BORON-ENHANCED BIOLOGICAL EFFECTIVENESS OF PROTON IRRADIATION: STRATEGY TO ASSESS THE UNDERPINNING MECHANISM.

Proton radiotherapy for the treatment of cancer offers an excellent dose distribution. Cellular experiments have shown that in terms of biological effects, the sharp dose distribution is further amplified, by as much as 75%, in the presence of boron. It is a matter of debate whether the underlying physical processes involve the nuclear reaction of 11B with protons or 10B with secondary neutrons, both producing densely ionizing short-ranged particles. Likewise, potential roles of intercellular communication or boron acting as a radiosensitizer are not clear. We present an ongoing research project based on a multiscale approach to elucidate the mechanism by which boron enhances the effectiveness of proton irradiation in the Bragg peak. It combines experimental with simulation tools to study the physics of proton-boron interactions, and to analyze intra- and inter-cellular boron biology upon proton irradiation.

EXPERIMENTAL IN VITRO DOSIMETRY OF 223RA AND 177LU.

Targeted alpha therapy with radionuclides undergoing multiple alpha-particle decays is a promising method of nuclear medicine. To study the effectiveness of alpha versus beta emitters, survival of DU145 prostate cancer cells exposed to 223Ra or 177Lu was assessed. Per decay, the cells were much more sensitive to the alpha than beta emitter. However, per unit dose the sensitivities would be comparable, contrary to the well-known evidence, if the decay energy were deposited within the sample completely and homogeneously. Measurements by Timepix detectors showed about three times higher counts of alpha particles above than below the sample. After the first alpha decay of 223Ra to 219Rn, this gas likely moves upwards and its subsequent three alpha decays occur in the upper part of the sample. Correct estimation of absorbed dose is a critical issue when analysing in vitro data and when translating their results to clinical applications.

RADIATION DAMAGE TO DNA PLASMIDS IN THE PRESENCE OF BOROCAPTATES.

Boron derivatives have great potential in cancer diagnostics and treatment. Borocaptates are used in boron neutron capture therapy and potentially in proton boron fusion therapy. This work examines modulation effects of two borocaptate compounds on radiation-induced DNA damage. Aqueous solutions of pBR322 plasmid containing increasing concentrations of borocaptates were irradiated with 60Co gamma rays or 30 MeV protons. Induction of single and double DNA strand breaks was investigated using agarose gel electrophoresis. In this model system, representing DNA without the intervention of cellular repair mechanisms, the boron derivatives acted as antioxidants. Clinically relevant boron concentrations of 40 ppm reduced the DNA single strand breakage seven-fold. Possible mechanisms of the observed effect are discussed.

VERTICAL DOSIMETRIC MEASUREMENT OF ELECTRIC DISCHARGE INDUCED AT LIGHTNING IMPULSE VOLTAGE GENERATOR.

This paper deals with the measurement of ionising radiation emitted in the vertical field of electric discharge generated between the tip and the grounded plate electrode. The generator set-up was for 400 consecutive negative discharges with voltage amplitude 0.9 MV, waveform shape 1.2/50 $\mu $s, discharge energy 80 kJ and electrodes 1 m apart. Thermoluminescent, passive detectors were used to avoid electromagnetic interference from the discharge. The detectors were placed at the electrodes and three vertical positions 1 m from the discharge axis. This work extends a previous study focused on dosimetry in the plane parallel to the ground. This experiment confirmed the presence of a photon/electron component at the electrode tip and 0.5 m vertical position at a distance of 1 m from the discharge axis. In addition, the presence of neutrons was found for new positions in the vertical axis and the ground electrode.

Assessment of DNA damage with an adapted independent reaction time approach implemented in Geant4-DNA for the simulation of diffusion-controlled reactions between radio-induced reactive species and a chromatin fiber.

