Biomedical subjects

I D Adler

Publications and source records attributed to I D Adler.

At least 37 records · Page 2Linked to original sources

PubMed1998

DNA adducts in mouse testis and lung after inhalation exposure to 1,3-butadiene.

1,3-Butadiene is a widely used industrial chemical and also an environmental contaminant. Recent findings have shown that butadiene can also be a male germ cell mutagen. In this study, DNA adduct formation in testis and lung has been explored by using N7-alkylated guanines as a marker of biological effective dose. The adducts measured were the four structurally different guanine N7-adducts alkylated by butadienemonoepoxide, the main metabolite of 1,3-butadiene. This study demonstrates the dose-dependent adduct formation in lung and testis. At lower exposures (50 and 200 ppm) the adduct levels were about the same in the two organs, but at 500 ppm the adduct level was significantly (p < 0.03) higher in testis than in lung. The enantiomeric composition of the adducts detected was also different. In lung, all 4 possible adducts were present (S-C-1" dominating, 49%), but in testis only two out of four adducts were detected (S-C-2" being the most abundant adduct, 71%). These novel observations indicate that the DNA repair is different in these two organs studied and that heritable genetic effects observed may be mediated through the DNA adducts.

Alkylation↗

PubMed1998

Dose response study for 1,3-butadiene-induced dominant lethal mutations and heritable translocations in germs cells of male mice.

Butadiene (BD) and its metabolites have extensively been studied in the EU sponsored research project "Multi-endpoint analysis of genetic damage induced by 1,3-butadiene and its major metabolites". Within this project a dominant lethal test and a heritable translocation test were performed with male mice to study the dose-response relationships for the respective endpoints. BD concentrations of 130 and 500 ppm were tested in the dominant lethal assay by exposing male mice on 6 h/day for five consecutive days resulting in doses of 3900 and 15,000 ppmh, respectively. Males were mated for four consecutive weeks at a ratio of 1:2 to untreated females. A positive dominant lethal effect was observed in the first mating week in the experiment with 15,000 ppmh but no dominant lethality was found with the lower dose of 3900 ppmh. The present dominant lethal data have to be viewed together with the data already published for a BD dose of 39,000 ppmh (1300 ppm at 6 h/day on 5 consecutive days) [1]. The main difference between results with the highest and the middle dose is that mating weeks one and two (sperm and late spermatids) showed an effect at 39,000 ppmh while only mating week one (sperm) showed an effect at 15,000 ppmh. In the heritable translocation assay, males mice were exposed with a BD dose of 15,000 ppmh and mated for one week to untreated females. Among 434 F1 offspring, we found 5 translocation carriers (1.15% vs. 0.05% in the historical control, p < 0.001). In the previous heritable translocation experiment with a BD dose of 39,000 ppmh of DB exposure, 2.7% of the offspring carried a reciprocal translocation [2]. These data can be used for quantification of genetic risk. The dose response for BD-induced heritable translocations in sperm and late spermatids of mice was linear (Y = 0.05 + 6.9 x 10(-5)X) and a doubling dose of 725 ppmh could be calculated.

Animals↗

PubMed1998

Genetic effects of 1,3-butadiene and associated risk for heritable damage.

