Most of the important biological effects associated with the exposure to ionizing radiations are mirrored at the chromosomal level. In all cases, changes in the levels of cytogenetic effects are associated with changes in absorbed dose, dose rate and radiation quality. Some of the complexities associated with the quantitative description of such changes in response can be circumvented by appealing to concepts embodied in what has been called the “mean inactivation dose”. Additional metrics designed to provide LET-dependent “signatures” of damage have been employed with moderate degrees of success. These, along with some alternative approaches, are discussed in an effort to stimulate discussion, and to further work leading to a better understanding of mechanisms involved in the production and significance of chromosome aberrations after exposure to ionizing radiations.

The development of effective biomarkers for detecting the magnitude of radiation exposure and resiliency of host response is crucial to identifying appropriate treatment strategies after radiation exposure. We hypothesized that the gastrointestinal resident bacteria would demonstrate predictable, dose-dependent changes after radiation exposure across two large animal models of acute radiation syndrome. Here, Göttingen minipigs (GMP) (n = 50) and rhesus macaques (n = 48) were exposed to five dose levels (resulting in mortality rates of 33–100% and 25–68.7%, respectively). Fecal samples taken prior to and after irradiation (day 0 for GMP; day 0, 3 and 14 for macaques) were used for 16S rRNA gene sequence amplicon high-throughput sequencing. Baseline gut microbiota profiles were dissimilar between GMP and macaques, however, radiation appeared to have similar effect at the phylum level, resulting in Bacteroidetes decrease and Firmicutes increase in both models. The abundance of the main Bacteroidetes genus (Bacteroides for GMP, Prevotella for macaques) was profoundly decreased by irradiation. Intracellular symbionts [Elusimicrobia in GMP, Treponema (Spirochaetes) in macaques] consistently increased after irradiation, suggesting their use as potential biomarkers of intestinal injury, and potential negative effect on health. Prevotella, Lactobacillus, Clostridium XIVa, Oscillibacter and Elusimicrobium/Treponema abundances were found to be very significantly correlated with radiation intensity. Furthermore, Prevotella, Enterorhabdus and Ruminococcus and Enterorhabdus maintenance was strongly associated with survival in GMP, while Prevotella, Oscillibacter and Treponema were strongly associated with survival and Streptococcus with death in macaques. Overall, we found that a wide range of gut bacterial genera known to be abundant in the human gut microbiota are excellent biomarkers of radiation intensity and resilience in animal models, and that detrimental effects can be monitored, and potentially prevented, by targeting selected genera.

There have been numerous published studies reporting on the extent of genetic damage observed in animal and human cells exposed in vitro and in vivo to non-ionizing radiofrequency fields (RF, electromagnetic waves that carry energy as they propagate in air and dense media). Overall, the data are inconsistent; while some studies have suggested significantly increased damage in cells exposed to RF energy compared to unexposed and/or sham-exposed control cells, others have not. Several variables in exposure conditions used in the experiments might have contributed to the controversy. In this comprehensive review, four specific quality control measures were used to determine the quality of 225 published studies in animal and human cells exposed in vitro and in vivo to RF energy, and the results from 2,160 tests with different sample sizes were analyzed. The four specific quality control measures were as follows: 1. “Blind” collection/analysis of the data to eliminate individual/observer “bias”; 2. Adequate description of “dosimetry” for independent replication/confirmation; 3. Inclusion of “positive controls” to confirm the outcomes; and 4. Inclusion of “sham-exposed controls” which are more appropriate to compare the data with those in RF exposure conditions. In addition, meta-analysis of the genetic damage in cells exposed to RF energy and control cells, thus far available in the RF literature database, was performed to obtain the “d” values, i.e., standardized mean difference between these two types of cells or the effect size. The relationship between d values and the above-mentioned quality control measures was ascertained. In addition, the correlation between the quality control measures and the conclusions reported in the publications (no significant difference between the cells exposed to RF energy and control cells; increased damage in former cells compared to the latter; increased, no significant difference and decreased damage in cells exposed to RF energy in the same experiment; or decreased damage in cells exposed to RF energy) was examined. The overall conclusions were as follows: 1. When all four quality control measures were mentioned in the publication, the d values were smaller compared to those when one or more quality control measures were not mentioned in the investigation; 2. Based on the inclusion of quality control measures, the weighted outcome in cells exposed to RF energy (d values) indicated a very small effect, if any; 3. The number of published studies reporting no significant difference in genetic damage of cells exposed to RF energy, compared to that of control cells, increased with increased number of quality control measures employed in investigations; 4. The number of published studies reporting increased genetic damage in cells exposed to RF energy decreased with increased number of quality control measures; and 5. There was a “bias” towards the publications reporting increased genetic damage in cells exposed to RF energy even with very small sample size. Overall, the results from this study underscore the importance of including quality control measures in investigations so that the resulting data are useful, nationally and internationally, in evaluating “potential” health risks from exposure to RF energy.

