Chromosome aberration frequencies and chromosome instability in mice after long-term exposure to low-dose-rate gamma-irradiation

Chronological changes of chromosome aberration rates related to accumulated doses in chronically exposed humans and animals at a low-dose-rate have not been well studied. C3H female specific pathogen-free mice (8 weeks of age) were chronically irradiated. Chromosome aberration rate in mouse splenocytes after long-term exposure to low-dose-rate (LDR) gamma-rays was serially determined by conventional Giemsa method. Incidence of dicentrics and centric rings increased almost linearly up to 8000 mGy following irradiation for about 400 days at a LDR of 20 mGy/day. Clear dose-rate effects were observed in the chromosome aberration frequencies between dose rates of 20 mGy/day and 200 Gy/day. Furthermore, the frequencies of complex aberrations increased as accumulated doses increased in LDR irradiation. This trend was also observed for the incidences of micronuclei and trisomies of chromosomes 5, 13 and 18 in splenocytes, detected by micronucleus assay and metaphase fluorescence in situ hybridization (FISH) method, respectively. Incidences of 2-4 micronuclei and trisomy increased in mouse splenocytes after irradiation of 8000 mGy at a LDR of 20 mGy/day. These complex chromosome aberrations and numerical chromosome aberrations seem to be induced indirectly after radiation exposure and thus the results indicate that continuous gamma-ray irradiation for 400 days at LDR of 20 mGy/day induced chromosomal instability in mice. These results are important to evaluate the biological effects of long-term exposure to LDR radiation in humans.     

Chromosome aberration frequencies produced by a 70-MeV proton beam

The effectiveness of a 70-MeV proton beam in the induction of chromosome aberrations was studied. We employed peripheral lymphocytes and analyzed the frequencies of dicentrics and rings after irradiation at doses ranging from 0.1 to 8.0 Gy at various depths within a Lucite phantom. The frequency of chromosome aberrations after irradiation with an unmodulated proton beam at 5 mm showed a dose-response relationship similar to that of 60Co gamma rays. However, irradiation at greater depths with the spread-out Bragg peak induced higher aberration frequencies at doses lower than those with gamma rays. Furthermore, the distribution curve of chromosome aberration frequencies as a function of depth was found to be slightly different from the physically measured depth-dose curve. With the spread-out Bragg peak the biological effects were more marked at greater depths, resulting in a distribution of relative biological effectiveness values. The results obtained from chromosome aberration analysis may not be related directly to those for the relationship between dose and cell killing. Slight differences in values for relative biological effectiveness due to the change of dose and site of proton beam irradiation may not be important for practical proton beam therapy, but may be important in the prevention of late radiation injuries.     

Chromosome aberration peristence in swine leukocytes following exposure to fission neutrons

Chromosome aberration rates in swine lymphocytes decreased rapidly within the first 24 h postirradiation. This initial loss coincided with a decrease in lymphocyte number and continued until lymphocyte numbers began to recover at 7 days after irradiation. Subsequent losses were less dramatic, and numbers of acentric fragments reached preirradiation levels by 56 days postexposure; rings and dicentrics persisted throughout the 84-day study. Mathematical models descrine the predicted aberration yield as a function of time after irradiation. The disappearance rate of aberrant lymphocytes after irradiation suggests that anerration persistence kinetics should be considered when using lymphocyte aberrations as biologic dosimeters.     

~(60)Coγ线引起上海真厉螨染色体畸变的研究

本文首次报道用~(6O)Coγ线照射一种革螨——上海真厉螨引起的染色体畸变的研究。用~(6O)Coγ线(剂量1—50Krad)照射雌性革螨,引起的染色体畸变类型有:染色体裂隙、断片、微小体、环形染色体、粉碎化和多倍体,染色体断片是最常见的畸变类型,并观察到微核的形成。染色体畸变率随照射剂量增加而增高,辐射剂量与畸变率之间存在密切相关(相关系数为0.85,P<0.025),配得曲线回归方程为Y=3.27+14.49lg(X+1)。     

