Reflections and meditations upon complex chromosomal exchanges.

The application of FISH chromosome painting techniques, especially the recent mFISH (and its equivalents) where all 23 human chromosome pairs can be distinguished, has demonstrated that many chromosome-type structural exchanges are much more complicated (involving more "break-rejoins" and arms) than has hitherto been assumed. It is clear that we have been greatly under-estimating the damage produced in chromatin by such agents as ionising radiation. This article gives a brief historical summary of observations leading up to this conclusion, and after outlining some of the problems surrounding the formation of complex chromosomes exchanges, speculates about possible solutions currently being proposed.

Effects of the K-shell X-ray absorption of phosphorus on the scission of the pentadeoxythymidylic acid.

The scission of pentadeoxythymidylic acid, d(pT)5, by monochromatic soft X-rays on (2153 eV) and below (2147 eV) the K-shell absorption peak of phosphorus was studied as a model of strand breakage in DNA. Samples dried on glass plates were irradiated by monochromatic soft X-rays in vacuo, and the products were analysed by HPLC. The main products, in ascending order of the retention time, were thymine, 5'-dTMP, d(pT)2, d(pT)3, d(pT)4, d(pTp) and three unknown products (UK 1, 2 and 3), which were presumed to be d(pTpTp), d(pTpTpTp) and d(pTpTpTpTp), respectively. No difference between 2147 and 2153 eV irradiation in the nature of the induced products was detected, indicating that the K-shell absorption of phosphorus and the following Auger process produced the same types of products as those produced by photoabsorption at the other shells of phosphorus and at other atoms. The cross-sections for the induction of products at 2153 eV were 3.3-4.0 times larger than those at 2147 eV, the ratios of these values being scattered around the ratio (3.65) of absorbance of the sample between 2153 and 2147 eV. The dependence on the X-ray energy, however, almost disappeared after conversion from exposure to absorbed dose; the ratios of the G-values (number of products per 100 eV) of the products were 0.92-1.11. Photoabsorption at the K-shell of phosphorus induced products comparable or slightly less effectively than photoabsorption at the others. These results indicate that the K-shell absorption of phosphorus and the following Auger process do not have any characteristic effect on strand breakage in dry DNA, either qualitatively or quantitatively.

[The radiation load of the Swiss population due to radiodiagnosis]. / Die Strahlenbelastung der Schweizer Bevölkerung infolge röntgendiagnostischer Unversuchungen

It is recommended that the so-called "genetically significant dose" (GSD) be applied as a quantitative index of the radiation exposure of the genetic material of a population, i.e. the dose which, if received by every member of the population, would be expected to produce the same total genetic injury to the population as do the actual doses received by the various individuals. The GSD of the Swiss population for the year 1971 was calculated on the basis of information received from physicians concerning approximately 60 000 patients. Exeripmental determination of the gonad dose was done in various hospitals with the aid of TLD dosimeter, and sensitive high-pressure and normal-pressure ionization chambers. The results show that the annual rate of increase of X-ray diagnostic examinations is 3-4% in Switzrland as elsewhere. The GSD for Switzerland amounted in 1971 to 42-43 mrad per person as compared with 22 mrad in 1957. The "annual mean marrow dose of a population" can be calculated from the single doses received from each part of active bone marrow irradiated in the course of an X-ray examination and, as in the case of the genetically significant dose, from the demographic figures mentioned. The annual mean marrow dose (1971) for the Swiss population was tentatively calculated using the values of the mean marrow dose per examination of the UK survey of 1957/58 and amounts to about 121 mrad. Barium meal contributes about 30% of the total, while intravenous pyelography, carium enema, lumbar spine and mass miniature radiographies each account for about 10%.

Mutagenic DNA repair in Escherichia coli. II. Factors affecting loss of photoreversibility of UV induced mutations.

The photoreversibility of UV-induced mutations to Trp+ in strain Escherichia coli WP2 uvrA trp (unable to excise pyrimidine dimers) was lost at different rates during incubation in different media. In Casamino acids medium after a short initial lag, photoreversibility was lost over about one generation time; in minimal medium with tryptophan, photoreversibility persisted for more than two generations; in Casamino acids medium with pantoyl lactone photoreversibility was lost extremely slowly. The rate of loss of photoreversibility was unaffected by UV dose in either Casamino acids medium or in minimal medium. The same eventual number of induced mutants was obtained when cells were incubated for two generations in any of the three media before being transferred to selective plates supplemented with Casamino acids. Thus in each the proportion of cells capable of giving rise to a mutant was the same and only the rate at which these cells did so during post-irradiation growth varied, suggesting that there might be a specific fraction of pyrimidine dimers at a given site capable of initiating a mutagenic repair event, and that the size of this fraction is dose dependent. Segregation experiments have shown that error-prone repair appears to occur once only and is not repeated in subsequent replication cycles, in contrast to (presumed error-free) recombination repair. The results are discussed in the light of current models of UV mutagenesis.

[Clonal analysis of the progeny of UV-irradiated Bacillus subtilis (uvr+ and uvr-) cells]. / Klonal'nyi analiz potomstva UF-obluchennykh kletok Bacillus subtilis (uvr+ i uvr-)

The revertants to adenine prototrophy or mutants to auxotrophy can be easily identified on synthetic media which are partly enriched by caseine hydrolysate and yeast extract. It is shown with the use of these media that 1.5% colonies formed by Bacillus subtilis cells of the original type (ade6 met5) have mutant clones which are initiated by spontaneous revertants to adenine prototrophy. These revertants arise in the time of division of cells in macrocolonies. After plating diluted suspension of irradiated cells those colonies which contain mutant clones formed by spontaneous revertants can be erroneously taken for mixed colonies formed by induced revertants. About 40% mutants to auxotrophy induced by high dose of UV-light in in uvr+ cells form pure mutant colonies. The same mutants, induced by uvr cells by five time less UV-dose, usually form mixed colonies.

Integration efficiency in DNA-induced transformation of Pneumococcus. II. Genetic studies of mutant integrating all the markers with a high efficiency.

Transformation of the pneumococcus mutant 401 by DNA's bearing the standard reference marker and several other markers belonging to two unlinked loci has shown that differences in the integration efficiencies of these markers were considerably reduced in this strain compared to the wild-type strain Cl(3). The sensitivities of mutant 401 to ultraviolet light and to X-ray irradiation are the same as those of Cl(3). However, in 401 all the markers tested are more resistant to inactivation as shown by transformation of 401 and Cl(3) by ultraviolet-irradiated DNA. The increase in resistance is greater for low efficiency (LE) markers than for high efficiency (HE) markers.-The decreased discrimination between LE and HE markers in strain 401 is not due to a mechanism related to modification of markers in the transforming DNA by the recipient cells, nor are the proteins inducing competence of the cells responsible for the differences in the integration efficiencies of various markers.-Genetic studies of the fate of recombinants as well as the measure of the amount of DNA taken up have shown that all the markers are integrated in strain 401 by the same recombination process, that specific to high efficiency markers.