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N M Pleskach

Publications and source records attributed to N M Pleskach.

At least 37 records · Page 2Linked to original sources
PubMed1995

[The retardation of DNA synthesis in adjacent replicon clusters in the lymphocytes of patients with Down's syndrome as a model of premature aging].

In peripheral blood lymphocytes taken from 7 patients with Down syndrome (DS) and 2 normal donors, using DNA fiber autoradiography, we estimated the rate of DNA-chain growth, which depends oil the number of simultaneously replicating adjacent replicon clusters, but not on the rate of fork movement. No difference was found in the rate of fork movement in these cells. 6 of 7 DS patients showed a significant reduction in the rate of DNA-chain growth as compared to that in normal control. Thus, a new molecular defect in DS lymphocytes has ben first revealed. Besides, new arguments have been provided in favour of genetic heterogeneity of this genetic disorder belonging to premature ageing diseases. Relation of molecular DNA replication defects with ageing is discussed.

Aging↗
PubMed1994

[The relation of the characteristics of DNA replication to cell sensitivity to ionizing radiation in xeroderma pigmentosum in form II (XP2CP)].

Using DNA fiber autoradiography, DNA replication in cultured fibroblasts, derived from normal donors, and from XPII patient with increased sensitivity to ionizing radiation was estimated. Here evidence is provided on the fact that the fork movement, significantly decreased in XPII cells before irradiation, remains the same after exposure to X-rays. The density of replicon clusters simultaneously operating in tandem groups, which is initially much less in XPII cells compared to normal cells, also remains unchanged after exposure to X-rays (5 Gy), since the inhibition of DNA replication occurs to individual replicons only. Our data suggest that the inhibition of DNA replication in normal cells throughout the whole cluster, that drastically reduces the rate of DNA-chain growth, may provide an additional time to restore damaged chromosomes, i. e. it is part of the cellular defence mechanism. It seems likely that XPII cells are deficient in such a defence mechanism.

Autoradiography↗
PubMed1993

[DNA replication in intact and X-ray-irradiated cells in the Cockayne syndrome].

Using DNA fiber autoradiography, an estimation was made of DNA replication in normal fibroblasts and in those derived from a patient with Cockayne syndrome. The rate of replication fork movement as well as the rate of DNA chain growth, dependent on the frequency of initiation sites in the adjacent clusters of replicons, did not differ in Cockayne syndrome cells, compared to cells of normal donors, either before or after exposure to ionizing radiation.

Autoradiography↗
PubMed1992

[The interrelation between changes in the structural organization of replicon clusters, a retarded fork displacement rate and the high level of spontaneous SCEs in form II of xeroderma pigmentosum].

A cytogenetic observation, that the sister chromatid exchanges (SCE) occur 3 times more frequently in a special form of xeroderma pigmentosum--XPII than in the norm, prompted a study of DNA replication in this rare disease. Using DNA fiber autoradiography, the rate of fork movement and the frequency of initiation in the adjacent clusters of replicons were estimated. The rate of fork movement was significantly slower than that in classical XP and in normal cells. Here evidence was provided on another defect in DNA replication in XPII that involves a significantly decreased number of simultaneously operating adjacent clusters of replicons, which results in a decreased rate of DNA chain-growth. According to the Painter replication model for SCE, the exchanges arise due to double-strand DNA breaks occurring on the border between two adjacent clusters, respectively, completely and partially replicated. A retarded fork-displacement rate together with a decreased rate of DNA-chain growth may cause this situation to persist longer than in the norm. Thus, our data provide a further support of the replication model for SCE. A similar combination of cytogenetic and molecular defects has been obtained earlier in the Bloom syndrome cells.

Autoradiography↗
PubMed1991

[The effect of staphylococcal alpha-toxin on DNA replication in human cells].

