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G Obe

Publications and source records attributed to G Obe.

At least 37 records · Page 2Linked to original sources
PubMed2000

Lipid peroxidation status, somatic mutations and micronuclei in peripheral lymphocytes: a case observation on a possible interrelationship.

A controlled dietary study was conducted in healthy female volunteers and reported elsewhere [1]. In a subset of samples four different biomarkers were analyzed: plasma malondialdehyde (MDA) levels and urinary 8-isoprostaglandin-F(2alpha) were measured as markers for lipid peroxidation. The frequency of hprt (hypoxanthine guanine phosphoribosyl transferase) mutants and micronuclei in peripheral blood lymphocytes were analyzed as indicators of genotoxic effects. One of the ten individuals showed extremely high background levels in all of the four endpoints measured. This case observation raises the possibility that life style factors and dietary habits affect the level of DNA reactive lipid peroxidation products, which in turn increase mutagenic and cytogenetic effects. A possible association between these biomarkers, particularly in relation to dietary fat intake and antioxidant status, should now be studied in a larger trial.

Adult↗
PubMed2000

Chromosomal aberrations of blood lymphocytes induced in vitro by radon-222 daughter alpha-irradiation.

Blood samples were irradiated in vitro with alpha-rays emitted from short-lived radon decay products dissolved in the culture medium at doses between 0.03 and 41.4 mGy. The data were collected from experiments conducted during the period 1984-1992 and comprise a total of about 64000 scored metaphases. For statistical reasons, only 60,022 metaphases were used for the subsequent analysis. The results for total chromosome aberrations and dicentrics indicate a linear dose dependence in the dose range above about 10 mGy, consistent with other experimental observations. At doses below about 10 mGy, aberration frequencies cannot be linearly extrapolated from higher doses, suggesting that there is no dependence on dose within a certain low-dose range. In addition, a statistically significant minimum has been observed at a dose of about 0.03 mGy, which is consistently lower than the related control values. The behavior of the aberration frequencies in the low-dose region seems to be influenced by the control values, which also depend on the environmental radiation burdens to the donors before blood sampling and thus were significantly affected by the Chernobyl fallout.

Alpha Particles↗
PubMed2000

Mechanisms of DNA double-strand break repair and their potential to induce chromosomal aberrations.

DNA double-strand breaks (DSB) are considered to be critical primary lesions in the formation of chromosomal aberrations. DSB may be induced by exogenous agents, such as ionizing radiation, but also occur spontaneously during cellular processes at quite significant frequencies. To repair this potentially lethal damage, eukaryotic cells have evolved a variety of repair pathways related to homologous and illegitimate recombination, also called non-homologous DNA end joining, which may induce small scale mutations and chromosomal aberrations. In this paper we review the major cellular sources of spontaneous DSB and the different homologous and illegitimate recombination repair pathways, with particular focus on their potential to induce chromosomal aberrations.

Animals↗
PubMed1999

Influence of sodium butyrate on the induction of radiation-induced chromosomal aberrations and sister chromatid exchanges in Chinese hamster ovary (CHO) cells.

CHO cells were pre-treated with sodium butyrate (SB) for 24 h and then X-irradiated in G1. Metaphases were scored for the induction of chromosomal aberrations and sister chromatid exchanges (SCEs). The data were compared with those obtained after irradiation of cells not pre-treated with SB and showed that SB has different effects on the endpoints examined. The frequencies of dicentric chromosomes were elevated and of small acentric rings (double minutes, DMs) reduced. These results are discussed to be a consequence of conformational changes in hyperacetylated chromatin which could lead to more interchromosomal and to less intrachromosomal exchanges. SB itself induces a few SCEs but suppresses the induction of SCEs by X-rays. We assume that a minor part of radiation induced SCEs are 'false' resulting from structural chromosomal aberrations, such as inversions, induced in G1. Inversions are the symmetrical counterparts of DMs. If inversions are suppressed by SB treatment to a similar extent as DMs a small reduction of SCEs by SB can be expected.

Animals↗
PubMed1999

X-irradiation of G1 CHO cells induces SCE which are both true and false in BrdU-substituted cells but only false in biotin-dUTP-substituted cells.