A summary of the results of the studies conducted in the EU Project "Multi-endpoint analysis of genetic damage induced by 1,3-butadiene and its major metabolites in somatic and germ cells of mice, rats and man" is presented. Results of the project are summarized on the detection of DNA and hemoglobin adducts, on the cytotoxic and clastogenic effects in somatic and germinal cells of mice and rats, on the induction of somatic mutations at the hprt locus of experimental rodents and occupationally exposed workers, on the induction of dominant lethal mutations in mice and rats, and on heritable translocations induced in mice, after exposure to butadiene (BD) or its major metabolites, butadiene monoepoxide (BMO), diepoxybutane (DEB) and butadiene diolepoxide (BDE). The primary goal of this project was to collect experimental data on the genetic effects of BD in order to estimate the germ cell genetic risk to humans of exposure to BD. To achieve this, the butadiene exposure are based on data for heritable translocations and bone marrow micronuclei induced in mice and chromosome aberrations observed in lymphocytes of exposed workers. A doubling dose for heritable translocations in human germ cells of 4900 ppm/h is estimated, which, assuming cumulative BD exposure over the sensitive period of spermatogenesis, corresponds to 5-6 weeks of continuous exposure at the workplace to 20-25 ppm. Alternatively, the rate of heritable translocation induction per ppm/h of BD exposure is estimated to be approximately 0.8 per million live born, compared to a spontaneous incidence of balanced translocations in humans of approximately 800 per million live born. These estimates have large confidence intervals and are only intended to indicate orders of magnitude of human genetic risk. These risk estimates are based on data from germ cells of BD-exposed male mice. The demonstration that clastogenic damage was induced by DEB in preovulatory oocytes at doses which were not ovotoxic implies that additional studies on the response of mammalian female germ cells to BD and its metabolites are needed. The basic assumption of the above genetic risk estimates is that experimental mouse data obtained after BD exposure can be extrapolated to humans. Several points exist in the present report and in the literature which contradict this assumption: (1) the level of BMO-hemoglobin adducts was significantly elevated in BD-exposed workers; however, it was considerably lower than would have been predicted from comparable rat and mouse exposures; (2) the concentrations of the metabolites DEB and BMO were significantly higher in mouse than in rat blood after BD exposure. Thus, while metabolism of BD is qualitatively similar in the two species, it is quantitatively different; (3) no increase of HPRT mutations was shown in 19 workers exposed on average to 1.8 ppm of BD, while in a different population of workers from a US plant exposed on average to 3.5 ppm of BD, a significant increase of HPRT variants was detected; and (4) data from cancer bioassays and cancer epidemiology suggest that rat is a more appropriate model than mouse for human cancer risk from BD exposure. However, the dominant lethal study in rats gave a negative result. At present, we do not know which BD metabolite(s) may be responsible for the genetic effects even though the bifunctional alkylating agent DEB is the most likely candidate for the induction of clastogenic events. Unfortunately, methods to measure DEB adducts in hemoglobin or DNA are only presently being developed. Despite these several uncertainties the use of the mouse genetic data is regarded as a justifiable and conservative approach to human genetic risk estimation given the considerable heterogeneity observed in the biotransformation of BD in humans.

Animals↗

PubMed1998

Trichlorfon exposure, spindle aberrations and nondisjunction in mammalian oocytes.

Consumption of trichlorfon-poisoned fish by women in a small Hungarian village has been associated with trisomy resulting from an error of meiosis II in oogenesis. We therefore examined mouse oocytes exposed for 3 h during fertilization to 50 microg/ml trichlorfon. Spindle morphology was not visibly altered by the pesticide. Chromosomes segregated normally at anaphase II with no induction of aneuploidy. However, formation of a spindle was disturbed in many oocytes resuming meiosis I in the presence of trichlorfon. In spite of the spindle aberrations and the failure of bivalents to align properly at the equator, oocytes did not become meiotically arrested but progressed to metaphase II. At this stage, spindles were highly abnormal, and chromosomes were often totally unaligned, unattached or dispersed on the elongated and disorganized spindle. By causing spindle aberrations and influencing chromosome congression, trichlorfon appears, therefore, to predispose mammalian oocytes to random chromosome segregation, especially when they undergo a first division and develop to metaphase II during exposure. This is the first case in which environmentally induced human trisomy can be correlated with spindle aberrations induced by chemical exposure. Our observations suggest that oocytes may not possess a checkpoint sensing displacement of chromosomes from the equator at meiosis I and may therefore be prone to nondisjunction.

Anaphase↗

PubMed1998

Induction of specific-locus and dominant lethal mutations in male mice by ifosfamide (Holoxan).

Ifosfamide induced dominant lethal mutations in spermatozoa of mice at doses of 200 and 300 mg/kg and in spermatids and spermatocytes at 600 mg/kg. The highest dose also induced specific-locus mutations in post-spermatogonial germ-cell stages of mice but not in spermatogonial stem cells. The nature of the induced mutations suggests they are intergenic. The spermatogenic specificity of ifosfamide in mouse germ cells is similar to that of the structurally related cytostatic drugs cyclophosphamide and trofosfamide. Due to the post-spermatogonial germ cell specificity of ifosfamide, the genetic risk is limited to a few weeks after exposure.

