Possible association of G6PC2 and MUC6 induced by low­dose­rate irradiation in mouse intestine with inflammatory bowel disease.

Although there are several types of radiation exposure, it is debated whether low­dose­rate (LDR) irradiation (IR) affects the body. Since the small intestine is a radiation­sensitive organ, the present study aimed to evaluate how it changes when exposed to LDR IR and identify the genes sensitive to these doses. After undergoing LDR (6.0 mGy/h) γ radiation exposure, intestinal RNA from BALB/c mice was extracted 1 and 24 h later. Mouse whole genome microarrays were used to explore radiation­induced transcriptional alterations. Reverse transcription­quantitative (RT­q) PCR was used to examine time­ and dose­dependent radiation responses. The histopathological status of the jejunum in the radiated mouse was not changed by 10 mGy of LDR IR; however, 23 genes were upregulated in response to LDR IR of the jejunum in mice after 1 and 24 h of exposure. Upregulated genes were selected to validate the results of the RNA sequencing analysis for RT­qPCR detection and results showed that only Na+/K+ transporting subunit α4, glucose­6­phosphatase catalytic subunit 2 (G6PC2), mucin 6 (MUC6) and transient receptor potential cation channel subfamily V member 6 levels significantly increased after 24 h of LDR IR. Furthermore, G6PC2 and MUC6 were notable genes induced by LDR IR exposure according to protein expression via western blot analysis. The mRNA levels of G6PC2 and MUC6 were significantly elevated within 24 h under three conditions: i) Exposure to LDR IR, ii) repeated exposure to LDR IR and iii) exposure to LDR IR in the presence of inflammatory bowel disease. These results could contribute to an improved understanding of immediate radiation reactions and biomarker development to identify radiation­susceptible individuals before histopathological changes become noticeable. However, further investigation into the specific mechanisms involving G6PC2 and MUC6 is required to accomplish this.

Quantitative and qualitative mutational impact of ionizing radiation on normal cells.

The comprehensive genomic impact of ionizing radiation (IR), a carcinogen, on healthy somatic cells remains unclear. Using large-scale whole-genome sequencing (WGS) of clones expanded from irradiated murine and human single cells, we revealed that IR induces a characteristic spectrum of short insertions or deletions (indels) and structural variations (SVs), including balanced inversions, translocations, composite SVs (deletion-insertion, deletion-inversion, and deletion-translocation composites), and complex genomic rearrangements (CGRs), including chromoplexy, chromothripsis, and SV by breakage-fusion-bridge cycles. Our findings suggest that 1 Gy IR exposure causes an average of 2.33 mutational events per Gb genome, comprising 2.15 indels, 0.17 SVs, and 0.01 CGRs, despite a high level of inter-cellular stochasticity. The mutational burden was dependent on total irradiation dose, regardless of dose rate or cell type. The findings were further validated in IR-induced secondary cancers and single cells without clonalization. Overall, our study highlights a comprehensive and clear picture of IR effects on normal mammalian genomes.

Low dose rate radiation impairs early follicles in young mice.

Low-dose radiation is generally considered less harmful than high-dose radiation. However, its impact on ovaries remains debated. Since previous reports predominantly employed low-dose radiation delivered at a high dose rate on the ovary, the effect of low-dose radiation at a low dose rate on the ovary remains unknown. We investigated the effect of low-dose ionizing radiation delivered at a low dose rate on murine ovaries. Three- and ten-week-old mice were exposed to 0.1 and 0.5 Gy of radiation at a rate of 6 mGy/h and monitored after 3 and 30 days. While neither body weight nor ovarian area showed significant changes, ovarian cells were damaged, showing apoptosis and a decrease in cell proliferation after exposure to 0.1 and 0.5 Gy radiation. Follicle numbers decreased over time in both age groups proportionally to the radiation dose. Younger mice were more susceptible to radiation damage, as evidenced by decreased follicles in 3-week-old mice after 30 days of 0.1 Gy exposure, while 10-week-old mice showed reduced follicles only following 0.5 Gy exposure. Primordial or primary follicles were the most vulnerable to radiation. These findings suggest that even low-dose radiation, delivered at a low dose rate, can adversely affect ovarian function, particularly in the early follicles of younger mice.

Biological dosimetry dose-response curves for residents living near nuclear power plants in South Korea.

The purpose of this study was to establish the dose-response curves for biological dosimetry of the Dong Nam Institute of Radiological and Medical Sciences to monitor radiation exposure of local residents in the vicinity of the nuclear power plant. The blood samples of five healthy volunteers were irradiated with gamma ray, and each sample was divided equally for analysis of chromosomal aberrations by Giemsa staining and three-color fluorescence in situ hybridization painting of the triplet (chromosomes #1, #2, and #4). The results of chromosomal aberrations followed the Poisson distribution in all individual and averaged data which include inter-individual variation in radiation susceptibility. Cytogenetics Dose Estimate Software version 5.2 was used to fit the dose-response curve and to determine the coefficients of linear-quadratic equations. The goodness of fit of the curves and statistical significance of fitted α and ß-coefficients were confirmed in both Giemsa-based dicentric analysis and FISH-based translocation analysis. The coefficients calculated from the five-donor average data were almost identical in both of the analyses. We also present the results that the dose-response curve for dicentric chromosomes plus fragments could be more effective for dose estimation following low-dose radiation accidents.

