Applicability of Scoring Calyculin A-Induced Premature Chromosome Condensation Objects for Dose Assessment Including for Radiotherapy Patients.

As an extension to a previous study, a linear calibration curve covering doses from 0 to 10 Gy was constructed and evaluated in the present study using calyculin A-induced premature chromosome condensation (PCC) by scoring excess PCC objects. The main aim of this study was to assess the applicability of this PCC assay for doses below 2 Gy that are critical for triage categorization. Two separate blind tests involving a total of 6 doses were carried out; 4 out of 6 dose estimates were within the 95% confidence limits (95% CL) with the other 2 just outside. In addition, blood samples from five cancer patients undergoing external beam radiotherapy (RT) were also analyzed, and the results showed whole-body dose estimates statistically comparable to the dicentric chromosome assay (DCA) results. This is the first time that calyculin A-induced PCC was used to analyze clinical samples by scoring excess objects. Although dose estimates for the pre-RT patient samples were found to be significantly higher than the mean value for the healthy donors and were also significantly higher than those obtained using DCA, all these pre-treatment patients fell into the same category as those who may have received a low dose (<1 Gy) and do not require immediate medical care during emergency triage. Additionally, for radiological accidents with unknown exposure scenario, PCC objects and rings can be scored in parallel for the assessment of both low- and high-dose exposures. In conclusion, scoring excess objects using calyculin A-induced PCC is confirmed to be another potential biodosimetry tool in radiological emergency particularly in mass casualty scenarios, even though the data need to be interpreted with caution when cancer patients are among the casualties.

International Comparison Exercise for Biological Dosimetry after Exposures with Neutrons Performed at Two Irradiation Facilities as Part of the BALANCE Project.

In the case of a radiological or nuclear event, biological dosimetry can be an important tool to support clinical decision-making. During a nuclear event, individuals might be exposed to a mixed field of neutrons and photons. The composition of the field and the neutron energy spectrum influence the degree of damage to the chromosomes. During the transatlantic BALANCE project, an exposure similar to a Hiroshima-like device at a distance of 1.5 km from the epicenter was simulated, and biological dosimetry based on dicentric chromosomes was performed to evaluate the participants ability to discover unknown doses and to test the influence of differences in neutron spectra. In a first step, calibration curves were established by irradiating blood samples with 5 doses in the range of 0-4 Gy at two different facilities in Germany (Physikalisch-Technische Bundesanstalt [PTB]) and the USA (the Columbia IND Neutron Facility [CINF]). The samples were sent to eight participating laboratories from the RENEB network and dicentric chromosomes were scored by each participant. Next, blood samples were irradiated with 4 blind doses in each of the two facilities and sent to the participants to provide dose estimates based on the established calibration curves. Manual and semiautomatic scoring of dicentric chromosomes were evaluated for their applicability to neutron exposures. Moreover, the biological effectiveness of the neutrons from the two irradiation facilities was compared. The calibration curves from samples irradiated at CINF showed a 1.4 times higher biological effectiveness compared to samples irradiated at PTB. For manual scoring of dicentric chromosomes, the doses of the test samples were mostly successfully resolved based on the calibration curves established during the project. For semiautomatic scoring, the dose estimation for the test samples was less successful. Doses >2 Gy in the calibration curves revealed nonlinear associations between dose and dispersion index of the dicentric counts, especially for manual scoring. The differences in the biological effectiveness between the irradiation facilities suggested that the neutron energy spectrum can have a strong impact on the dicentric counts.

Editor's Choice - Estimated Radiation Dose to the Operator During Endovascular Aneurysm Repair.

OBJECTIVE: To estimate operator organ doses from fluoroscopically guided infrarenal endovascular aneurysm repair (EVAR) procedures, using the detailed exposure information contained in radiation dose structured reports. METHODS: Conversion factors relating kerma area product (PKA) to primary operator organ doses were calculated using Monte Carlo methods for 91 beam angles and seven x-ray spectra typical of clinical practice. A computer program was written, which selects the appropriate conversion factor for each exposure listed in a structured report and multiplies it by the respective PKA. This system was used to estimate operator doses for 81 EVAR procedures for which structured reports were available. The impact of different shielding scenarios and variations in operator position was also investigated. RESULTS: Without any shielding, the median estimated effective dose was 113 µSv (interquartile range [IQR] 71, 252 µSv). The highest median organ doses were for the colon (154 µSv, IQR 81, 343) and stomach (133 µSv, IQR 76, 307). These dose estimates represent all exposures, including fluoroscopy and non-fluoroscopic digital acquisitions. With minimal shielding provided by 0.25 mm of Pb covering the torso and upper legs, the effective dose was reduced by a factor of around 6. With additional shielding from ceiling and table shields, a 25 to 50 fold reduction in dose is achievable. Estimated doses were highest where the primary beam was pointed directly away from the operator. CONCLUSION: The models suggest that with optimal use of shielding, operator doses can be reduced to levels equivalent to one to two days of natural background exposure and well below statutory dose limits.

