The action of low doses of gamma radiation on mammalian cells]

Effect of gamma irradiation in low doses (10, 20, 40 and 50 cGy) on HeLa cells was studied. The survival of cells exposed to the irradiation in the dose of 50 cGy was decreased while it remained unchanged in cells irradiated in the dose of 10-40 cGy and their descendants. Nonetheless, their survival following an additional treatment with a mixture of cytosine arabinoside and hydroxyurea was reduced. It was suggested that the genome stability was diminished in irradiated cells and their descendants.     

Controversial cytogenetic observations in mammalian somatic cells exposed to radiofrequency radiation

During the years 1990-2003 a large number of investigations were conducted using rodents, cultured rodent and human cells, and freshly collected human blood lymphocytes to determine the genotoxic potential of exposure to radiofrequency (RF) radiation. The results of most of these studies (58%) did not indicate increased damage to the genetic material (assessed from DNA strand breaks, incidence of chromosomal aberrations, micronuclei and sister chromatid exchanges) in cells exposed to RF radiation compared to sham-exposed and/or unexposed cells. Some investigations (23%) reported an increase in such damage in cells exposed to RF radiation. The observations from other studies (19%) were inconclusive. This paper reviews the investigations published in scientific journals during 1990-2003 and attempts to identify probable reason(s) for the conflicting results. Recommendations are made for future research to address some of the controversial observations.     

The dose dependence of cytogenetic damages and the adaptive response of mammalian cells under the action of ionizing radiation at low doses

The dose-effect dependence of cytogenetic damage after single dose irradiation in the dose range of 0.1-2 Gy and the adaptive response after double-dose irradiation were studied on Chinese hamster and human melanoma cells in culture. The non-linear dose dependencies were found for the induction of chromosome aberrations with decrease in cell radiosensitivity in the definite dose range. This decrease started at 10 and 20 cGy for melanoma and Chinese hamster cells respectively. The maximal adaptive response was induced at 1 cGy for melanoma cells and at 20 cGy for Chinese hamster cells. It can be supposed that the same inducible repair processes are responsible for non-linearity of dose-effect curves and induction of the adaptive response. These processes are similar in mechanisms and different in quantitative proportion for different cell types.     

DNA damage in Chinese hamster cells repeatedly exposed to low doses of X-rays

In a recent paper we reported the results of an experiment carried out by analysing chromosomal damage in Chinese hamster (CHO) cells exposed to low doses of X-rays. The present investigation was undertaken in order to validate those results using a different approach, the single cell gel electrophoresis assay (comet assay) immediately after irradiation. Cells were cultured during 14 cycles, irradiation treatment was performed once per cycle when the cells were at 90-95% of confluence. Doses of 2.5, 5.0 and 10.0 mSv were used. Sequential irradiation of CHO cells induced a decrease of cells without migration and an increase of cells showing DNA damage with the three doses employed. Significant increases of low-level damaged cells (p < 0.001) were found for the 14 exposures when compared to controls except for the first irradiations with 2.5 and 10 mSv, respectively. No significant increase of the frequency of cells with severe damage was observed in any case. These findings could be explained by assuming a complex interactive process of cell recovery, DNA damage and repair together with the induction of genomic instability, the incidence of bystander effects as well as some kind of radioadaptative response of the cells. If these phenomena are limited to the cell line employed deserves further investigation.     

Controversial cytogenetic observations in mammalian somatic cells exposed to extremely low frequency electromagnetic radiation: a review and future research recommendations

During the years 1990-2003, a large number of investigations were conducted using animals, cultured rodent and human cells as well as freshly collected human blood lymphocytes to determine the genotoxic potential of exposure to nonionizing radiation emitted from extremely low frequency electromagnetic fields (EMF). Among the 63 peer reviewed scientific reports, the conclusions from 29 studies (46%) did not indicate increased damage to the genetic material, as assessed from DNA strand breaks, incidence of chromosomal aberrations (CA), micronuclei (MN), and sister chromatid exchanges (SCE), in EMF exposed cells as compared with sham exposed and/or unexposed cells, while those from 14 investigations (22%) have suggested an increase in such damage in EMF exposed cells. The observations from 20 other studies (32%) were inconclusive. This study reviews the investigations published in peer reviewed scientific journals during 1990-2003 and attempts to identify probable reason(s) for the conflicting results. Recommendations are made for future research to address some of the controversial observations.     

