Intercomparison of translocation and dicentric frequencies between laboratories in a follow-up of the radiological accident in Estonia

Purpose : To perform an interlaboratory comparison of FISH chromosome painting and to study the time-course of translocations and dicentrics in three accident victims exposed to radiation. Also, to use the data in the validation of the FISH technique as a retrospective dosimeter. Materials and methods : Twelve blood samples were collected during 4 years from three subjects exposed to radiation in an accident in Estonia in 1994 involving γ -radiation from a 137Cs source. Two of the subjects were exposed during approximately 7 h, both receiving a protracted dose of about 1 Gy and also localized exposure. The third subject received a protracted whole-body dose of 2.7Gy during 4 weeks as well as a short-term partialbody dose. Preparations from 48-h metaphase cultures were painted by the FISH technique using routine methods and probe cocktails in four laboratories. Samples from each subject were analysed in two different laboratories that used different combinations of whole chromosome probes. The PAINT nomenclature was applied when recording chromosome aberrations. Results : The intercomparison of FISH analysis data showed reasonable similarities between laboratories, the largest discrepancy being 21% in the frequency of two-way translocations in subject 3. Half-time calculations, based on combined data sets from two laboratories, showed that dicentrics decreased rapidly with half-times of approximately 2 years. In all cases, the initial dicentric yields were lower than the initial translocation yields. During the 4-year follow-up, the frequencies of all translocations in cells containing only simple rearrangements fell on average to about 65% of their initial value. Two-way translocations were slightly more persistent than all translocations. The average halftime was about 8 years for two-way translocations and around 6 years for all translocations. Cells containing complex rearrangements were few in number and they disappeared with time. In general, the inclusion of complex cells caused a more rapid fall in aberration yield. Conclusions : In general, the results imply that relatively consistent scoring data were obtained with different chromosome painting protocols. They also support the idea that the reduction of translocations with time is associated with partial-body irradiation.     

Time-course of translocation and dicentric frequencies in a radiation accident case

PURPOSE: To assess the persistence of exchange aberrations measured by FISH chromosome painting after accidental radiation exposure. MATERIALS AND METHODS: Chromosome analyses were carried out in peripheral lymphocytes of a 13-year-old boy exposed to protracted low dose-rate whole-body and short-time partial-body irradiation from a radiation accident in Estonia in 1994. Up to November 1998, the frequencies of translocations and dicentrics were periodically measured using FISH chromosome painting of the target chromosomes 1, 4 and 12, with a simultaneous pancentromeric probe. RESULTS: For the yields of dicentrics, an expected rapid temporal decline was found with a half-time of 14.2+/-1.9 months. The yields of reciprocal translocations also revealed a gradual but significant reduction with a half-time of 51.7+/-12.7 months. CONCLUSION: An unchanged temporal persistence of so-called stable translocations cannot be assumed. Any significant reduction of this aberration type with time obviously limits the application of FISH-based translocation measurements for reliable long-term biodosimetry after combined protracted whole-body and partial-body radiation exposure.     

Netherlands Radiobiological Society

Purpose : To assess the persistence of exchange aberrations measured by FISH chromosome painting after accidental radiation exposure. Materials and methods : Chromosome analyses were carried out in peripheral lymphocytes of a 13-year-old boy exposed to protracted low dose-rate whole-body and short-time partial-body irradiation from a radiation accident in Estonia in 1994. Up to November 1998, the frequencies of translocations and dicentrics were periodically measured using FISH chromosome painting of the target chromosomes 1, 4 and 12, with a simultaneous pancentromeric probe. Results : For the yields of dicentrics, an expected rapid temporal decline was found with a half-time of 14.2 ±1.9 months. The yields of reciprocal translocations also revealed a gradual but significant reduction with a half-time of 51.7 ±12.7 months. Conclusion : An unchanged temporal persistence of so-called stable translocations cannot be assumed. Any significant reduction of this aberration type with time obviously limits the application of FISH-based translocation measurements for reliable long-term biodosimetry after combined protracted whole-body and partial-body radiation exposure.     