Comparison of three inbred lines of mice by overall and genetic radiosensitivity

The radiosensitivities of three mouse strains (BALB/cLacY, C3H/SnY, and 101/HY) have been compared using the following parameters: survival after irradiation at a dose of 6-7 Gy, chromosome aberration frequency in bone marrow cells after irradiation at a dose of 1.5 Gy, and the change in testis weight and frequency of abnormal sperm heads (ASHs) after irradiation at doses from 0.5 to 4 Gy. Strain BALB/c is the most radiosensitive with respect to the survival and chromosome aberration frequency in the bone marrow but the most resistant with respect to the change in testis weight and the frequency of abnormal sperm heads. Strain 101/HY was the most resistant with respect to survival and chromosome aberration frequency in bone marrow after irradiation but the most radiosensitive with respect to testis damage.     

Comparison of Three Inbred Mouse Strains with Respect to General and Genetic Radiosensitivity

The radiosensitivities of three mouse strains (BALB/cLacY, C3H/SnY, and 101/HY) have been compared using the following parameters: survival after irradiation at a dose of 6–7 Gy, chromosome aberration frequency in bone marrow cells after irradiation at a dose of 1.5 Gy, and the change in testis weight and frequency of abnormal sperm heads (ASHs) after irradiation at doses from 0.5 to 4 Gy. Strain BALB/c is the most radiosensitive with respect to the survival and chromosome aberration frequency in the bone marrow but the most resistant with respect to the change in testis weight and the frequency of abnormal sperm heads. Strain 101/HY was the most resistant with respect to survival and chromosome aberration frequency in bone marrow after irradiation but the most radiosensitive with respect to testis damage.     

The relationship between DNA and chromosome damage after bleomycin treatment: dose-response measurements

The molecular basis for chromosome aberration formation has been studied using the sensitive techniques of premature chromosome condensation and DNA alkaline elution. The dose response of Chinese hamster ovary cells to bleomycin treatment at the DNA and chromosome levels was compared. Each DNA elution curve showed a 2-component profile, with a more sensitive component apparent at low doses. The chromosome aberration curves also exhibited a 2-component profile when determined in G2-PCC; however, this phenomenon was less apparent when chromosome damage was enumerated in mitotic figures. These results suggest that differential sensitivity to bleomycin exists within the cellular chromatin. The effect of dose rate on aberration formation was examined by administering bleomycin at 2 concentrations that, with different treatment times, yielded equivalent amounts of DNA damage. The chromatid exchange rate was independent of dose rate, suggesting that rapidly repaired DNA lesions are not involved in the formation of exchanges.     

激光辐照玉米诱发同工酶及染色体变异的研究

利用ＣＯ２激光辐照玉米种子,分析了酯酶同工酶（ＥＳＴ）和过氧化物酶同工酶（ＰＯＤ）及根尖细胞染色体变异,探讨了苗期损伤与染色体和同工酶变异的关系。结果表明：ＣＯ２激光辐照可诱发ＥＳＴ和ＰＯＤ活性和种类的变化,ＥＳＴ３和ＰＯＤ３随辐照剂量增加活性逐渐减弱以致消失,在根尖细胞中可观察到微核、染色体落后和染色体桥等变异,其畸变率的剂量效应可用Ｙ＝ａ＋ｂｘ来描述。Ｍ１种子发芽率、苗高与同工酶变化大小表现出     

A comparative assessment of the changes in absorbed neutron doses and in the frequency of chromosome aberrations in samples of human blood lymphocytes by the depth of the water phantom during irradiation by the biomedical BR-10 reactor beam

Distribution of the chromosome aberration frequency in human blood lymphocyte samples and absorbed doses have been compared by the water phantom depth during irradiation with 1.5 Gy neutrons (mean energy of 0.85 MeV). There is a good concordance of their depth distribution. The half-fall layer of the absorbed dose within the tissue-equivalent medium is similar (approximately 5 cm) with both measurements done. The aberration frequency in the biological samples placed outside the radiation field in the phantom increases which indicates that the neutron been bounds are indistinct upon passing the tissue-equivalent medium.     

X-ray-induced chromosome aberrations in mouse dictyate oocytes. I. Time and dose relationships.

