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Independent forms of potentially lethal damage fixed in plateau-phase Chinese hamster cells by postirradiation
Abstract:
Plateau-phase Chinese V79 hamster cells were sequentially treated after exposure to gamma rays in medium made hypertonic by the addition of sodium chloride (370 mM) and with various concentrations of 9-beta-D-arabinofuranosyladenine (araA) to study their combined effect on fixation of potentially lethal damage (PLD). A 10-min treatment in hypertonic medium fixed an extensive amount of PLD and caused a decrease in D0 from 1.8 to 1.2 Gy without significantly affecting Dq. Subsequent treatment with araA caused further fixation of PLD but resulted in a specific, concentration-dependent reduction in Dq from 4.9 to 1.6 Gy after a 4-h exposure to 150 microM araA. A 30-min treatment in hypertonic medium reduced not only Do (from 1.8 to 1.0 Gy) but also Dq (from 4.9 to 2.7 Gy). Subsequent treatment with araA in this case affected only the residual shoulder, reducing it to 1.6 Gy after a 4-h treatment with 100 microM araA, a value similar to that obtained after treatment with araA of cells exposed to salt for only 10 min. When the repair of PLD fixed by a 10-min treatment with salt was measured by delaying its postirradiation application in the presence of various amounts of araA, a small decrease in the repair rate was observed but no significant effect on the relative increase in survival. Qualitatively similar results were obtained for repair of PLD sensitive to araA after a 10-min treatment in hypertonic medium. These results suggest the radiation induction of forms of PLD with different sensitivity to fixation by postirradiation treatments. araA is proposed to fix a form of PLD termed alpha-PLD, the repair of which takes place within 4-6 h and which causes the formation of the shoulder in the survival curve of cells plated immediately after irradiation. Short treatments in hypertonic medium (less than 10 min) are proposed to fix a form of PLD termed beta-PLD, the repair of which takes place within 1 h and leads to restoration of the slope to values equal to those obtained in the survival curve of cells plated immediately after irradiation. However, longer treatments in hypertonic medium also affect Dq and thus also alpha-PLD. Repair of beta-PLD was not significantly affected by araA and repair of alpha-PLD was not significantly affected by short hypertonic treatment, thus indicating the independence of the two forms of PLD.(ABSTRACT TRUNCATED AT 400 WORDS)