Correction: Belchior et al. Repair Kinetics of DSB-Foci Induced by Proton and α-Particle Microbeams of Different Energies. Life 2022, 12, 2040.

In the original publication [...].

Repair Kinetics of DSB-Foci Induced by Proton and α-Particle Microbeams of Different Energies.

In this work, the induction and repair of radiation-induced 53BP1 foci were studied in human umbilical vein endothelial cells irradiated at the PTB microbeam with protons and α-particles of different energies. The data were analyzed in terms of the mean number of 53BP1 foci induced by the different ion beams. The number of 53BP1 foci found at different times post-irradiation suggests that the disappearance of foci follows first order kinetics. The mean number of initially produced foci shows the expected increase with LET. The most interesting finding of this work is that the absolute number of persistent foci increases with LET but not their fraction. Furthermore, protons seem to produce more persistent foci as compared to α-particles of even higher LET. This may be seen as experimental evidence that protons may be more effective in producing severe DNA lesions, as was already shown in other work, and that LET may not be the best suited parameter to characterize radiation quality.

The role of the construction and sensitive volume of compact ionization chambers on the magnetic field-dependent dose response.

PURPOSE: The magnetic-field correction factors k B , Q of compact air-filled ionization chambers have been investigated experimentally and using Monte Carlo simulations up to 1.5 T. The role of the nonsensitive region within the air cavity and influence of the chamber construction on its dose response have been elucidated. MATERIALS AND METHODS: The PTW Semiflex 3D 31021, PinPoint 3D 31022, and Sun Nuclear Cooperation SNC125c chambers were studied. The k B , Q factors were measured at the experimental facility of the German National Metrology Institute (PTB) up to 1.4 T using a 6 MV photon beam. The chambers were positioned with the chamber axis perpendicular to the beam axis (radial); and parallel to the beam axis (axial). In both cases, the magnetic field was directed perpendicular to both the beam axis and chamber axis. Additionally, the sensitive volumes of these chambers have been experimentally determined using a focused proton microbeam and finite element method. Beside the simulations of k B , Q factors, detailed Monte Carlo technique has been applied to analyse the secondary electron fluence within the air cavity, that is, the number of secondary electrons and the average path length as a function of the magnetic field strength. RESULTS: A nonsensitive volume within the air cavity adjacent to the chamber stem for the PTW chambers has been identified from the microbeam measurements and FEM calculations. The dose response of the three investigated ionization chambers does not deviate by more than 4% from the field-free case within the range of magnetic fields studied in this work for both the radial and axial orientations. The simulated k B , Q for the fully guarded PTW chambers deviate by up to 6% if their sensitive volumes are not correctly considered during the simulations. After the implementation of the sensitive volume derived from the microbeam measurements, an agreement of better than 1% between the experimental and Monte Carlo k B , Q factors for all three chambers can be achieved. Detailed analysis reveals that the stem of the PTW chambers could give rise to a shielding effect reducing the number of secondary electrons entering the air cavity in the presence of magnetic field. However, the magnetic field dependence of their path length within the air cavity is shown to be weaker than for the SNC125c chamber, where the length of the air cavity is larger than its diameter. For this chamber it is shown that the number of electrons and their path lengths in the cavity depend stronger on the magnetic field. DISCUSSION AND CONCLUSION: For clinical measurements up to 1.5 T, the required k B , Q corrections of the three chambers could be kept within 3% in both the investigated chamber orientations. The results reiterate the importance of considering the sensitive volume of fully guarded chambers, even for the investigated compact chambers, in the Monte Carlo simulations of chamber response in magnetic field. The resulting magnetic field-dependent dose response has been demonstrated to depend on the chamber construction, such as the ratio between length and the diameter of the air cavity as well as the design of the chamber stem.

Three-dimensional characterization of the active volumes of PTW microDiamond, microSilicon, and Diode E dosimetry detectors using a proton microbeam.

PURPOSE: The purpose of this work is the three-dimensional characterization of the active volumes of commercial solid-state dosimetry detectors. Detailed knowledge of the dimensions of the detector's active volume as well as the detector housing is of particular interest for small-field photon dosimetry. As shown in previous publications from different groups, the design of the detector housing influences the detector signal for small photon fields. Therefore, detailed knowledge of the active volume dimension and the surrounding materials form the basis for accurate Monte Carlo simulations of the detector. METHODS: A 10 MeV proton beam focused by the microbeam system of the Physikalisch-Technische Bundesanstalt was used to measure two-dimensional response maps of a synthetic diamond detector (microDiamond, type 60019, PTW Freiburg) and two silicon detectors (microSilicon, type 60023, PTW Freiburg and Diode E, type 60017, PTW Freiburg). In addition, the thickness of the active volume of the new microSilicon was measured using the method developed in a previous study. RESULTS: The analysis of the response maps leads to active area of 1.18 mm2 for the Diode E, 1.75 mm2 for the microSilicon, and 3.91 mm2 for the microDiamond detector. The thickness of the active volume of the microSilicon detector was determined to be (17.8 ± 2) µm. CONCLUSIONS: This study provides detailed geometrical data of the dosimetric active volume of three different solid-state detector types.