Mutagenic adaptive response to high-LET radiation in human lymphoblastoid cells exposed to low doses of heavy-ion radiation

Adaptive response (AR) and bystander effect are two important phenomena involved in biological responses to low doses of ionizing radiation (IR). Furthermore, there is a strong interest in better understanding the biological effects of high-LET radiation. We previously demonstrated the ability of low doses of X-rays to induce an AR to challenging heavy-ion radiation [8]. In this study, we assessed in vitro the ability of priming low doses (0.01Gy) of heavy-ion radiation to induce a similar AR to a subsequent challenging dose (1-4Gy) of high-LET IR (carbon-ion: 20 and 40keV/μm, neon-ion: 150keV/μm) in TK6, AHH-1 and NH32 cells. Our results showed that low doses of high-LET radiation can induce an AR characterized by lower mutation frequencies at hypoxanthine-guanine phosphoribosyl transferase locus and faster DNA repair kinetics, in cells expressing p53.     

Differentiation and delayed cell death in embryonal stem cells exposed to low doses of ionising radiation

Embryonal stem cells have been used to study the effects of environmentally relevant doses of radiation on cell death and differentation. The ES cells were found to have a greater than 60% chance of surviving the traversal of a single alpha-particle, the lowest possible dose of high linear energy transfer radiation a cell may receive. The ES cells appeared to possess the cell cycle checkpoints believed to prevent the transmission of the radiation damage. However, delayed effects were observed in the progeny. An increased incidence of apoptosis and haempoietic differentiation capacity was found to persist in the ES cell population over many cell divisions. Since both cell death and differentiation are known to play a key role in tissue kinetics, an ES cell model will provide a valuable and versatile cell system for studying the role of cell death and differentiation in the pathology of radiogenic diseases.     

DNA damage in cells exposed to ionizing radiation

Ionizing radiation induces variety of structural lesions in DNA of irradiated organisms. Their formation depends largely on the degree of cell oxygenation, the level of endogenous antioxidants, on DNA-protein complexes and compactization of DNA in the chromatin and activity of DNA repair systems. All ionizing radiation-induced DNA lesions can arbitrarily be divided into two groups. Group 1 includes singly damaged sites (single-sites): base modification, single-strand breaks, alkaline-labile sites (including a basic sites). Group 2 contains: locally multiply damaged sites (clustered lesions), double-strand breaks, intermolecular cross-links. The yields of lesions of group 2 increases with high linear energy transfer of radiation and these lesions play a dominant role in the radiation death, formation of chromosome and gene mutations, cell transformation.     

Cytogenetic effects in children exposed to low doses of ionizing radiation

The obtained data indicate that frequencies of different types of cytogenetic anomalies in investigated children groups living in radionuclide contaminated territories and children irradiated in utero have complicated patterns. The frequency of chromosomal anomalies in the investigated groups of children exceeds the average population level. At the same time, no statistically significant differences in frequencies of various types aberrations between groups of children were revealed.     

DNA damage and repair in mammalian cells exposed to p-hydroxyphenylpyruvic acid

Tyrosinemia type 1 (HT1) is an autosomal recessive disorder of the tyrosine metabolism in which the fumarylacetoacetate hydrolase enzyme is defective. This disease is clinically heterogeneous and a chronic and acute form is discerned. Characteristic of the chronic form is the development of cellular hepatocarcinoma. Although p-hydroxyphenylpyruvic acid (pHPPA) is used as one of the diagnostic markers of this disease, it was suggested that it is unlikely to be involved in the pathophysiology of HT1 as it is present in other disorders that does not have hepatorenal symptoms. It was the aim of this study to investigate the possible effect of pHPPA on DNA damage and repair in mammalian cells. The comet assay was used to establish the genotoxicity of pHPPA in human peripheral blood lymphocytes and isolated rat hepatocytes after their exposure to pHPPA. At first glance the damage to DNA caused by pHPPA seemed reparable in both cell types, however, after challenging the DNA repair capacity of metabolite-treated cells with treatment with H(2)O(2), a marked impairment in the DNA repair capability of these cells was observed. We suggest that the main effect of pHPPA is the long-term impairment of the DNA repair machinery rather than the direct damage to DNA and that this effect of pHPPA, together with the other characteristic metabolites, e.g., FAA and MAA, causes cellular hepatocarcinoma to develop in the chronic form of HT1.     

