Biodosimetry for high dose accidental exposures by drug induced premature chromosome condensation (PCC) assay

Balakrishnan, S. A.; Shirsath, Kapil; Bhat, Nagesh; Anjaria, K.B.

doi:10.1016/j.mrgentox.2010.03.008

Cited by 57 publications

(33 citation statements)

References 20 publications

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“…It has been reported that the Poisson distribution applies for whole-irradiated blood, even after high-LET radiation exposure, using the PCC protocol employed in this study, i.e. with the inclusion of Colcemid [9–10], while non-Poisson distributions are more frequently reported using the PCC protocol without Colcemid treatment [11, 20]. By flow cytometry it has been reported that Colcemid treatment in PCC cultures accumulates metaphases, which increase the proportion of G2/M PCC cells [8].…”

Section: Discussionmentioning

confidence: 99%

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Assessment of simulated high-dose partial-body irradiation by PCC-R assay

Romero

García

Lamadrid

et al. 2013

Journal of Radiation Research

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The estimation of the dose and the irradiated fraction of the body is important information in the primary medical response in case of a radiological accident. The PCC-R assay has been developed for high-dose estimations, but little attention has been given to its applicability for partial-body irradiations. In the present work we estimated the doses and the percentage of the irradiated fraction in simulated partial-body radiation exposures at high doses using the PCC-R assay. Peripheral whole blood of three healthy donors was exposed to doses from 0–20 Gy, with 60Co gamma radiation. To simulate partial body irradiations, irradiated and non-irradiated blood was mixed to obtain proportions of irradiated blood from 10–90%. Lymphocyte cultures were treated with Colcemid and Calyculin-A before harvest. Conventional and triage scores were performed for each dose, proportion of irradiated blood and donor. The Papworth's u test was used to evaluate the PCC-R distribution per cell. A dose-response relationship was fitted according to the maximum likelihood method using the frequencies of PCC-R obtained from 100% irradiated blood. The dose to the partially irradiated blood was estimated using the Contaminated Poisson method. A new D0 value of 10.9 Gy was calculated and used to estimate the initial fraction of irradiated cells. The results presented here indicate that by PCC-R it is possible to distinguish between simulated partial- and whole-body irradiations by the u-test, and to accurately estimate the dose from 10–20 Gy, and the initial fraction of irradiated cells in the interval from 10–90%.

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Section: Discussionmentioning

confidence: 99%

“…These and others factors, such as the dose rate, number of cells scored, etc. may determine the differences in coefficients for the linear relationship usually obtained for the PCC-R assay after high doses [8, 9, 11, 20]. …”

Section: Discussionmentioning

confidence: 99%

Assessment of simulated high-dose partial-body irradiation by PCC-R assay

Romero

García

Lamadrid

et al. 2013

Journal of Radiation Research

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“…The immediate induction of PCC in interphase lymphocytes allows rapid scoring of radiationinduced chromosomal aberrations, but the difficulty of the technique and subsequent analysis of the data restrict its use. Various endpoints have been recommended: the frequency of chromosome rings (Lamadrid et al 2007), the mixture of the total number of chromosomes and the G2-PCC index (Gotoh et al 2005), the dimension ratio between the shortest and longest chromosome (Gotoh & Durante 2006), and the yield of additional chromosome pieces (Balakrishnan et al 2010). The frequency of PCC rings has been reported to increase for doses of up to 20 Gy, indicating its suitability for dose assessment after high-dose radiation exposure (Puig et al 2013).…”

Section: Discussionmentioning

confidence: 99%

An alternative approach for the induction of premature chromosome condensation in human peripheral blood lymphocytes using mitotic Akodon cells

Sekaran

Ricoul

Brochard

et al. 2019

International Journal of Radiation Biology

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Purpose: The premature chromosome condensation (PCC) technique is used to study exposure to external radiation through the determination of chromosome fragments observed in interphase cells. The presence of large telomeric signals in CHO cells interferes with the detection of PCC fragments and the identification of dicentric chromosomes. We present an improved method for the fusion of G0-lymphocytes with mitotic Akodon cells (few chromosomes and weakly-staining telomeric sequences) to induce PCC in combination with rapid quantification of dicentric chromosomes and centric rings as an alternative to the classical CHO cell fusion technique. Materials and methods: Whole blood from three healthy volunteers was c-irradiated with 0, 2, or 4 Gy. Following a 24 h incubation post-exposure at 37 C, chromosome spreads of isolated lymphocytes were prepared by standard PCC procedures using mitotic Akodon cells. Results: The percentage of scorable fusions, measured by telomere/centromere (T/C) staining, for Akodon-induced PCC was higher than that for CHO-induced PCC, irrespective of radiation exposure. Importantly, both techniques gave the same result for biodosimetry evaluation. Conclusion: The mitotic Akodon cell-induced PCC fusion assay, in combination with the scoring of dicentric chromosomes and rings by T/C staining of G0-lymphocytes is a suitable alternative for fast and reliable dose estimation after accidental radiation exposure. ARTICLE HISTORY

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“…It is, however, difficult or even impossible to obtain sufficient number of chromosomes from cells exposed high dose of ionizing irradiation, because these cells arrest in S or G2 and do not enter in mitosis. As drug-induced PCC allows us to obtain chromosomes in cells even irradiated 40 Gy dose of γ-rays (Gotoh and Asakawa 1996;Wang et al 2009;Balakrishnan et al 2010;Lindholm et al 2010), we used to calyculin A induced PCC method to visualize the chromosome aberration in G2 cells after γ-irradiation, but we irradiated the cells with more milder dose of 10 Gy of γ-rays as not give severe damages to the cells. Figure 4 (reproduced from Molecular Biology International, Special Issue on "DNA in 3R: Repair, Replication, and Recombination", Gotoh and Asakawa 2011, in press) shows the chromosome painting of human #4 chromosomes against calyculin A induced prematurely condensed chromosomes (PCCs) of PBLs at 48 hours after 10 Gy of γ-rays irradiation.…”

Section: Determination Of Irradiation Dose For Effective Detection Ofmentioning

confidence: 99%

Visualize Dynamics of Chromosome Structure Formation and DNA Repair/Recombination Coupled With DNA Replication: Tight Coupled Role of DNA Replication in Chromosome Compaction and DNA Recombination

Gotoh¹

2011

Fundamental Aspects of DNA Replication

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Biodosimetry for high dose accidental exposures by drug induced premature chromosome condensation (PCC) assay

Cited by 57 publications

References 20 publications

Assessment of simulated high-dose partial-body irradiation by PCC-R assay

Assessment of simulated high-dose partial-body irradiation by PCC-R assay

An alternative approach for the induction of premature chromosome condensation in human peripheral blood lymphocytes using mitotic Akodon cells

Visualize Dynamics of Chromosome Structure Formation and DNA Repair/Recombination Coupled With DNA Replication: Tight Coupled Role of DNA Replication in Chromosome Compaction and DNA Recombination

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