The influence of Staphylococcus alpha-toxin has been investigated on the duration of S-phase of lymphocyte mitotic cycle and on DNA replication in human fibroblasts in vitro. The duration of the S-phase of lymphocytes was measured by counting labeled metaphases and by making replication curves. Alpha-toxin in a dose of 3 micrograms/ml enhances the onset of S-phase, which is inhibited at a dose of 33 micrograms/ml of alpha-toxin. The action of alpha-toxin resulted in a decreased rate of replication fork and in a progressive activation of replicon groups. This effect was most prominent at 33 micrograms/ml of alpha-toxin. The data obtained allow to suggest that immunodeficiency of the second order, so characteristic of the staphylococcal sepsis, may be due, in many respects, to suppression of DNA replication.

Autoradiography↗
PubMed1991

[The relationship between the radiosensitivity of DNA replicative synthesis and the formation of sister chromatid exchanges].

It has been found that irradiation in doses 0.5-2.0 Gy does not enhance the frequency of sister chromatid exchanges in cells of patients with Down's syndrome and ataxia-telangiectasia compared to the normal cells. In the case of ataxia, this phenomenon was accompanied with radioresistant replicative DNA synthesis, whereas in two cases of Down's syndrome the replicative DNA synthesis was found to be as radiosensitive as in the norm. According to these data, the mechanism of sister chromatid exchanges proposed in our previous publication (Pleskach et al., 1988) seems to be rather doubtful.

Ataxia Telangiectasia↗
PubMed1990

Sister chromatid exchanges and inhibition of DNA synthesis in irradiated human cells.

X-ray irradiation inhibits DNA synthesis and enhances the frequency of sister chromatid exchanges (SCE) in normal human lymphocytes. On the contrary, cells from patients with Down's syndrome, Xeroderma pigmentosum (form II) and progeria, characterized by radioresistant DNA synthesis, do not show such an increase in SCE frequency. We suggest that radiation-induced SCE frequency is a result of inhibition of DNA replication, rather than a direct damage of chromosomes by ionizing radiation. It is in agreement with Painter's /13/ hypothesis according to which SCE are formed due to asynchronous completion of replication in contiguous replicon clusters. So, probability of SCE formation is the more the lower is rate of replication. Thus, the extent of radiation damage cannot be measured directly by the SCE frequency.

DNA↗
PubMed1990

[The cytogenetic action of antilymphocyte globulin on the lymphocytes of bronchial asthma patients].

Chromosomal aberration levels and frequencies of sister chromatid exchanges (SCE) in peripheral blood lymphocytes were investigated for patients with bronchial asthma treated with antilymphocyte globulin (ALG). The significant elevation of chromosomal aberration levels and SCE frequencies were revealed as compared to the healthy controls. The significant differences in chromosomal aberration levels were not observed before and after the courses of ALG therapy. At the same time the SCE frequencies appeared to be significantly decreased after the course of ALG therapy. The similar effects were observed in studies in vitro. The data obtained may suggest that ALG at the used doses does not cause any significant cytogenetic damages in the examined patients.

Antilymphocyte Serum↗
PubMed1989

[The action of ionizing radiation on DNA replication in the cells of a healthy human subject and of a patient with Down's syndrome].

Using DNA fiber autoradiography we have revealed a new defect earlier unknown in Down's syndrome but analogous to that earlier shown by the authors in AT and basal cell nevus. This syndrome involves a significantly decreased number of simultaneously operating groups of replicons compared to that in normal cells., the rate of fork movement and the fusion of neighbouring units in the group being unchanged. Ionizing radiation (5 Gy) does not change the rate of DNA chain growth in the cells derived from the affected individuals, however, it significantly reduces this parameter in normal cells due to inhibition of replicon initiation in the whole clusters. Thus, radioresistant DNA synthesis in chromosomal instability syndromes may be explained by some defect in DNA replication in unirradiated cells analogous to that in irradiated normal cells.

Autoradiography↗
PubMed1989

[Disruption of DNA replication in non-irradiated cells in basal cell nevus syndrome and the effect of ionizing radiation].

