Molecular-biochemical and cytogenetic analyses were made on blood cells of 17 radiation accident victims who, from 1.7 to 43.8 years previously, had suffered acute radiation sickness (ARS) ranging from severity grades I to IV. Molecular-biochemical data were obtained with patients' leukocytes and with mononuclear cells on their oxidative status by a) the level of an anion-radical O2*- in the 3-(4,5-dimethyl-2-thiazol-2-yl)-2,5-diphenyl-2-tetrazolium bromide (MTT) test and b) the sum of reactive oxygen species in the 2,7-dichlorodihydrofluorescein diacetate (DCFDA) test together with a test for DNA strand unwinding in alkaline conditions by measurements of fluorescence intensity of ethidium bromide. Each parameter was measured in freshly sampled cells and during a 5-hour incubation as absolute means on 5 measurements and in % change from the initial values. Cytogenetic data were obtained from the standard metaphase preparations scored for routine unstable chromosomal aberrations (us-CA)--dicentrics; and stable aberrations (sCA)--translocations. The latter ones were assayed by the FISH method using whole chromosome 2, 4 and 12 fluorescent probes and scaled up to genome equivalence. For all patients reduced oxidative status of about 25-30% was obtained by the MTT-test (p < 0.005), and the DCFDA-test (p < 0.027). The yield of usCA depended on the time after irradiation with higher yields associated with the shorter postirradiation times and reducing almost to expected background frequencies. The yield of sCA was high for all patients, correlating with the severity of ARS whilst the molecular-biochemical parameters showed no relationship with ARS. A correlation was observed between parameters of oxidative status and % of cells with usCA: by the MTT-test r = from 0.50 up to 0.61 (p = from 0.06 up to 0.003), but by the DCFDA-test the strength of correlation was smaller: r = from 0.38 up to 0.48. An inverse correlation was found between initial oxidative state of mononuclear cells and the frequency of CA in lymphocytes. Similarly a marked inverse correlation between degree of DNA unwinding by the ethidium bromide assay on leukocytes and sCA in lymphocytes was also noted. The feasibility of radiation-induced delayed genomic instability in vivo for humans long time after irradiation is discussed.
|Number of pages||17|
|Journal||Radiatsionnaia biologiia, radioecologiia / Rossiǐskaia akademiia nauk|
|Publication status||Published - Jul 2006|