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G Stephanou

Publications and source records attributed to G Stephanou.

16 recordsLinked to original sources

Biomarkers of genotoxicity of urban air pollution. Overview and descriptive data from a molecular epidemiology study on populations exposed to moderate-to-low levels of polycyclic aromatic hydrocarbons: the AULIS project.

Epidemiologic studies indicate that prolonged exposure to high pollution levels is associated with increased risk of cancer, especially lung cancer. However, under conditions of moderate or low air pollution, epidemiologic evidence does not permit reliable conclusions. Biomarker-based population studies may serve as complementary tools providing a better understanding of the relative contribution of ambient atmospheric pollution to the overall genotoxic burden suffered by city dwellers. However, past efforts to apply biomarkers to studies of low levels exposure to urban air pollution have given inconclusive results, partly because of the absence of adequate data on personal exposure, covering a time-window which is appropriate for the biomarkers being examined, as well as a battery of biomarkers reflecting different stages of the carcinogenic process. In the present paper, the potential of biomarker-based population studies to aid the assessment of the genotoxic and carcinogenic effects of urban air pollution is reviewed by reference to the achievements and limitations of earlier reported studies. The design and methodology adopted in a recently completed large-scale population study, carried out in the context of the European Union Environment and Climate Programme, known by the short name of AULIS project, is discussed and descriptive statistics of the main findings of the project are presented. These findings indicate that for cohorts suffering moderate-to-low exposures to airborne particulate-bound polycyclic aromatic hydrocarbons (PAHs), no simple correlation with biomarkers of genotoxicity existed and suggest that additional factors made a significant contribution to the overall genotoxic burden.

Adolescent↗

Evaluation and characterization of micronuclei induced by the antitumour agent ASE [3beta-hydroxy-13alpha-amino-13, 17-seco-5alpha-androstan-17-oic-13, 17-lactam-p-bis(2-chloroethyl)amino phenylacetate] in human lymphocyte cultures.

3beta - Hydroxy - 13alpha - amino - 13, 17 - seco - 5alpha - androstan - 17 -oic-13,17-lactam-p-bis(2-chloroethyl)amino phenylacetate (ASE) is a homo-aza-steroidal ester of p-bis(2-chloroethyl) amino phenyl acetic acid and has been shown to display antineoplastic, mutagenic and genotoxic activity. In the present study an effort has been made to evaluate the ability of ASE to induce micronuclei (MN) in human lymphocytes treated in vitro using the cytokinesis-block assay. Lympocytes were treated with different concentrations of ASE (0.1, 0.25, 0.5, 1, 2.5, 5, 10 and 20 microg/ml) at two different cell culture times, 21 and 41 h after culture initiation. ASE treatment lasted until cell harvest, for 51 and 31 h, respectively. Two types of cultures were used, whole blood and isolated lymphocyte cultures. The content of induced MN was identified by FISH analysis, using an alpha-satellite DNA probe, in binucleate cells. Our results suggest that ASE is capable of increasing MN frequencies in human lymphocytes under both culture conditions. This increase is related to the concentration in a linear dose-dependent manner and is also dependent on the duration of treatment. FISH analysis has shown that the induced MN resulted mainly from breakage events. Additionally, a weak aneugenic effect was found at the higher concentrations in whole blood cultures as well as in isolated lymphocyte cultures. Cytotoxic effects of ASE were observed under both cell culture conditions with a linear dose-dependent relationship according to CBPI evaluation and were more pronounced in isolated lymphocyte cultures.

Adult↗

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Journal Article↗

Chromosomal aberrations, sister-chromatid exchanges, cells with high frequency of SCE, micronuclei and comet assay parameters in 1, 3-butadiene-exposed workers.

The association of occupational exposure to 1,3-butadiene (BD) and induction of cytogenetic damage in peripheral lymphocytes was studied in 19 male workers from a monomer production unit and 19 control subjects from a heat production unit. The exposure to BD was measured by passive personal monitors. The following biomarkers were used: chromosomal aberrations (CA), sister chromatid exchanges (SCE), cells with a high frequency of SCE (HFC), micronuclei, comet assay parameters like tail length (TL) and percentage of DNA in tail [T (%)] and polymorphisms of GSTM1 and GSTT1 genotypes. BD exposure with a median value of 0.53 mg/m3 (range: 0.024-23.0) significantly increased (a) the percentage of cells with chromosomal aberrations in exposed vs. control groups (3.11% vs. 2.03%, P<0.01), (b) the frequency of SCE per cell (6.96 vs. 4.87, P<0.001), and (c) the percentage of HFC (19.9% vs. 4.1%, P<0.001). BD exposure had no significant effects on formation of micronuclei and on comet assay parameters. Effect of smoking was observed only for HFC in BD-exposed group. GSTM1 genotype affected chromosomal aberrations in exposed group, while GSTT1 genotype affected chromosomal aberrations in controls. No effect of GSTM1 or GSTT1 genotypes was observed on any other biomarkers used.

