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A series of hydrogenated isoindolone derivatives structurally related to cytochalasin B has been synthesized and their ability to induce binucleation in the human lymphocytes were tested. All compounds were found able to inhibit cell cytokinesis at different extent in the range (3.12-25 mumol/l) respect to the negative control; however the highest percentage of binucleated cells is induced by Cyt B.

The human lymphocyte micronucleus (MN) test combined with fluorescence in situ hybridization (FISH) of a centromeric probe is considered a useful screening assay to distinguish between clastogenic and aneugenic agents. Four suspected aneuploidy-inducing chemicals, acetaldehyde (AA), diethylstilbestrol (DES), diethylstilbestrol dipropionate (DESdp) and griseofulvin (GF), have been evaluated with the assay. All compounds induced a significant increase of MN at all doses tested. After the application of the FISH technique with a pancentromeric DNA sequence, DES, DESdp and GF showed a statistically significant increase in the percentage of positive signals compared with the control culture. GF induced the highest percentage of centromere-positive MN observed to date (>90% on average). AA did not show a significant difference in the percentage of centromere-positive MN. The results indicate that in human lymphocytes DES, DESdp and GF act primarily as aneugens, while AA seems capable of causing both chromosome breakage and aneuploidy.

Occupational exposure of floriculturists is characterized by alternating periods of intense pesticide spraying and reduced or no activity. Induction of sister chromatid exchanges (SCE), structural chromosome aberrations (CA) and micronuclei (MN) was investigated in peripheral lymphocytes of a group of 23 Italian floriculturists and 22 matched controls. Blood sampling was performed during and one month after the end of intensive pesticide treatments, in order to cover a period of high and low exposure, respectively. Each donor was genotyped for glutathione S-transferase M1 (GSTM1), T1 (GSTT1), and N-acetyltransferase 2 (NAT2), three polymorphic genes involved in xenobiotic metabolism, to assess their potential role in individual genotoxic response to the pesticide exposure. No effect of the pesticide exposure on the cytogenetic parameters were detected. Smoking, however, was found to increase SCE levels. The only significant influence of phenotype composition on cytogenetic response was an increase in SCE levels in the GSTT1 positive individuals compared with the GSTT1 nulls (P=0.02). This finding was, however, based on only four GSTT1 null donors (n=41 for GSTT1 positive donors). In addition, a possible interaction was observed between smoking and GSTM1 genotype in the CA assay, GSTM1 null smokers, earlier reported to have an elevated risk for lung cancer, showing higher CA frequencies than GSTM1 positive smokers.

Three vanadium salts, vanadyl sulfate (SVO5), sodium orthovanadate (Na3VO4) and ammonium metavanadate (NH4VO3), were tested for induction of genotoxic effects in bone marrow of mice following intragastric treatment. Micronucleus (MN) induction in polychromatic erythrocytes (PCEs), structural (sCA) and numerical (nCA) chromosome aberrations in bone marrow cells were evaluated. The micronucleus test, performed at different harvesting times (0-72 h), was found to be positive for all compounds tested. In contrast, except for vanadyl sulfate, no difference was found between controls and treated animals in the sCA test performed 24 and 36 h after treatment. At the same sampling intervals, second metaphases (M II) were positively scored for nCA induction for all three vanadium salts. In addition, the frequency of hypoploid and hyperploid cells was shown to be statistically different from the control value. Polyploid cells were also induced by all compounds, but their frequency was not statistically significant. The positive results obtained by nCA analysis support the finding of a significant presence of types of micronuclei that are probably aneuploidy-related. This finding was further supported by the successful classification of such micronuclei on the basis of shape and size according to Tinwell and Ashby (1991) during microscope analysis.

Vanadium salts have been shown to be aneuploidogenic in human lymphocyte cultures. In particular, increases in the frequency of chromosome satellite associations and a high proportion of induced micronuclei with centromeric signals seem to be connected with chromosome malsegregation mechanisms in which acrocentric chromosomes may be involved. Our aim was to assess the contribution of these chromosomes to the formation of vanadium-induced micronuclei by applying the fluorescence in situ hybridization technique to the human lymphocyte micronucleus assay. Whole blood cultures were treated after 24 h with 0, 10, 40, and 80 microM sodium orthovanadate or vanadyl sulfate and harvested at 72 h; vinblastine, 20 ng/ml, was used as a reference compound. The slides were then hybridized with biotin-labeled beta-satellite DNA probes specific for all human acrocentric chromosomes. After chemical treatment, the percentage of micronuclei with fluorescent signals was found to be statistically higher than that in control cultures, whereas vinblastine induced only a slight increase in micronuclei.