PURPOSE: Simulation of indirect damage originating from the attack of free radical species produced by ionizing radiation on biological molecules based on the independent pair approximation is investigated in this work. In addition, a new approach, relying on the independent pair approximation that is at the origin of the independent reaction time (IRT) method, is proposed in the chemical stage of Geant4-DNA. METHODS: This new approach has been designed to respect the current Geant4-DNA chemistry framework while proposing a variant IRT method. Based on the synchronous algorithm, this implementation allows us to access the information concerning the position of radicals and may make it more convenient for biological damage simulations. Estimates of the evolution of free species as well as biological hits in a segment of DNA chromatin fiber in Geant4-DNA were compared for the dynamic time step approach of the step-by-step (SBS) method, currently used in Geant4-DNA, and this newly implemented IRT. RESULTS: Results show a gain in computation time of a factor of 30 for high LET particle tracks with a better than 10% agreement on the number of DNA hits between the value obtained with the IRT method as implemented in this work and the SBS method currently available in Geant4-DNA. CONCLUSION: Offering in Geant4-DNA more efficient methods for the chemical step based on the IRT method is a task in progress. For the calculation of biological damage, information on the position of chemical species is a crucial point. This can be achieved using the method presented in this paper.

Independent reaction times method in Geant4-DNA: Implementation and performance.

PURPOSE: The simulation of individual particle tracks and the chemical stage following water radiolysis in biological tissue is an effective means of improving our knowledge of the physico-chemical contribution to the biological effect of ionizing radiation. However, the step-by-step simulation of the reaction kinetics of radiolytic species is the most time-consuming task in Monte Carlo track-structure simulations, with long simulation times that are an impediment to research. In this work, we present the implementation of the independent reaction times (IRT) method in Geant4-DNA Monte Carlo toolkit to improve the computational efficiency of calculating G-values, defined as the number of chemical species created or lost per 100 eV of deposited energy. METHODS: The computational efficiency of IRT, as implemented, is compared to that from available Geant4-DNA step-by-step simulations for electrons, protons and alpha particles covering a wide range of linear energy transfer (LET). The accuracy of both methods is verified using published measured data from fast electron irradiations for • OH and e aq - for time-dependent G-values. For IRT, simulations in the presence of scavengers irradiated by cobalt-60 γ-ray and 2 MeV protons are compared with measured data for different scavenging capacities. In addition, a qualitative assessment comparing measured LET-dependent G-values with Geant4-DNA calculations in pure liquid water is presented. RESULTS: The IRT improved the computational efficiency by three orders of magnitude relative to the step-by-step method while differences in G-values by 3.9% at 1 µs were found. At 7 ps, • OH and e aq - yields calculated with IRT differed from recent published measured data by 5% ± 4% and 2% ± 4%, respectively. At 1 µs, differences were 9% ± 5% and 6% ± 7% for • OH and e aq - , respectively. Uncertainties are one standard deviation. Finally, G-values at different scavenging capacities and LET-dependent G-values reproduced the behavior of measurements for all radiation qualities. CONCLUSION: The comprehensive validation of the Geant4-DNA capabilities to accurately simulate the chemistry following water radiolysis is an ongoing work. The implementation presented in this work is a necessary step to facilitate performing such a task.

INVESTIGATION OF NUCLEAR EMULSIONS IN TERMS OF NEUTRON DOSIMETRY.

Neutron detection using nuclear emulsions can offer an alternative in personal dosimetry. The production of emulsions and their quality have to be well controlled with respect to their application in dosimetry. Nuclear emulsions consist mainly of gelatin and silver halide. Gelatin contains a significant amount of hydrogen, which can be used for fast neutron detection. The addition of B-10 in the emulsion is convenient for thermal neutron detection. In this paper, standard nuclear emulsions BR-2 and nuclear emulsions BR-2 enriched with boron produced at the Slavich Company, Russia, were applied for evaluation of fast and thermal neutron fluences. The results were obtained by calculation from the presumed emulsion composition without prior calibration. Evidence that nuclear emulsions used in the experiment are suitable for neutron dosimetry is provided.

RADIATION-INDUCED PLASMID DNA DAMAGE: EFFECT OF CONCENTRATION AND LENGTH.

Plasmid DNA is commonly used as a simpler substitute for a cell in studies of early effects of ionizing radiation because it allows to determine yields of primary DNA lesions. Experimental studies often employ plasmids of different lengths, in different concentrations in the aqueous solution. Influence of these parameters on the heavy-ion induced yields of primary DNA damage has been studied, using plasmids pUC19 (2686 bp), pBR322 (4361 bp) and pKLAC2 (9107 bp) in 10 and 50 ng/µl concentration. Results demonstrate the impact of plasmid length, while no significant difference was observed between the two concentrations. The uncertainty of the results is discussed.

