No induction of chromosomal aberrations in Chinese hamster ovary cells by chrysophanol.

Chrysophanol is an anthraquinone which occurs in several herbal drugs, e.g. senna, a commonly used laxative. As there are only limited data on its clastogenic potential we have investigated its capability to cause chromosomal aberrations in the Chinese hamster ovary cell assay. There were no significant increases in chromosomal aberrations when chrysophanol was tested up to its limit of solubility with or without metabolic activation. We conclude that chrysophanol had no clastogenic activity under the conditions described.

Fluorescence in situ hybridisation with chromosome-specific centromeric probes: a sensitive method to detect aneuploidy.

Cytochalasin B-blocked binucleate human lymphocytes from female donors have been used to measure micronucleus induction and other aneuploidy events after treatment with colchicine, vinblastine or carbendazim. For the aneuploidy events, centromeric probes for 6 selected chromosomes (1, 8, X, 11, 17, 18) were used to measure chromosome loss, addition and non-disjunction in the interphase nuclei of these binucleate cells. The chromosomes were probed in pairs using Cy-3 (red) and FITC (green) labels for the 2 different centromeric regions. For colchicine, the total non-disjunction frequencies for chromosomes 1 and 8 were similar to the total micronucleus frequencies, but were detected as significant at lower concentrations. For vinblastine (chromosomes 1 and 8) and carbendazim (all 6 chromosomes) the frequencies of non-disjunction far exceeded (7 and > 2-fold, respectively) the peak frequencies of micronucleus induction. Although most chromosomes exhibited similar sensitivity in all the aneuploidy events measured, there was an indication that chromosome X was more than susceptible to non-disjunction than the other chromosomes. We believe that measurement of non-disjunction in binucleate human lymphocytes using chromosome specific centromeric probes offers a sensitive method for detection of aneuploidy and is particularly appropriate for the establishment of thresholds.

Aroclor-1254-induced rat-liver S9 causes chromosomal aberrations in CHO cells but not human lymphocytes: a role for active oxygen?

Incubation of CHO cells with Aroclor-1254 induced S9 mix gave higher than normal levels of chromosomal aberrations (CA), sometimes achieving exceptional levels (e.g. up to 40 CA, minus gaps, per 100 cells against a normal range of 0-10). The same batches of S9 gave normal CA frequencies in human lymphocytes from whole blood cultures, normal mutation frequencies in bacterial reversion (Ames) tests and mammalian cell hgprt mutation tests, and normal levels of repair in scheduled DNA synthesis tests. As cytochrome P-450 enzymes, in the absence of substrate, will cycle electrons and could produce active oxygen species (AOS), which are known to be clastogenic, this was investigated. Preliminary studies showed the complete S9 mix was required for the high levels of CA, and that CA levels could be dramatically reduced by co-incubation with catalase or vitamin E. Interestingly, uninduced S9 and phenobarbitone/beta-naphthoflavone induced S9 mixes did not cause high levels of CA, and presumably have different isozymes of P-450 that do not generate AOS as readily. These observations suggest that CHO cells are sensitive to AOS which are inactivated by blood components. This may have implications for the choice of cell system in routine clastogenicity testing of novel chemicals.

Normal chromosomal aberration frequencies in peripheral lymphocytes of healthy human volunteers exposed to a maximum daily dose of paracetamol in a double blind trial.

Paracetamol (acetaminophen) has been examined for mutagenic potential in numerous studies: gene mutation tests consistently gave negative results while in vitro chromosomal aberration tests showed equally consistently positive effects. In vivo studies for chromosome breaking activity gave clearly negative, equivocal or weakly positive results. In particular two reports have indicated that human volunteers taking a maximum daily dose of paracetamol (3 x 1000 mg over 8 h) exhibited significantly elevated frequencies of chromatid breaks in their peripheral lymphocytes 24 h later. In the one study evaluating the time course, levels returned to normal between 3 and 7 days later. We performed a carefully controlled double-blind study in which volunteers were pre-screened for normal liver function, they all were non-smoking and their diet and environmental exposures were controlled during the study. Cell-cycle kinetics were monitored and paralleled and a placebo group was included. Although a larger number of cells than in the other studies was analysed we were unable to reproduce their findings. No significant increases in structural chromosome aberrations (CA) were found either when the paracetamol group (male, female or both) post-dosing values were compared with pre-dosing values, or when treated groups at any sampling time were compared with the placebo groups. There was not even any evidence that individuals responded to the clastogenic potential of paracetamol or that a group response may have been masked by non-responders. In conjunction with the recently published results of the NTP bioassay, showing no carcinogenic activity in mice and no carcinogenic activity in rats except an increase of mononuclear cell leukaemia in female rats which is of doubtful relevance, the study presented here argues that paracetamol does not pose an unacceptable (if any) genotoxic/carcinogenic risk to man.

