RENEB Inter-Laboratory Comparison 2021: Inter-Assay Comparison of Eight Dosimetry Assays.

Tools for radiation exposure reconstruction are required to support the medical management of radiation victims in radiological or nuclear incidents. Different biological and physical dosimetry assays can be used for various exposure scenarios to estimate the dose of ionizing radiation a person has absorbed. Regular validation of the techniques through inter-laboratory comparisons (ILC) is essential to guarantee high quality results. In the current RENEB inter-laboratory comparison, the performance quality of established cytogenetic assays [dicentric chromosome assay (DCA), cytokinesis-block micronucleus assay (CBMN), stable chromosomal translocation assay (FISH) and premature chromosome condensation assay (PCC)] was tested in comparison to molecular biological assays [gamma-H2AX foci (gH2AX), gene expression (GE)] and physical dosimetry-based assays [electron paramagnetic resonance (EPR), optically or thermally stimulated luminescence (LUM)]. Three blinded coded samples (e.g., blood, enamel or mobiles) were exposed to 0, 1.2 or 3.5 Gy X-ray reference doses (240 kVp, 1 Gy/min). These doses roughly correspond to clinically relevant groups of unexposed to low exposed (0-1 Gy), moderately exposed (1-2 Gy, no severe acute health effects expected) and highly exposed individuals (>2 Gy, requiring early intensive medical care). In the frame of the current RENEB inter-laboratory comparison, samples were sent to 86 specialized teams in 46 organizations from 27 nations for dose estimation and identification of three clinically relevant groups. The time for sending early crude reports and more precise reports was documented for each laboratory and assay where possible. The quality of dose estimates was analyzed with three different levels of granularity, 1. by calculating the frequency of correctly reported clinically relevant dose categories, 2. by determining the number of dose estimates within the uncertainty intervals recommended for triage dosimetry (±0.5 Gy or ±1.0 Gy for doses <2.5 Gy or >2.5 Gy), and 3. by calculating the absolute difference (AD) of estimated doses relative to the reference doses. In total, 554 dose estimates were submitted within the 6-week period given before the exercise was closed. For samples processed with the highest priority, earliest dose estimates/categories were reported within 5-10 h of receipt for GE, gH2AX, LUM, EPR, 2-3 days for DCA, CBMN and within 6-7 days for the FISH assay. For the unirradiated control sample, the categorization in the correct clinically relevant group (0-1 Gy) as well as the allocation to the triage uncertainty interval was, with the exception of a few outliers, successfully performed for all assays. For the 3.5 Gy sample the percentage of correct classifications to the clinically relevant group (≥2 Gy) was between 89-100% for all assays, with the exception of gH2AX. For the 1.2 Gy sample, an exact allocation to the clinically relevant group was more difficult and 0-50% or 0-48% of the estimates were wrongly classified into the lowest or highest dose categories, respectively. For the irradiated samples, the correct allocation to the triage uncertainty intervals varied considerably between assays for the 1.2 Gy (29-76%) and 3.5 Gy (17-100%) samples. While a systematic shift towards higher doses was observed for the cytogenetic-based assays, extreme outliers exceeding the reference doses 2-6 fold were observed for EPR, FISH and GE assays. These outliers were related to a particular material examined (tooth enamel for EPR assay, reported as kerma in enamel, but when converted into the proper quantity, i.e. to kerma in air, expected dose estimates could be recalculated in most cases), the level of experience of the teams (FISH) and methodological uncertainties (GE). This was the first RENEB ILC where everything, from blood sampling to irradiation and shipment of the samples, was organized and realized at the same institution, for several biological and physical retrospective dosimetry assays. Almost all assays appeared comparably applicable for the identification of unexposed and highly exposed individuals and the allocation of medical relevant groups, with the latter requiring medical support for the acute radiation scenario simulated in this exercise. However, extreme outliers or a systematic shift of dose estimates have been observed for some assays. Possible reasons will be discussed in the assay specific papers of this special issue. In summary, this ILC clearly demonstrates the need to conduct regular exercises to identify research needs, but also to identify technical problems and to optimize the design of future ILCs.