The 60Co gamma ray dose-response for chromosomal aberrations in human lymphocytes analysed by FISH; applicability to biological dosimetry

Purpose : To investigate the in vitro dose-response for 60Co irradiated human lymphocytes assayed by FISH, and to consider how this may be applied to retrospective dosimetry. Method : Blood was irradiated with doses in the range 0.25-4.0Gy. Cultured lymphocytes were scored for all stable and unstable aberrations involving painted chromosomes 2, 3 and 5 and, in addition, all unstable aberrations in the counterstained chromosomes. A pancentromeric probe was included. Results : The relative numbers of painted and full genome dicentrics agreed well with the Lucas hypothesis for calculating genome equivalence. The involvement of each painted chromosome in exchanges agreed with their relative arm lengths. The doseresponse relationship fitted well to the linear quadratic model; Y (0.9 10 2)D (6.5 10 2)D2 where D is the dose in Gy for the incidence Y, of all one plus two-way translocations in all cells corrected for genome equivalence. Complex rearrangements also became more frequent with increasing dose. A correlation was noted between the distributions of dicentrics and translocations among the cells and this was entirely due to complexes. Conclusions : For retrospective dosimetry it is recommended to use an in vitro dose-response for apparently simple translocations in stable (Cs) cells. To date, acute linear yield coefficients from FISH data carry statistical uncertainties too large for useful application to retrospective dosimetry of persons exposed to chronic or low doses. As an interim measure it is suggested that one may derive a linear term from full genome dicentrics corrected by a factor representing the translocation to dicentric ratio.     

The 60Co gamma ray dose-response for chromosomal aberrations in human lymphocytes analysed by FISH; applicability to biological dosimetry.

PURPOSE: To investigate the in vitro dose-response for 60Co irradiated human lymphocytes assayed by FISH, and to consider how this may be applied to retrospective dosimetry. METHOD: Blood was irradiated with doses in the range 0.25-4.0 Gy. Cultured lymphocytes were scored for all stable and unstable aberrations involving painted chromosomes 2, 3 and 5 and, in addition, all unstable aberrations in the counterstained chromosomes. A pancentromeric probe was included. RESULTS: The relative numbers of painted and full genome dicentrics agreed well with the Lucas hypothesis for calculating genome equivalence. The involvement of each painted chromosome in exchanges agreed with their relative arm lengths. The dose-response relationship fitted well to the linear quadratic model; Y=(0.9 x 10(-2))D+(6.5 x 10(-2))D2 where D is the dose in Gy for the incidence Y, of all one plus two-way translocations in all cells corrected for genome equivalence. Complex rearrangements also became more frequent with increasing dose. A correlation was noted between the distributions of dicentrics and translocations among the cells and this was entirely due to complexes. CONCLUSIONS: For retrospective dosimetry it is recommended to use an in vitro dose-response for apparently simple translocations in stable (Cs) cells. To date, acute linear yield coefficients from FISH data carry statistical uncertainties too large for useful application to retrospective dosimetry of persons exposed to chronic or low doses. As an interim measure it is suggested that one may derive a linear term from full genome dicentrics corrected by a factor representing the translocation to dicentric ratio.     

Reciprocal translocations as an indicator for radiation exposures in the low dose range.

A few years back, the frequency of dicentrics was determined shortly after exposure in five accidentally exposed radiation workers. In all cases the observed dicentric yield was significantly higher in comparison with the background level, and the resulting estimated whole body doses lay between 0.2 and 0.3 Gy. Now, a number of years later (1 to 11 years), the frequency of translocations has been determined by means of the FISH technique. Chromosomes 2, 4 and 8 were painted. The measured translocation frequency lay, however, within the range of the spontaneous variation between individuals. No radiation exposure could, therefore, be proven. In two further cases, the dicentrics were determined by means of the conventional Giemsa staining technique, and the translocations by means of chromosome painting carried out on the same blood samples, which were taken about 3 and 10 years, respectively, after the radiation exposure. The dose estimates obtained on the basis of dicentric frequency using the Qdr method, and on the basis of FISH detected translocations, are compared.     