Animals↗

PubMed1998

Colcemid and econazole do not show aneugenicity in male mouse germ cells.

Colcemid (COM) and econazole (EZ) were tested for induction of mitotic arrest and C-mitotic effects in mouse bone marrow cells and for meiotic delay and induction of hyperploidy in mouse spermatocytes after single injection. Doses of 1 and 3 mg/kg COM were used and bone marrow and spermatocytes were sampled at 2, 6, 10, 14 and 18 h. For EZ a dose of 120 mg/kg and intervals of 6, 10, 14 and 18 h were chosen. At 2 h after COM treatment of bone marrow cells the mitotic index and C-mitotic effect were highest, then both decreased from 6 to 18 h. At 18 h the mitotic indices decreased to or significantly below the control level at doses of 1 and 3 mg/kg respectively, whereas the corresponding frequencies of C-mitotic cells were still significantly higher than in the controls. EZ also induced mitotic arrest and C-mitotic effects in mouse bone marrow cells. In contrast to COM, the effects of EZ were highest at 18 h after treatment. COM and EZ caused disturbances in progression from the first to second meiotic division, however, after COM treatment the ratios of MMII to MMI were significantly below the control. In contrast, after EZ treatment the ratios were significantly higher than in the controls. Such differences may result from the different mechanisms by which COM and EZ act on cell cycle progression. COM and EZ did not induce non-disjunction under the experimental conditions. However, EZ induced structural chromosomal aberrations.

Animals↗

PubMed1998

[Studies of meiotic origin of the extra chromosome 21 in Down syndromes detected by using (GT)n polymorphic DNA markers].

The polymorphics of two pericentric (GT)n sequences on the long arm of human chromosome 21 have been analyzed after PCR amplification, PAGE and Ag-staining for the first time in 50 Chinese Han people, and were used to detect meiotic origin of the extra chromosome 21 in Down syndromes. Six and 5 alleles were found in Chinese Han people for D21S215 and D21S120, respectively, with observed heterozygosities of 0.68 and polymorphic information content PIC, 0.67 and 0.65. For 17 Down syndromes whose parental origin of the extra chromosome 21 were known, meiotic origin of the extra chromosome 21 were determined in 16 cases, with 7 and 4 maternal meiosis I and II nondisjunction, 2 and 3 paternal meiosis I and II, respectively. The possible biological significance of the study on origin of the extra chromosome 21 has been discussed.

Chromosomes, Human, Pair 21↗

PubMed1997

Evaluation of the mutagenicity of acetochlor to male rat germ cells.

Male rat dominant lethal (DL) assays conducted on the herbicide acetochlor are described. Single dose studies conducted at the maximum tolerated dose (MTD, < or = 1000 mg/kg) produced no effects on any of the DL assay parameters at any of the ten weekly sampling periods. It is concluded that acetochlor is non-mutagenic to rat germ cells. Due to initial limited knowledge of the MTD of acetochlor it was also evaluated in the DL assay at a dose level of 2000 mg/kg. At this high dose level severe bodyweight loss and some deaths occurred among the treated animals. In addition, reduced implantations and reduced pregnancy rates were observed at the third sampling period (18-25 days post dosing) in the absence of an increase in early post-implantation deaths. These results indicated that the use of supra-MTD doses of acetochlor had reduced the fertility of the treated males leading to the production of a pseudo-DL assay response, as alerted to and defined by Ehling. Although several such pseudo-DL assay responses have been described, none have been explained mechanistically. It was therefore decided to pursue the effects seen in the DL assay when using supra-MTD doses of acetochlor. Ova analysis of female rats mated with male rats exposed to 2000 mg/kg acetochlor revealed unfertilized ova at the critical third sampling time. Normal fertilization of ova was observed at the first and fifth sampling period and, for a dose of 200 mg/kg acetochlor, at the third sampling period. The magnitude and temporal nature of these effects confirmed the induction of a pseudo-DL assay response, and studies were then undertaken to probe its genesis. Rats treated with 2000 mg/kg acetochlor had normal testicular and epididymal pathology and normal sperm numbers and sperm motility at the critical third sampling period. Despite a small reduction in testicular and epididymal glutathione levels 12 h after exposure to 2000 mg/kg acetochlor, testicular LDH and LDH-X enzyme levels were unaffected. Further, no reduction in the level of free sulphydryl groups (-SH) were observed in epididymal caput sperm heads isolated 0.5, 7 or 14 days after treatment of male rats with 2000 mg/kg acetochlor. The only sperm parameter affected by treatment with 2000 mg/kg acetochlor was an increase in epididymal cauda sperm with head abnormalities. The non-specific nature of this effect was considered inadequate to explain fully the high dose fertility effects seen in the DL assays, which therefore remain unexplained. The present data establish that acetochlor is non-mutagenic to rat germ cells. They also confirm the importance of segregating mutagenic and fertility effects in the DL assay, and emphasize the need for appropriate dose-setting studies prior to the conduct of rodent genetic toxicity assays.