Reactions of mammalian nerve cells to small doses of radiation

In experiments with rat brain slices it was shown that a radiation-induced short-term increase in spontaneous neuron activity was mainly a function of dose rate. Pulsed X-radiation (pulse length of 2 X 10(-8) s, doses of 3 X 10(-5) to 6 X 10(-4) Gy) caused the most pronounced reactions that were almost completely prevented by caffeine, euphylline, and norepinephrine (10(-4) to 10(-3) M).     

The role of ADP-ribosylation processes in the repair of cytogenetic damage in mammalian cells

Nicotinamide (NA) was shown to increase the yield of chromosome aberrations in irradiated Chinese hamster cells. The effect was observed with all doses used (1-4 Gy) and in all phases of the cell cycle; it was maximum as cells transferred from S to G1 phase. The modification of radiation-induced aberrations was more pronounced in the chromatid deletions and in exchanges. The combined action of NA and caffeine showed a synergism. It is assumed that NA inhibits reparation in a different way than caffeine does.     

Long-term mutagenic effect in various types of somatic cells in monkeys exposed to low doses of penetrating radiation

High level of chromosome aberrations has been determined in different types of somatic cells of monkeys which were subjected to general daily repeated low-capacity radiation (0.049-0.0078 Gy per a day) and accumulated summary doses of about 8.26-36.77 Gy. Long persistence of cytogenetic irradiation aftereffect (15-19 years of observation) has been shown. Differences are revealed in the frequency and types of chromosome aberrations in tissues with different level of cell proliferation: in bone marrow, peripheral blood lymphocytes and epithelial renal cells of irradiated monkeys.     

The genetic basis of resistance to ionising radiation damage in cultured mammalian cells

To test the genetic similarity of independently-isolated hamster cell mutants sensitive to ionising radiation, these were fused in pairs and the hybrids exposed to X-rays. Some mutants (irs1, irs3, xrs-1, XR-1, BLM2) were found to complement all others tested for radiosensitivity in hybrids, and are therefore in separate genetic groups. The mutants irs2 and V-E5, both isolated from V79 cells, did not complement and therefore belong to the same group. Another pair, EM7 and irs1SF, formed hybrids with intermediate levels of survival between mutant and wild-type. However, the parental cells fused to irs1SF also showed intermediate sensitivity, suggesting a semi-dominant mutant phenotype rather than a lack of complementation. Crosses of some of these hamster mutants to the radiosensitive mouse mutant M10 showed clear complementation (irs1 x M10, irs2 x M10) but for others the complementation did not greatly exceed the sensitivity of one (irs3 x M10) or both mutants (XR-1 x M10). Taken with our previously-published data, these results show that there are at least 8 genetic groups determining resistance to ionising radiation damage in rodent cells.     

The observation of mitotic division aberrations in mammalian cells exposed to chemical and radiation treatments

The fidelity of chromosome segregation and the maintenance of the integrity of the chromosome karyotype of eukaryotiic cells is dependent upon the synthesis and functioning of division-related structures such as the nuclear spindle and events such as the attachment of chromosomes to the spindle and their subsequent movement to the poles of the dividing cell. Chemical and physical treatment which modify the synthesis and functioning of division-related events may potentially lead to the production of cells with abnormal chromosome numbers (of both whole chromosome sets and of individual chromosomes).

The ability of environmental agents to modify division-related structures in mammalian cells has been assessed by morphological examination of exposed mitotically dividing cells using staining techniques which identify spindle and chromosome structure and by the analysis of the characteristics of microtubule polymerisation in vitro. Such techniques have been used to identify the spindle-modifying effects of chemicals such as the synthetic hormone diethylstilboestrol and modifications of chromosome to spindle attachment in cells exposed to both UV- and X-irradiation. Such modifications of cell-division-related activities may lead to alterations in the fidelity of division events leading to the production of chromosomally abnormal daughter cells with aneuploid or polyploid karyotypes.     