Effect of americium-241 alpha-particles on the dose-response of chromosome aberrations in human lymphocytes analysed by fluorescence in situ hybridization

PURPOSE: To evaluate by the fluorescent in-situ hybridization (FISH) technique the dose-response and intercellular distribution of alpha-particle-induced chromosome aberrations. In particular, the validity of using the yield of characteristic types of chromosome abnormalities in stable cells as quantitative indicators for retrospective dose reconstruction has been evaluated. MATERIAL AND METHODS: Monolayers of human peripheral lymphocytes were exposed at doses from 0.02 to 1 Gy to alpha-particles emitted from a source of americium-241. The most probable energy of the alpha-particles entering the cells was 2.7 MeV. FISH painting was performed using DNA probes for chromosomes 2, 4 and 8 in combination with a pan-centromeric probe. In complete first-division cells, identified by harlequin staining, aberrations involving painted target chromosomal material were recorded as well as aberrations involving only unpainted chromosomal material. RESULTS: In total, the percentage of complex aberrations was about 35% and no dose dependence was observed. When complex-type exchanges were reduced to simple base types, the different cell distributions were clearly over-dispersed, and the linear coefficients of the dose-effect curves for translocations were significantly higher than for dicentrics. For past dose reconstruction, only a few complex aberrations were in stable cells. The linear coefficient obtained for transmissible aberrations in stable cells was more than seven times lower than that obtained in all analysed cells, i.e. including unstable cells. CONCLUSION: FISH-based analysis of complex rearrangements allows discrimination between partial-body exposures to low-linear energy transfer radiation and high-linear energy transfer exposures. In assessing past or chronic exposure to alpha-particles, the use of a dose-effect curve obtained by FISH-based translocation data, which had not excluded data determined in unstable cells, would underestimate the dose. Insertions are ineffective biomarkers because their frequency is too low.     

Distribution of radiation-induced exchange aberrations in all human chromosomes

PURPOSE: To investigate the DNA-proportional distribution of radiation-induced chromosome aberrations for all chromosomes of a male and a female human karyotype. MATERIALS AND METHODS: Metaphases were prepared from whole blood cultures obtained from two healthy donors and set up after irradiation with 3 Gy 220 kV X-rays. Single whole-chromosome FISH painting simultaneously with pancentromeric DNA painting were performed separately for each chromosome of the human karyotype. One thousand exclusively first-division metaphases were analysed per chromosome and donor. After statistical analysis, the data obtained were compared with theoretically expected values. RESULTS: All aberration types (translocations, dicentrics) showed deviations from a DNA-proportional distribution. For both donors, chromosomes 2 and 3 exhibited significantly less and chromosome 4 more symmetrical translocations than expected. Chromosomes 15 and 22 showed more symmetrical translocations than predicted for one of the two donors. Less dicentrics than expected became apparent for chromosomes 2, 3 and 18, while more dicentrics were seen for chromosomes 15, 16 and 17. Moreover, chromosomes 4, 14 and 22 showed a significant deviation from the theoretically expected 1:1 ratio of the yields of symmetrical translocations to the yields of dicentrics. CONCLUSION: The results from the whole-chromosome complement in two different donors confirmed published data from the analysis of single chromosomes that some human chromosomes were not involved in radiation-induced dicentrics and symmetrical translocations proportional to their DNA content. This must be taken into account if chromosome subsets for dose reconstruction are selected or if whole genomic frequencies have to be calculated from partial genome analysis.     

Comparison of X-ray dose-response curves obtained by chromosome painting using conventional and PAINT nomenclatures

PURPOSE: The compare the suitability of PAINT and conventional nomenclature systems for the construction of chromosome aberration dose-effect curves for X-rays using FISH techniques, and to compare these curves with those based on solid-stained dicentrics analysed in first division metaphases by the FPG technique. MATERIALS AND METHODS: Blood samples were irradiated at 0.1, 0.25, 0.50, 0.75, 1, 1.5, 2, 3, 4 and 5 Gy 180 kV X-rays. FISH painting was performed using probes for chromosomes 1, 4 and 11 in combination with a pan-centromeric probe. RESULTS: Translocations showed a higher background frequency than dicentrics. This influences the ratio of translocations:dicentrics at the lower doses and the uncertainties of dose-effect curves for translocations. The dose-effect curves for dicentrics obtained by FISH and solid stain were in close agreement. CONCLUSION: For short-term biological dosimetry purposes by FISH, the use of dic(BA) (PAINT nomenclature) or total dicentrics (conventional nomenclature) should give similar dose estimates. For dose reconstruction, the use of total or complete translocations result in similar uncertainties.     