Dicentric chromosome assay using a deep learning-based automated system.

The dicentric chromosome assay is the "gold standard" in biodosimetry for estimating radiation exposure. However, its large-scale deployment is limited owing to its time-consuming nature and requirement for expert reviewers. Therefore, a recently developed automated system was evaluated for the dicentric chromosome assay. A previously constructed deep learning-based automatic dose-estimation system (DLADES) was used to construct dose curves and calculate estimated doses. Blood samples from two donors were exposed to cobalt-60 gamma rays (0-4 Gy, 0.8 Gy/min). The DLADES efficiently identified monocentric and dicentric chromosomes but showed impaired recognition of complete cells with 46 chromosomes. We estimated the chromosome number of each "Accepted" sample in the DLADES and sorted similar-quality images by removing outliers using the 1.5IQR method. Eleven of the 12 data points followed Poisson distribution. Blind samples were prepared for each dose to verify the accuracy of the estimated dose generated by the curve. The estimated dose was calculated using Merkle's method. The actual dose for each sample was within the 95% confidence limits of the estimated dose. Sorting similar-quality images using chromosome numbers is crucial for the automated dicentric chromosome assay. We successfully constructed a dose-response curve and determined the estimated dose using the DLADES.

Differential Effects of Low and High Radiation Dose Rates on Mouse Spermatogenesis.

The adverse effects of radiation are proportional to the total dose and dose rate. We aimed to investigate the effects of radiation dose rate on different organs in mice. The mice were subjected to low dose rate (LDR, ~3.4 mGy/h) and high dose rate (HDR, ~51 Gy/h) radiation. LDR radiation caused severe tissue toxicity, as observed in the histological analysis of testis. It adversely influenced sperm production, including sperm count and motility, and induced greater sperm abnormalities. The expression of markers of early stage spermatogonial stem cells, such as Plzf, c-Kit, and Oct4, decreased significantly after LDR irradiation, compared to that following exposure of HDR radiation, in qPCR analysis. The compositional ratios of all stages of spermatogonia and meiotic cells, except round spermatid, were considerably reduced by LDR in FACS analysis. Therefore, LDR radiation caused more adverse testicular damage than that by HDR radiation, contrary to the response observed in other organs. Therefore, the dose rate of radiation may have differential effects, depending on the organ; it is necessary to evaluate the effect of radiation in terms of radiation dose, dose rate, organ type, and other conditions.

An intercomparison exercise to compare scoring criteria and develop image databank for biodosimetry in South Korea.

PURPOSE: Mutual cooperation of biodosimetry laboratories is required for dose assessments of large numbers of people with potential radiation exposure, as in mass casualty accidents. We launched an intercomparison exercise to validate the performance of biodosimetry laboratories in South Korea. MATERIALS AND METHODS: Participating laboratories shared metaphase images from dicentric chromosome assays (DCAs) and fluorescence in situ hybridization (FISH)-based translocation assays, which were evaluated based on their own scoring protocols. RESULTS: Overall, the coefficient of variation among three laboratories was less than 10% for counting scorable metaphases and chromosomal aberrations. However, there was variation in the interpretation of the International Atomic Energy Agency guidelines for selecting scorable metaphases and identifying chromosomal aberrations. In a technical workshop, scoring discrepancies were extensively discussed in order to harmonize biodosimetry protocols in Korea. In addition, metaphase images with agreement among all participating laboratories were compiled into an image databank, which can be used for education and training of scorers. CONCLUSIONS: These findings and exercises may improve the accuracy of dose assessment, as well as increase the capacity for biodosimetry in South Korea.

Feasibility Study on Automatic Interpretation of Radiation Dose Using Deep Learning Technique for Dicentric Chromosome Assay.

