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T I Matula

Publications and source records attributed to T I Matula.

At least 19 recordsLinked to original sources
PubMed2003

Analysis of pCU1 replication origins: dependence of oriS on the plasmid-encoded replication initiation protein RepA.

The broad-host-range replicon of the plasmid pCU1 has three origins of vegetative replication called oriB, oriS, and oriV. In the multi-origin replicon, individual origins can distinguish among replication factors provided by the host. It has been found that during replication in Escherichia coli polA(-) host, oriS was the only active origin of a mutant pCU1 derivative bearing a mutation in the gene encoding replication initiation protein RepA. To further investigate the capacity of oriS to function in an E. coli polA(-) host we constructed a number of clones of the basic replicon of pCU1 containing oriS as the only replication origin. An oriS construct created with pUC18 could transform the polA(-) strain when RepA was supplied in trans. When the oriS region (between nucleotides 290 and 832) was ligated to an antibiotic resistance Omega fragment, the construct could be recovered as a plasmid from polA(+) strain if functional RepA was provided in trans. Our results therefore indicate that the basic replicon of pCU1, containing oriS as the sole origin, does require RepA to initiate plasmid replication in E. coli

Bacterial Proteins↗
PubMed1998

In vitro investigation of toxaphene genotoxicity in S. typhimurium and Chinese hamster V79 lung fibroblasts.

The polychlorinated pesticide toxaphene has been identified as a persistent environmental contaminant and is of particular concern in the Great Lakes and Arctic regions of Canada. Inconsistencies in published in vitro genotoxicology studies have hindered risk assessments of toxaphene exposure. When toxaphene mutagenicity was re-evaluated in the Ames Salmonella/microsome assay at 10-10,000 microg/plate, a dose-dependent increase in His revertants occurred in all five strains of S. typhimurium tested (TA97, TA98, TA100, TA102 and TA104) with higher mutation frequencies observed in the absence of S9 metabolic activation. However, the mutagenic potential of toxaphene was relatively low with concentrations greater than 500 microg/plate required to induce mutation. Toxaphene genotoxicity was also examined in a mammalian system using Chinese hamster V79 lung fibroblasts with metabolic activation provided by human HepG2 hepatoma cells. Genotoxicity of 1-10 microg/ml toxaphene was examined by measuring the frequency of sister chromatid exchange (SCE) and mutation induction at the hypoxanthine guanine phosphoribosyl transferase (HGPRT) gene locus. Although small increases in SCE were observed at toxic concentrations of toxaphene approaching the LD50 (10 microg/ml), they were not found to be statistically significant relative to control. Toxaphene was also unable to induce HGPRT mutagenesis at the concentrations tested. These results show that while toxaphene is a weak, direct-acting mutagen in the Ames Salmonella Test, convincing evidence of dose-dependent SCE induction and mutagenicity at the HGPRT gene locus could not be demonstrated in V79 cells.

Animals↗
PubMed1997

Mutagenic heterocyclic aromatic amines (HAAs) in 'processed food flavour' samples.