ICRP workshop on the review and revision of the system of radiological protection: a focus on research priorities-feedback from the international community.

In September 2022, the International Commission on Radiological Protection (ICRP) organised a workshop in Estoril, Portugal, on the 'Review and Revision of the System of Radiological Protection: A Focus on Research Priorities'. The workshop, which was a side event of the European Radiation Protection Week, offered an opportunity to comment on a recent paper published by ICRP on areas of research to support the System of Radiological Protection. Altogether, about 150 individuals participated in the workshop. After the workshop, 16 of the 30 organisations in formal relations with ICRP provided written feedback. All participants and organisations followed ICRP's view that further research in various areas will offer additional support in improving the System in the short, medium, and long term. In general, it was emphasised that any research should be outcome-focused in that it should improve protection of people or the environment. Many research topics mentioned by the participants were in line with those already identified by ICRP in the paper noted above. In addition, further ideas were expressed such as, for example, that lessons learned during the COVID-19 pandemic with regards to the non-radiological social, economic and environment impacts, should be analysed for their usefulness to enhance radiological protection, and that current protection strategies and application of current radiological protection principles may need to be adapted to military scenarios like those observed recently during the military conflict in the Ukraine or the detonation of a nuclear weapon. On a broader perspective, it was discussed how radiation research and radiological protection can contribute towards the Sustainable Development Goals announced by the United Nations in 2015. This paper summarises the views expressed during the workshop and the major take home messages identified by ICRP.

Sensitivity and latency of ionising radiation-induced cataract.

When managed with appropriate radiation protection procedures, ionising radiation is of great benefit to society. Opacification of the lens, and vision impairing cataract, have recently been recognised at potential effects of relatively low dose radiation exposure, on the order of 1 Gy or below. Within the last 10 years, understanding of the effects of low dose ionising radiation on the lens has increased, particularly in terms of DNA damage and responses, and how multiple radiation or other events in the lens might contribute to the overall risk of cataract. However, gaps remain, not least in the understanding of how radiation interacts with other risk factors such as aging, as well as the relative radiosensitivity of the lens compared to tissues of the body. This paper reviews the current literature in the field of low dose radiation cataract, with a particular focus on sensitivity and latency.

A faster and easier biodosimetry method based on calyculin A-induced premature chromosome condensation (PCC) by scoring excess objects.

Dicentric analysis and the ring PCC assay as established biodosimetry methods both have limitations in the estimation of absorbed doses in suspected overexposure cases between 5 and 10 Gy. The proposed method based on calyculin A-induced PCC overcomes these limitations by scoring excess objects as the endpoint. This new scoring method can potentially serve as a faster and up-scalable approach that complements the existing methods with higher accuracy at different dose ranges. It can also potentially be performed by less skilled workers when no automated system is available in mass casualty emergency cases to assist with the triage of patients. Additionally, it offers the possibility to further reduce the sample size and PCC induction time. In this pilot study, a calibration curve for excess objects was constructed using the new scoring method for the first time and a blind validation test composed of three unknown doses was carried out. Almost all the dose estimates were within the 95% confidence limits of the actual test doses by scoring only 50-100 PCC spreads. This method was found to be more accurate than ring PCC for doses below 10 Gy.

Accidental neutron exposure in a medical setting: a case study.

In May 2016, a new linear accelerator (Linac) was installed at a hospital oncology department. A team of individuals supervised the installation, including a Radiation Oncologist who acted as an independent observer to the installation, calibration, beam data collection and shielding measurements. In order to ensure the shielding was correct, a licensed representative of the Turkish Atomic Energy Authority carried out formal measurements of the gamma and neutron dose rates at a variety of locations in and around the Linac facility. At 18 MV, the maximum neutron dose rate was 172µSv h-1and the maximum gamma dose rate was approximately 2µSv h-1(ambient dose equivalent in both cases), significantly higher than the expected and local background doses. As the neutron dose rates in particular were so high, it was concluded that the shielding was not sufficient, potentially due to an inadequate design. In order to rule out overexposure during the installation, biological dosimetry was carried out for a number of the individuals involved. The estimated doses were closely aligned with the doses measured using commercially available neutron dosemeters and were also within the tolerance dose ranges estimated using Monte Carlo simulations, which also supported the investigation. The results underline the need for careful planning before and after installation of new radiation exposure facilities, especially high MV Linac operation for which photo-neutrons might need to be mitigated. The results clearly indicate the importance of such checks, in addition to demonstrating the relevance of biological dosimetry supported by modelling strategies complex or unclear exposure scenarios.