Air Pollutants↗

Micronucleus induction in somatic cells of mice as evaluated after 1,3-butadiene inhalation.

The effect of different 1,3-butadiene (BD) inhalation doses, 130, 250, and 500 ppm, on somatic cells of mice was studied. Two different cell populations with diverse replicative and differentiative activities, namely splenocytes and peripheral blood reticulocytes, were examined and micronucleus (MN) frequencies were estimated. In splenocytes, different postinhalation time intervals were studied with regard to MN induction and characterisation. BD was found to be clastogenic by inducing increased micronucleus frequencies in both cell compartments and also to induce cytotoxicity at the highest level of exposure. In mouse splenocytes, BD has also shown a weak aneugenic effect at a short time interval after the exposure. Postinhalation time influences the induction of chromosome damage in stimulated splenocytes treated in vivo, since MN frequency decreases with time; in addition, BD has shown its aneugenic and cytotoxic potential only at 2 days after exposure.

Animals↗

Effects of cetirizine dihydrochloride on human lymphocytes in vitro: micronucleus induction. Evaluation of clastogenic and aneugenic potential using CREST and FISH assays.

Cetirizine dihydrocloride, a widely administered antiallergic drug with the amine piperazine in its molecule, was studied as to its ability to cause micronucleus formation in human lymphocyte cultures treated in vitro. Peripheral lymphocytes from four different donors were cultured and treated with different concentrations of the compound. Cetirizine dihydrocloride was shown to induce enhanced micronucleus frequency in a dose-dependent manner, although lymphocytes from the different donors showed different susceptibilities to the compound. The content of induced micronuclei was investigated in one of the four donors by two independent assays, CREST (the application of antikinetochore antibodies) and FISH (fluorescence in situ hybridization) on cytochalasin B-formed binucleated cells. It was shown that the induced micronuclei resulted from breakage events as well as chromosome loss, thus characterizing cetirizine dihydrocloride as both clastogen and aneugen. Since our results were derived only from in vitro experiments, we believe that an extensive in vivo study is necessary before drawing conclusions as to the effects of cetirizine dihydrochloride in patients.

Adult↗

Induction of micronuclei and sister chromatid exchange in mouse splenocytes after exposure to the butadiene metabolite 3,4-epoxy-1-butene.

3,4-Epoxy-1-butene (EB) is one of the main metabolites of 1,3 butadiene, a widely used industrial chemical. The mutagenic potential of 1,3 butadiene and its metabolites have been studied in different test systems. In this work the genotoxic effects of EB were studied by estimating micronuclei (MN) and sister chromatid exchange (SCE) frequencies in stimulated mouse splenocytes. Mice were treated in vivo with various doses of EB (24.4, 48.8 and 73.2 mg/kg). The antikinetochore antibody technique (CREST) was also applied to MN in cytokinesis blocked cells to investigate any possible aneugenic effect. Both MN and SCE frequencies increased after EB treatment. The induced MN resulted mainly from acentric fragments but a weak aneugenic effect was found as well. Cytotoxic effects of EB were observed at the highest dose. The above results, in combination with others on the effect of 1,3 butadiene and its metabolites in somatic and germ cells of mouse and rat as well as in somatic human cells, form a part of the information needed for application of the parallelogram approach and extrapolation to human risk.

Animals↗

m-N-N-bis(2-chloroethyl)aminocinnamic acid and four new homo-aza-steroidal esters induce chromosomal abnormalities and affect protein synthesis in human lymphocytes in vitro.

The alkylating agent m-N,N-bis(2-chloroethyl)aminocinnamic acid (m-ACA) and four new homo-aza-steroidal esters were studied for their ability to induce chromosomal abnormalities and to affect protein synthesis in human lymphocytes in vitro. A mitotic index reduction and an increase in the total number of aberrations were observed. Analysis of chromosomal abnormalities has shown that these are mainly chromatid breaks. A decrease in protein synthesis was also observed that seems to fit with the order of activity of the above compounds reflected in the induction of chromosomal aberrations. The observation that protein synthesis and the induction of chromosomal aberrations are affected by these chemicals may reflect interactions between these molecules and DNA that result in structural chromosome changes and decreased protein synthesis.

