Sister-chromatid exchanges in human lymphocytes treated with 4-chloromethylbiphenyl and benzyl chloride

At non-inhibitory concentrations, 4CMB and BC both induced small, but dose-related increases in SCE. The dose-response curves gave a highly significant correlation for 4CMB (r=0.996, P<0.001) and a significant correlation for BC (r=0.757, P<0.05) which reflects the relative activities of these substances in bacterial mutation tests.     

The activity of 4CMB, 4HMB and BC in Saccharomyces cerevisiae JD1

3 structurally related compounds, 4-chloromethylbiphenyl (4CMB), 4-hydroxymethylbiphenyl (4HMB), and benzyl chloride (BC) were assayed for their ability to induce mitotic gene conversion in stationary phase cultures of the yeast, Saccharomyces cerevisiae JD1. This strain allows gene conversion to be scored at 2 independent loci, trp 5 and his 4.The results reported in this paper indicate that both 4CMB and BC are genetically active in yeast, producing dose-related increases in mitotic gene conversion at both the loci tested; 4HMB showed no such activity. At high survival levels 4CMB and BC showed comparable activity. However, as toxicity increased BC showed much more potent convertogenic activity, whereas with 4CMB a reduction in induced gene conversion was observed. The presence of a microsomal activation system derived from the livers of Aroclor-induced male rats did not significantly affect the activity of any of the compounds.     

UKEMS trial: Bacterial mutation tests of 4-chloromethylbiphenyl, 4-hydroxymethylbiphenyl,and benzyl chloride,using E. coli WP2uvrA(pKM101) and S. typhimurium TA98 and TA100

4CMB, 4HMB and BC were assayed in plate tests, using E. coli WP2uvrA(pKM101), and S. typhimurium TA98 and TA100, in the presence or absence of microsomal activation. 4CMB was also assayed in fluctuation tests using E. coli WP2uvrA(pKM101). 4HMB was uniformly negative, and 4CMB was mutagenic to all 3 strains. BC was negative in TA98 and positive in TA100 and WP2uvrA(pKM101). The presence or absence of S9 made no substantial difference to the mutagenicity of 4CMB or BC.     

An assay for mutagenic activity of 4CMB, 4HMB and BC,using the “microtitre” fluctuation test

4CMB, 4HMB and BC were assayed for mutagenic activity using the ‘microtitre’ bacterial fluctuation test without metabolic activation. 4CMB was positive in strains of Salmonella typhimurium detecting both base-substitution and frameshift mutation. BC was weakly positive only in the strain which detects base-substitution mutation. 4HMB was negative in both strains. 4CMB and 4HMB were equally toxic to the strains, whilst BC was comparatively less toxic.     

Fluctuation test data on 4-chloromethylbiphenyl (4CMB), 4-hydroxymethylbiphenyl (4HMB) and Benzyl Chloride (BC) using Salmonella typhimurium TA98 and TA100

Bacterial fluctuation tests (Green et al., 1976) were performed both with and without metabolic activation using the ‘Ames’ Salmonella typhimurium strains TA98 and TA100 (Ames et al., 1975) to assay the mutagenic potential of 4CMB, 4HMB and BC.4CMB and 4HMB were tested on the same occasion. However, 4CMB was only compared to BC in one assay. The results also show an independent test of BC.     

A comparison of the response to 4CMB, 4HMB and BC of 5 yeast strains differing in their radiosensitivities

The 3 test compounds 4CMB, 4HMB and BC were assayed for their genotoxicity using stationary phase cultures of 5 yeast strains which differ in their mutagen sensitivity. It was found that 4HMB produced no differences in survival between the 5 strains whereas 4CMB and BC caused more lethality in the triple rad strain than the other 4 strains. The results indicate that both BC and 4CMB are capable of inducing DNA damage which results in cell lethality in the repair-deficient triple mutant.     

UKEMS trial compounds: In vitro bacterial mutagenicity

4CMB, 4HMB and BC were examined in the Ames test using Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100. 4CMB was mutagenic for all of the indicator strains, 4HMB was inactive and BC was weakly mutagenic for TA100 only.     