CR10-A PUBLIC DATABASE OF COSMIC RADIATION MEASUREMENTS AT AVIATION ALTITUDES OF ABOUT 10 KM.

Long-term measurements using silicon radiation spectrometer Liulin on board commercial aircraft have been performed since 2001; results were put into a new database, which covers more than 4500 flights with more than 130 000 measurements. Methodology and tools were developed to normalize the data with respect to latitude and altitude and thus enable comparison with other radiation detectors and with model calculations. This capability is demonstrated using data from the neutron monitor at Lomnický stít. Instead of providing data files for individual measurement period, two software solutions are delivered. First is a web-based user interface for visualizing and downloading arbitrary time window of interest from the database hosted at http://cr10.odz.ujf.cas.cz. The second is a set of interactive Python notebooks available at GitHub. Those implement the calibration, normalization and visualization methods-so the outputs can be tailored to user needs. The software and data are provided under GNU/CC license.

HOW DETECTION OF PLASMID DNA FRAGMENTATION AFFECTS RADIATION STRAND BREAK YIELDS.

A compromised detection of radiation-induced plasmid DNA fragments results in underestimation of calculated damage yields. Electrophoretic methods are easy and cheap, but they can only detect a part of the fragments, neglecting the shortest ones. These can be detected with atomic force microscopy, but at the expense of time and price. Both methods were used to investigate their capabilities to detect the DNA fragments induced by high-energetic heavy ions. The results were taken into account in calculations of radiation-induced yields of single and double strand breaks. It was estimated that the double strand break yield is twice as high when the fragments are at least partially detected with the agarose electrophoresis, compared to when they were completely omitted. Further increase by 13% was observed when the measured fragments were corrected for the fraction of the shortest fragments up to 300 base pairs, as detected with the atomic force microscopy. The effect of fragment detection on the single strand break yield was diminished.

Extremely rapid isotropic irradiation of nanoparticles with ions generated in situ by a nuclear reaction.

Energetic ions represent an important tool for the creation of controlled structural defects in solid nanomaterials. However, the current preparative irradiation techniques in accelerators show significant limitations in scaling-up, because only very thin layers of nanoparticles can be efficiently and homogeneously irradiated. Here, we show an easily scalable method for rapid irradiation of nanomaterials by light ions formed homogeneously in situ by a nuclear reaction. The target nanoparticles are embedded in B2O3 and placed in a neutron flux. Neutrons captured by 10B generate an isotropic flux of energetic α particles and 7Li+ ions that uniformly irradiates the surrounding nanoparticles. We produced 70 g of fluorescent nanodiamonds in an approximately 30-minute irradiation session, as well as fluorescent silicon carbide nanoparticles. Our method thus increased current preparative yields by a factor of 102-103. We envision that our technique will increase the production of ion-irradiated nanoparticles, facilitating their use in various applications.

Dosimetry as a Catch in Radiobiology Experiments.

Experimental radiobiological studies in which the effects of ionizing radiation on a biological model are examined often highlight the biological aspects while missing detailed descriptions of the geometry, sample and dosimetric methods used. Such omissions can hinder the reproducibility and comparability of the experimental data. An application based on the Geant4 simulation toolkit was developed to design experiments using a biological solution placed in a microtube. The application was used to demonstrate the influence of the type of microtube, sample volume and energy of a proton source on the dose distribution across the sample, and on the mean dose in the whole sample. The results shown here are for samples represented by liquid water in the 0.4-, 1.5- and 2.0-ml microtubes irradiated with 20, 30 and 100 MeV proton beams. The results of this work demonstrate that the mean dose and homogeneity of the dose distribution within the sample strongly depend on all three parameters. Furthermore, this work shows how the dose uncertainty propagates into the scored primary DNA damages in plasmid DNA studies using agarose gel electrophoresis. This application is provided freely to assist users in verifying their experimental setup prior to the experiment.

Length computation of irradiated plasmid DNA molecules.