Patient radiation dose from x-ray guided endovascular aneurysm repair: a Monte Carlo approach using voxel phantoms and detailed exposure information.

Endovascular aneurysm repair (EVAR) is a well-established minimally invasive technique that relies on x-ray guidance to introduce a stent through the femoral artery and manipulate it into place. The aim of this study was to estimate patient organ and effective doses from EVAR procedures using anatomically realistic computational phantoms and detailed exposure information from radiation dose structured reports (RDSR). Methods: Lookup tables of conversion factors relating kerma area product (PKA) to organ doses for 49 different beam angles were produced using Monte Carlo simulations (MCNPX2.7) with International Commission on Radiological Protection (ICRP) adult male and female voxel phantoms for EVAR procedures of varying complexity (infra-renal, fenestrated/branched and thoracic EVAR). Beam angle specific correction factors were calculated to adjust doses according to x-ray energy. A MATLAB function was written to find the appropriate conversion factor in the lookup table for each exposure described in the RDSR, perform energy corrections and multiply by the respective exposure PKA. Using this approach, organ doses were estimated for 183 EVAR procedures in which RDSRs were available. A number of simplified dose estimation methodologies were also investigated for situations in which RDSR data are not available. Results: Mean estimated bone marrow doses were 57 (range: 2-247), 86 (2-328) and 54 (8-250) mGy for infra-renal, fenestrated/branched and thoracic EVAR, respectively. Respective effective doses were 27 (1-208), 54 (1-180) and 37 (5-167) mSv. Dose estimates using non-individualised, average conversion factors, along with those produced using the alternative Monte Carlo code PCXMC, yielded reasonably similar results overall, though variation for individual procedures could exceed 100% for some organs. In conclusion, radiation doses from x-ray guided endovascular aneurysm repairs are potentially high, though this must be placed in the context of the life sparing nature and high success rate for this procedure.

Quantities for assessing high doses to the body: a short review of the current status.

Tissue reactions are the most clinically significant consequences of high-dose exposures to ionizing radiation. However, there is currently no universally recognized or recommended dose quantity that can be used to assess generalized risks to individuals following whole body exposures in the high-dose range. This is particularly problematic in emergency response situations, for example, following external exposures of large numbers of individuals: in attempts to relate the triage dosemeter absorbed dose to the risk to the individual, such that a 'dose' may subsequently be reported to medical professionals, it is necessary to first agree on the quantity to be reported. The current paper presents a brief review of the likely scenarios and emergency dosimetry techniques that require such a quantity, before examining the biological constraints and requirements that might underpin any future definition. The aim of this work is to outline the arguments for developing a commonly agreed dose quantity for reporting high-dose radiation exposures.

Quantities for assessing high photon doses to the body: a calculational approach.

Tissue reactions are the most clinically significant consequences of high-dose exposures to ionising radiation. However, currently there is no universally recognized dose quantity that can be used to assess and report generalised risks to individuals following whole body exposures in the high-dose range. In this work, a number of potential dose quantities are presented and discussed, with mathematical modelling techniques employed to compare them and explore when their differences are most or least manifest. The results are interpreted to propose the average (D GRB) of the absorbed doses to the stomach, small intestine, red bone marrow, and brain as the optimum quantity for informing assessments of risk. A second, maximally conservative dose quantity (D Max) is also suggested, which places limits on any under-estimates resulting from the adoption of D GRB. The primary aim of this work is to spark debate, with further work required to refine the final choice of quantity or quantities most appropriate for the full range of different potential exposure scenarios.

Chromosome analysis in a case of a plutonium contaminated wound.

Chromosome analysis of peripheral blood lymphocytes was undertaken over a 10 year period following an intake of plutonium through a hand wound. Frequencies of cells with unstable complex aberrations remained high throughout this time, probably reflecting direct exposure of lymphocytes as they passed plutonium which had transferred to regional lymph nodes. Analysis at the final sampling time also revealed cells with stable aberrations at a much higher frequency relative to the number of unstable cells than expected from direct exposure, and is therefore most likely to be reflecting exposure to lymphocyte precursor cells from plutonium that has become deposited on bone surfaces.