A multidisciplinary study on a general population exposed to vehicle exhaust was undertaken in Pisa in 1991. Environmental factors such as air pollution and those associated with lifestyle were studied. Meanwhile, biological and medical indicators of health condition were investigated. Chromosomal aberrations, sister chromatid exchanges (SCEs), and micronuclei in lymphocytes were included for the assessment of the genotoxic risk. Because of the large number (3800) of subjects being investigated, standardization of protocols was compulsory. The results on data reproducibility are reported. To assess the reliability of the protocol on a large scale, the population of Porto Tolle, a village located in northeast Italy, was studied and compared to a subset of the Pisa population. Preliminary results showed that probable differences between the two populations and individuals were present in terms of SCE frequencies. The study was potentially able to detect the effects of several factors such as age, smoking, genetics, and environment. The in vitro treatment of lymphocytes with diepoxybutane confirmed the presence of more responsive individuals and permitted us to investigate the genetic predisposition to genetic damage. The possible influence of environmental factors was studied by correlation analyses with external exposure to air pollutants as well as with several lifestyle factors.

Bacitracin, an antibiotic widely utilized in clinical and veterinary use, was tested on murine erythroleukemia (MEL) cells. Tests were performed to evaluate the capacity of the drug to interfere with erythroid differentiation. Cells were exposed to a single treatment in S phase at sublethal doses of bacitracin. Two responses were found depending on the drug concentration. At higher concentrations (25 micrograms/ml and 250 ng/ml) a reduction in number of differentiating cells was observed but the kinetics of the process remained unchanged. At lower concentrations (from 2.5 ng/ml to 2.5 fg/ml) a dramatic alteration of the dynamic of differentiation was found. These two responses are related to different activities of the DNA repair mechanisms. Higher doses of bacitracin stimulate repair while lower concentrations are not able to active repair, as demonstrated by tests with hydroxyurea. The bacitracin-induced damage can be considered a stable genetic and/or epigenetic alteration, as demonstrated by the high frequency of mutant clones isolated from low-dose treated cells. The suitability of MEL cells system in evaluating genotoxicity of drugs for veterinary use is underlined.

The micronucleus test in peripheral blood lymphocytes was employed in the cytogenetic monitoring of children with acute lymphocytic leukemia (ALL), who had undergone chemotherapy and radiotherapy. Patients were treated with a variety of drugs, which included vincristine, methotrexate, daunomycin and prednisone; they also underwent cranial irradiation at the end of the first intensive phase of therapy. The first group under study consisted of 15 subjects on therapy, who showed a marked increase in micronucleated lymphocytes (mean: 19.96 +/- 12.96%) as a consequence of treatment compared with the control group (mean: 3.67 +/- 1.55%), while lower average values were obtained from 15 other subjects at the end of treatment (mean: 13.16 +/- 8.44%). A group of 6 patients was monitored during the entire period of therapy, namely at diagnosis, after 3 months of therapy, throughout maintenance therapy and at the end of it. The whole treatment lasted about 2 years. The results revealed a marked increase in basal micronucleus frequency, due to therapy: the micronucleated lymphocyte frequency remained significantly high throughout the treatment for almost all patients. These data clearly suggest the validity of the methodology in pointing out the role played by antileukemic agents in inducing somatic genetic damage.

A series of aliphatic epoxides were tested for their ability to induce forward mutations in the yeast Schizosaccharomyces pombe. For all the compounds under study, a linear dose-response relationship was found, and the ranking of the relative specific activity was: epichlorohydrin greater than ethylene oxide greater than glycidol greater than 1,2-epoxybutane greater than 1,1,1-trichloropropylene oxide greater than propylene oxide greater than 2,3-epoxybutane. The influence of the metabolic conversion of the epoxides by the mouse-liver S9 fraction was also investigated. In such conditions, except for the 2,3-epoxybutane, the genetic activity of epoxides seems to be reduced.

In vivo and in vitro methodologies that have employed the yeast Schizosaccharomyces pombe as genetic indicator have been utilized to investigate the mutagenicity of two trichloroethylene (TCE) samples of pure and technical grade. Mutagenicity assays were also performed on two stabilizers contained in the technical grade sample: epichlorohydrin and 1,2-epoxybutane. In the in vitro studies a metabolic conversion system was supplied by liver homogenate (S-9) from mice and rats untreated and pretreated with phenobarbital and/or beta-naphthoflavone. Up to highly toxic doses of TCE were applied to growing and stationary-phase yeast cells. In the in vivo studies two different host-mediated assays, intrasanguineous and intraperitoneal methodologies, were performed on different mice breeds treated by oral administration. Epichlorohydrin and epoxybutane were tested singly or combined in a mixture of the same ratio as in the technical grade TCE sample. Both TCE samples gave negative results for in vivo and in vitro assays, whereas the two contaminants were found mutagenic only in vitro. The high toxicity of the technical TCE sample did not allow us to reach concentrations containing effective levels of its two additives.