Genetic activities of 4-chloromethylbiphenyl,the 4-hydroxy derivative and benzyl chloride in the soma and germ line of Drosophila melanogaster

The genetic activities of 4CMB, 4HMB and BC were assayed as regards the induction of somatic alterations in gene expression on an unstable w⁺ locus with an intragenic TE and all the simultaneously induced germinal mutations on the X-chromosome carrying this locus. The compounds were applied topically in solution at equimolar doses on late embryos and newly hatched larvae. The somatic events were scored as aberrantly pigmented eye sectors in the emerging adult males and the germinal mutations in their F₂ progeny, according to the Muller-5 technique.The somatic events were expressed as red or white mosaic eye sectors; the former could be an outcome of the repression or deletion of the zeste-regulatory proximal subunits of w⁺ locus, and the latter generally attributable to deletions (w⁻) within its structural part. All 3 compounds were effective in the induction of red sectors at the higher tested doses (0.5–2.0 mM) and the level of this activity was virtually the same for 4CMB and 4HMB, but was 2-fold higher for BC. In contrast, the frequencies of the simultaneously scored white sectors were not raised significantly above the controls with 4CMB, but showed decisive increases above this level with both 4HMB and BC.The germinal X-chromosome mutations (recessive lethals and visibles) were only induced at the highest tested dose (2.0 mM), and their frequencies were virtually the same for all 3 compounds reaching a common level of about 0.6%, which is some 3-fold the normal control level for the test system. Specific-locus mutability at the TE w⁺ was suggestively positive only with BC.     

Bacterial mutagenicity tests on 4-chloromethylbiphenyl and 2 structural analogues

4CMB, 4HMB and BC were tested in 5 strains of S. typhimurium and 2 strains of E. coli without S9. 4HMB was negative in all strains. 4CMB was a strong positive mutagen in TA1535, TA1537, TA1538, TA98, TA100 and WP2uvrA⁻(pKM101), and BC was a weak mutagen in TA100 and WP2uvrA⁻(pKM101). Positivity was determined as a dose response over 3 or more points, in repeat experiments, giving a significant correlation coefficient.     

The effects of 4CMB, 4HMB and BC on SCE,chromosome aberration and point mutation in cultured Chinese hamster ovary cells

Cultured Chinese hamster ovary cells (CHO-KI-BH₄) were treated for 2 h with 4CMB, 4HMB and BC, in the absence of any exogenous metabolic activation system. The cells were subjected to tests for survival, sister-chromatid exchange, chromosome aberration and mutation to thioguanine resistance.4HMB had no effect in any test at concentrations up to 100 μg/ml. 4CMB was slightly more toxic than BC. Both 4CMB and BC induced SCE and chromosome aberrations, but the effects were more marked with BC. With 4CMB, SCE increased with dose only up to about 7 μg/ml and then levelled off. A weak mutagenic effect was observed with both BC and 4CMB, but in each case, the response reached a peak and was not evident at higher doses.     

The induction of mitotic gene conversion in the yeast,Saccharomyces cerevisiae JD1 by 4-chloromethylbiphenyl (4CMB), Benzyl Chloride (BC) and 4-hydroxymethylbiphenyl (4HMB)

The induction of mitotic gene conversion by 4CMB, BC and 4HMB was studied in both log-phase and stationary-phase cultures of the yeast, Saccharomyces cerevisiae JD1. Assays were performed both in the presence and in the absence of S9 microsomal fraction obtained from a liver homogenate from rats pretreated with Aroclor 1254.Exposure of both stationary-phase and log-phase cultures to 4CMB and BC resulted in an increase in mitotic gene conversion, both in the presence and in the absence of a microsomal activation system; the magnitude of response was greater in stationary-phase cultures. 4HMB did not increase the gene conversion frequency in log-phase or stationary-phase cultures.     

Testing for mitotic crossingover and induced aneuploidy using Aspergillus nidulans as part of the UKEMS test programme

4CMB and BC were shown to induce mitotic crossingover in Aspergillus nidulans, whereas no such activity was shown in the presence of 4HMB. None of the 3 test compounds were able to induce chromosome aneuploidy.     

Assay of the induction of mitotic crossing-over and aneuploidy in yeast by BC, 4CMB and 4HMB

Both BC and 4CMB but not 4HMB were shown to be capable of inducing mitotic crossing-over in exponential phase cells of the D6 strain of the yeast Saccharomyces cerevisiae. In contrast, none of the 3 test compounds were capable of inducing mitotic chromosome aneuploidy.     

Drosophila,sex-linked recessive lethal tests with 4-chloromethylbiphenyl and benzyl chloride

4CMB failed to induce sex-linked recessive lethal mutations in Drosophila melanogaster, by feeding to larvae or adults, or by injecting adults. BC was negative by larval feeding.     

Chromosome analysis of cultured rat-liver epithelial cells (RL4) treated with 4-chloromethylbiphenyl, 4-hydroxymethylbiphenyl and benzyl chloride

Cultured rat-liver epithelial cells, RL₄, were exposed to maximum concentrations of 24 μg/ml 4CMB, 100 μg/ml 4HMB and 30 μg/ml BC. 4CMB and BC induced highly significant numbers of aberrations; 4HMB did not cause an increase in aberration levels.     