Animals↗

PubMed1997

Induction of specific-locus and dominant lethal mutations in male mice by 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU).

1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) induced dominant lethal and specific-locus mutations in male mice. For both compounds the germ cell stage sensitive to the induction of dominant lethal mutations was dose dependent. A dose of 5 mg BCNU per kg b.wt. induced dominant lethal mutations primarily in spermatocytes, whereas higher doses of BCNU induced dominant lethals in spermatids and spermatocytes. Following doses of 5 and 10 mg CCNU per kg b.wt. dominant lethals were induced in spermatids and spermatocytes similar to the results for higher doses of BCNU. Higher dose exposure to BCNU and CCNU was associated with dominant lethals expressed as pre-implantation loss (reduction in total number of implants). In addition, higher doses of CCNU showed a cytotoxic effect in differentiating spermatogonia. Both compounds induced specific-locus mutations in post-spermatogonial germ cell stages of mice. However, CCNU increased also the specific-locus mutation frequency in spermatogonia in two out of three experiments. We conclude in analogy with criteria developed by IARC, that BCNU and CCNU are potential human mutagens.

Animals↗

PubMed1997

Quantitative and qualitative studies of micronucleus induction in mouse erythrocytes using flow cytometry. I. Measurement of micronucleus induction in peripheral blood polychromatic erythrocytes by chemicals with known and suspected genotoxicity.

Quantitative and qualitative aspects of the in vivo micronucleus-inducing potential of five chemicals were studied using flow cytometric enumeration of micronucleated polychromatic peripheral blood erythrocytes in mice. The chemicals were hydroquinone, vinblastine sulphate, chloral hydrate (tested in two different mouse strains), 5-bromo-2-deoxyuridine and 2-chlorobenzylidene malonitrile. Repeat samplings of peripheral blood were made at 0, 24, 40, 48 and 72 h and for low doses of 5-bromo-2-deoxyuridine 96 h after i.p. treatment. The agents hydroquinone (lowest effective dose 25 mg/kg), vinblastine sulphate (lowest effective dose 0.05 mg/kg) and 5-bromo-2-deoxyuridine (lowest effective dose 200 mg/kg) gave rise to significant increases in the frequencies of micronucleated polychromatic erythrocytes. No significant induction of micronucleated polychromatic erythrocytes by 2-chlorobenzylidene malonitrile or chloral hydrate was found. The frequencies of induced micronucleated polychromatic erythrocytes peaked at 40 h after hydroquinone treatment, at 48 h after vinblastine treatment and at 72 h after 5-bromo-2-deoxyuridine treatment with evident dose-dependent differences in the kinetics of the induction of micronucleated polychromatic erythrocytes. The mean relative Hoechst 33342 fluorescence of the populations of induced micronucleated polychromatic erythrocytes was used as an indicator of the DNA content of induced micronuclei. These values were found to be in agreement with the presumed mechanisms of micronucleus induction for hydroquinone, vinblastine sulphate and 5-bromo-2-deoxyuridine. Flow cytometric enumeration of micronucleated polychromatic erythrocytes in peripheral blood is an efficient method for the study of in vivo micronucleus induction, combining rapid analysis and high sensitivity with information on possible mechanisms of micronucleus induction. The method also allows a substantial reduction in the number of animals needed.