Zero-inflated regression models for radiation-induced chromosome aberration data: A comparative study.

Within the field of cytogenetic biodosimetry, Poisson regression is the classical approach for modeling the number of chromosome aberrations as a function of radiation dose. However, it is common to find data that exhibit overdispersion. In practice, the assumption of equidispersion may be violated due to unobserved heterogeneity in the cell population, which will render the variance of observed aberration counts larger than their mean, and/or the frequency of zero counts greater than expected for the Poisson distribution. This phenomenon is observable for both full- and partial-body exposure, but more pronounced for the latter. In this work, different methodologies for analyzing cytogenetic chromosomal aberrations datasets are compared, with special focus on zero-inflated Poisson and zero-inflated negative binomial models. A score test for testing for zero inflation in Poisson regression models under the identity link is also developed.

Verification by the FISH translocation assay of historic doses to Mayak workers from external gamma radiation.

The aim of this study was to apply the fluorescence in situ hybridization (FISH) translocation assay in combination with chromosome painting of peripheral blood lymphocytes for retrospective biological dosimetry of Mayak nuclear power plant workers exposed chronically to external gamma radiation. These data were compared with physical dose estimates based on monitoring with badge dosimeters throughout each person's working life. Chromosome translocation yields for 94 workers of the Mayak production association were measured in three laboratories: Southern Urals Biophysics Institute, Leiden University Medical Center and the former Health Protection Agency of the UK (hereinafter Public Health England). The results of the study demonstrated that the FISH-based translocation assay in workers with prolonged (chronic) occupational gamma-ray exposure was a reliable biological dosimeter even many years after radiation exposure. Cytogenetic estimates of red bone marrow doses from external gamma rays were reasonably consistent with dose measurements based on film badge readings successfully validated in dosimetry system "Doses-2005" by FISH, within the bounds of the associated uncertainties.

radir package: an R implementation for cytogenetic biodosimetry dose estimation.

The Bayesian framework has been shown to be very useful in cytogenetic dose estimation. This approach allows description of the probability of an event in terms of previous knowledge, e.g. its expectation and/or its uncertainty. A new R package entitled radir (radiation inverse regression) has been implemented with the aim of reproducing a recent Bayesian-type dose estimation methodology. radir adopts the method of dose estimation under the Poisson assumption of the responses (the chromosomal aberrations counts) for the required dose-response curve (typically linear or quadratic). The individual commands are described in detail and relevant examples of the use of the methods and the corresponding radir software tools are given. The suitability of this methodology is highlighted and its application encouraged by providing a user-friendly command-type software interface within the R statistical software (version 3.1.1 or higher), which includes a complete manual.

AOP report: Development of an adverse outcome pathway for deposition of energy leading to cataracts.

Cataracts are one of the leading causes of blindness, with an estimated 95 million people affected worldwide. A hallmark of cataract development is lens opacification, typically associated not only with aging but also radiation exposure as encountered by interventional radiologists and astronauts during the long-term space mission. To better understand radiation-induced cataracts, the adverse outcome pathway (AOP) framework was used to structure and evaluate knowledge across biological levels of organization (e.g., macromolecular, cell, tissue, organ, organism and population). AOPs identify a sequence of key events (KEs) causally connected by key event relationships (KERs) beginning with a molecular initiating event to an adverse outcome (AO) of relevance to regulatory decision-making. To construct the cataract AO and retrieve evidence to support it, a scoping review methodology was used to filter, screen, and review studies based on the modified Bradford Hill criteria. Eight KEs were identified that were moderately supported by empirical evidence (e.g., dose-, time-, incidence-concordance) across the adjacent (directly linked) relationships using well-established endpoints. Over half of the evidence to justify the KER linkages was derived from the evidence stream of biological plausibility. Early KEs of oxidative stress and protein modifications had strong linkages to downstream KEs and could be the focus of countermeasure development. Several identified knowledge gaps and inconsistencies related to the quantitative understanding of KERs which could be the basis of future research, most notably directed to experiments in the range of low or moderate doses and dose-rates, relevant to radiation workers and other occupational exposures.

In vitro study of radiosensitivity in colorectal cancer cell lines associated with Lynch syndrome.