Lectinhistochemical study of cell surface alterations in mouse embryos exposed to low radiation doses

Summary The peroxidase-coupled Dolichos biflorus agglutinin (DBA) served as a marker for cell surface alterations in embryonic mouse tissues exposed to low-dose radiation during the early organogenesis (day 9 post conception). In unirradiated embryos, DBA bound selectively to various organ primordia, depending on their differentiation state. The auditory vesicles and the developing blood vessels were the only tissues staining strongly with the lectin. The vascular endothelia also showed the highest radiosensitivity, expressed by the maximal reaction already at 12.5 cGy. Marked surface changes as well were registered in the basal part of the Rathke's pocket and in the roof of the diencephalon. After exposure to 25 cGy, a transient amplification of the reaction as compared with 12.5 cGy occurred firstly in the Rathke's pocket, then in the infundibulum and in auditory vesicles. The most distinct effects were achieved with 50 cGy. Remarkable is the prompt rise of the DBA-affinity in a narrow area of the myelencephalon, and subsequently also in the roof of the diencephalon. Furthermore, the infundibulum and the Rathke's pocket, both anlagen of the pituitary gland, bound heavily DBA during the entire examination period of 24 h. The present study demonstrated the oustanding suitability of theDolichos biflorus agglutinin for histochemical detection of subtle cellular alterations by small radiation doses.     

The micronucleus assay as a biological dosimeter in hospital workers exposed to low doses of ionizing radiation

The health risk associated with low levels of ionizing radiation is still a matter of debate. A number of factors, such as non-target effects, adaptive responses and low-dose hypersensitivity, affect the long-term outcome of low-dose exposures. Cytogenetic bio-dosimetry provides a measure of the absorbed dose, taking into account the individual radiation sensitivity. The aim of the present study is to evaluate the value of the micronucleus (MN) test as a bio-dosimeter in hospital workers exposed to low doses of ionizing radiation. Blood samples were obtained from 30 subjects selected among workers exposed to X- and gamma-radiation, and 30 controls matched for sex, age and smoking from the same hospital. Micronucleus frequencies were analyzed by use of the cytokinesis-block method. The MN frequency was compared among the groups considering the confounding factors and the length of employment. No increase in the number of bi-nucleated cells with MN (BNMN), but a significant increase in the number of mono-nucleated cells with micronuclei (MOMN) was observed in exposed subjects compared with the controls. The relationship between MN frequency and accumulated dose (mSv) was evaluated. The length of employment did not affect the extent of MN frequency, but an increase of BNMN and MOMN cells was observed based on the accumulated radiation dose. Our study shows the sensitivity of the MN test in the detection of cytogenetic effects of cumulative exposure levels, suggesting the potential usefulness of this assay in providing a biological index in medical surveillance programs.     

Cytogenetic effects of low doses of radiation in mammalian cells: analysis of the hypersensitivity phenomenon and induced resistance

The induction of cytogenetic damage after irradiation of chinese hamster cells and human melanoma cells within a dose range 1-200 cGy was studied. The anaphase and metaphase analysis of chromosome damage and micronuclei test were applied. The hypersensitivity (HRS) at doses below 20 cGy and the increased radio-resistence at higher doses (IR) were shown with all cytogenetic critheria for both cell lines. The phenomenon of HRS/IR was reproduced in synchronic as well as in a synchronic population of chinese hamster cells. This fact shows that HRS was caused by high radiosensitivity of all cells and can not be explained by any differential sensitivity of cells in different phase of the cell cycle. So it was supposed that the increasing radio-resistence is determined by the inclusion of the inducible repair processes in all cells. This conclusion consents with the facts, that there was no evidence of HRS on dose-effect curves and that some parts of pre-existent damage was repaired after preliminary irradiation with low doses (1-20 cGy) which induce repair processes. It can be concluded that same inducible repair processes an analogous in mechanisms underlying in the base of HRS/IR phenomenon and adaptive response.