Animals↗

PubMed1997

Quantitative and qualitative studies of micronucleus induction in mouse erythrocytes using flow cytometry. II. Analysis of micronuclei of aneugenic and clastogenic origin by dual-colour FISH on populations of bone marrow PCEs flow sorted on the basis of their relative DNA content.

Simultaneous dual-colour fluorescent in situ hybridization (FISH) with the major and minor mouse satellite probes was performed on flow-sorted fractions of micronuclei with defined Hoeschst 33342 fluorescence intensities, reflecting their relative DNA content. The purpose of the study was to investigate the relationship between probe binding and the size of micronuclei induced by a set of chemicals with different mechanisms of micronucleus induction. The agents studied were vinblastine, cyclophosphamide, hydroquinone and 5-bromo-2-deoxyuridine. The Hoechst 33342 fluorescence distributions of induced micronuclei differ markedly between cyclophosphamide, vinblastine and hydroquinone. While the distributions for cyclophosphamide and vinblastine agree well with those obtained for other model aneugens and clastogens, hydroquinone shows a distribution with characteristics similar to both clastogens and aneugens. A comparison between the Hoechst 33342 fluorescence distributions of hydroquinone-induced micronuclei positive for both major and minor probes and those positive for the major probe only indicates that the former consist of whole chromosomes, while the latter originate from chromosome fragments. The absolute frequency and Hoechst 33342 fluorescence distribution of major-only-positive micronuclei induced by vinblastine indicates that this model aneugen also shows a certain in vivo clastogenic potential. Micronuclei induced by 5-bromo-2-deoxyuridine indicate that this agent is a clastogen, preferentially inducing pericentric breaks, but may also to some extent be aneugenic.

Animals↗

PubMed1997

Extruded micronuclei induced by colchicine or acrylamide contain mostly lagging chromosomes identified in paintbrush smears by minor and major mouse DNA probes.

In the mouse bone marrow micronucleus assay, it was studied whether micronuclei (MN) could be expelled from polychromatic erythrocytes (PCE) in a similar way to the main nucleus. To avoid the disrupting centrifugation step of the conventional bone marrow preparation procedure, the paintbrush technique was used in the present experiments. With May-Grunwald-Giemsa staining of paintbrush slides, 5 % of the colchicine (COL)-induced MN were found attached to the outside membranes of PCE and were regarded as extruded. Of the acrylamide (AA)-induced MN, 22% were extruded. After fluorescence in situ hybridization (FISH) of a total of 300 MN per chemical treatment with the mouse minor and major satellite DNA probes, 9.7% MN were extruded in the COL group and 8.3% MN were extruded in the AA group. FISH showed that 76% of the retained COL-induced MN were signal-positive, indicating that they contained entire chromosomes. With AA, 29% minor-positive and 28.3 % major-positive retained MN were found, confirming its known clastogenicity. However, the observed frequency of signal-positive MN (1.7 MNPCE(pos)/ 1000 PCE) in the AA group was about three times higher than in the control (0.5 MNPCE(pos)/1000 PCE) which indicates that AA has aneugenic potential. FISH analysis of the extruded MN showed 72-100% major as well as minor signals. It is concluded that expelled MN contain mostly entire chromosomes.

Acrylamide↗

PubMed1997

In vitro and in vivo mutagenicity of the butadiene metabolites butadiene diolepoxide, butadiene monoepoxide and diepoxybutane.

Three metabolites of 1,3-butadiene, namely butadiene diolepoxide, butadiene monoepoxide and diepoxybutane, were tested in the bacterial mutation assay using Salmonella typhimurium strain TA100 with and without metabolic activation (S9 mix). All three compounds showed a mutagenic response. The bifunctional epoxide was more effective than the diolepoxide which was more effective than the monoepoxide. Toxicity appeared to follow the ranking of the chemicals for their mutagenic potency. The monoepoxide and the diolepoxide were also tested for induction of micronuclei in mouse bone marrow erythrocytes and for dominant lethal mutation induction in postmeiotic male mouse germ cells. The effects of the diepoxide in both in vivo tests have been published earlier. In the micronucleus assay, the three metabolites gave a positive response whereby the diepoxide was more effective than the monoepoxide which was more effective than the diolepoxide. In contrast to the diepoxide which was positive at a dose as low as 36 mg/kg, the monoepoxide and the diol did not show an induction of dominant lethal effects up to doses of 120 and 240 mg/kg, respectively. It is concluded that the metabolites were mutagenic in bacteria without metabolic activation and clastogenic in mouse bone marrow; only the bifunctional diepoxide, however, was active in postmeiotic male mouse germ cells.

