PMID- 26520380 OWN - NLM STAT- MEDLINE DCOM- 20160209 LR - 20161126 IS - 1879-3592 (Electronic) IS - 1383-5718 (Linking) VI - 793 DP - 2015 Nov TI - Triage biodosimetry using centromeric/telomeric PNA probes and Giemsa staining to score dicentrics or excess fragments in non-stimulated lymphocyte prematurely condensed chromosomes. PG - 107-14 LID - S1383-5718(15)00153-9 [pii] LID - 10.1016/j.mrgentox.2015.06.013 [doi] AB - The frequency of dicentric chromosomes in human peripheral blood lymphocytes at metaphase is considered as the "gold-standard" method for biological dosimetry and, presently, it is the most widely used for dose assessment. Yet, it needs lymphocyte stimulation and a 2-day culture, failing the requirement of rapid dose estimation, which is a high priority in radiation emergency medicine and triage biodosimetry. In the present work, we assess the applicability of cell fusion mediated premature chromosome condensation (PCC) methodology, which enables the analysis of radiation-induced chromosomal aberrations directly in non-stimulated G0-lymphocytes, without the 2-day culture delay. Despite its advantages, quantification of an exposure by means of the PCC-method is not currently widely used, mainly because Giemsa-staining of interphase G0-lymphocyte chromosomes facilitates the analysis of fragments and rings, but not of dicentrics. To overcome this shortcoming, the PCC-method is combined with fluorescence in situ hybridization (FISH), using simultaneously centromeric/telomeric peptide nucleic acid (PNA)-probes. This new approach enables an accurate analysis of dicentric and centric ring chromosomes, which are formed within 8h post irradiation and will, therefore, be present in the blood sample by the time it arrives for dose estimation. For triage biodosimetry, a dose response curve for up to 10Gy was constructed and compared to that obtained using conventional metaphase analysis with Giemsa or centromeric/telomeric PNA-probes in metaphase. Since FISH is labor intensive, a simple PCC-method scoring Giemsa-stained fragments in excess of 46 was also assessed as an even more rapid approach for triage biodosimetry. First, we studied the rejoining kinetics of fragments and constructed a dose-response curve for 24h repair time. Then, its applicability was assessed for four different doses and compared with the PCC-method using centromeric/telomeric PNA-probes, through the evaluation of speed of analysis and minimum number of cells required for dose estimation and categorization of exposed individuals. CI - Copyright (c) 2015 Elsevier B.V. All rights reserved. FAU - Karachristou, Ioanna AU - Karachristou I AD - Laboratory of Health Physics, Radiobiology & Cytogenetics, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", Athens, Greece. FAU - Karakosta, Maria AU - Karakosta M AD - Laboratory of Health Physics, Radiobiology & Cytogenetics, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", Athens, Greece. FAU - Pantelias, Antonio AU - Pantelias A AD - Laboratory of Health Physics, Radiobiology & Cytogenetics, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", Athens, Greece. FAU - Hatzi, Vasiliki I AU - Hatzi VI AD - Laboratory of Health Physics, Radiobiology & Cytogenetics, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", Athens, Greece. FAU - Karaiskos, Pantelis AU - Karaiskos P AD - Medical Physics Laboratory, Medical School, University of Athens, Greece. FAU - Dimitriou, Panagiotis AU - Dimitriou P AD - Medical Physics Laboratory, Medical School, University of Athens, Greece. FAU - Pantelias, Gabriel AU - Pantelias G AD - Laboratory of Health Physics, Radiobiology & Cytogenetics, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", Athens, Greece. FAU - Terzoudi, Georgia I AU - Terzoudi GI AD - Laboratory of Health Physics, Radiobiology & Cytogenetics, Institute of Nuclear & Radiological Sciences & Technology, Energy & Safety, National Centre for Scientific Research "Demokritos", Athens, Greece. Electronic address: gterzoudi@rrp.demokritos.gr. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150625 PL - Netherlands TA - Mutat Res Genet Toxicol Environ Mutagen JT - Mutation research. Genetic toxicology and environmental mutagenesis JID - 101632149 RN - 0 (Azure Stains) RN - 0 (DNA Probes) RN - 0 (Peptide Nucleic Acids) SB - IM MH - Azure Stains MH - Cells, Cultured MH - Centromere/*genetics MH - *Chromosome Aberrations MH - DNA Probes/genetics MH - Humans MH - In Situ Hybridization, Fluorescence MH - Lymphocytes/cytology/*radiation effects MH - Peptide Nucleic Acids/*genetics MH - Radiometry/*methods MH - Resting Phase, Cell Cycle MH - Telomere/genetics MH - Triage/methods OTO - NOTNLM OT - Cell fusion OT - Centromere/telomere staining OT - Excess PCC fragments OT - PNA-FISH OT - Premature chromosome condensation OT - Triage biodosimetry EDAT- 2015/11/02 06:00 MHDA- 2016/02/10 06:00 CRDT- 2015/11/02 06:00 PHST- 2015/06/15 00:00 [received] PHST- 2015/06/17 00:00 [accepted] PHST- 2015/11/02 06:00 [entrez] PHST- 2015/11/02 06:00 [pubmed] PHST- 2016/02/10 06:00 [medline] AID - S1383-5718(15)00153-9 [pii] AID - 10.1016/j.mrgentox.2015.06.013 [doi] PST - ppublish SO - Mutat Res Genet Toxicol Environ Mutagen. 2015 Nov;793:107-14. doi: 10.1016/j.mrgentox.2015.06.013. Epub 2015 Jun 25.