Application of EPR dosimetry in bone for ex vivo measurements of doses in radiotherapy patients.

In the present study, bone samples from three patients treated in radiotherapy facilities in Poland were used for the determination of doses absorbed during radiotherapy. The samples were obtained during surgical treatments of patients performed due to medical indications. For the purpose of retrospective dosimetry, sensitivity of the radiation-induced EPR signal was individually calibrated in the samples by re-irradiation of the samples with known doses. The doses reconstructed in bones extracted within 6 months after irradiation were consistent with those calculated by treatment planning systems. The dose reconstructed in the bone removed 6 y after radiotherapy was ∼14% lower than the calculated one.

The effect of dose and water treatment on EPR signals in irradiated fingernails.

Fast and precise retrospective dosimetry is crucial in making decisions about medical procedures and safety measures in radiation accidents. Electron paramagnetic resonance (EPR) spectroscopy has a potential as one of available biodosimetry methods for use in victims of such incidents. In this study, authors present the findings on EPR dosimetry in fingernails. Authors describe changes of EPR signals in unirradiated and irradiated nails in time after cutting and the effect of water on the mechanically induced and radiation-induced EPR signals measured ex vivo in the fingernails. The effect of dose on amplitude of the EPR signal was measured in nails that were soaked for 10 min in water after their irradiation. The obtained dose-response curves, which reflect changes in concentration of the radiation-induced RIS5 radicals, reach their maximum for doses of 40-60 Gy.

The effect of dose on light-sensitivity of radicals in alanine EPR dosimeters.

In this work we present a study of light-induced effects on free radicals and their transformations in gamma-irradiated pure L-alanine and in commercially available alanine detectors: rods, pellets and films. Samples irradiated to doses from 2 Gy to 4000 kGy were exposed to light from a fluorescent lamp and to ordinary daylight. The observed changes in EPR spectra of the samples were analyzed with regard to their intensity and shape. The shape analysis was based on numerical decomposition of the measured spectra into model spectra reflecting contributions of R1, R2 and R3 radical populations in the samples. The illumination of alanine dosimeters resulted in significant decrease of the central EPR line and was accompanied by distinct variations in the shape of EPR spectra. The rate of light-induced decay in spectra amplitude was found to be dependent on dose of ionizing radiation--the sensitivity to light was decreasing with increase in dose in all detectors in the 2-5x10(5) Gy dose range. The exposure of gamma-irradiated (to 300 Gy) alanine to normal, diffused daylight resulted in decay of the signal amplitude at rate about 0.5% per week. It was shown that decay in the R1 component was responsible for the observed reduction of the spectra amplitude. The observed increase in R2 contributions in samples exposed to light confirmed a hypothesis of R-->R2 radical transformations promoted by visible light. The reported effects indicate a necessity of protection of irradiated dosimeters from their prolonged exposure to light.

Lessons of the 3rd international intercomparison on EPR dosimetry with teeth: similarities and differences of two successful techniques.

Despite the considerable improvement in accuracy in comparison with previous intercomparison programmes, the outcome of the recent 3rd International Intercomparison on EPR Tooth Dosimetry has demonstrated that performance of various protocols practised in different laboratories significantly varies. SCRM and MUG took part in this intercomparison with their own versions of EPR dosimetry protocols, demonstrating the good correlation between reconstructed and nominal doses (best result for SCRM and fourth best for MUG) and the lowest both absolute and relative mean deviations from the nominal doses. Although the general results of the 3rd Intercomparison are being discussed elsewhere in this issue by Wieser et al., this presentation is focused on the discussion of the common features of the two techniques, which may have an effect on good performance in dose reconstruction. In addition to the mthods of analysis of the intercomparison results, as used in Wieser et al., SCRM and MUG studied the influence of an additional factor--the selection of the standard of the native signal--on the quality of the dose reconstruction.

