Different allele frequencies in Trypanosoma brucei brucei and Trypanosoma brucei gambiense populations.

Restriction fragment length polymorphism (RFLP) has been analysed in Trypanosoma brucei DNA following hybridization with different DNA probes. This polymorphism seems to be due to allelic variation, and not to variation between sequence duplicates, since the genomic environment of the probed polymorphic fragments is conserved over considerable distances. In an analysis of 35 non-gambiense stocks, we found different combinations of homozygotes and heterozygotes for the four RFLP probes used, in keeping with previous observations that genetic reassortment occurs in T.

The genome and the antigen gene repertoire of Trypanosoma brucei gambiense are smaller than those of T. b. brucei.

The amount of nuclear DNA of Trypanosoma brucei gambiense is only 70% of that of T. b. brucei.

Trypanosome hybrids generated in tsetse flies by nuclear fusion.

Genetic exchange may occur between two particular Trypanosoma brucei clones simultaneously transmitted by the same tsetse fly. We report here that this exchange takes place in the fly, through nuclear fusion. The resulting hybrids appear to be sub-tetraploid, some particular DNA sequences from one of the parental stocks being lost before enough cloned hybrid trypanosomes could be harvested for DNA analysis.

Chromosome aberration induction in human diploid fibroblast and epithelial cells.

The relative sensitivity of cultured human fibroblasts and epithelial cells to radiation-induced chromosomal aberrations was investigated. Lung fibroblast and kidney epithelial cells from the same fetus were compared, as were skin fibroblasts and epithelial keratinocytes from the same foreskin sample. After exposure of proliferating fetal cells to 1.

Evidence for haploidy in metacyclic forms of Trypanosoma brucei.

The parasitic flagellate Trypanosoma brucei undergoes a series of morphologic and metabolic changes during its passage in the digestive organs of its insect vector, a Glossina or tsetse fly. This morphogenesis ends by the differentiation, in the salivary gland of the fly, of the metacyclic form, which will be transmitted in the bloodstream of the mammalian host. On the basis of DNA microfluorometric measurements, we propose that these metacyclic trypanosomes have a haploid amount of DNA, compared to that of bloodstream forms and also of the proventricular forms, which initiate the invasion of the salivary glands.

The effect of caffeine on X-ray-induced mitotic delay in normal human and ataxia-telangiectasia fibroblasts.

We previously showed that radiation-sensitive fibroblasts from ataxia-telangiectasia (A-T) patients sustain less G2 delay after X-irradiation than normal fibroblasts (Scott and Zampetti-Bosseler, 1982). Caffeine is known to reduce the amount of X-ray-induced delay in various mammalian cell types. Painter and Young (1980) proposed that A-T cells have an altered chromatin structure, similar to that of caffeine-treated normal cells and that this results in a failure of A-T cells to delay their progression through the cell cycle to allow time for DNA repair.

The response of normal and ataxia-telangiectasia cells to bleomycin: relationships between chromosome damage, cell cycle delay and cell killing.

In agreement with our earlier observation (Scott and Zampetti-Bosseler, 1982) on X-irradiated normal and ataxia-telangiectasia (A-T) fibroblasts, we now report that after bleomycin or neocarzinostatin treatment also, A-T cells exhibit less G2 delay than normal cells. We confirm that A-T cells sustain more chromosome damage and lethality than normal cells after bleomycin. These observations support the hypothesis (Painter and Young, 1980) that A-T cells are defective in the recognition of certain lesions which normally lead to delays in progression through the cell cycle, during which they are repaired, and which, if unrepaired, lead to cell-lethal chromosome damage.

Relationships between chromosome damage, cell cycle delay and cell killing induced by bleomycin or X-rays.

The extent of mitotic delay and chromosome aberration induction by X-rays and bleomycin has been compared in normal human foetal fibroblasts at doses giving approximately equal levels of cell killing, assayed as colony-forming ability. Bleomycin induced much less G2 delay and chromosome damage than X-rays. We conclude that the major mechanism of cell killing by bleomycin does not involve chromosome damage but the cells pass through a number of division cycles before dying and a common DNA lesion is involved in G2 delay and chromosome damage.

Cell cycle dependence of mitotic delay in X-irradiated normal and ataxia-telangiectasia fibroblasts.

The delay in progression of X-irradiated cells through the cell cycle, which is more pronounced in normal (N) than in A-T fibroblasts, is greatest for cells in G2 at the time of irradiation. The greater effect of radiation on the initiation of DNA synthesis in N than in A-T cells is reflected in the shape of the percent labelled mitosis curves after 3H-thymidine treatment. The duration of the S phase in unirradiated A-T cells is greater than in N cells.

UV-irradiation of related mouse hybrid cells: similar increase in capacity to replicate intact minute-virus-of-mice but differential enhancement of survival of UV-irradiated virus.

3 hybrid cell lines between mouse fibroblasts (A9) and mouse lymphoma cells (L5178YS) were compared with regard to the ability of UV-pre-irradiated cells to replicate intact (unirradiated) Minute-virus-of-Mice (MVM) and to reactivate UV-irradiated MVM. UV irradiation of cells before virus infection enhanced their ability to plaque intact virus (Enhanced Capacity) to a similar extent in the 3 hybrid cell lines. However, pretreatment of cells with UV radiation enhanced the survival of UV-damaged virus (Enhanced Reactivation) in only 1 of these hybrids.

Cell death, chromosome damage and mitotic delay in normal human, ataxia telangiectasia and retinoblastoma fibroblasts after x-irradiation.

We recently showed (Scott and Zampetti-Bosseler 1980) that X-ray sensitive mouse lymphoma cells sustain more chromosome damage, mitotic delay and spindle defects than X-ray resistant cells. We proposed that (a) chromosome aberrations contribute much more to lethality than spindle defects, and (b) that DNA lesions are less effectively repaired in the sensitive cells and give rise to more G2 mitotic delay and chromosome aberrations. Our present results on human fibroblasts with reported differential sensitivity to ionizing radiation (i.

The relationship between cell killing, chromosome aberrations, spindle defects and mitotic delay in mouse lymphoma cells of differential sensitivity to X-rays.

The ultrasensitivity of a subline of L5178Y mouse lymphoma cells to X-rays was thought to result from chromosome structural aberrations which are much more frequent in these cells than in radiation-resistant cells derived from them (Scott, Fox and Fox 1974). However, Ehmann, Nagasawa, Peterson and Lett (1974) in time-lapse photography studies of the sensitive line, concluded that the induction of multipolar mitoses by X-rays might be a more important mechanism of cell killing than chromosome aberrations. We have now shown that at survival levels above about 20 per cent, chromosome structural aberrations which lead to bridges and fragments at anaphase are about four times more frequent than spindle defects.