Increased sensitivity of cell strains from Cockayne's syndrome to sister-chromatid-exchange induction and cell killing by UV light.

Ultraviolet light-induced sister-chromatid exchanges (SCE) and cell killing were investigated in 4 fibroblast cell strains from patients with the sun-sensitive disease Cockayne's syndrome (CS). All 4 CS cell strains proved to be hypersensitive to UV for both of these end-points, but no close correlation between levels of SCE and lethality was observed. Cell strains from two individuals heterozygous for CS were indistinguishable from wild-type.

Safety evaluation of esperase.

Esperase is a proteolytic enzyme preparation that can be used as a processing aid in the food industry. The following studies were performed to establish safety for the consumer: oral toxicity study (13 wk) in the rat; teratogenicity study in the rat; gene mutation assays in Salmonella typhimurium and mammalian cells in vitro, and chromosome aberration assay in vitro. General toxicity was low; the effects seen were attributed to proteolytic activity and the loading with sodium chloride. Neither of these factors will be relevant to consumers of the processed food. There was no evidence of effects on pregnancy outcome or mutagenic potential. When these results are considered together with existing knowledge of the production organism and the chemical and microbiological characterization of the enzyme preparation, they indicate that Esperase will be safe for its intended application in food processing.

Mutagenic and clastogenic properties of (3-[3-(6-benzoyloxy-3-cyano-2- pyridyloxycarbonyl) benzoyl]-1-ethoxymethyl-5-fluorouracil) (BOF-A2), a new 5-fluorouracil derivative with anti-tumour activity.

As part of the safety assessment of (3-[3-(6-benzoyloxy-3-cyano-2- pyridyloxycarbonyl)benzoyl]-1-ethoxymethyl-5-fluorouracil) (BOF-A2), a new 5-fluorouracil (5-FU) derivative with anti-tumour activity, its potential genotoxicity was studied in 3 different tests. BOF-A2 was negative in a reverse mutation (Ames) test in strains of S. typhimurium and E. coli. BOF-A2 induced chromosomal aberrations in Chinese hamster cells in vitro especially in the presence of exogenous metabolic activation, and was clastogenic in vivo, inducing micronuclei in mouse bone marrow. The clastogenic activity of BOF-A2 was similar to that of 5-FU.

On the types of chromosomal aberrations induced by 8-methoxypsoralen.

When activated by near-ultraviolet light, 8-methoxypsoralen can react with pyrimidine bases to produce mono-adducts in DNA. Upon further irradiation these mono-adducts can be converted to interstrand crosslinks, but if the re-irradiation is carried out in the absence of unbound 8-methoxypsoralen, no new mono-adducts can be formed. The effects of re-irradiation are, therefore, a consequence of the conversion of mono-adducts into crosslinks. Here we report the types of chromosomal aberrations produced by re-irradiation and, hence, by DNA crosslinks. Our results demonstrate that crosslinks induce a wide variety of chromosomal aberrations in the first division after treatment. In addition, crosslinks are shown to induce new aberrations in second-division cells, a result which shows that the crosslink or some lesion derived from it survives at least one round of DNA replication.

The use of two-dimensional electrophoresis to detect mutations induced in mouse spermatogonia by ethylnitrosourea.

Two-dimensional electrophoresis should, in theory, be a suitable method for the measurement of induced mutation rates in the germ cells of mice. Not only can the polypeptide products of a large number of genes be resolved on a single gel but the detection of mutations which lead to proteins with altered electrophoretic properties (but not necessarily altered function) is possible. Our attempts to apply two-dimensional electrophoresis to the detection of mutation in vivo have involved three stages: (i) the rapid production of gels of high resolution and reproducibility; (ii) the identification of eight interstrain protein variants and demonstration of their simple genetic basis; and (iii) a pilot experiment using the powerful germ-cell mutagen ethylnitrosourea. It was found that although interstrain protein variants could be detected and shown to be inherited in a codominant manner, induced variants were rarely detected even on high quality gels. Only 2 variants were detected among 67 offspring of male mice treated with 150 mg/kg ethylnitrosourea. This represented a mutation rate of 0.88 X 10(-4) mutations per locus per gamete.

Medication induced gingival overgrowth

A number of idiopathic, pathological and pharmacological reactions may result in an overgrowth of the gingiva. This review concentrates on those overgrowths associated with various pharmacological agents. The pharmaco-kinetics and side effects of each drug associated with gingival overgrowth are discussed along with the clinical and histological features and treatment. By examining the possible pathogeneses for these overgrowths we propose a unifying hypothesis for the causation based around inhibition of apoptosis and decreased collagenase activity modulated by cytoplasmic calcium