The SCE-test is widely used in genetic toxicology and therefore knowledge of the contribution of BrdU to the formation of spontaneous and induced SCE is of great importance. The present study was undertaken to analyse the role of BrdU in X-ray-induced SCE. If SCE resulted from inversions, rings and double minutes (RDM) would be the asymmetrical counterparts of SCE and should therefore have the same frequencies. Dose-effect relationships of SCE and RDM show that the frequencies of SCE are much higher than those of RDM. We conclude that only a few SCE may represent inversions. In a second set of experiments, endoreduplications were induced in cells irradiated either before or after labelling with BrdU. Analysis of SCE in endoreduplicated chromosomes allows the discrimination of the cell cycle in which they originated. The results show that SCE are only induced in the first cell cycle following irradiation of BrdU-substituted cells, indicating that labelling with BrdU is a necessary prerequisite for the formation of SCE. In order to test this directly, radiation-induced SCE frequencies were studied in cells prelabelled with BrdU or biotin-dUTP in a third set of experiments. The structure of biotin-dUTP suggests that, in contrast to BrdU, it does not give rise to radicals during irradiation. Significantly lower frequencies of SCE were observed in biotin-dUTP-substituted cells than in BrdU-labelled cells. Calculations show that nearly all SCE induced in biotin-dUTP-labelled chromosomes can be explained by chromosomal aberrations (false SCE). In contrast to this, most SCE induced by X-rays in BrdU-labelled cells are not due to chromosomal aberrations, but result from S-dependent lesions (true SCE). This clearly points towards radiation damage in BrdU-moieties as the source of DNA lesions leading to SCE.

Animals↗
PubMed1999

Analysis of X-ray-induced aberrations in human chromosome 5 using high-resolution multicolour banding FISH (mBAND).

Peripheral lymphocytes were exposed to 4 Gy X-rays and aberrations were analysed in human chromosome 5 using high-resolution multicolour banding fluorescence in-situ hybridization (mBAND). This method is suited to detect simple and complex aberrations including peri- and paracentric inversions and exchanges between both chromosomes 5. Additionally, breakpoints carr be assigned to specific regions in chromosome 5. Quantitative relationships of induced aberration types are discussed.

Chromosome Aberrations↗
PubMed1999

Sister chromatid differentiation with biotin-dUTP.

The method of choice to differentiate sister chromatids is to incorporate BrdU in replicating DNA. The disadvantage of BrdU is that its spontaneous or induced radicalization may itself lead to sister chromatid exchanges. Biotin-labelled dUTP is a widely used thymidine analogon for labelling isolated DNA. Its chemical structure suggests that, in contrast to BrdU, it does not give rise to radical formation. We electroporated proliferating Chinese hamster ovary (CHO) cells in the presence of biotin-dUTP which was subsequently detected in metaphase cells with TRITC-conjugated avidin. Microscopic analysis of second mitoses after labelling revealed a clear differential staining of sister chromatids. Thus substitution of thymidine with biotin-dUTP is another method to analyse SCE.

Animals↗
PubMed1999

Chromosomal alterations in peripheral lymphocytes of female cabin attendants.

PURPOSE: To analyse lymphocyte chromosomes from female cabin attendants flying the North Atlantic route. MATERIALS AND METHODS: Peripheral lymphocytes from 58 cabin attendants and 29 controls, all female, were cultured and analyzed for structural chromosomal aberrations and sister chromatid exchanges (SCE). Cytogenetic data from 56 age-matched females from Berlin investigated between 1994 and 1996 as part of routine medical checks were used as an additional control group. RESULTS: The mean frequencies of dicentric and ring chromosomes per 1000 cells amount to 1.3+/-0.2 in cabin attendants, 1.4+/-0.2 in ground personnel and 1.3+/-0.2 in the Berlin control group. Sister chromatid exchanges were found at mean frequencies of 7.0+/-0.1, 6.8+/-0.1 and 6.8+/-0.2 per cell respectively in the three groups. CONCLUSIONS: When compared with controls, cabin attendants have no elevation of the frequencies of dicentric and ring chromosomes and SCE.

Adolescent↗
PubMed1999

Manned missions to Mars and chromosome damage.

PURPOSE: To estimate the frequencies of dicentric chromosomes in peripheral lymphocytes of astronauts on missions to Mars. MATERIALS AND METHODS: Calculations of expected frequencies of dicentrics are based on dose estimations and lymphocyte biology. RESULTS: Frequencies of dicentrics are predicted to be 10-40 times higher than the background level. CONCLUSIONS: Stressing the importance of careful mission planning and optimized shielding of transfer vehicle to Mars.

Astronauts↗
PubMed1999

Comparison of AluI-induced frequencies of dicentrics and translocations in human lymphocytes by chromosome painting.