Antineoplastic Agents, Alkylating↗

A comparative study on the effect of MNU on human lymphocyte cultures in vitro evaluated by O6-mdG formation, micronuclei and sister chromatid exchanges induction.

N-Nitroso-compounds are a large group of chemicals present in a number of environmental sources and many of them are mutagens as well as carcinogens in experimental animals. Among the known N-nitroso-compounds, N-nitroso-N-methylurea (MNU) is a strong mutagen. In this study an effort has been made to compare the ability of MNU to methylate the O6-guanosine site in DNA and to induce micronuclei and sister chromatid exchanges in human lymphocyte cultures in vitro. To quantitate O6-methyldeoxyguanosine (O6-mdG) a highly sensitive immunoassay, immuno-slot-blot (ISB), has been used. For the evaluation of micro nuclei (MN) the cytokinesis block micronucleus method has been used. Different concentrations (75, 100, 125 micrograms/ml) were tested. At the highest concentration tested for the MN induction, 125 micrograms/ml, the occurrence of binucleates and micro nuclei is higher than twice in relation to control and a reduction in NDI is also observed. The same concentrations were used for the estimation of sister chromatid exchanges (SCEs) induction. The mean number of SCEs at 125 micrograms/ml is almost three times that of the control level. The concentrations tested for the quantitation of O6-mdG were 200, 300 and 400 micrograms/ml and this was done because for the test system we used and for the given experimental conditions the first indication of O6-mdG formation was at 200 micrograms/ml. Our results show that methylation of O6-guanosine increases with concentration and at 400 micrograms/ml the concentration of O6-mdG is 5.83 fmol/microgram DNA, while at the control level it is 2.40 fmol/microgram DNA. Since O6-mdG formation is observed in higher concentrations than those of MN and SCE induction it would be interpreted that O6-mdG levels are not correlated with the studied cytogenetic effects although one has to take into consideration the total promutagenic lesions in DNA, induced by MNU, as well as AGT repair activity.

Carcinogens↗

Biological effect monitoring in industrial workers from the Czech Republic exposed to low levels of butadiene.

Blood samples were collected twice (in 1993 and 1994) from 19 workers exposed to 1,3-butadiene and 19 matched controls. Three exposed and three control subjects were the same in 1993 and 1994. Personal passive dosimetry was performed in 1993 and twice in 1994 on the day preceding blood sampling. Mean exposure level in 1994 was 1.76 +/- 4.20 ppm (S.D.) and individual exposure levels ranged between 0.012 ppm (detection limit) and 19.77 ppm. Using the clonal assay, geometric mean of hprt mutant frequencies adjusted for cloning efficiency, age and smoking were, respectively, 7.85 (+/- 7.09) x 10(-6) and 10.14 (+/- 9.16) x 10(-6) in pooled (1993 plus 1994) exposed and control subjects. The difference was not statistically significant indicating that 1,3-butadiene did not induce a detectable increase in mutations at the hprt locus. A similar result was obtained for the 1994 subjects alone. There was no difference between adjusted geometric mean mutant frequencies of exposed and unexposed non-smokers or between exposed and unexposed smokers. Analysis of chromosomal aberrations in lymphocytes from 1994 subjects indicated that the percentage of aberrant cells was significantly enhanced in exposed subjects. In 1993 (data not shown), it was impossible to demonstrate a significant increase of aberrant cells in subjects exposed to 1,3-butadiene. Frequencies of micronuclei in cytochalasin-B blocked binucleate lymphocytes in exposed and unexposed 1994 subjects were not significantly different. This was also the case for earlier samples analyzed in the same plant. Using the comet assay for 1994 subjects, no statistically significant difference was found between the whole group of exposed and unexposed subjects. This was true for both the comet tail length and the percentage of DNA in the tail. In exposed smokers, however, the comet tail length was significantly longer than in unexposed smokers. Unexpectedly, in unexposed smokers the tail length was significantly shorter than in unexposed non-smokers. It was also unexpected that the percentage of DNA in the comet tail was significantly lower in exposed non-smokers than in unexposed non-smokers.

Adult↗

Human cytogenetic biomonitoring of occupational exposure to 1,3-butadiene.