Comparison of X-ray dose-response curves obtained by chromosome painting using conventional and PAINT nomenclatures

Purpose: The compare the suitability of PAINT and conventional nomenclature systems for the construction of chromosome aberration dose-effect curves for X-rays using FISH techniques, and to compare these curves with those based on solid-stained dicentrics analysed in first division metaphases by the FPG technique. Materials and methods: Blood samples were irradiated at 0.1, 0.25, 0.50, 0.75, 1, 1.5, 2, 3, 4 and 5Gy 180kV X-rays. FISH painting was performed using probes for chromosomes 1, 4 and 11 in combination with a pan-centromeric probe. Results: Translocations showed a higher background frequency than dicentrics. This influences the ratio of translocations:dicentrics at the lower doses and the uncertainties of dose-effect curves for translocations. The dose-effect curves for dicentrics obtained by FISH and solid stain were in close agreement. Conclusion: For short-term biological dosimetry purposes by FISH, the use of dic (BA) (PAINT nomenclature) or total dicentrics (conventional nomenclature) should give similar dose estimates. For dose reconstruction, the use of total or complete translocations result in similar uncertainties.     

Distribution of radiation‐induced exchange aberrations in all human chromosomes

Purpose: To investigate the DNA‐proportional distribution of radiation‐induced chromosome aberrations for all chromosomes of a male and a female human karyotype.

Materials and methods: Metaphases were prepared from whole blood cultures obtained from two healthy donors and set up after irradiation with 3?Gy 220?kV X‐rays. Single whole‐chromosome FISH painting simultaneously with pancentromeric DNA painting were performed separately for each chromosome of the human karyotype. One thousand exclusively first‐division metaphases were analysed per chromosome and donor. After statistical analysis, the data obtained were compared with theoretically expected values.

Results: All aberration types (translocations, dicentrics) showed deviations from a DNA‐proportional distribution. For both donors, chromosomes 2 and 3 exhibited significantly less and chromosome 4 more symmetrical translocations than expected. Chromosomes 15 and 22 showed more symmetrical translocations than predicted for one of the two donors. Less dicentrics than expected became apparent for chromosomes 2, 3 and 18, while more dicentrics were seen for chromosomes 15, 16 and 17. Moreover, chromosomes 4, 14 and 22 showed a significant deviation from the theoretically expected 1:1 ratio of the yields of symmetrical translocations to the yields of dicentrics.

Conclusion: The results from the whole‐chromosome complement in two different donors confirmed published data from the analysis of single chromosomes that some human chromosomes were not involved in radiation‐induced dicentrics and symmetrical translocations proportional to their DNA content. This must be taken into account if chromosome subsets for dose reconstruction are selected or if whole genomic frequencies have to be calculated from partial genome analysis.     

用FISH技术对忻州事故宫内受照者进行回顾性剂量重建

目的 对忻州事故中宫内受照者"京"进行回顾性剂量重建.方法 受照后16年取"京"及其母亲"芳"的外周静脉血,用常规方法分析非稳定性染色体畸变,用CB法分析双核淋巴细胞微核;用全染色体探针的FISH方法分析染色体易位,参照本实验室建立的全基因组易位率与吸收剂量之间的剂量-效应曲线进行剂量估算.参照"芳"的剂量校正系数推算"京"在宫内的受照剂量.结果 受照16年后未观察到两位受照者的外周血非稳定性染色体畸变,淋巴细胞微核率在正常范围内.参照本实验室建立的吸收剂量与全基因组易位率之间的剂量-效应曲线,"芳"在16年后用FISH方法估算的剂量为0.76(0.41～1.00)Gy,而"芳"在事故后短期内估算的生物剂量为2.30(2.07～2.50)Gy,其剂量校正系数为3.03;"京"在16年后估算的剂量为0.61(0.44～0.86)Gy,推算"京"在宫内受照的剂量为1.85(1.33～2.61)Gy.结论 16年后用FISH方法对宫内受照者进行剂量重建,参照受照母亲的剂量校正系数,可推算出胎儿宫内受照的大致剂量,为临床提供依据.     

Analysis of chromosome aberrations by FISH and Giemsa assays in lymphocytes of cancer patients undergoing whole-body irradiation: comparison of in vivo and in vitro irradiation.

