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E Nasonova

Publications and source records attributed to E Nasonova.

15 recordsLinked to original sources
PubMed2006

Prediction of dose response for radiation induced exchange aberrations taking cell cycle delays into account.

Chromosomal aberrations (CAs) are regarded as one of the most sensitive biological indicators of genetic alterations. The aberration frequency is routinely determined in the first metaphase. Yet, the data interpretation can be complicated due to radiation induced mitotic delays. To investigate the effect of delays on CA frequency in the first mitosis, human lymphocytes were irradiated with X rays and Giemsa detectable CAs were measured at different sampling times. Besides, a computer simulation was performed reproducing the main effects under investigation, that is, CA induction and cell progression through the mitotic cycle. The CA formation model takes into account the structural organisation of interphase chromosomes in a lymphocyte nucleus, DNA double-strand break (DSB) induction and their rejoining/misrejoining. Lymphocyte transition through the cell cycle was simulated by a Monte Carlo technique. The delay was proposed to result from DNA DSBs. The predicted ratios of first/second/third cycle metaphases agree with the experimental data for control and irradiated samples. Both experimental and calculated CA frequencies in the first mitosis were nearly time-independent. This was proposed to result from de-synchronisation of the lymphocyte population.

Cell Cycle↗
PubMed2005

Chromosome aberration yields and apoptosis in human lymphocytes irradiated with Fe-ions of differing LET.

In the present paper the relationship between cell cycle delays induced by Fe-ions of differing LET and the aberration yield observable in human lymphocytes at mitosis was examined. Cells of the same donor were irradiated with 990 MeV/n Fe-ions (LET=155 keV/micrometers), 200 MeV/n Fe-ions (LET=440 keV/micrometers) and X-rays and aberrations were measured in first cycle mitoses harvested at different times after 48-84 h in culture and in prematurely condensed G2-cells (PCCs) collected at 48 h using calyculin A. Analysis of the time-course of chromosomal damage in first cycle metaphases revealed that the aberration frequency was similar after X-ray irradiation, but increased two and seven fold after exposure to 990 and 200 MeV/n Fe-ions, respectively. Consequently, RBEs derived from late sampling times were significantly higher than those obtained at early times. The PCC-data suggest that the delayed entry of heavily damaged cells into mitosis results especially from a prolonged arrest in G2. Preliminary data obtained for 4.1 MeV/n Cr-ions (LET=3160 keV/micrometers) revealed, that these delays are even more pronounced for low energy Fe-like particles. Additionally, for the different radiation qualities, BrdU-labeling indices and apoptotic indices were determined at several time-points. Only the exposure to low energy Fe-like particles affected the entry of lymphocytes into S-phase and generated a significant apoptotic response indicating that under this particular exposure condition a large proportion of heavily damaged cells is rapidly eliminated from the cell population. The significance of this observation for the estimation of the health risk associated with space radiation remains to be elucidated.

Adult↗
PubMed2005

Correlation between mitotic delay and aberration burden, and their role for the analysis of chromosomal damage.

The aim was to investigate further the relationship between radiation-induced mitotic delay and the expression of chromosome damage in V79 cells. Recently published data on the time-course of chromosome aberrations in V79 first-cycle metaphases after exposure to 10.4 MeV u(-1) Ar ions (LET = 1226 keV microm(-1)) were supplemented and reanalysed. A statistical analysis of the distribution of aberrations among cells was performed. Furthermore, cells were grouped into subpopulations carrying 0, 1 -2, 3-4, 5- 6 and 7 or more aberrations. Then, based on the mitotic index, the flux of each subgroup through the first mitosis was determined and the average entrance time to mitosis was estimated. For comparison, the flux of aberrant V79 cells generated by X-irradiation was analysed. Analysis of the Ar ion data revealed that the flux of each subpopulation through the first mitosis is strongly affected by its aberration burden, i.e. a positive correlation between the mitotic delay and the number of aberrations carried by a cell was observed. The distribution of aberrations among cells could be well described by Neyman-type A statistics; the corresponding fit parameters also reflect the damage-dependent mitotic delay. Interestingly, comparison of the flux of Ar ion and X-ray-irradiated V79 cells through mitosis revealed (1) that a direct correlation exists between the number of aberrations carried by a cell and its average entrance time to mitosis, and (2) that this effect is independent of the linear energy transfer. The role of these observations for radiation cytogenetics is discussed.

Animals↗
PubMed2004

Cell cycle arrest and aberration yield in normal human fibroblasts. I. Effects of X-rays and 195 MeV u(-1) C ions.