Analysis of DNA fiber autoradiograms from basal cell nevus syndrome (BCNS) skin fibroblasts has revealed for the first time a new defect in DNA replication earlier unknown in other chromosomal instability syndromes, that involves a significantly decreased rate of DNA-chain growth in unirradiated cells. Here we present evidence that the defect may be due to a marked reduction in number of simultaneously operating groups of replicons compared to that in normal cells, the rate of fork movement and the fusion of neighbouring units in the group remaining unchanged. Radioresistant DNA synthesis was observed in the BCNS cells. The exposure of cells derived from normal donor to gamma-rays at a dose of 5 Gy reduces the number of simultaneously operating groups of replicons to the level occurring in unirradiated BCNS cells, the rate of folk movement being unchanged in both cell types. However, the incidence of fusion between neighbouring units within the group is lower in the cells exposed to gamma-rays, due perhaps to a radiation-induced lesion in the group. Thus, ionizing radiation reduces the rate of DNA synthesis to the same level, however from different initial levels. Our data suggest that the phenomenon of radioresistant DNA synthesis may be explained by the presence of the initial defect in DNA replication in BCNS or any other chromosomal instability disorders.

Adult↗
PubMed1988

[Sister chromatid exchanges, during x-irradiation, in the blood lymphocytes of patients with hereditary diseases and radioresistant DNA synthesis].

X-ray irradiation induced sister chromatid exchanges (SCE) in blood lymphocytes from patient with Down's syndrome and adult progeria (in both the cases radioresistant DNA synthesis takes place). In these diseases, likely as upon form II of xeroderma pigmentosum (the replicative DNA synthesis is radioresistant), X-ray irradiation lowers the rate of SCE compared with that in the control, then the SCE rate rises with the increase in radiation dose, reaching the rate of SCE in non-irradiated cells. In normal lymphocytes (in which ionizing radiation inhibits the replicative synthesis of DNA) the rate of SCE rises with the rise of radiation dose. Thus, the rate of SCE in X-ray irradiated lymphocytes is in reverse dependence with radioresistance of replicative synthesis of DNA. The data obtained are explained in accordance with the replicative hypothesis of the SCE nature (Painter, 1980a): in cells of patients with Down's syndrome, xeroderma pigmentosum form II and progeria of adults the time of existence of partly replicated clusters of replicons is decreased due to radioresistant replicative synthesis of DNA, but the presence of partly replicated clusters of replicons is necessary for SCE formation. Therefore the rate of SCE in X-irradiated cells of these patients decreases.

Adolescent↗
PubMed1987

Sister-chromatid exchanges in a special form of xeroderma pigmentosum (form II).

The frequency of sister-chromatid exchanges (SCE) was studied in peripheral blood lymphocytes from a xeroderma pigmentosum (form II, XPII) patient. The cells were irradiated with UV or X-rays. In some experiments novobiocin (NB), inhibitor of topoisomerase II, or caffeine (CA), inhibitor of DNA repair were added to the cultures. The level of spontaneous SCE in the patient's lymphocytes was found to be significantly increased in comparison to that in the cells from normal donors. The inhibitors and UV-light caused a rise in the frequency of SCE in the cells taken from normal donors and except for NB, in the lymphocytes from the patient XPII. X-Rays did not increase SCE frequency in normal lymphocytes and lowered it in the patient's cells. SCE frequency rose when inhibitors of DNA replication and repair were used in combination with mutagens.

Caffeine↗
PubMed1986

Radioresistant DNA synthesis in cells of patients showing increased chromosomal sensitivity to ionizing radiation.

The rate of DNA synthesis after gamma-irradiation was studied either by analysis of the steady-state distribution of daughter [3H]DNA in alkaline sucrose gradients or by direct assay of the amount of [3H]thymidine incorporated into DNA of fibroblasts derived from a normal donor (LCH882) and from Down's syndrome (LCH944), Werner's syndrome (WS1LE) and xeroderma pigmentosum (XP2LE) patients with chromosomal sensitivity to ionizing radiation. Doses of gamma-irradiation that markedly inhibited the rate of DNA synthesis in normal human cells caused almost no inhibition of DNA synthesis in the cells from the affected individuals. The radioresistant DNA synthesis in Down's syndrome cells was mainly due to a much lower inhibition of replicon initiation than that in normal cells; these cells were also more resistant to damage that inhibited replicon elongation. Our data suggest that radioresistant DNA synthesis may be an intrinsic feature of all genetic disorders showing increased radiosensitivity in terms of chromosome aberrations.