PURPOSE: To study the cytogenetic effects of fractionated radiotherapy in peripheral blood lymphocytes of five cancer patients. In vitro experiments were performed in parallel using the same dose range and a comparison was made of the induced frequencies of stable and unstable chromosome aberrations. The object was to clarify the use of an in vitro calibration curve for immediate and retrospective dosimetry in cases of radiation accidents. MATERIALS AND METHODS: Patients were exposed to 60Co gamma-rays at a single dose of 11.5 cGy each day up to a total dose of 57.5 cGy, given in 5 days. For measurement of chromosome aberrations, blood was collected from patients before irradiation and after each exposure. Blood taken before treatment was used as a control and for in vitro irradiation experiments in the dose range 8-50 cGy. Chromosome aberration frequency (stable as well as unstable) was determined using fluorescence in situ hybridization (FISH) assay with specific DNA libraries for chromosomes 1, 4 and 8 and a pancentromertic probe for the whole genome. Giemsa-stained preparations were used to score unstable aberrations following in vivo and in vitro exposure. RESULTS: A linear dose-response curve was determined for both dicentrics and translocations. The in vivo frequency of translocations was higher than for dicentrics. Dose-response curves generated for translocations following in vivo and in vitro irradiation yielded similar frequencies. In contrast, for dicentrics, in vitro irradiation yielded a higher frequency when compared with data generated following in vivo exposure. CONCLUSIONS: For dose reconstruction purposes, translocations frequency seems to be a more adequate end-point than the scoring of dicentrics. The established in vitro calibration curve for dicentrics may underestimate absorbed radiation dose in cases of protracted exposure.     

Evaluation of spectral karyotyping (SKY) in biodosimetry for the triage situation following gamma irradiation

PURPOSE: Biological dosimetry in an acute triage situation of radiation exposure is traditionally performed by scoring unstable dicentric chromosomal aberrations after conventional Giemsa staining, and more recently also by detection of chromosomal translocations after chromosome painting by fluorescence in situ hybridization (FISH). By spectral karyotyping (SKY) each chromosome can be painted in an individual colour, permitting the scanning for structural aberrations throughout the genome in each individual metaphase. Here we have evaluated the performance of SKY analysis in a simulated triage situation after gamma irradiation. MATERIALS AND METHODS: Peripheral leukocytes were irradiated by 60Co (0 - 5 Gy) and analysed by SKY, Giemsa staining and FISH painting of chromosomes 1, 2, and 3. RESULTS: At 1 Gy and higher doses, dicentric aberrations (Dic+) as well as classical one- and two-way translocations were found in increasing and dose-dependent frequencies by SKY. The frequency of dicentrics detected by Giemsa was found to be significantly higher than the total aberrations detected by SKY (p<0.001), but did not differ significantly from that of FISH painting. The difference was mainly attributable to the low sensitivity of SKY to detect Dic+ following frequent lack of acentric fragments with matching chromosomal composition. CONCLUSIONS: The findings anticipate that radiation induced chromosomal aberrations may be more complex than expected from conventional and single chromosome painting analyses. While conventional Giemsa staining was found to be the method of choice for the triage situation, it is expected that extended SKY analysis will add to the knowledge of underlying mechanisms for irradiation associated chromosomal aberrations.     

Analysis of chromosome aberrations by FISH and Giemsa assays in lymphocytes of cancer patients undergoing whole-body irradiation: comparison of in vivo and in vitro irradiation

Purpose : To study the cytogenetic effects of fractionated radiotherapy in peripheral blood lymphocytes of five cancer patients. In vitro experiments were performed in parallel using the same dose range and a comparison was made of the induced frequencies of stable and unstable chromosome aberrations. The object was to clarify the use of an in vitro calibration curve for immediate and retrospective dosimetry in cases of radiation accidents. Materials and methods : Patients were exposed to 60 Co γ-rays at a single dose of 11.5 cGy each day up to a total dose of 57.5 cGy, given in 5 days. For measurement of chromosome aberrations, blood was collected from patients before irradiation and after each exposure. Blood taken before treatment was used as a control and for in vitro irradiation experiments in the dose range 8-50 cGy. Chromosome aberration frequency (stable as well as unstable) was determined using fluorescence in situ hybridization (FISH) assay with specific DNA libraries for chromosomes 1, 4 and 8 and a pancentromertic probe for the whole genome. Giemsa-stained preparations were used to score unstable aberrations following in vivo and in vitro exposure. Results : A linear dose-response curve was determined for both dicentrics and translocations. The in vivo frequency of translocations was higher than for dicentrics. Dose-response curves generated for translocations following in vivo and in vitro irradiation yielded similar frequencies. In contrast, for dicentrics, in vitro irradiation yielded a higher frequency when compared with data generated following in vivo exposure. Conclusions : For dose reconstruction purposes, translocations frequency seems to be a more adequate end-point than the scoring of dicentrics. The established in vitro calibration curve for dicentrics may underestimate absorbed radiation dose in cases of protracted exposure.     