The Third International Intercomparison on EPR Tooth Dosimetry: part 2, final analysis.

The objective of the Third International Intercomparison on EPR Tooth Dosimetry was to evaluate laboratories performing tooth enamel dosimetry <300 mGy. Final analysis of results included a correlation analysis between features of laboratory dose reconstruction protocols and dosimetry performance. Applicability of electron paramagnetic resonance (EPR) tooth dosimetry at low dose was shown at two applied dose levels of 79 and 176 mGy. Most (9 of 12) laboratories reported the dose to be within 50 mGy of the delivered dose of 79 mGy, and 10 of 12 laboratories reported the dose to be within 100 mGy of the delivered dose of 176 mGy. At the high-dose tested (704 mGy) agreement within 25% of the delivered dose was found in 10 laboratories. Features of EPR dose reconstruction protocols that affect dosimetry performance were found to be magnetic field modulation amplitude in EPR spectrum recording, EPR signal model in spectrum deconvolution and duration of latency period for tooth enamel samples after preparation.

The 3rd international intercomparison on EPR tooth dosimetry: Part 1, general analysis.

The objective of the 3rd International Intercomparison on Electron Paramagnetic Resonance (EPR) Tooth Dosimetry was the evaluation of laboratories performing tooth enamel dosimetry below 300 mGy. Participants had to reconstruct the absorbed dose in tooth enamel from 11 molars, which were cut into two halves. One half of each tooth was irradiated in a 60Co beam to doses in the ranges of 30-100 mGy (5 samples), 100-300 mGy (5 samples), and 300-900 mGy (1 sample). Fourteen international laboratories participated in this intercomparison programme. A first analysis of the results and an overview of the essential features of methods applied in different laboratories are presented. The relative standard deviation of results of all methods was better than 27% for applied doses in the range of 79-704 mGy. In the analysis of the unirradiated tooth halves 8% of the samples were identified as outliers with additional absorbed dose above background dose.

EPR study of light illumination effects on radicals in gamma-irradiated L-alanine.

Exposure of gamma-irradiated L-alanine samples to sunlight and to light from a regular, fluorescent lamp resulted in significant changes in their EPR resonance patterns, both to spectral shapes and intensities. The experimental EPR spectra were numerically decomposed into three components reflecting contributions of three different radicals (R1-R3) generated by ionizing radiation in alanine. The light exposure caused a decay of the measured EPR signal intensity. For similar light intensities and exposure times the decay was much more pronounced in samples illuminated by sunlight than in samples illuminated by the fluorescent lamp. In both cases light-induced decay of R1 radicals was observed. Sunlight illumination resulted in a moderate decay of R2 radicals and in a doubling of the R3 radical population. On the other hand, fluorescent light caused a significant increase of R2 radicals and did not change the amount of R3 radicals. A quantitative analysis of the variations of the three radical contributions to the total EPR spectra upon fluorescent light exposure suggests a net R1-->R2 free radical transformation. These effects of light on the alanine dosimetric signal should be taken into account in dosimetry protocols, assuring protection of alanine dosimeters from extended exposure to light.

The effect of high-linear energy transfer ions on the electron paramagnetic resonance signal induced in alanine.

Microcrystalline samples of L-alanine irradiated with energetic high-LET cobalt and iron ions had different EPR spectra compared to alanine samples irradiated with low-LET photons. The differences in the shapes of the EPR spectra and their dependence on the microwave power are related to the differences in the microwave power saturation of the radicals induced by the various types of ionizing radiation. The changes in the shape of the EPR spectra, which were caused by increasing microwave power, were more pronounced in samples irradiated with low-LET radiation than with high-LET particles. This effect showed a long-term stability and can be used to monitor radiation quality.

The effects of boron on the electron paramagnetic resonance spectra of alanine irradiated with thermal neutrons.