Cell Cycle↗
PubMed1986

[Spontaneous and induced sister chromatid exchanges in the blood lymphocytes of healthy persons and of xeroderma pigmentosum patients exposed to the inhibitors of DNA repair and replication caffeine, 3-methoxybenzamide and novobiocin].

The frequency of sister chromatid exchanges (SCEs), both spontaneous and induced by UV-light, X-rays, mitomycin C and ethylmetansulphonate (EMS), has been investigated in cultured human peripheral blood lymphocytes. Besides, frequency of spontaneous and induced SCEs was studied under the action of the inhibitors of topoisomerase II, polymerase poly(ADP-ribose), and DNA repair, i. e. novobiocin, 3-metoxybenzamide, and caffeine, respectively. It is shown that the base-line SCEs in lymphocytes of the patient with xeroderma pigmentosum II (XP2LE) is dramatically higher compared to that in normal and pigmented xerodermoid cells (XP3LE). The above inhibitors of DNA synthesis and repair enhance the rate of spontaneous SCEs in normal, XP2LE and XP3LE cells. UV-, X-ray and chemical mutagens induced an increased frequency of SCEs in these cells. Simultaneous treatment with mutagenes and inhibitors of DNA synthesis and DNA repair enhanced the rate of SCEs in lymphocytes of healthy donors and in the XP3LE patient. The frequency of the XP2LE cells. Novobiocin, 3-MBA and caffeine significantly decreased the frequency of SCEs in mitomycin C- and EMS-treated XP2LE lymphocyte, which nevertheless was much higher than that in normal cells treated with the same agents.

Adult↗
PubMed1985

[Radioresistant DNA synthesis in the fibroblasts of a Down's syndrome patient].

DNA synthesis after gamma-irradiation was analysed either by direct assay of the amount of 3H-Td incorporated into DNA of fibroblasts derived from normal donor and from a patient with Down's syndrome, or by analysis of the steady-state distribution of 3H-DNA in alkaline sucrose gradients. Doses of gamma-radiation that markedly inhibited the rate of DNA synthesis in normal human cells caused almost no inhibition in fibroblasts of the patient with Down's syndrome. The radioresistant DNA synthesis in cells of this patient was mainly due to a much less inhibition of replicon initiation than that in normal cells. Thus, in the case of Down's syndrome, the cells fail to go through the delays during which DNA lesions can be repaired, unlike the situation being the case in normal cells.

Cells, Cultured↗
PubMed1985

[Occurrence and elimination of single- and double-stranded DNA breaks in the fibroblasts of xeroderma pigmentosum patients exposed to gamma radiation].

A defect in ability to rejoin gamma-induced single-strand DNA breaks, earlier found in the lymphocytes of a patient with form II of xeroderma pigmentosum, has been also shown for cultured skin fibroblasts. In successive subcultivation the ability to rejoin DNA breaks gradually increases. Elimination of double-strand DNA breaks occurs with the same fullness and rate as in the cells of patients with classic form of XP and healthy donors. The subcultivation does not influence this process. Possible causes of the phenomena discovered are discussed.

Cells, Cultured↗
PubMed1982

[Cellular DNA repair, proliferative activity and biochemical characteristics in the human premature aging syndrome (progeria)].

Fibroblasts of a patient with the Hutchinson-Gilford progeria show a decreased proliferative activity in culture and run through no more than 19 subcultivations. Progeria cells rejoin gamma-induced single strand DNA breaks to the same extent and with the same rate as do normal cells. Spontaneous and induced by X-ray chromosome aberration frequency in progeria cells does not differ from that in normal cells. The activity of DNA-polymerases alpha, beta and gamma in different way depends on culture conditions in progeria and normal cells, but no expressed deficiency of these enzymes was found in progeria cells. The activity of easy-soluble arginine-specific proteases in progeria fibroblasts is sharply decreased, and sensitivity of proteins in these cells to trypsin-catalyzed hydrolysis is significantly increased compared to that of normal cells. Defect in the turnover of intracellular proteins is considered to be a reason of a rapid accumulation of altered proteins in cycling cells and a possible reason of accelerated ageing of progeria cells in vitro.

Cell Division↗