用FISH技术对忻州事故宫内受照者进行回顾性剂量重建

目的 对忻州事故中宫内受照者"京"进行回顾性剂量重建。方法 受照后16年取"京"及其母亲"芳"的外周静脉血,用常规方法分析非稳定性染色体畸变,用CB法分析双核淋巴细胞微核;用全染色体探针的FISH方法分析染色体易位,参照本实验室建立的全基因组易位率与吸收剂量之间的剂量-效应曲线进行剂量估算。参照"芳"的剂量校正系数推算"京"在宫内的受照剂量。结果 受照16年后未观察到两位受照者的外周血非稳定性染色体畸变,淋巴细胞微核率在正常范围内。参照本实验室建立的吸收剂量与全基因组易位率之间的剂量-效应曲线,"芳"在16年后用FISH方法估算的剂量为0.76(0.41～1.00)Gy,而"芳"在事故后短期内估算的生物剂量为2.30(2.07～2.50)Gy,其剂量校正系数为3.03;"京"在16年后估算的剂量为0.61(0.44～0.86)Gy,推算"京"在宫内受照的剂量为1.85(1.33～2.61)Gy。结论 16年后用FISH方法对宫内受照者进行剂量重建,参照受照母亲的剂量校正系数,可推算出胎儿宫内受照的大致剂量,为临床提供依据。     

Collaborative exercise on the use of FISH chromosome painting for retrospective biodosimetry of Mayak nuclear-industrial personnel

PURPOSE: To investigate within the framework of a multilaboratory study the suitability of FISH chromosome painting to measure so-called stable translocations in peripheral lymphocytes of Mayak nuclear-industrial workers (from the Southern Urals) and their use for retrospective biodosimetry. MATERIALS AND METHODS: Chromosime analyses were carried out from 69 workers who had received protracted occupational radiation exposures (0.012-6.065 Gy) up to approximately 40 years before blood sampling. Twenty-one unexposed people living in the same area were controls. A multicolour FISH-painting protocol with the target chromosomes 1, 4 and 8 simultaneously with a pancentromeric probe was used to score potentially transmissible chromosome-type aberrations (reciprocal translocations 2B and related 'one-way' patterns I-III according to the S&S classification). RESULTS: Individual biodosimetry estimates were obtained in terms of these potentially long-term surviving aberration types based on the linear component of a low dose-rate gamma-ray calibration curve produced using identical staining and scoring protocols. For comparison, the workers personal and total background doses were converted to red bone marrow doses. The estimated doses were mainly lower than would be predicted by the calibration curve, particularly at accumulated higher dose levels. CONCLUSIONS: Owing to the limited life-time of circulating T-lymphocytes, the long-term persistence of translocations in vivo requires the assumption of a clonal repopulation of these naturally senescing cells from the haemopoietic stem cell compartments. Obviously such a replacement cannot be fully achieved, leading to a temporal decline even of the yield of transmissible aberrations types. Assuming further a highly selective capacity of stem cells against any type of chromosomal damage and the fact that one must rely on partial genome findings, the potential of FISH chromosome painting for retrospective dose reconstruction is probably limited to a decade or so after high-level protracted radiation exposure.     