Structural chromosome aberrations were analyzed in superovulated metaphase-I oocytes of the mouse, Mus musculus, at various times after a single acute dose of 200 R of X-rays. The aberrations seen were of the chromatid type, i.e., chromatid interchanges, isochromatid deletions and chromatid deletions. The aberration frequency was low during the interval 24 h to 5 days between irradiation and ovulation; peak frequency was reached when irradiation was given 14 days prior to ovulation. A dose-response study was made 14 days prior to ovulation at doses of 50, 100, 200, 300 and 400 R. A curve of these data indicated that a significant two-track component was present for both interchanges and deletions. Centromere staining revealed that symmetrical and asymmetrical interchanges occurred at approximately equal frequency and also that the asymmetrical equivalent of crossing-over was induced at a measurable frequency.     

Structural and numerical chromosome aberration inducers in liver micronucleus test in rats with partial hepatectomy

The liver micronucleus test is an important method to detect pro-mutagens such as active metabolites not reaching bone marrow due to their short lifespan. We have already reported that dosing of the test compound after partial hepatectomy (PH) is essential to detect genotoxicity of numerical chromosome aberration inducers in mice [Mutat. Res. 632 (2007) 89-98]. In naive animals, the proportion of binucleated cells in rats is less than half of that in mice, which suggests a species difference in the response to chromosome aberration inducers. In the present study, we investigated the responses to structural and numerical chromosome aberration inducers in the rat liver micronucleus test. Two structural chromosome aberretion inducers (diethylnitrosamine and 1,2-dimethylhydrazine) and two numerical chromosome aberration inducers (colchicine and carbendazim) were used in the present study. PH was performed a day before or after the dosing of the test compound in 8-week old male F344 rats and hepatocytes were isolated 4 days after the PH. As a result, diethylnitrosamine and 1,2-dimethylhydrazine, structural chromosome aberration inducers, exhibited significant increase in the incidence of micronucleated hepatocyte (MNH) when given either before and after PH. Colchicine and carbendazim, numerical chromosome aberration inducers, did not result in any toxicologically significant increase in MNH frequency when given before PH, while they exhibited MNH induction when given after PH. It is confirmed that dosing after PH is essential in order to detect genotoxicity of numerical chromosome aberration inducers in rats as well as in mice. Regarding the species difference, a different temporal response to colchicine was identified. Colchicine increased the incidence of MNH 4 days after PH in rats, although such induction in mice was observed 8-10 days after PH.     

Periodic variations of spontaneous chromosome aberration rate in human blood lymphocytes

Spontaneous chromosome aberration rate in human blood lymphocytes was studied using the data collected during 30 years. Seasonal variation was found. An absolute maximum of chromosome aberration rate was observed in winter and a local maximum, in summer. In spring and autumn, this value decreased. There was a statistically significant trend towards higher frequencies of aberrant metaphases during the period studied. A periodic pattern of changes in the chromosome aberration frequency was found. The data fitted a sinusoid with a period of 4.5 years.     

Variation of Chromosome Aberration Rate in the Culture of Human Blood Lymphocytes with Time

Spontaneous chromosome aberration rate in human blood lymphocytes was studied using the data collected during 30 years. Seasonal variation was found. An absolute maximum of chromosome aberration rate was observed in winter and a local maximum, in summer. In spring and autumn, this value decreased. There was a statistically significant trend towards higher frequencies of aberrant metaphases during the period studied. A periodic pattern of changes in the chromosome aberration frequency was found. The data fitted a sinusoid with a period of 4.5 years.     

重铬酸钾对蚕豆根尖细胞致畸效应的研究

以蚕豆根尖为材料,研究重铬酸钾对蚕豆根尖细胞的致畸效应。采用蚕豆根尖细胞的微核试验和染色体畸变试验方法,以不同浓度的重铬酸钾为诱变剂,测定蚕豆根尖细胞的微核率和染色体畸变率。结果表明:重铬酸钾能诱发较高频率的微核率,即在一定浓度范围内,其微核率随重铬酸钾处理浓度的升高而增加,但高于一定浓度后反而呈下降趋势;不同浓度的重铬酸钾均使蚕豆根尖细胞有丝分裂指数增大;重铬酸钾还能诱导蚕豆根尖细胞产生较高频率的染色体畸变,且产生多种类型的染色体畸变。结论是重铬酸钾对蚕豆根尖细胞具有明显的致畸效应。     