The association of occupational exposure to 1,3-butadiene and chromosomal damage in peripheral blood lymphocytes was studied in 40 workers from two production facilities. Control persons, 30 in all, were chosen from other departments of the same plants, and they were roughly matched for age and smoking habits. The exposure levels to ambient butadiene were measured both by personal sampling using diffuse monitors and by stationary sampling at production and handling sites. Chromosome aberrations (CA), micronuclei (MN) and sister-chromatid exchanges (SCE) in peripheral lymphocytes were analyzed as markers of exposure. Smoking had a slight effect on the frequency of MN, and the mean frequency of SCEs was also higher in smokers than in non-smokers. No effect of smoking, however, was seen in relation to chromosomal aberrations. No exposure related effects were seen in any of the three cytogenetic endpoints in either of the butadiene production plants, representing typical low (below 3 ppm) exposure levels of the butadiene manufacturing industry.

Adult↗

Altered protein synthesis rate in ovaries of D. melanogaster caused by new antitumour alkylating agents.

1. The effect of two homo-aza-steroidal esters with antineoplastic activity, namely 3 beta-hydroxy-13 alpha-amino-13,17-seco-5 alpha-androstan-17-oic-13,17-lactam-p-bis(2-chloroethyl)aminoph enoxyacetate (NSC 294859) and 3 beta-hydroxy-13 alpha-amino-13,17-seco-5 alpha-androstan-17-oic-13,17-lactam-p-bis(2-chloroethyl)aminoph enylacetate (ASE) on protein synthesis rate was studied in ovaries of Drosophila melanogaster females. 2. Two different concentrations for each compound were examined. 3. Both esters containing the same alkylating agent have been shown to decrease protein synthesis in relation to control.

Alkylating Agents↗

Genotoxic effect of the antitumour agent homo-aza-steroidal ester of p-bis-(2-chloroethyl)aminophenoxy acetic acid (NSC 294859) tested in the Drosophila wing somatic mutation and recombination test.

The genotoxic activity of the new antineoplastic alkylating agent 3 beta-hydroxy-13 alpha-amino-13,17-seco-5 alpha-androstan-17-oic-13,17- lactam-p-bis-(2-chloroethyl) aminophenoxyacetate (NSC 294859) was studied in the somatic mutation and recombination test (SMART) in Drosophila melanogaster. The drug was administered to larvae which were transheterozygous for two recessive wing cell markers mwh and flr3. Several concentrations were tested and two of them showed a clear induction of mosaic spots, expressing the mutant phenotypes. Multiple wing hair (mwh) spots were the main type observed but twin spots and flares (flr3) spots were also detected. We conclude that the compound induces both mutations and somatic recombination in the SMART test.

Animals↗

Heat shock phenomena in Aspergillus nidulans. II. Combined effect of heat and bleomycin to heat shock protein synthesis, survival rate and induction of mutations.

The combined action of hyperthermia and Bleomycin on Aspergillus nidulans was studied at three different levels: mycelial protein synthesis, spore viability and induction of mutations. It was found that Bleomycin treatment of preincubated mycelia during the heat shock enhances the incorporation of 35S-methionine into heat shock bands. Furthermore, simultaneous treatment with hyperthermia (43 degrees C) and Bleomycin results in greater cytotoxic activity in spores and in a higher induction rate of point mutations.

Aspergillus nidulans↗

Heat shock phenomena in Aspergillus nidulans. I. The effect of heat on mycelial protein synthesis.

Heat shock was found to induce characteristic changes in the pattern of protein synthesis in Aspergillus nidulans as analysed by SDS-polyacrylamide gel electrophoresis. Six to seven new bands were found to show increased incorporation to 35S-methionine at 43 degrees C compared to 37 degrees C, the standard temperature for this organism. The heat shock response of five different strains of A. nidulans was examined. This comparative study showed that these strains (haploids and diploids) show exactly the same set of heat shock proteins.

Aspergillus nidulans↗

Effects of cetirizine dihydrochloride on human lymphocytes in vitro: evaluation of chromosome aberrations and sister chromatid exchanges.

The ability of cetirizine dihydrochloride, an antihistaminic agent, to induce chromosome aberrations as well as sister chromatid exchanges (SCEs) was evaluated in human lymphocyte cultures treated in vitro. The following concentrations were tested: 25, 50, 75, 100 and 200 micrograms/ml. The results of our study revealed that cetirizine dihydrochloride is capable of inducing chromosome aberrations, at least at the higher concentrations studied, 100 and 200 micrograms/ml. The majority of aberrations was of chromatid type. Cetirizine is also a weak inducer of SCEs. Further studies are now warranted in order to define the in vivo cytogenetic activity of cetirizine in humans.

Cells, Cultured↗