PURPOSE: To examine the relationship between cell proliferation and the expression of chromosomal damage in normal human skin fibroblasts after X-ray and particle irradiation. MATERIALS AND METHODS: Confluent G0/G1 AG1522B cells were exposed to X-rays or 195MeV u(-1) C ions with a linear energy transfer of 16.6 keV microm(-1) in the dose range 1-4 Gy. Directly after irradiation, cells were reseeded at a low density in medium containing 5-bromo-2'-deoxyuridine. At multiple time points post-irradiation, the cumulative BrdU-labelling index, mitotic index and aberration frequency were measured. Based on these data, the total amount of damage induced within the entire cell population was estimated by means of mathematical analysis. RESULTS: Both types of radiation exposure exert a pronounced effect on the cell cycle progression of fibroblasts. They result in delayed entry of cells into S-phase and into the first mitosis, and cause a dramatic reduction in mitotic activity. Measurement of chromosomal damage in first-cycle cells at multiple time points post-irradiation shows that the frequencies of aberrant cells and aberrations increase with time up to twofold for the lower doses. However, for the higher doses, this effect is less pronounced or even disappears. When the data for the whole cell population are analysed, it becomes evident that only a few damaged fibroblasts can progress to the first mitosis, a response attributable at least in part to a long-term arrest of injured cells in the initial G0/G1-phase. As observed in other investigations, the effectiveness of 195 MeV u(-1) C ions was similar or slightly higher than X-rays for all endpoints studied leading to a relative biological effectiveness in the range 1.0-1.4. CONCLUSIONS: Cell cycle arrests affect the aberration yield observable in normal human fibroblasts at mitosis. The data obtained for the cell population as a whole reveal that injured cells are rapidly removed from the mitotically active population through a chronic cell cycle arrest, which is consistent with other studies that indicate that this response is a specific strategy of fibroblasts to minimize the fixation and propagation of genetic alterations.

Apoptosis↗
PubMed2004

Cytogenetic effects of densely ionising radiation in human lymphocytes: impact of cell cycle delays.

The classical cytogenetic assay to estimate the dose to which an individual has been exposed relies on the measurement of chromosome aberrations in lymphocytes at the first post-irradiation mitosis 48 h after in vitro stimulation. However, evidence is accumulating that this protocol results in an underestimation of the cytogenetic effects of high LET radiation due to a selective delay of damaged cells. To address this issue, human lymphocytes were irradiated with C-ions (25-mm extended Bragg peak, LET: 60-85 keV/ micro m) and aberrations were measured in cells reaching the first mitosis after 48, 60, 72 and 84 h and in G2-phase cells collected after 48 h by calyculin A induced premature chromosome condensation (PCC). The results were compared with recently published data on the effects of X-rays and 200 MeV/u Fe-ions (LET: 440 keV/ micro m) on lymphocytes of the same donor (Ritter et al., 2002a). The experiments show clearly that the aberration yield rises in first-generation metaphase (M1) with culture time and that this effect increases with LET. Obviously, severely damaged cells suffer a prolonged arrest in G2. The mitotic delay has a profound effect on the RBE: RBE values estimated from the PCC data were about two times higher than those obtained by conventional metaphase analysis at 48 h. Altogether, these observations argue against the use of single sampling times to quantify high LET induced chromosomal damage in metaphase cells.

Adult↗
PubMed2002

Effect of LET on the yield and quality of chromosomal damage in metaphase cells: a time-course study.

PURPOSE: To investigate further the effect of linear energy transfer (LET) on the yield and quality of aberrations at different post-irradiation sampling times. MATERIALS AND METHODS: V79 G(1)-cells were exposed to either 10.6 MeV u-1 Ne ions (360 keV microm-1) or 11.1 MeV u-1 Kr ions (3980 keV microm-1) and chromosomal damage was measured in metaphase cells at several 2-h sampling intervals up to 30 h post-irradiation. To differentiate between cells in the first and second post-irradiation cycle, the fluorescence-plus-Giemsa technique was applied. RESULTS: In both experiments, an increase in the yield of aberrant cells as well as the number of aberrations per cell was observed in first- and second-cycle metaphases. Yet, the increase in the number of aberrations per cells was more pronounced for Kr ions and at comparable fluences Kr ions produced more aberrations than Ne ions. Because no sampling time was representative for the whole cell population, the total amount of Ne and Kr ion-induced chromosomal damage was determined by means of a mathematical approach and used for the comparison of data. Furthermore, in accordance with previous studies, LET-dependent changes in the spectrum of aberration types were detected, i.e. with increasing LET a higher fraction of chromatid-type aberrations was observed, although cells had been exposed in G1. In addition, more chromosomal breaks and less exchange-type aberrations were found. CONCLUSIONS: The observation that cell-cycle progression is related to the amount of aberrations harboured by a cell demonstrates that the routinely applied method to measure aberration frequencies in metaphase cells at only one post-irradiation sampling time will unavoidably result in an under- or overestimation of the cytogenetic effects of particles. Consequently, for a meaningful quantification of chromosomal damage, multiple fixation regimes should be used so that the complete time-course of aberrations can be taken into account. Moreover, to avoid bias, all aberration types should be recorded and included in the analysis since the aberration spectrum changes with LET.