Animals↗

PubMed1997

[Studies on meiotic delay and aneuploidies induced by colcemid (COM) in male mouse germ cells].

Colcemid (COM) was tested for induction of meiotic delay and aneuploidies in meiotic metaphase II (MMII) of male(101/E 1 XC 3 H/E 1)F 1 mice post single intraperitoneum (i.p.) injection. The dose of 1 mg/kg of COM was used and sampled at 2, 6, 10, 14 and 18 h after COM treatment. The number of MMII and MMI were the highest at 2 and 6 h repectively, and decreased rapidly with lengthening of COM treatment and reached the lowest at 14 h; then increased. However, the ratio of MMII to MMI was always significantly higher than in solvent control at every sample interval. Under our experimental conditions, COM did not show aneugenicity in male mouse germ cells. The possible mechanisms by which COM caused meiotic delay and reasons that COM did not induce aneuploidy in MMII were discussed.

Aneuploidy↗

PubMed1996

Analysis of micronuclei induced by 1,3-butadiene and its metabolites using fluorescence in situ hybridization.

In our previous study, micronuclei (MN) were induced in bone marrow cells of mice following inhalation exposure to 1300 ppm of 1,3-butadiene (BD) for 6 h per day on 5 consecutive days, and in splenocytes of mice and rats treated intraperitoneally with 80 mg/kg 1,2-epoxybutene (EB) and 30 mg/kg 1,2,3,4-diepoxybutane (DEB), respectively. In the present study, the nature of MN induced by BD, EB and DEB was analyzed by means of fluorescence in situ hybridization (FISH) using mouse minor satellite DNA and rat satellite I DNA as probes. Percentages of MN with centromere signals (MN+) measured following exposures to BD, EB and DEB indicate that these agents are predominantly clastogens. Frequencies of MN+ per 1000 cells suggest that BD, EB and DEB are not only strong clastogens, but also weak aneugens in mice. The weak aneugenic effect of EB and DEB was not observed in rats. Analysis of the number of centromere signals in individual MN, and the size distribution of MN with centromere signals in EB- and DEB-treated animals, and in animals exposed to the positive controls diethylstilbestrol (DES) and mitomycin C (MMC) led to the following conclusions: (1) analysis of MN for the number of centromere signals may be a useful indicator for identifying chemicals with aneugenic properties; (2) there is no correlation between the size of MN and their origin (i.e., chromosome loss/gain or fragment).

Animals↗

PubMed1996

Synthesis report of the step project detection of germ cell mutagens.

The project 'Detection of Germ Cell Mutagens' was designed with three major goals: (1) Detection and characterization of germ-cell mutagens; (2) standardization and validation of new germ-cell tests; and (3) development of a data base on germ-cell mutagenicity. All three goals were achieved. The classical germ-cell tests were applied to characterize the genetic effects of acrylamide (AA), 1,3-butadiene (BD), trophosphamide (TP) and urethane (UR). All but UR were found to cause heritable genetic damage. The experimental data obtained for AA and BD were the basis for genetic risk evaluations during the EC/US Workshop on Risk Assessment 'Human Genetic Risk from Exposure to Chemicals, Focusing on the Feasibility of the Parallelogram Approach'. Nine chemicals were employed to validate the spermatid micronucleus assay with mice and rats: AA, BD and its metabolites 1,2-epoxybutene-3 and 1,2:3,4-diepoxybutane, chlorambucil, mitomycin C, methylnitrosourea, TP and UR. The spermatid micronucleus test was combined with micronucleus tests in somatic cells such as bone marrow or peripheral blood erythrocytes, and splenocytes which allowed a comparison of effects in somatic and germinal cells. Improvements of the spermatid micronucleus test included BrdU-labelling of premeiotic S-phase for the determination of stage sensitivity and fluorescence in situ hybridization with pancentromeric DNA-probes to distinguish between clastogenic and aneugenic events. The results indicate that the spermatid micronucleus test with its improvements is an adequate procedure to detect germ-cell clastogenicity and to compare the activity of chemicals in different tissues and between species, i.e., rats and mice. Other germ cell methods under study were the flow cytometric measurement of testicular sperm DNA and the cytogenetic analysis of preimplantation embryos for chromosomal aberrations and micronuclei. The collection of a reliable germ-cell data base was accomplished through a critical evaluation of the literature and with the data obtained in the present project. Remarkable concordance between responses of germ cell tests to chemical mutagens was the most striking conclusion to be drawn from the present data base.