Compromised detection of short DNA fragments can result in underestimation of radiation-induced clustered DNA damage. The fragments can be detected with atomic force microscopy (AFM), followed by image analysis to compute the length of plasmid molecules. Plasmid molecules imaged with AFM are represented by open or closed curves, possibly with crossings. For the analysis of such objects, a dedicated algorithm was developed, and its usability was demonstrated on the AFM images of plasmid pBR322 irradiated with 60Co gamma rays. The analysis of the set of the acquired AFM images revealed the presence of DNA fragments with lengths shorter than 300 base pairs that would have been neglected by a conventional detection method.

Proton-induced direct and indirect damage of plasmid DNA.

Clustered DNA damage induced by 10, 20 and 30 MeV protons in pBR322 plasmid DNA was investigated. Besides determination of strand breaks, additional lesions were detected using base excision repair enzymes. The plasmid was irradiated in dry form, where indirect radiation effects were almost fully suppressed, and in water solution containing only minimal residual radical scavenger. Simultaneous irradiation of the plasmid DNA in the dry form and in the solution demonstrated the contribution of the indirect effect as prevalent. The damage composition slightly differed when comparing the results for liquid and dry samples. The obtained data were also subjected to analysis concerning different methodological approaches, particularly the influence of irradiation geometry, models used for calculation of strand break yields and interpretation of the strand breaks detected with the enzymes. It was shown that these parameters strongly affect the results.

Comparison of cosmic rays radiation detectors on-board commercial jet aircraft.

Aircrew members and passengers are exposed to increased rates of cosmic radiation on-board commercial jet aircraft. The annual effective doses of crew members often exceed limits for public, thus it is recommended to monitor them. In general, the doses are estimated via various computer codes and in some countries also verified by measurements. This paper describes a comparison of three cosmic rays detectors, namely of the (a) HAWK Tissue Equivalent Proportional Counter; (b) Liulin semiconductor energy deposit spectrometer and (c) TIMEPIX silicon semiconductor pixel detector, exposed to radiation fields on-board commercial Czech Airlines company jet aircraft. Measurements were performed during passenger flights from Prague to Madrid, Oslo, Tbilisi, Yekaterinburg and Almaty, and back in July and August 2011. For all flights, energy deposit spectra and absorbed doses are presented. Measured absorbed dose and dose equivalent are compared with the EPCARD code calculations. Finally, the advantages and disadvantages of all detectors are discussed.

Clustered DNA damage on subcellular level: effect of scavengers.

Clustered DNA damages are induced by ionizing radiation, particularly of high linear energy transfer (LET). Compared to isolated DNA damage sites, their biological effects can be more severe. We investigated a clustered DNA damage induced by high LET radiation (C 290 MeV u(-1) and Fe 500 MeV u(-1)) in pBR322 plasmid DNA. The plasmid is dissolved in pure water or in aqueous solution of one of the three scavengers (coumarin-3-carboxylic acid, dimethylsulfoxide, and glycylglycine). The yield of double strand breaks (DSB) induced in the DNA plasmid-scavenger system by heavy ion radiation was found to decrease with increasing scavenging capacity due to reaction with hydroxyl radical, linearly with high correlation coefficients. The yield of non-DSB clusters was found to occur twice as much as the DSB. Their decrease with increasing scavenging capacity had lower linear correlation coefficients. This indicates that the yield of non-DSB clusters depends on more factors, which are likely connected to the chemical properties of individual scavengers.

Lethal events in V79 cells irradiated by low-energy protons and correlations with distribution patterns of energy deposition, radical concentration and DNA damage.

Published survival data of V79 cells irradiated by 0.5-5.0 MeV (7-40 keV/microm) protons have been analyzed with a detailed radiobiological model to estimate the per-track yields of lethal lesions. Their correlations with distribution patterns of deposited energy, radical concentrations and with the yields of specific classes of DNA damage have been studied. The observed correlations indicate a potential interpretation of DNA damage lethal for the cell and the initial physical and chemical processes leading to such damage.

Theoretical modelling of radiolytic damage of free DNA bases and within DNA macromolecule.

The base damages in living cell are the most frequent product of deleterious effect of ionizing radiation. Experimental yields of modified bases determined in free bases and in the double stranded DNA show significant differences in the yields of stable products of radiolytic attack. An attempt to explain these differences is made with the help of theoretical calculations. The relative probabilities of reactions of the most important radiolytic product, OH(*) radical, with individual nucleobases, nucleosides and short DNA oligomers are calculated and compared to available experimental yields of base damages.

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.