From Energy Deposition of Ionizing Radiation to Cell Damage Signaling: Benchmarking Simulations by Measured Yields of Initial DNA Damage after Ion Microbeam Irradiation.

Advances in accelerator technology, which have enabled conforming radiotherapy with charged hadronic species, have brought benefits as well as potential new risks to patients. To better understand the effects of ionizing radiation on tumor and surrounding tissue, it is important to investigate and quantify the relationship between energy deposition at the nanometric scale and the initial biological events. Monte Carlo track structure simulation codes provide a powerful tool for investigating this relationship; however, their success and reliability are dependent on their improvement and development accordingly to the dedicated biological data to which they are challenged. For this aim, a microbeam facility that allows for fluence control, down to one ion per cell nucleus, was used to evaluate relative frequencies of DNA damage after interaction between the incoming ion and DNA according to radiation quality. Primary human cells were exposed to alpha particles of three different energies with respective linear energy transfers (LETs) of approximately 36, 85 or 170 keV·µm-1 at the cells' center position, or to protons (19 keV·µm-1). Statistical evaluation of nuclear foci formation (53BP1/γ-H2AX), observed using immunofluorescence and related to a particle traversal, was undertaken in a large population of cell nuclei. The biological results were adjusted to consider the factors that drive the experimental uncertainties, then challenged with results using Geant4-DNA code modeling of the ionizing particle interactions on a virtual phantom of the cell nucleus with the same mean geometry and DNA density as the cells used in our experiments. Both results showed an increase of relative frequencies of foci (or simulated DNA damage) in cell nuclei as a function of increasing LET of the traversing particles, reaching a quasi-plateau when the LET exceeded 80-90 keV·µm-1. For the LET of an alpha particle ranging from 80-90 to 170 keV·µm-1, 10-30% of the particle hits did not lead to DNA damage inducing 53BP1 or γ-H2AX foci formation.

Determination of the active volumes of solid-state photon-beam dosimetry detectors using the PTB proton microbeam.

PURPOSE: This study aims at the experimental determination of the diameters and thicknesses of the active volumes of solid-state photon-beam detectors for clinical dosimetry. The 10 MeV proton microbeam of the PTB (Physikalisch-Technische Bundesanstalt, Braunschweig) was used to examine two synthetic diamond detectors, type microDiamond (PTW Freiburg, Germany), and the silicon detectors Diode E (PTW Freiburg, Germany) and Razor Diode (Iba Dosimetry, Germany). The knowledge of the dimensions of their active volumes is essential for their Monte Carlo simulation and their applications in small-field photon-beam dosimetry. METHODS: The diameter of the active detector volume was determined from the detector current profile recorded by radially scanning the proton microbeam across the detector. The thickness of the active detector volume was determined from the detector's electrical current, the number of protons incident per time interval and their mean stopping power in the active volume. The mean energy of the protons entering this volume was assessed by comparing the measured and the simulated influence of the thickness of a stack of aluminum preabsorber foils on the detector signal. RESULTS: For all detector types investigated, the diameters measured for the active volume closely agreed with the manufacturers' data. For the silicon Diode E detector, the thickness determined for the active volume agreed with the manufacturer's data, while for the microDiamond detectors and the Razor Diode, the thicknesses measured slightly exceeded those stated by the manufacturers. DISCUSSION: The PTB microbeam facility was used to analyze the diameters and thicknesses of the active volumes of photon dosimetry detectors for the first time. A new method of determining the thickness values with an uncertainty of ±10% was applied. The results appear useful for further consolidating detailed geometrical knowledge of the solid-state detectors investigated, which are used in clinical small-field photon-beam dosimetry.

Analysis of Radiation-Induced Chromosomal Aberrations on a Cell-by-Cell Basis after Alpha-Particle Microbeam Irradiation: Experimental Data and Simulations.