The effects of boric acid admixture on the intensity and line structure of EPR spectra of free radicals produced in alanine by thermal neutrons are presented. The EPR signal enhancement, up to a factor of 40 depending on the boron concentration, is related to additional energy deposition in alanine crystals by the disintegration products resulting from the capture of a thermal neutron by boron, 10B(n,alpha)7Li. The changes in the shape of the EPR spectra observed by changing the microwave power are due to the differences in the microwave power saturation of the free radicals produced by a low-LET radiation and those produced by the high-LET components of the radiation after the neutron capture reaction.

The effects of dose and radiation quality on the shape and power saturation of the EPR signal in alanine.

Variations in the shape and the power saturation of EPR spectra of L-alanine irradiated with photons, electrons, neutrons and protons are reported. It is shown that the ratio of the intensities of the satellite lines attributable to "spin flips" and the central line depend on the EPR microwave power, and it is proposed as a quantitative measure of the signal saturation effect. Dependence of this ratio on the microwave power is affected by the radiation quality and for doses in excess of 10 kGy by the absorbed dose. At high doses of low-LET radiation these changes are attributed to a high local density of free radicals, while for low doses of high-LET radiation these are due to changes induced in the crystal lattice. Consequently, the conventional peak-to-peak amplitude measurement of the EPR signal intensity is inaccurate when used for high doses and for comparison between radiations with different beam quality. The possibility to determine radiation quality from an EPR measurement is discussed.

Energy response of agar-alanine free radical dosimetry to therapeutic electron beams.

Calculations of the energy response of an agar-alanine phantom dosimeter (AAPD) to electrons in the energy range of 0.15-20 MeV together with experimental results at 16 MeV are presented. It is shown that the sensitivity of the EPR dosimeter (EPR signal/Gray) is independent of alanine crystal size and varies less than 2%, in the electron energy range indicated. Thus, the measured free radical density distribution may be used directly as an indication of the absorbed dose distribution in an irradiated phantom.

Dose enhancement in buildup region by lead, aluminum, and lucite absorbers for 15 MVp photon beam.

Changes in dose distributions in buildup region resulting from the presence of lead, aluminum, and lucite absorbers above the surface of a polystyrene phantom were evaluated. The surface dose, as a function of the absorber thickness, is presented as well as the influence of the air gap between the lead absorber and the phantom surface. It has been found that the surface dose does not depend on absorber thickness for absorbers thicker than the range of secondary electrons in the absorber material (after corrections for the attenuation of the primary beam in the absorber). Similarly, the depth dose curves in the phantom were elevated only at depths lower than the range of secondary electrons in the phantom. The applicability of the presented data in clinical radiotherapy is discussed.

The energy response of agar-alanine phantom dosimeter to gamma radiation.

Calculations of the energy response of an electron paramagnetic resonance (EPR) signal induced by gamma radiation in an agar-alanine phantom dosimeter are presented. Theoretically calculated slopes of the EPR signal calibration lines are comparable with those obtained experimentally for low-(50 kVp), medium-(662 keV), and high-(15 MVp) energy photons. The sensitivity of the phantom dosimeter (EPR signal amplitude/Gray) varies less than 2% within the 150- to 20-MeV energy range. For energies above 150 keV, the influence of variations in the size of alanine crystals is negligible.

Continuous three-dimensional radiation dosimetry in tissue-equivalent phantoms using electron paramagnetic resonance in L-alpha-alanine.

A new tissue-equivalent phantom material has been developed which also acts as a dosimeter. The new phantom material has a similar elemental composition to that of soft tissue and has a density 1.1 g/cm3. The phantom has an agar-gel base, and contains crystallized L-alpha-alanine which traps radiation-induced free radicals. Samples from the phantom were analyzed by an electron paramagnetic resonance (EPR) spectrometer and the intensity of the EPR signal was related to the absorbed dose. When calibrated, the phantom material acts as a dosimeter, with applications in radiation therapy.