Animals↗
PubMed2001

Analysis of Ar-ion and X-ray-induced chromatin breakage and repair in V79 plateau-phase cells by the premature chromosome condensation technique.

PURPOSE: The premature chromosome condensation technique has been used to compare chromatin breakage and repair in noncycling V79 cells following high and low LET radiation. MATERIALS AND METHODS: Plateau-phase V79 cells were exposed to graded doses of low energy Ar ions (LET 1233 keV/microm) and X-rays. Cells were fused to mitotic V79 cells immediately after exposure to examine initial chromatin breakage or after various time intervals of post-irradiation incubation to investigate the kinetics of chromatin break rejoining as well as the fraction of unrejoined fragments. RESULTS AND CONCLUSIONS: For both radiation qualities an average initial number of about 2.4 excess PCC fragments per cell per Gy was found increasing linearly with dose. The distributions of PCC chromosomes plus excess fragments among cells followed Poisson statistics after X-ray irradiation, while an overdispersion of the frequencies was observed after Ar-irradiation indicating that a single particle traversal through a cell nucleus can produce multiple chromatin lesions. Moreover, for both radiation types the rejoining of excess fragments has been examined. Both data sets could be fitted well to first-order kinetics with a single component. Despite similar rates of rejoining cellular repair was noticeably less effective for Ar ions than for X-rays. While after 10 h of post-irradiation incubation 60% of Ar ion induced excess fragments remained unrejoined, only 14% of X-ray-induced lesions were not rejoined. Furthermore, comparison of the residual number of excess PCC fragments with recently published data on the yield of chromosome aberrations in first post-irradiation metaphases shows that for both radiation types more aberrations are detected in interphase than in metaphase cells. Yet, for comparable doses this difference is more pronounced for Ar ions indicating that scoring of high LET induced aberrations in metaphase cells might result in a significant underestimation of the produced damage.

Animals↗
PubMed2001

Mathematical models of radiation-induced mitotic delay: time course analysis and statistics of lesions.

Detailed investigations of high and low LET radiation induced chromosome aberrations in various mammalian cell lines have shown that the registered yield of aberrations depends on cell cycle progression delays. The effect of radiation on the cell kinetics can be analyzed in terms of kinetic growth models. The method yields the number of aberrant cells and the number of aberrations as totals obtained after integration over given time-interval.

Animals↗
PubMed2001

High-LET-induced chromosomal damage: time-dependent expression.

Chromosome aberrations are routinely analysed in metaphase cells at one sampling time post-irradiation. Yet, accumulating evidence shows that radiation-induced cycle perturbations and mitotic delay influence the yield of aberrations detectable in mitosis. In extended time-course studies a drastic increase in the number of aberrations with sampling time has been observed after particle irradiation, while after the exposure to sparsely ionizing radiation a less pronounced effect has been found. This difference in the time-course of chromosomal damage is particularly important for the determination of accurate RBE values. As will be discussed, meaningful RBE values for particles can only be obtained, if cells are analysed at multiple sampling times and the complete time-course of aberrations is considered. Otherwise, particle-induced damage will be over- or underestimated. Moreover, depending on the cell system chosen for the analysis, factors like the loss of damaged cells due to apoptosis or a permanent cell cycle arrest complicate the determination of accurate RBE values based on chromosome data.

Animals↗
PubMed2000

High-LET-induced chromosome aberrations in V79 cells analysed in first and second post-irradiation metaphases.

PURPOSE: As an extension of previous studies, the time-course of high-LET-induced chromosomal damage was investigated in first- and second-cycle V79 Chinese hamster cells. MATERIALS AND METHODS: Cells were exposed in G1 to 10.4 MeV/u Ar ions (LET = 1226 keV/microm) and chromosomal damage was measured at 2h sampling intervals between 10 h and 34 h after irradiation. To distinguish between cells in different post-irradiation cycles, the fluorescence-plus-Giemsa technique was applied. RESULTS: For first- and second-generation cells, the number of aberrant metaphases and aberrations per metaphase were found to increase markedly with sampling time, demonstrating that cell cycle progression was delayed according to the number of lesions carried by the cell. To account for the time-dependent expression of chromosomal damage a mathematical approach was used based on the integrated flux of aberrant cells entering mitosis. Moreover, the analysis of Ar ion-induced chromosome lesions confirmed that high-LET radiation results in specific changes in the spectrum of aberration types. In particular, an increased rate of chromatid-type aberrations as well as a high frequency of chromosomal breaks was found, although the cells were exposed in G1. CONCLUSIONS: Due to the fact that cells collected at one sampling time are not representative of the entire population, the complete time-course of chromosomal damage has to be taken into account for the determination of a meaningful RBE value. Otherwise, the analysis of chromosomal damage can result in a pronounced over- or underestimation of the RBE depending on the subpopulation of cells entering mitosis at that particular sampling time.