There is a continued need for further clarification of various aspects of radiation-induced chromosomal aberration, including its correlation with radiation track structure. As part of the EMRP joint research project, Biologically Weighted Quantities in Radiotherapy (BioQuaRT), we performed experimental and theoretical analyses on chromosomal aberrations in Chinese hamster ovary cells (CHO-K1) exposed to α particles with final energies of 5.5 and 17.8 MeV (absorbed doses: ∼2.3 Gy and ∼1.9 Gy, respectively), which were generated by the microbeam at the Physikalisch-Technische Bundesanstalt (PTB) in Braunschweig, Germany. In line with the differences in linear energy transfer (approximately 85 keV/µm for 5.5 MeV and 36 keV/µm for 17.8 MeV α particles), the 5.5 MeV α particles were more effective than the 17.8 MeV α particles, both in terms of the percentage of aberrant cells (57% vs. 33%) and aberration frequency. The yield of total aberrations increased by a factor of ∼2, although the increase in dicentrics plus centric rings was less pronounced than in acentric fragments. The experimental data were compared with Monte Carlo simulations based on the BIophysical ANalysis of Cell death and chromosomal Aberrations model (BIANCA). This comparison allowed interpretation of the results in terms of critical DNA damage [cluster lesions (CLs)]. More specifically, the higher aberration yields observed for the 5.5 MeV α particles were explained by taking into account that, although the nucleus was traversed by fewer particles (nominally, 11 vs. 25), each particle was much more effective (by a factor of ∼3) at inducing CLs. This led to an increased yield of CLs per cell (by a factor of ∼1.4), consistent with the increased yield of total aberrations observed in the experiments.

53BP1 and MDC1 foci formation in HT-1080 cells for low- and high-LET microbeam irradiations.

An improved assessment of the biological effects and related risks of low doses of ionizing radiation is currently an important issue in radiation biology. Irradiations using microbeams are particularly well suited for precise and localized dose depositions, whereas recombinant cell lines with fluorescent proteins allow the live observation of radiation-induced foci. Living cells of the fibrosarcoma cell line HT-1080 stably expressing 53BP1 or full-length reconstituted MDC1 fused to Green Fluorescent Protein (GFP) were irradiated with protons and α-particles of linear energy transfers (LETs) of 15 and 75 keV/µm, respectively. Using a microbeam, the irradiations were carried out in line patterns, which facilitated the discrimination between undefined background and radiation-induced foci. As expected, foci formation and respective kinetics from α-particle irradiations with a high LET of 75 keV/µm could be detected in a reliable manner by both fusion proteins, as reported previously. Colocalization of γ-H2AX foci confirmed the DSB nature of the detected foci. As a novel result, the application of protons with low LET of 15 keV/µm generated 53BP1- and MDC1-mediated foci of almost equal size and slightly different kinetics. This new data expands the capability of 53BP1 and wild-type MDC1 on visible foci formation in living cells after irradiation with low-LET particles. Furthermore, the kinetics in HT-1080 cells for α-particle irradiation show a delay of about 20 s for 53BP1 foci detection compared to wild-type MDC1, confirming the hierarchical assembly of both proteins. Preliminary data for proton irradiations are shown and also these indicate a delay for 53BP1 versus MDC1.

The role of nonhomologous end joining and homologous recombination in the clonogenic bystander effects of mammalian cells after exposure to counted 10 MeV protons and 4.5 MeV alpha-particles of the PTB microbeam.

We have studied the dependence of clonogenic bystander effects on defects in the pathways of DNA double-strand break (DSB) repair and on linear energy transfer (LET). The single-ion microbeam of the Physikalisch-Technische Bundesanstalt (PTB) was used to irradiate parental Chinese hamster ovary cells or derivatives deficient in nonhomologous end joining (NHEJ) or homologous recombination (HR) in the G1-phase of the cell cycle. Cell nuclei were targeted with 10 MeV protons (LET = 4.7 keV/microm) or 4.5 MeV alpha-particles (LET = 100 keV/microm). During exposure, the cells were confluent, allowing signal transfer through both gap junctions and diffusion. When all cell nuclei were targeted with 10 MeV protons, approximately exponential survival curves were obtained for all three cell lines. When only 10% of all cell nuclei were targeted, a significant bystander effect was observed for parental and HR-deficient cells, but not for NHEJ-deficient cells. For all three cell lines, the survival data after exposure of all cell nuclei to 4.5 MeV alpha-particles could be fitted by exponential curves. When only 10% of all cell nuclei were targeted, significant bystander effects were obtained for parental and HR-deficient cells, whereas for NHEJ-deficient cells a small, but significant, bystander effect was observed only at higher doses. The data suggest that bystander cell killing is a consequence of un- or misrejoined DSB which occur in bystander cells during the S-phase as a result of the processing of oxidative bistranded DNA lesions. The relative contributions of NHEJ and HR to the repairing of DSB in the late S/G2-phase may affect clonogenic bystander effects.