4-chloromethylbiphenyl (4CMB), benzyl chloride (BC) and 4-hydroxymethylbiphenyl (4HMB): Reverse mutation tests with Salmonella typhimurium

In this study 4CMB was shown to be a strong, direct-acting, mutagen for S. typhimurium strains TA1538, TA1537, TA98 and TA100. However, for strain TA1535 the compound was only weakly mutagenic. No conclusive evidence of mutagenic activity was seen in tests with BC or 4HMB.     

The mutagenic activity of 4-chloromethylbiphenyl (4CMB) and Benzyl Chloride (BC) in the bacterial/microsome assay

The mutagenic activity of 4CMB was investigated in agar layer cultures of Salmonella typhimurium TA1535, TA1537, TA1538, TA98 and TA100, and Escherichia coli WP2 and WP2 uvrA. The mutagenic activity of BC was investigated in the Salmonella strains only. Assays were performed both in the absence and in the presence of S9 microsomal fraction obtained from a liver homogenate from rats pretreated with Aroclor 1254.     

Dose–response assessment of metal toxicity upon indigenous Thiobacillus thiooxidans BC1

This study provides a first attempt from a toxicological perspective to put forward, in general terms and explanations, the toxicity series of Cd(II), Cu(II) and Zn(II) to Thiobacillus thiooxidans BC1. Sulphur oxidation and sulphuric acid production are strongly related to microbial growth at pH less than 4. Dose–response analysis on chronic and acute toxicity (e.g. EC₂₀, median effective dose EC₅₀ and slope factor B) of divalent cadmium, copper and zinc cations suggests a toxicity series of Cu>Cd>>Zn to T. thiooxidans BC1. Zn(II) is termed non-toxic and the maximum treatment concentrations of Cd(II) and Cu(II) are approximately 300, 400 mg/l, respectively. This assessment clearly indicates viable operation ranges of metal bioleaching for mine wastewater treatment, suggesting a technological feasibility of biotreatment using acidophilic thiobacilli T. thiooxidans BC1.     

Impact of baseline telomere length on survival and chemotherapy related toxicity in breast cancer patients receiving (neo)adjuvant anthracycline containing chemotherapy

PurposeThe aim of this study is to assess baseline mean leukocyte telomere length (TL) as a potential predictive factor for chemotherapy toxicity and a prognostic marker for long-term outcome in early breast cancer (BC) patients.Methods445 BC patients were selected, diagnosed between 2007 and 2010 with early BC and treated with (neo)adjuvant fluorouracil, epirubicin and cyclophosphamide (FEC) or with FEC and Docetaxel (FEC-D). RT-qPCR was performed on germline DNA samples collected at diagnosis before any treatment, to measure mean leukocyte TL. Uni- and multivariable logistic regression or Cox proportional hazard regression analyses were carried out to assess correlation between baseline TL and toxicity parameters (derived from the medical chart) or longer-term outcome.ResultsBaseline TL correlated with age as expected (p = 0.005), but not with febrile neutropenia (n = 97), left ventricular ejection fraction >10% decrease (n = 17) nor other toxicity endpoints measured (all p > 0.05). TL was neither associated with overall survival, breast cancer specific survival or distant disease-free survival (all p > 0.05).ConclusionsBaseline TL is not associated with chemotherapy-related toxicity nor long-term outcome in BC patients.     

HER2 Ile655Val and PTEN IVS4 polymorphisms in patients with breast cancer

Although HER2/PTEN pathway is commonly disrupted in cancer, association of HER2 and PTEN polymorphisms with breast cancer (BC) remains controversial. We investigated the HER2 Ile655 Val and PTEN IVS4 polymorphisms in patients with BC in Turkish population. HER2 Ile655Val (rs 1136201) and PTEN IVS4 (rs 3830675) polymorphisms were determined using polymerase chain reaction-based restriction fragment length polymorphism (PCR–RFLP) in blood samples of 118 BC patients and 118 age-matched healthy controls. We found that the frequency of the Ile/Val genotype of HER2 Ile655Val gene was significantly higher in BC patients (p < 0.009; OR: 1,983 95 % CI: 1.181—3.328). The presence of ATCTT insertion (+/+) genotype at downstream of exon 4 in intron 4 of PTEN IVS4 gene was also associated with 1.83 fold decreased risk of BC development (p < 0.033; OR: 1.83, 95 % CI: 1.11—3.03). Analysis on clinico-pathological parameters showed neither HER2 Ile655Val nor PTEN IVS4 genotypes were not associated with any of the variables (p > 0.05).In conclusion, our findings suggest that the Ile/Val genotype of HER2 and ATCTT insertion (+/+) genotype of PTEN IVS4 gene may play an important role as genetic markers for breast cancer risk, but both genes genotypes may not be useful for predicting tumor prognosis in Turkish population.