Acrylamide↗

PubMed1996

The detection and evaluation of aneugenic chemicals.

Although aneuploidy makes a significant contribution to both somatic and inherited disease the mechanisms by which environmental chemicals may induce numerical chromosome aberrations are only poorly defined. The European Union Project was aimed to further our understanding of those chemical interactions with the components of the mitotic and meiotic cell division cycle which may lead to aneuploidy and to characterise the parameters such as cellular metabolism which may influence the activity of aneugenic chemicals. C-mitosis can be induced by the highly lipophilic polychlorinated biphenyl and the completion of mitosis and cleavage can be modified by agents which deplete cellular levels of reduced glutathione. Modifications of the fidelity of chromosome segregation were produced by inhibiting the functioning of topoisomerase II during chromatid separation. In contrast, the modification of centromere integrity resulted in chromosome breakage as opposed to disturbance of segregation. Modifiers of tubulin assembly and centriolar functioning in somatic cells such as acrylamide, vinblastine and diazepam reproduced their activity in rodent bone marrow and male germ cells. The analysis of chromosome malsegregation in Aspergillus nidulans by a structurally related series of halogenated hydrocarbons was used to develop a QSAR model which had high predictive value for the results of fungal tests for previously untested related chemicals. Metabolic studies of potential aneugens in genetically engineered human lymphoblastoid cells demonstrated the detoxification of the aneugenic activity of chloral hydrate and the activation of 2,3-dichlorobutane, 1,1,2-trichloroethane and trichloroethylene by Phase I biotransforming enzymes. Cell transformation studies in Syrian hamster dermal cultures using a panel of 22 reference and or potential aneugens indicated that 15 of the 22 produced positive results following single exposures. Five of the aneugens which were negative following single exposures produced positive results where cultures were continuously exposed for up to 6 weeks to low concentrations following a single non-transforming exposure to the mutagen dimethyl sulphate. The transformation studies indicate that a significant proportion of chemical aneugens are potential complete carcinogens and/or co-carcinogens. To optimise the enumeration of chromosomes following exposure to potential chemical aneugens whole chromosome paints and centromere specific probes suitable for use in fluorescence in situ hybridisation (FISH) were developed for the rat, mouse and Chinese hamster and selected human probes evaluated for their suitability for routine use. Molecular chromosome probes were used to develop protocols for enumerating chromosomes in metaphase cells and centromeres and micronuclei in interphase cells. The analysis of segregation of specific centromeres in binucleate cells following cytochalasin B treatment was shown to be a potentially valuable system for characterising non-disjunction following chemical exposure. Whole chromosome paints and centromere specific probes were used to demonstrate the presence of dose-response thresholds following treatment with a reference panel of spindle inhibiting chemicals. These data indicate that the FISH technology is suitable for evaluating the relative hazards of low-dose exposures to aneugenic chemicals.

Aneuploidy↗

PubMed1996

IPCS harmonization of methods for the prediction and quantification of human carcinogenic/mutagenic hazard, and for indicating the probable mechanism of action of carcinogens.

A flow chart is presented as a recommended sequence of tests to predict the carcinogenic hazard, and to predict and quantify the mutagenic hazard to germ cells of chemicals to humans. Ten associated principles of testing for these endpoints are also suggested. These recommendations are the result of a meeting convened under the auspices of the International Programme on Chemical Safety (IPCS), as part of their project on 'Harmonization of Approaches to the Assessment of Risk from Exposure to Chemicals'. The meeting was held at Carshalton, Surrey, from 13-17 February 1995.

Animals↗