Introduction: Lynch syndrome patients have an inherited predisposition to cancer due to a deficiency in DNA mismatch repair (MMR) genes which could lead to a higher risk of developing cancer if exposed to ionizing radiation. This pilot study aims to reveal the association between MMR deficiency and radiosensitivity at both a CT relevant low dose (20 mGy) and a therapeutic higher dose (2 Gy). Methods: Human colorectal cancer cell lines with (dMMR) or without MMR deficiency (pMMR) were analyzed before and after exposure to radiation using cellular and cytogenetic analyses i.e., clonogenic assay to determine cell reproductive death; sister chromatid exchange (SCE) assay to detect the exchange of DNA between sister chromatids; γH2AX assay to analyze DNA damage repair; and apoptosis analysis to compare cell death response. The advantages and limitations of these assays were assessed in vitro, and their applicability and feasibility investigated for their potential to be used for further studies using clinical samples. Results: Results from the clonogenic assay indicated that the pMMR cell line (HT29) was significantly more radio-resistant than the dMMR cell lines (HCT116, SW48, and LoVo) after 2 Gy X-irradiation. Both cell type and radiation dose had a significant effect on the yield of SCEs/chromosome. When the yield of SCEs/chromosome for the irradiated samples (2 Gy) was normalized against the controls, no significant difference was observed between the cell lines. For the γH2AX assay, 0, 20 mGy and 2 Gy were examined at post-exposure time points of 30 min (min), 4 and 24 h (h). Statistical analysis revealed that HT29 was only significantly more radio-resistant than the MLH1-deficient cells lines, but not the MSH2-deficient cell line. Apoptosis analysis (4 Gy) revealed that HT29 was significantly more radio-resistant than HCT116 albeit with very few apoptotic cells observed. Discussion: Overall, this study showed radio-resistance of the MMR proficient cell line in some assays, but not in the others. All methods used within this study have been validated; however, due to the limitations associated with cancer cell lines, the next step will be to use these assays in clinical samples in an effort to understand the biological and mechanistic effects of radiation in Lynch patients as well as the health implications.

CytoBayesJ: software tools for Bayesian analysis of cytogenetic radiation dosimetry data.

A number of authors have suggested that a Bayesian approach may be most appropriate for analysis of cytogenetic radiation dosimetry data. In the Bayesian framework, probability of an event is described in terms of previous expectations and uncertainty. Previously existing, or prior, information is used in combination with experimental results to infer probabilities or the likelihood that a hypothesis is true. It has been shown that the Bayesian approach increases both the accuracy and quality assurance of radiation dose estimates. New software entitled CytoBayesJ has been developed with the aim of bringing Bayesian analysis to cytogenetic biodosimetry laboratory practice. CytoBayesJ takes a number of Bayesian or 'Bayesian like' methods that have been proposed in the literature and presents them to the user in the form of simple user-friendly tools, including testing for the most appropriate model for distribution of chromosome aberrations and calculations of posterior probability distributions. The individual tools are described in detail and relevant examples of the use of the methods and the corresponding CytoBayesJ software tools are given. In this way, the suitability of the Bayesian approach to biological radiation dosimetry is highlighted and its wider application encouraged by providing a user-friendly software interface and manual in English and Russian.

Manual versus automated γ-H2AX foci analysis across five European laboratories: can this assay be used for rapid biodosimetry in a large scale radiation accident?

The identification of severely exposed individuals and reassurance of the 'worried well' are of prime importance for initial triage following a large scale radiation accident. We aim to develop the γ-H2AX foci assay into a rapid biomarker tool for use in accidents. Here, five laboratories established a standard operating procedure and analysed 100 ex vivo γ-irradiated, 4 or 24h incubated and overnight-shipped lymphocyte samples from four donors to generate γ-H2AX reference data, using manual and/or automated foci scoring strategies. In addition to acute, homogeneous exposures to 0, 1, 2 and 4Gy, acute simulated partial body (4Gy to 50% of cells) and protracted exposures (4Gy over 24h) were analysed. Data from all laboratories could be satisfactorily fitted with linear dose response functions. Average yields observed at 4h post exposure were 2-4 times higher than at 24h and varied considerably between laboratories. Automated scoring caused larger uncertainties than manual scoring and was unable to identify partial exposures, which were detectable in manually scored samples due to their overdispersed foci distributions. Protracted exposures were detectable but doses could not be accurately estimated with the γ-H2AX assay. We conclude that the γ-H2AX assay may be useful for rapid triage following a recent acute radiation exposure. The potentially higher speed and convenience of automated relative to manual foci scoring needs to be balanced against its compromised accuracy and inability to detect partial body exposures. Regular re-calibration or inclusion of reference samples may be necessary to ensure consistent results between laboratories or over long time periods.