60Coγ射线辐照花魔芋球茎的早期诱变效应研究

本试验开展了花魔芋早期诱变效应研究。通过^60Coγ射线辐照后的植株苗期发育观察和根尖细胞学检测分析,结果表明魔芋辐射诱变效应显著：辐照诱发核畸变和染色体畸变,且变异频率与剂量呈二次曲线关系;低剂量时对细胞分裂有刺激作用;辐照抑制芽体发育、苗期生长,抑制效应随剂量增加而加大,直至产生致死作用。依据花魔芋的辐射敏感性,建立了魔芋辐照诱变体系,以催芽球茎为诱变材料,诱变剂量范围为0Gy～50Gy,剂量率为1Gy／min,中等适宜剂量为7Gy～10Gy,致死剂量50Gy。     

The role of short-lived DNA lesions in the production of chromosome-exchange aberrations

Human lymphocytes were treated with combined UVC radiation and X-rays or they were X-irradiated and incubated for 60–90 min in the presence of DNA-repair inhibitor ara-C. The X-ray induced chromosome exchange aberration yield was enhanced both by UVC and ara-C by approximately a factor of two in the linear (low dose) portion of the dose-response curve. The enhancement was small in the dose squared (high dose) portion where previous dose-fractionation experiments have shown that X-ray-induced lesions leading to aberrations exist for several hours. The yield of aberrations in lymphocytes incubated after irradiation in the presence of ara-C reaches a saturation level almost immediately after irradiation (5–15 min). These cytogenetic observations together with a previous finding (Holmberg and Strausmanis, 1983) give direct and indirect evidence that the enhanced aberration yield is due to short-lived DNA breaks formed immediately after X-irradiation.

Measurements on the repair kinetics of the DNA breaks induced by X-irradiation show that ara-C strongly impairs the repair of short-lived X-ray-induced DNA breaks. It was also observed that the DNA breaks generated after UVC irradiation occur almost immediately after irradiation and the level of these transient DNA breaks reaches saturation even for short incubation times. Thus, the repair of these breaks can compete with the repair of short-lived X-ray-induced DNA-breaks in combined irradiation with UVC and X-rays.

The experimental results can be explained on the assumption that X-ray-induced aberrations originate from exchange complexes formed in interactions between both short-lived DNA breaks. The short-lived DNA breaks give rise to exchange complexes mainly within single ionization tracks where the DNA breaks are close together. The time between irradiation and exchange complex formation is of the order of 5–15 min within such a track, and short-lived breaks might be repaired before complexes have been formed. If the DNA repair of these breaks is delayed by UVC or ara-C treatment this results in a higher probability of exchange-complex formation. In contrast, interactions between breaks in different tracks originate from long-lived DNA breaks and the probability for complex formation from these breaks is not markedly affected by UVC or ara-C.     

The mutagenic activity of aflatoxin B1 in the Cricetulus griseus hamster and Macaca mulatta monkey

Chromosome aberrations were scored in bone marrow cells of Cricetulus griseus hamsters and Macaca mulatta monkeys given a single i.p. injection of aflatoxin B1 (AFB1). The mutagenic activity of AFB1 was assessed by the percentage of cells bearing aberrations and by the total frequency of chromosome and chromatid breaks. Chinese hamsters were treated with five different doses of AFB1 ranging from 1 microgram to 5 mg/kg (LD50/30 = 12.2 mg/kg) and the aberration yields at each AFB1 dose level tested were determined at 24 h intervals for 5 consecutive days. Compared to controls the increase in the two types of chromosome abnormalities was significant in all tests. At 5 mg/kg of AFB1 the tests were carried out over a period of 92 days to assure the analysis of aberration yields with time. All chromosome aberration assays conducted during this period showed significant increases in the frequencies of aberrant cells and chromosome and chromatid breaks in comparison to controls. Macaque monkeys were treated in the same fashion using 0.1 and 1.0 mg/kg of AFB1 and the dynamics of chromosome aberration yields was analyzed for a period of 730 days. Similarly as in the case of Chinese hamsters the percentage of cells with aberrations and the frequency of chromosome and chromatid breaks were always higher in this period than the control value. Long-term aberration yield data obtained experimentally were expressed in the form of analytical curves which allowed to establish the time when the yields of aberrant cells reached their maxima and when they returned to the control level. In both animal species tested the courses of analytical curves had a similar dynamics. Factors that might be responsible for a long-term persistence and relatively great fluctuations of the chromosome aberration yields encountered after a single injection of AFB1 are discussed in detail.     