It has been shown repeatedly that following irradiation of human lymphocytes in the G0 stage, more translocations are induced than dicentrics. To check the role of DNA double-strand breaks (DSB) alone for the induction of symmetrical and asymmetrical chromosome aberrations, the frequencies of induced exchange aberrations by the restriction enzyme AluI were analyzed. The enzyme was introduced into cells using the pellet pipetting technique. Frequencies of induced translocations and dicentrics were determined using a chromosome painting assay with chromosome-specific DNA libraries for chromosomes 1, 4 and X (representing 16.8% of the human genome). The number of translocations detected was approximately 3-fold higher than the number of dicentrics, indicating that the increased frequency of translocations compared with dicentrics found in irradiated human lymphocytes does not result from DNA lesions other than DSB but from differential processing of DSB.

Centromere↗
PubMed1999

Analysis of inversions and sister chromatid exchanges in chromosome 3 of human lymphocytes exposed to X-rays.

It has been shown repeatedly that exposure of G(1) cells unifilarily labelled with 5-bromodeoxyuridine (BrdU) to X-rays leads to sister chromatid exchanges (SCE) when the cells are allowed to grow for one further cycle in the absence of BrdU. It has been suggested that damage induced by ionizing radiation does not lead to 'true' SCE and that the observed SCE are 'false', resulting from structural chromosomal aberrations, especially interstitial inversions. We used a painting probe for the p14 region of human chromosome 3 and anti-BrdU antibodies to analyse the frequency of radiation-induced SCE in that chromosome. This method allowed us to discriminate between para- and pericentric 'true' and 'false' SCE. Our results indicate that most radiation-induced SCE do not result from inversions.

Adolescent↗
PubMed1999

Cytogenetic investigations in flight personnel.

During long-distance flights at high altitudes flight personnel are exposed to cosmic radiation. In order to determine whether there are biological effects of such low dose radiation exposure in aircrew, chromosomal aberrations were investigated in 59 female cabin attendants and a matched control group of 31 members of station personnel. The mean number of dicentric chromosomes amounts to 1.3 (95% CI 1.0-1.6) per 1000 cells in cabin attendants and 1.4 (95% CI 1.0-1.9) per 1000 cells in controls. In an additional control group of 56 female clerks from Berlin the mean frequency of dicentric chromosomes was 1.3 (95% CI 1.0-1.6). Neither in dicentric frequency and distribution nor in other aberrations was a significant difference between the groups of flight and station personnel found. The high frequency of multi-aberrant cells was remarkable in flight personnel as well as in station personnel. The reason for this phenomenon is unknown and needs further investigation.

Adult↗
PubMed1998

Localization of chromosome breakpoints: implication of the chromatin structure and nuclear architecture.

Restriction endonucleases and ionizing radiations have been extensively used to study the origin of chromosomal aberrations. Although a non-random distribution of chromosome breakpoints induced by these agents has been claimed by several authors, the significance of the chromatin structure and nuclear architecture in the localization of breakpoints is still not well understood. Breakpoint patterns produced by endonucleases targeted to specific genome sequences or by ionizing radiations could provide additional evidence to clarify this point. Results obtained from the localization of breakpoints induced by AluI, BamHI or DNase I as well as by neutrons or gamma-rays in G-banded Chinese hamster ovary (CHO) chromosomes are presented. AluI and BamHI were electroporated into CHO cells either during the G1 or S-phase of the cell cycle. A co-localization of breakpoints was found with a preferential occurrence in G-light bands independent of the cell cycle stage in which aberration production took place. Since AluI and BamHI recognition sequences are partitioned in the housekeeping and tissue-specific subgenomes respectively, we postulated that nuclease sensitive sites in active chromatin could be the main targets for the induction of breakpoints by these endonucleases. This assumption is supported by the finding that DNase I-induced breakpoint patterns in CHO cells are similar to those produced by AluI and BamHI. Digestion of fixed CHO chromosomes with these endonucleases induced G-banding suggesting a higher sensitivity of G-light chromatin. For comparison purposes, CHO cells were irradiated with neutrons or gamma-rays and breakpoints localized in G-banded chromosome aberrations. A higher occurrence of breakpoints in G-light bands was also observed. We detected seven breakage-prone G-light bands that were preferentially damaged by the three endonucleases and by both types of radiation. These results emphasize the possible implication of the chromatin structure and the nuclear architecture in the localization of chromosome breakpoints induced by endonucleases, neutrons and gamma-rays.