The interpretation of radiation dose is an important procedure for both radiological operators and persons who are exposed to background or artificial radiations. Dicentric chromosome assay (DCA) is one of the representative methods of dose estimation that discriminates the aberration in chromosomes modified by radiation. Despite the DCA-based automated radiation dose estimation methods proposed in previous studies, there are still limitations to the accuracy of dose estimation. In this study, a DCA-based automated dose estimation system using deep learning methods is proposed. The system is comprised of three stages. In the first stage, a classifier based on a deep learning technique is used for filtering the chromosome images that are not appropriate for use in distinguishing the chromosome; 99% filtering accuracy was achieved with 2,040 test images. In the second stage, the dicentric rate is evaluated by counting and identifying chromosomes based on the Feature Pyramid Network, which is one of the object detection algorithms based on deep learning architecture. The accuracies of the neural networks for counting and identifying chromosomes were estimated at over 97% and 90%, respectively. In the third stage, dose estimation is conducted using the dicentric rate and the dose-response curve. The accuracies of the system were estimated using two independent samples; absorbed doses ranging from 1- 4 Gy agreed well within a 99% confidential interval showing highest accuracy compared to those in previous studies. The goal of this study was to provide insights towards achieving complete automation of the radiation dose estimation, especially in the event of a large-scale radiation exposure incident.

Gonad dose assessment in paediatric kidney nuclear medicine test using Monte Carlo simulation.

In this study, we evaluated the effect of radiation dose on gonads during paediatric kidney nuclear medicine tests. Using Monte Carlo simulations, the distribution and effects of radiation were physically evaluated by displaying the distribution path of the source in the human body over time. In particular, the evaluation of doses in children, who are sensitive to radiation during nuclear medicine tests that use internal exposure among several types of medical exposures, was conducted to obtain data for the management of medical exposures. Our results indicated that under normal kidney function, the dose received by the target kidney was 0.430 mGy/mCi, which is ~6% higher than the dose suggested by the International Commission on Radiation Protection (ICRP). Furthermore, when kidney function was compromised, the dose estimated was 0.726 mGy/mCi, which is ~2% lower than the dose suggested by the ICRP. In the male and female gonads, namely the testicles and ovaries, the doses received were 0.359 mGy/mCi and 0.394 mGy/mCi, respectively, under normal kidney function. Similarly, under abnormal kidney function, the doses ranged from 0.187 to 0.353 mGy/mCi and 0.238 to 0.388 mGy/mCi in the male and female gonads, respectively.

The resveratrol analogue, HS­1793, enhances the effects of radiation therapy through the induction of anti­tumor immunity in mammary tumor growth.

Radiotherapy can induce the infiltration of immune suppressive cells which are involved in promoting tumor progression and recurrence. A number of natural products with immunomodulating abilities have been gaining attention as complementary cancer treatments. This attention is partly due to therapeutic strategies which have proven to be ineffective as a result of tumor­induced immunosuppressive cells found in the tumor microenvironment. The present study investigated whether HS­1793, a resveratrol analogue, can enhance the antitumor effects by inhibiting lymphocyte damage and immune suppression by regulatory T cells (Tregs) and tumor­associated macrophages (TAMs), during radiation therapy. FM3A cells were used to determine the role of HS­1793 in the radiation­induced tumor immunity of murine breast cancer. HS­1793 treatment with radiation significantly increased lymphocyte proliferation with concanavalin A (Con A) stimulation and reduced the DNA damage of lymphocytes in irradiated tumor­bearing mice. The administration of HS­1793 also decreased the number of Tregs, and reduced interleukin (IL)­10 and transforming growth factor (TGF)­ß secretion in irradiated tumor­bearing mice. In addition, HS­1793 treatment inhibited CD206+ TAM infiltration in tumor tissue when compared to the controls or irradiation alone. Mechanistically, HS­1793 suppressed tumor growth via the activation of effector T cells in irradiated mice. On the whole, the findings of the present study reveal that HS­1793 treatment improves the outcome of radiation therapy by enhancing antitumor immunity. Indeed, HS­1793 appears to be a good therapeutic candidate for use in combination with radiotherapy in breast cancer.

Low-dose irradiation promotes Rad51 expression by down-regulating miR-193b-3p in hepatocytes.

Current evidence indicates that there is a relationship between microRNA (miRNA)-mediated gene silencing and low-dose irradiation (LDIR) responses. Here, alterations of miRNA expression in response to LDIR exposure in male BALB/c mice and three different types of hepatocytes were investigated. The miRNome of the LDIR-exposed mouse spleens (0.01 Gy, 6.5 mGy/h) was analyzed, and the expression of miRNA and mRNA was validated by qRT-PCR. Western blotting, chromatin immunoprecipitation (ChIP), and luciferase assays were also performed to evaluate the interaction between miRNAs and their target genes and to gain insight into the regulation of miRNA expression. The expression of miRNA-193b-3p was down-regulated in the mouse spleen and liver and in various hepatocytes (NCTC, Hepa, and HepG2 cell lines) in response to LDIR. The down-regulation of miR-193b-3p expression was caused by histone deacetylation on the miR-193b-3p promoter in the HepG2 cells irradiated with 0.01 Gy. However, the alteration of histone deacetylation and miR-193b-3p and Rad51 expression in response to LDIR was restored by pretreatment with N-acetyl-cyctein. In conclusion, we provide evidence that miRNA responses to LDIR include the modulation of cellular stress responses and repair mechanisms.