Several investigators performing bone marrow transplantation studies have previously reported sporadic increases in mortality that were associated with pronounced swelling in the face, head and neck of mice. Over the past few years, we and others have noted an increasing number of experiments in which mice that have received total-body irradiation (TBI) or partial-body irradiation (PBI) develop swollen muzzles, drastic thickening of the upper lip and redness, bruising and/or swelling around the nose and muzzle and sometimes over the top of the head. We refer to this rapid and extreme swelling after irradiation as swollen muzzle syndrome (SMS). The development of SMS postirradiation is associated with morbidity that occurs earlier than would be expected from the traditional hematopoietic acute radiation syndrome (HARS), and has impeded studies in several laboratories attempting to evaluate medical countermeasures (MCM) against radiation. However, little has been done to characterize this somewhat unpredictable radiation effect. To investigate the cause and etiology of SMS, data from three different laboratories collected over a seven-year period from 100 MCM 30-day survival studies using mice from different vendors were retrospectively analyzed to determine the time of onset, progression and incidence of SMS in male and female mice exposed to various doses of ionizing radiation. An additional study compared incidence and etiology of SMS in mice from two different vendors (identified as vendors A and B) after exposure to the LD_50/30 (X rays). Mice presenting with SMS, as well as non-SMS (irradiated) control mice, were necropsied to determine microbial status of the blood, heart, spleen, liver, kidney and muzzle tissue. Only mice from vendor A (20%) developed SMS. While the number of bacterial species isolated from various tissues of SMS and non-SMS mice was not different, the number of tissues positive for bacteria was significantly greater in SMS mice. At least one tissue in 83% of SMS mice from vendor A tested positive for Streptococcus agalactiae [group B beta Streptococcus (GBS)], compared to 25% of non-SMS mice from vendor A, and 0% of non-SMS mice from vendor B. In addition, all mice from vendor A with SMS had at least one tissue with >10 ⁴ CFU/g, with GBS as the predominant bacterium, compared to only 25% of non-SMS mice from vendor A, and 0% of non-SMS mice from vendor B. The incidence and magnitude of GBS growth in cultures correlated with the onset of SMS; the earliest and heaviest infections occurred in mice presenting with SMS on days 5–6 postirradiation. The majority of SMS mice (5 out of 6) had positive blood cultures, with the same bacterial strain isolated from other tissues, suggesting systemic translocation via the bloodstream. We propose that testing of mice and the identification of the microorganisms frequently associated with SMS may provide guidance for selection of antimicrobials for use by other investigators in studies evaluating potential MCM, and for the ordering, handling and care of immunodeficient mice or mice that are to be rendered immunodeficient after acute irradiation.

Pharmacologic ascorbate (P-AscH^–) is emerging as a promising adjuvant for advanced pancreatic cancer. P-AscH^– generates hydrogen peroxide (H₂O₂), leading to selective cancer cell cytotoxicity. Catalytic manganoporphyrins, such as MnT4MPyP, can increase the rate of oxidation of P-AscH^–, thereby increasing the flux of H₂O₂, resulting in increased cytotoxicity. We hypothesized that a multimodal treatment approach, utilizing a combination of P-AscH^–, ionizing radiation and MnT4MPyP, would result in significant flux of H₂O₂ and pancreatic cancer cytotoxicity. P-AscH^– with MnT4MPyP increased the rate of oxidation of P-AscH^– and produced radiosensitization in all pancreatic cancer cell lines tested. Three-dimensional (3D) cell cultures demonstrated resistance to P-AscH^–, radiation or MnT4MPyP treatments alone; however, combined treatment with P-AscH^– and MnT4MPyP resulted in the inhibition of tumor growth, particularly when also combined with radiation. In vivo experiments using a murine model demonstrated an increased rate of ascorbate oxidation when combinations of P-AscH^– with MnT4MPyP were given, thus acting as a radiosensitizer. The translational potential was demonstrated by measuring increased ascorbate oxidation ex vivo, whereby MnT4MPyP was added exogenously to plasma samples from patients treated with P-AscH^– and radiation. Combination treatment utilizing P-AscH^–, manganoporphyrin and radiation results in significant cytotoxicity secondary to enhanced ascorbate oxidation and an increased flux of H₂O₂. This multimodal approach has the potential to be an effective treatment for pancreatic ductal adenocarcinoma.