Animals↗
PubMed1998

Thiopronin reduces the frequencies of neocarzinostatin-induced chromosomal aberrations and sister chromatid exchanges in Chinese hamster ovary cells.

Chinese hamster ovary (CHO) cells were treated in the G1 phase of the cell cycle with different concentrations of neocarzinostatin (NCS) alone or in combination with N-(2-mercaptopropionyl)-glycine (thiopronin; TP). TP reduces the frequencies of NCS-induced chromosomal aberrations (CA) and of sister chromatid exchanges (SCE) significantly when added to the cultures simultaneously (TPsim), 1 min (TP1) or 10 min (TP10) after the addition of NCS. The addition of TP 30 min (TP30) or 60 min (TP60) after NCS reduces the frequencies of SCE, but not of CA. Our results indicate that the induction of CA and SCE by NCS is partially based on different mechanisms.

Animals↗
PubMed1998

Application of chromosome painting to clastogenicity testing in vitro.

To maximise sensitivity, protocols for testing chemicals in chromosomal aberration assays in vitro are designed so that cells are sampled when the peak frequency of aberrations might be expected to occur. They are not designed to measure the frequency of aberrations in cells which survive. Only chromosomal aberrations which are heritable, however, can have any relevance to human health, but the detection of those aberrations most likely to be tolerated (inversions, reciprocal translocations) is notoriously difficult with conventional light microscopy. Current protocol design is justified by arguing that the presence of structural aberrations of any type at early times after treatment indicates a risk that a proportion of aberrations will persist and be maintained in the population. Chromosome painting allows reciprocal exchanges to be relatively easily measured and permits the validity of these assumptions to be tested. To date, the kinetics of induction and dose-response relationships of reciprocal translocations induced by chemicals have been little investigated. We compared the frequency of chromosome-type aberrations in human lymphocytes following treatment with two powerful clastogens, streptonigrin and Trenimon, using conventional staining techniques and chromosome painting. The results show that although reciprocal translocations can be shown to arise and persist in treated populations of human lymphocytes for several days following treatment, their frequency is very low, even at concentrations where large amounts of chromosomal damage are induced, indicating that, at present, the value of using chromosome painting as an adjunct to traditional clastogenicity testing is limited.

Chromosome Aberrations↗
PubMed1998

Intrachromosomal localization of aberration breakpoints induced by neutrons and gamma rays in Chinese hamster ovary cells.

Chromosome breakpoints induced by neutrons or gamma rays in Chinese hamster ovary cells were mapped to Giemsa-light or Giemsa-dark bands or to band junctions. Radiation-induced breakpoints were found to be distributed nonrandomly according to chromosome or band length. More than 60% of the breakpoints were localized in G-light bands. A group of 13 bands which corresponded to only 7% of the total chromosome length contained 22% of the breakpoints produced by neutrons and 14% of those induced by gamma rays. Seven of these 13 bands are also preferentially damaged by AluI, BamHI and DNase I as reported previously. The results indicate that chromatin and nuclear structure may play a role in the distribution of breakpoints produced by ionizing radiation and endonucleases.

Animals↗
PubMed1997

Malignant transformation by cyclin E and Ha-Ras correlates with lower sensitivity towards induction of cell death but requires functional Myc and CDK4.

We demonstrate in this paper that the G1 phase specific cell cycle regulator cyclin E is able to provoke focus formation when cotransfected with activated Ha-ras into primary rat embryo fibroblasts (REFs). Cyclin E/Ha-ras transformed cells are highly tumorigenic in synergeneic rats, are able to form colonies in soft agar and show protection towards apoptosis upon serum starvation or DNA damage compared to cells transformed by the combination of Myc, cyclin D1 or SV40 large T-antigen and Ha-ras. Lines that were established after cyclin E/Ha-ras or cyclin D1/Ha-ras transformation contain a large percentage of polyploid cells. This was not observed in cells transformed with other oncoproteins and Ha-ras pointing to an involvement of D- and E type cyclins in genomic instability. The cyclin dependent kinase inhibitors p21 and p27 but also p16 completely abrogate focus formation by cyclin E and Ha-ras suggesting that the oncogenic activity of cyclin E still requires functional G1 specific cyclin/CDK complexes. Moreover, inhibition of Myc function also blocks the oncogenic activity of cyclin E indicating a requirement of Myc for cyclin E function. The findings presented here demonstrate that cyclin E can act as an oncoprotein with a potential involvement in genomic instability and the prevention of cell death. Our data also present more evidence for a strict functional interdependency between G1 cyclin/CDK complexes and c-Myc.

Animals↗