Cell type-specific quantitative predictions of radiation-induced chromosome aberrations: a computer model approach

A quantitative computer model was applied to simulate the three-dimensional (3D) spatial organization of chromatin in human cell nuclei under defined conditions of virtual irradiation to explore the implications of spatial organization on chromosome aberrations. To calibrate the virtual irradiation algorithm, a dose-dependent spectrum of radiation-induced chromosome aberrations such as dicentrics, translocations and centric rings was calculated for low-LET radiation doses ranging from 0.5 to 5 Gy. This was compared with the results from experimental studies. While the dose-response curves calculated from model simulations agree well with experimental dose-response curves for dicentrics and translocations, centric rings are significantly more frequent in the model simulation than in experiments despite taking into account exclusive arm territories in the applied Spherical 1 Mbp Chromatin Domain (SCD) computer model explicitly. Taking into account the non-random positioning of chromosome territories observed in lymphocyte cell nuclei (a so-called gene density-correlated arrangement of chromosome territories), aberration frequencies were calculated with the calibrated irradiation algorithm to investigate the impact of chromosome territory neighborhood effects (proximity effects). The absolute frequencies of pairwise exchanges agree well with those found in an experimental study. In conclusion, the results obtained using the computer model approach presented here based on only a few adjustable parameters correlated well with those of experimental studies of chromosome aberration frequencies. Thus the model may be a useful tool in radiation-induced cancer risk estimates in combination with epidemiological studies.     

低水平辐射诱导的细胞遗传学适应性反应

先用0.01GY x-射线(剂量率:0.01GY/分)体外照射人、兔外周血,经不同时间后再用1.5GY X-射线(0.44GY/分)照射,发现在G_0、G_1、S和G_2期受0.01GY X-射线照射后再给大剂量照射者,其染色体畸变率明显低于单纯受1.5GY X-射线照射组(P<0.01)。这一适应性反应能持续3个细胞周期,在接受小剂量照射后超过3个细胞周期再受大剂量照射者,染色体畸变率未见减少。若在第三细胞周期以后再次给予小剂量照射,可再次诱导适应性反应。用小鼠整体小剂量照射后骨髓细胞和生殖细胞亦出现这种适应性反应。另外也探讨了不同剂量和不同剂量率的预先照射对适应性反应的影响。     

Elimination of X-ray-induced chromosomal aberrations in the progeny of female mice

Female NMRI mice were irradiated with various doses of X-rays and induced chromosome aberrations were scored in MII oocytes (Dosage: 0.222, 0.666, 2 and 6 Gy). After irradiation with 2 Gy, early zygotes were examined in the 2-cell stage; additional dominant lethals were counted and surviving embryos were examined after 13.5 days of pregnancy. 87.2% of the MII oocytes showed structural chromosomal aberrations after irradiation with 2 Gy. Surviving embryos, however, failed to show any increase in the aberration rate. This result points to (almost) complete elimination of genetically damaged oocytes and zygotes already before birth. In addition to the structural aberrations, aneuploidies were induced. Most of them, however, were hypoploidies. Hence, the study confirmed the well-known susceptibility of oocytes around the time of fertilization for induced chromosome loss. Induced hyperploidies, however, were very rare. Evidence for induction of meiotic non-disjunction was weak. In surviving embryos, no increase in numerical aberrations, either hypoploid or hyperploid was discovered. The significance of these data for the prediction of chromosomal damage due to to ionizing radiation in humans is discussed. Recent risk estimates of UNSCEAR and other agencies represent very cautious upper levels.