In this study, we sought to determine how diffusionweighted imaging (DWI) and proton magnetic resonance spectroscopy (¹H-MRS) features are associated with histopathological results, and explored the cellular mechanisms of DWI and ¹H-MRS in early radiosensitivity of transplanted liver tumors. VX2 tumors were implanted into the hind leg muscles of 60 New Zealand White Rabbits. All rabbits were randomly divided into ten subgroups according to treatment: irradiated or nonirradiated and according to different times postirradiation. Magnetic resonance scanning was then performed one day before irradiation and on days 1, 3, 5 and 7 postirradiation. Differences in tumor volume, apparent diffusion coefficient (ADC) value, choline/creatine ratio and lipid/creatine ratio, and their associations with histopathological findings, were assessed. Tumor volumes in the irradiated groups were smaller than control values, while ADC values increased gradually with time postirradiation; choline/creatine ratios were reduced while lipid/creatine ratios were larger compared to control values. Bax protein levels after irradiation increased with time. Interestingly, the ADC value and Bax-positive grade showed the same increasing trend (r = 0.900, P < 0.001). Additionally, choline/creatine and lipid/creatine ratios were respectively significantly associated with Bax-positive grade. Furthermore, significant associations of tumor volume with ADC value, choline/creatine ratio and lipid/creatine ratio were observed. These findings demonstrated that ADC value, choline/creatine ratio and lipid/creatine ratio, indicators of early radiosensitivity, are related to cell apoptosis.

Plutonium is a radiologically significant alpha-particle emitter. The potential for adverse health effects from internal exposures due to plutonium intakes has been recognized since the 1940s. The workforce of the Sellafield nuclear facility (Cumbria, UK), includes one of the world's most important groups of plutonium-exposed workers for studying the potential health risks of this internal exposure. However, for several hundred workers employed at the start of plutonium work at the facility (1952–1963), historical monitoring records based on measurements of urinary excretion of plutonium are not sufficiently reliable to provide the accurate and unbiased exposure assessments needed for epidemiological studies. Consequently, these early workers have had to be excluded from such studies, significantly reducing their power. We constructed a population-specific quantitative job exposure matrix (JEM) to estimate the average intakes of “typical plutonium workers” in this period, from 1952–1963, and assessed its validity and sensitivity to exposure assessment decisions. We conducted internal cross-validation using an a priori 10% extracted sample to evaluate reliability of estimates, explored JEM sensitivity to assumptions in the exposure assessment, and assessed the impact of uncertainty in urinalysis measurements on the precision of annual intake estimates using Markov Chain Monte Carlo (MCMC) methodology. Pairwise correlations (R_P) of estimated (JEM) and measured (10% sample) annual intakes were moderate to high (R_P > 0.4) for 10 out of 13 JEM groups, while absolute differences were <20% for 11 out of 13 JEM groups. There was little evidence of a temporal trend in correlations (P = 0.13) or absolute differences (P = 0.34). The median JEM-derived cumulative intake of 95.2 (IQR, 55.0–130.0) Bq was comparable to those based on alternative assumptions in the exposure assessment (median range, 95.2–100.0 Bq; 75th percentiles, 130.0–146.0 Bq). Measurement error simulation resulted in a 40–60% reduced median cumulative intake but higher maximum cumulative intakes. The JEM finds a balance between reliability and precision that makes it useful for epidemiological purposes and is relatively insensitive to specific choices in the exposure assessment. This JEM will allow the inclusion of workers with longest follow-up and who could not be included up until now in epidemiological studies without introducing significant bias.