Animals↗
PubMed1998

Induction of chromosomal damage in CHO-K1 cells and their repair-deficient mutant XRS5 by X-ray and particle irradiation.

The cytogenetic effects of X-rays and Au ions were investigated in repair-proficient CHO-K1 cells and their radiosensitive mutant strain xrs5, which shows a defect in the rejoining of DNA double-strand breaks. Both cell lines were synchronized by mitotic shake off, irradiated in G1-phase with either 250 kV X-rays or 780 MeV/u Au ions (LET: 1150 keV/micrometer) and chromosome aberrations were analyzed in first post-irradiation metaphases. Isoeffective doses of X-rays for the induction of aberrant cells and aberrations per cell were about 14 times lower for xrs5 than for CHO-K1 cells. After high LET radiation the difference in the cytogenetic response of both cell lines was drastically diminished. Furthermore, the analysis of the aberration types induced by sparsely and densely ionizing radiation showed for both cell lines specific changes in the spectrum of aberration types as LET increases. The experimental results are discussed with respect to the different types of lesions induced by sparsely and densely ionizing radiation.

Animals↗
PubMed1997

Study of Molecular Karyotypes in Amoeboaphelidium protococcarum,the Endotrophic Parasite of Chlorophycean alga Scenedesmus

On the basis of the pulsed field gel electrophoresis(PFGE) technique, molecular karyotypes in Amoeboaphelidiumprotococcarum (the endotrophic parasite of chlorophycean algae, whichcombines protozoon- and fungus-type characters) were determined. Molecularkaryotypes in the strains X1, X5, and X31, which differ in the host range andoriginate from Western, Central, and Eastern Euro-Asia respectively,demonstrate the intrageneric polymorphism-from 7 to 13 chDNA bands-with theestimated molecular size of 0.3-2.2 Mb were resolved, and the genome sizewas determined to be between 7.4 and 10.2 Mb. The molecular karyotype patternin A. protococcarum is different from that in Protozoa. The low-gradedistinction in molecular karyotypes between X1 and X5, on one hand, andhigh-grade distinction between X1/X5 and X31, on the other hand, makes itpossible to place X31 in a separate taxon.

Journal Article↗
PubMed1996

Comparison of chromosomal damage induced by X-rays and Ar ions with an LET of 1840 keV/micrometer in G1 V79 cells.

Synchronous V79 Chinese hamster cells were exposed in G1 to either X-rays or 4.6 MeV/u Ar-ions (LET = 1840 keV/micrometer) and the induction of chromosomal damage was measured at five sampling times ranging from 14 to 30 h after treatment. To distinguish between cells in the first and second post-irradiation cycle the fluorescence-plus-Giemsa technique was applied. The experiment showed that the time-course of the appearance of damaged cells was markedly influenced by radiation-induced cell cycle delays and depended on both radiation quality and dose. The yield of aberrant metaphases and the number of aberrations per metaphase was found to increase with sampling time, but this increase was more pronounced for Ar ions. These differences in yield-time profiles of X-ray and Ar ion induced chromosomal damage are particularly important for an accurate determination of the RBE for particles. Our data clearly indicate that meaningful RBEs can only be obtained if chromosomal damage is analysed at several post-irradiation sampling times and the complete time-course of the expression of chromosomal damage is taken into account. Besides these quantitative differences, differences in the spectrum of chromosomal lesions were observed for X-rays and Ar ions. Following particle exposure more breaks and less exchange-type aberrations were formed compared with X-irradiation and, despite irradiation in G(1), a significant number of chromatid-type aberrations occurred in Ar-irradiated samples. The experimental results are interpreted on the basis of the different pattern of energy deposition by sparsely and densely ionizing radiation. In addition, a statistical analysis based on the Neyman type A distribution is performed, which takes into account the specific stochastic properties of particle irradiation.

Animals↗
PubMed1994

Influence of radiation quality on the expression of chromosomal damage.

The amount of chromosomal damage induced in synchronous V79 cells by either 250 kV X-rays or 4.6 MeV/u Ar ions (LET: 1850 keV/microns) was determined at five successive sampling times. The experiments show that the time course of the appearance of damaged cells is strongly influenced by radiation-induced cell cycle perturbations and mitotic delay and depends on radiation quality and dose. The yield of chromosomal damage was found to increase with sampling time, but this increase was more pronounced for Ar ions. Because of the observed differences in the yield time profiles induced by sparsely and densely ionizing radiation the contribution of each sample to the overall damage was considered, i.e. the total (time integrated) amount of damage was determined. The obtained data are interpreted in terms of differences in the spatial energy deposition by sparsely and densely ionizing radiation.

Animals↗