiation quality depends not only on LET but also on dose, biological system and effect, and irradiation conditions. There is no data available to answer the question: which LET is suitable to improve the biological therapeutic gain of carbon ions? Here, three different LET values of 290 MeV/u carbon ions were selected, and the relative biological effectiveness was compared between tumour-growth retardation and skin damage using a murine transplantable tumour. Larger RBE values for tumours after than the skin type were obtained when carbon ions of intermediate LET were delivered daily for 2 to 5 fractions. The biological therapeutic gain would be high for the carbon ion SOBP if the number of fractions were correctly selected in clinical trials.     

Radiation-induced gamma-H2AX in mammalian cells irradiated with a synchrotron X-ray microbeam

In order to study the radiobiological effects from low dose radiation, a cell irradiation system using synchrotron X-ray microbeam has been developed, by which cells can be recognised individually and irradiated one by one with the desired dose of monochromatic X rays. The minimum beam sizes obtained are 2 microm with the focusing optics and 5 microm square with the non-focused beam, and the beam size can be changed easily with a high-precision slit in the case of a non-focused beam. Human fibroblast cells were individually irradiated with this system, and immunostained by gamma-H2AX antibody to visualise the DNA damage. Most of the fluorescent foci were observed in a localised area in cell nuclei, the size of which was almost the same as the beam size.     

Radiation measured for ISS-Expedition 12 with different dosimeters

Radiation in low Earth orbit (LEO) is mainly from Galactic Cosmic Rays (GCR), solar energetic particles and particles in South Atlantic Anomaly (SAA). These particles’ radiation impact to astronauts depends strongly on the particles’ linear energy transfer (LET) and is dominated by high LET radiation. It is important to investigate the LET spectrum for the radiation field and the influence of radiation on astronauts. At present, the best active dosimeters used for all LET are the tissue equivalent proportional counter (TEPC) and silicon detectors; the best passive dosimeters are thermoluminescence dosimeters (TLDs) or optically stimulated luminescence dosimeters (OSLDs) for low LET and CR-39 plastic nuclear track detectors (PNTDs) for high LET. TEPC, CR-39 PNTDs, TLDs and OSLDs were used to investigate the radiation for space mission Expedition 12 (ISS-11S) in LEO. LET spectra and radiation quantities (fluence, absorbed dose, dose equivalent and quality factor) were measured for the mission with these different dosimeters. This paper introduces the operation principles for these dosimeters, describes the method to combine the results measured by CR-39 PNTDs and TLDs/OSLDs, presents the experimental LET spectra and the radiation quantities.     

Energy dependence of TLD-300 response from 6 keV up to 1250 keV

The energy dependence of the response function S(E), of CaF2:Tm (TLD-300) thermoluminescent material (3.2 x 3.2 x 0.90 mm3 ribbons) was measured from 6 keV up to 1250 keV (60Co) using 55Fe, 137Cs and 60Co gamma ray sources as well as X ray ISO reference beams. To the best knowledge of the authors the data for energies below 45 keV are reported for the first time. The S(E) function was also calculated in the energy range from 1 keV up to 20 MeV and compared with the experimental data relative to the total glow curve. The comparison shows good agreement from 1250 keV down to 40 keV. At lower energy the discrepancy increases. This is interpreted in terms of the X ray attenuation within the thickness of TLD-300. Finally, measurements point out the effects of different reading and annealing treatments on the measured function S(E).     

Inactivation cross sections for mammalian cells exposed to charged particles: a phenomenological approach

Published data on inactivation of V79 cells irradiated with monoenergetic proton and ion beams (He, C, O, Ne) have been analysed. Values for RBE alpha, RBE10% and the inactivation cross section sigma have been evaluated in the LET range between 5 and 400 keV.micron-1. RBE against LET curves and inactivation cross sections against LET and against Z*2/beta 2 curves have been studied in a comparative approach with respect to the different ion types. RBE-LET curves depend strongly on the type of ion for LET > 30 keV.micron-1. At LET < 30 keV.micron-1 and low doses protons show the greatest effectiveness; at LET > 30 keV.micron-1 and high doses He ions provide the most effective radiation. Apart from protons, separation among the various ion curves is less marked in the sigma against Z*2/beta 2 plot than in the sigma against LET plot. sigma against Z*2/beta 2 curves for ions with 2 < or = Z < or = 10 and 200 < Z*2/beta 2 < 1500 show a common trend independent of Z and are well represented by a linear relationship.     

Significance of 8-oxoG in the spectrum of DNA damages caused by ionising radiation of different quality

The yields and composition of DNA damages caused by ionising radiation depends on radiation quality. With increasing light energy transfer (LET), the proportion of isolated DNA damages with respect to cluster damaged sites decreases. Non-double strand break complex damages are induced by gamma radiation in mammalian cells at least four times more frequently that prompt DSB. The most important product of oxidative damage to DNA bases is 8-oxo-7,8-dihydroguanine (8-oxoG). The modelling of DNA damage induced by ionising radiation of different qualities was performed to assess frequencies and composition of complex damages containing 8-oxoG. The occurrence of clusters containing 8-oxoG increases from 6 to 11% for LET in the range 0.4-160 keV microm(-1). Distributions of single strand break (SSB) on opposite DNA strand around induced 8-oxoG have similar shape for different ionising radiations, but differ in their occurrence in the whole spectrum of DNA damages. The most probable configuration is a strand break localised at position +/-3 bases from 8-oxoG.     

Chromosome aberration induction is dependent on the spatial distribution of energy deposition through a cell nucleus

The importance of the spatial distribution of energy deposition through the nucleus in determining the resultant chromosome rearrangements was investigated using fluorescent in situ hybridisation technique following either uniform or partial irradiation of HF19 human fibroblast cells with low-LET 1.5 keV ultrasoft X-rays. Irradiations were performed with and without a copper irradiation mask with a Poisson distribution of micron-sized holes immediately below the irradiation dish and the results are compared with previous results obtained following exposure to a Poisson distribution of alpha particles. For the same radiation quality, the spatial distribution of energy deposition within the nucleus was found to be important in determining the ultimate biological response, with an increased ratio of complex-to-simple aberrations observed for partial compared to uniform irradiation. Comparisons between low-LET ultrasoft X-rays and high-LET alpha particles indicate that the sub-micron clustering of damage along the alpha particle track is more important than just the total number of double-strand breaks produced.     

Active dosimetry on recent space flights

The radiation exposure inside the spacecraft in low earth orbit was investigated with a telescope based on two silicon planar detectors during three NASA shuttle-to-MIR missions (inclination 51.6 deg, altitude about 380 km). Count and dose rate profiles were measured, as well as separate linear energy transfer (LET) spectra, for the galactic cosmic rays (GCR) and the trapped radiation encountered in the South Atlantic Anomaly (SAA). Effective quality factors are deduced from the converted LET spectra (in water) in the range 0.1-120 keV micrometer-1 according to ICRP 60. Measured mission averaged dose rates in silicon are in the range 98-108 microGy d-1 and 137-178 microGy d-1 for the GCR and SAA contributions, respectively. The deduced effective quality factors are 2.95-3.29 (GCR) and 1.18-1.25 (SAA), resulting in mission averaged dose equivalent rates of 631-716 microSv d-1 for the comparable three missions.