Follow-up of stable chromosomal aberrations in gamma-ray irradiated non-human primates

PURPOSE: The purpose of this study was to examine a new approach to retrospective biological dosimetry, by using a long-term animal model to determine the stability of translocation frequency after in vivo irradiation. While the frequency of dicentrics is known to decrease over time, the persistence of more stable chromosomal aberrations such as translocations could be useful if their stability were definitively proved. MATERIALS AND METHODS: Four monkeys (Macaca fascicularis) were exposed to two different doses of ionizing radiation: 2 Gy whole body irradiation for two and 4 Gy for two others. Blood samples were obtained at various times after irradiation. Both total and two-way translocations were detected by fluorescence in situ hybridization. Translocations were scored in stable cells, that is, those without dicentrics, rings or fragments. The course of translocation frequency was analysed at four time-points: one hour (H1), 2 months (M2), 10 months (M10) and 31 months (M31) after irradiation. RESULTS: We observed two separate trends in translocation frequency: Total translocation frequency decreased slightly in animals irradiated with a dose of 2 Gy, while two-way translocation frequency was relatively stable in all irradiated animals. CONCLUSIONS: We confirmed the long-term stability of translocations and found that it seems to depend on the type of the translocation recorded. Overall translocations were stable for up to 31 months regardless of dose, but two-way translocations were more stable than those that were non-reciprocal, especially in stable cells.     

A comparative study on potential cytogenetic fingerprints for radiation LET in human lymphocytes

PURPOSE: To carry out a comparative study on potential cytogenetic fingerprints for radiation LET in human metaphase lymphocytes. MATERIALS AND METHOD: Human lymphocytes were irradiated in vitro with 3.0 Gy 60Co gamma-rays, 0.9 Gy 3H beta-rays or 0.2 Gy 2.7 Mev neutrons. Detailed chromosome aberrations were analysed by combined FISH with pan-telomere staining and specific whole-chromosome painting (1, 2 and 4). Total chromosome translocations and insertions were also analysed by multicolour whole-chromosome painting (chromosomes 1, 2 and 4 orange, chromosomes 3, 5 and 6 green). RESULTS: Among the six proposed radiation cytogenetic fingerprints, the ratio of total simple translocations to insertions (I-ratio), showed the largest difference between low-LET 60Co gamma-ray and high-LET neutron radiation. The ratios of complete exchanges to incomplete rejoinings [S(I)-ratio] and dicentrics to interstitial deletions (H-ratio), showed a similar significant difference between low- and high-LET radiation. The ratios of centric rings to interstitial deletion (G-ratio) showed a trend of LET-related difference, but the difference was not significant in this data set. The ratios of dicentrics to centric rings (F-ratio) and apparent complete exchanges to hidden complete exchanges [S(II)-ratio], showed no difference between low- and high-LET radiation. In the 1426 radiation-induced chromosome aberrations observed after 52 h culture, evidence for sister-chromatid fusion but not telomere addition was found. CONCLUSION: Pan-telomere staining plus specific whole chromosome painting allows simultaneous and objective detection of complete or incomplete chromosome exchanges and interstitial or terminal deletions in human peripheral lymphocytes. Of the six proposed cytogenetic ratios, the I-ratio is the most effective cytogenetic fingerprint for distinguishing low-LET from high-LET radiation in human metaphase human lymphocytes.     

Translocation frequencies measured in patients one year after radioactive iodine therapy for thyrotoxicosis

Purpose : To investigate the incidence of translocations induced by iodine-131 therapy in thyrotoxicosis patients 1 year after the administration of the radiolabelled compound. Materials and methods : Tricolour FISH with whole-chromosome-specific probes for chromosomes 2, 4 and 8 was used for scoring translocations. From the genomic translocation frequencies, derived using the Lucas formula, equivalent whole-body doses were calculated, based on the in vitro 60 Co γ-ray dose-response curve. Results : A total of 101 translocations were observed in 4864 metaphases, 63% being of the two-way type. In the control group used for obtaining dose-response data, nine translocations were observed in 5278 metaphases, 55% being two-way translocations. No correlation was found between the observed frequency of translocations and administered radioactivity. Using the in vitro dose-response, an estimated average dose for the group of nine patients of 0.79 ±0.22Gy was obtained. Compared with frequencies following the assumption that the involvement of a particular chromosome in a two-break exchange-type aberration is proportional to its DNA content, chromosome 4 was more frequently involved and chromosomes 2 and 8 less frequently involved in chromosomal rearrangements. Conclusion : This study shows that 131 I therapy for thyrotoxicosis patients induced translocations, especially in chromosome 4, which could be detected 1 year after the administration of the radiolabelled compound.