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• W2126876913 abstract "ABSTRACT A quantitative method of replicon analysis of DNA fibre autoradiographs has been used to study the relationship between mean rate of DNA chain growth (R) and distance between adjacent replicons (ID) in fibroblasts from cancer-prone conditions. Results are expressed in terms of the mean linear regression R = δ + (K.ID) 10−2. When replicon behaviour was examined in cells from patients with ataxia telangiectasia, basal cell naevus and Bloom’s syndromes grown at high density after 48 h in culture, no significant differences could be found in replicon behaviour between these syndromes and normal cultures. How ever when Bloom’s cells were grown at low density and examined 24 h earlier, the mean rate of chain growth R was reduced compared to normal cells at the same density. Both cell types at high densities at 24 h showed equal but lower R values than at 48 h after plating the cultures. The lower rate of chain growth in Bloom’s was accompanied by a longer S-period and cell cycle. Studies of cell proliferation kinetics using consecutive mitoses after bromodeoxyuridine (BUdR) incorporation and harlequin banding showed that Bloom’s cells at low cell density require a longer period to recover a normal cell cycle length after plating than do normal cells at the same density. Plating at higher densities and using conditioned media shorten the recovery period in Bloom’s cells, and when foetal calf serum/MEM is replaced by human AB serum/McCoy 5a medium as the growth media, cell cycle behaviour of low density Bloom’s and normal cells are equal at a much earlier time. It is concluded that the slow rate of DNA chain growth in Bloom’s cells is an artefact introduced by culture conditions and also may be present in normal cells at an earlier period. The behaviour of replicons during this recovery period appears to be similar in Bloom’s and normal cells except for the time lag. As recovery proceeds, the DNA chain growth in the associated replicon pairs recover progressively. This alters both the mean R value from 0·4 to 0·8 μm/min, the slope of the regression K from &lt; 1·0 1·0 ∼ 1·0 while the distance between initiation sites (ID) remains constant throughout. Pretreatment of all cultures with fluorodeoxyuridine (FUdR) produced the same differential effect on release from DNA synthesis inhibition, that is a similar increase in the activation of normally inactive replicons and a slightly slower rate of chain growth over all replicons. No evidence of a substance released by Bloom’s cells in culture capable of increasing the sister-chromatid frequency in normal cells could be found. Since SCE frequencies were found to increase with fixation time after BUdR introduction it is concluded that some of the reported changes could be due to differences in cell cycle kinetics brought about by the different media conditions." @default.
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• W2126876913 date "1979-12-01" @default.
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• W2126876913 title "Quantitative replicon analysis of DNA synthesis in cancer-prone conditions and the defects in Bloom's syndrome" @default.
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• W2126876913 doi "https://doi.org/10.1242/jcs.40.1.125" @default.
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