The risk of developing radiation-induced lung cancer differs between different strains of mice, but the underlying cause of the strain differences is unknown. Strains of mice also differ in how quickly they repair radiation-induced DNA double-strand breaks (DSBs). We assayed mouse strains from the CcS/Dem recombinant congenic strain set for their efficacy in repairing DNA DSBs during protracted irradiation. We measured unrepaired γ-H2AX radiation-induced foci (RIF), which persisted after chronic 24-h gamma irradiation, as a surrogate marker for repair efficiency in bronchial epithelial cells for 17 of the CcS/Dem strains and the BALB/c founder strain. We observed a very strong correlation (R² = 79.18%, P < 0.001) between the level of unrepaired RIF and radiogenic lung cancer incidence measured in the same strains. Interestingly, spontaneous levels of foci in nonirradiated mice also showed good correlation with lung cancer incidence when incidence data from male and female mice were combined. These results suggest that genetic differences in DNA repair capacity largely account for differing susceptibilities to radiationinduced lung cancer among CcS/Dem mouse strains, and that high levels of spontaneous DNA damage are also a relatively good marker of cancer predisposition. In a smaller pilot study, we found that the repair capacity measured in peripheral blood leucocytes also correlated well with radiogenic lung cancer susceptibility, raising the possibility that the assay could be used to detect radiogenic lung cancer susceptibility in humans.

Our understanding of radiation-induced cellular damage has greatly improved over the past few decades. Despite this progress, there are still many obstacles to fully understand how radiation interacts with biologically relevant cellular components, such as DNA, to cause observable end points such as cell killing. Damage in DNA is identified as a major route of cell killing. One hurdle when modeling biological effects is the difficulty in directly comparing results generated by members of different research groups. Multiple Monte Carlo codes have been developed to simulate damage induction at the DNA scale, while at the same time various groups have developed models that describe DNA repair processes with varying levels of detail. These repair models are intrinsically linked to the damage model employed in their development, making it difficult to disentangle systematic effects in either part of the modeling chain. These modeling chains typically consist of track-structure Monte Carlo simulations of the physical interactions creating direct damages to DNA, followed by simulations of the production and initial reactions of chemical species causing so-called “indirect” damages. After the induction of DNA damage, DNA repair models combine the simulated damage patterns with biological models to determine the biological consequences of the damage. To date, the effect of the environment, such as molecular oxygen (normoxic vs. hypoxic), has been poorly considered. We propose a new standard DNA damage (SDD) data format to unify the interface between the simulation of damage induction in DNA and the biological modeling of DNA repair processes, and introduce the effect of the environment (molecular oxygen or other compounds) as a flexible parameter. Such a standard greatly facilitates intermodel comparisons, providing an ideal environment to tease out model assumptions and identify persistent, underlying mechanisms. Through inter-model comparisons, this unified standard has the potential to greatly advance our understanding of the underlying mechanisms of radiation-induced DNA damage and the resulting observable biological effects when radiation parameters and/or environmental conditions change.

To date, the response activated in melanocytes by repeated genotoxic insults from radiotherapy has not been explored. We hypothesized that the molecular pathways involved in the response of melanocytes to ionizing radiation and ultraviolet radiation (UVR) are similar. Skin punch biopsies, not sunexposed, were collected from prostate cancer patients before, as well as at 1 and 6.5 weeks after daily doses of 0.05–1.1 Gy. Interfollicular melanocytes were identified by ΔNp63- and eosin-periodic acid Schiff staining. Immunohistochemistry and immunofluorescence were performed to detect molecular markers of the melanocyte lineage. Melanocytes were negative for ΔNp63, and the number remained unchanged over the treatment period. At radiation doses as low as 0.05 Gy, melanocytes express higher protein levels of microphthalmia-associated transcription factor (MITF) and Bcl-2. Subsets of MITF- and Bcl-2-negative melanocytes were identified among interfollicular melanocytes in unexposed skin; the cell number in both subsets was reduced after irradiation in a way that indicates low-dose hyperradiosensitivity. A corresponding increase in MITF- and Bcl-2-positive cells was observed. PAX3 and SOX10 co-localized to some extent with MITF in unexposed skin, more so with radiation exposure. Low doses of ionizing radiation also intensified c-KIT and DCT staining. Nuclear p53 and p21 were undetectable in melanocytes. Apoptosis and proliferation could not be observed. In conclusion, undifferentiated interfollicular melanocytes were identified, and responded with differentiation in a hypersensitive manner at 0.05 Gy doses. Radioresistance regarding cell death was maintained up to fractionated doses of 1.1 Gy, applied for 7 weeks. The results suggest that the initial steps of melanin synthesis are common to ionizing radiation and UVR, and underline the importance of keratinocyte-melanocyte interaction behind hyperpigmentation and depigmentation to radiotherapy.