Eight samples of 'processed food flavours' (PFFs), chosen from five different categories, were analysed for their mutagenic activity using the Ames Salmonella assay, and also for the presence of eight heterocyclic aromatic amines (HAAs), namely 2-amino-3-(trideuteromethyl)imidazo[4,5-f]quinoline (IQ), 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (7,8-DiMeIQx), 2-amino-3,7,8-trimethylimidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx), 2-amino-I-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) using liquid chromatography and mass spectrometry (LC/MS). The isolation of HAAs was based on sequential liquid-liquid extraction procedures of samples at both acidic and basic pH values. The recoveries and the clean-up were monitored by introduction of quality control samples and by spiking with three tri-deuterated standards of HAAs. Although the results for the mutagenicity assay were comparable by testing less-purified and highly-purified extracts, the analysis for identification and quantification of HAAs by LC/MS required highly purified concentrates. Four samples had little or no mutagenic activity and these results were in agreement with their LC/MS results: they had no detectable levels (detection limits 1-3 ppb) of any of the HAAs monitored. The mutagenic activity of one sample was in complete agreement with the quantification of HAAs by LC/MS. Two samples produced strong mutagenic responses (3115 and 2664 revertants/g). In one sample, LC/MS analysis revealed the presence of 9.6 ppb IQ, whereas LC/MS of the other could not confirm the presence of any of the eight HAAs monitored. Two samples produced mild mutagenic activity (204 and 160 revertants/g), but relatively elevated concentrations of IQ (6.7 and 6.8 ng/g) by LC/MS. The extracts from all samples were tested for their modifying effects on mutagenicity of four HAAs. The discrepancy between the Ames test and the LC/MS analysis of some samples indicates several possibilities, such as the presence of some other HAAs, of their isomers or of other mutagens. In addition, the presence of mutagen modifiers (inhibitors or synergists) was observed in most samples. The results indicate that although chemical tests (e.g. LC/MS) can provide quantitative data for the HAAs monitored, the Ames mutagenicity test should also be conducted to determine the mutagenic activities of PFFs, in order to assess their health risk potential.

Amines↗
PubMed1997

Heterocyclic aromatic amine content in pre-processed meat cuts produced in Canada.

In an ongoing survey, the presence of heterocyclic aromatic amines (HAAs) was determined in processed, ready-to-eat meat products sold as 'meat cuts'. HAAs are a group of recently recognized mutagenic/carcinogenic contaminants in foods that are produced during the heat processing of meat. 16 samples of meat cuts (e.g. turkey breast, salami, chicken loaf, cooked ham, all beef meat, pepperoni, etc.), randomly purchased from supermarkets and specialty food stores in the Ottawa area, were analysed for the presence of eight HAAs. The isolation of HAAs was based on sequential liquid-liquid extraction procedures of the samples at both acidic and basic pH values. The mutagenic activity of these samples was determined using the Ames/Salmonella microsome assay with the strain TA98 plus rat liver S-9 metabolic activation. The mutagenicity of these samples ranged from undetectable to slightly active. The highest mutagenic activity, 141 induced revertants/g, was found in a smoked turkey breast sample. 11 samples were not mutagenic, including two that indicated a tendency for inhibition of the spontaneous revertants. The remaining four samples exhibited very low mutagenic activity. For chemical analysis, the extracts were purified with two solid phase extraction cartridges. Quantitative analysis was performed by using liquid chromatography for separation and mass spectrometry for detection. With the exception of trace amounts (0.4 ng/g) of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) in the sample with highest mutagenic activity, the chemical analysis did not detect the presence of any of the eight most frequently found HAAs in fried or broiled meat products. These data suggest that consumption of meat cuts does not present a serious health risk from HAA-type contaminants.

Amines↗
PubMed1996

Ensuring recovery of intact RNA from rat pancreas.

The isolation of intact RNA from rat pancreas is compromised by autolysis and by the presence of endogenous ribonucleases. In order to ameliorate recovery we systematically investigated available RNA extraction methods and paid particular attention to the influence of frozen storage and ribonuclease inhibition strategies on overall yield and quality of RNA. Modifications to the basic procedure of Chomczynski and Sacchi (1987) are described which allow, reproducibly, to obtain rat pancreatic RNA suitable for Northern blot hybridization, RT-PCR, and differential display analysis.

Animals↗
PubMed1996

The effect of teas on the in vitro mutagenic potential of heterocyclic aromatic amines.

Water extracts of eight brands (five types: 'green', 'black', 'oolong', decaffeinated and instant) of common teas (derived from Camellia sinensis) and infusions of six randomly selected herbal teas were examined for inhibitory or potentiating effects on the mutagenicity of eight heterocyclic aromatic amines (HAA) using the Ames Salmonella typhimurium TA98 and S-9 assay. HAA, produced in foods during regular heat processing of meat, exhibit mutagenic/carcinogenic activities. Tea extracts from C. sinensis displayed very potent antimutagenic effects against most HAA: total or substantial inhibition of mutagenic activity of the eight HAA was obtained with extracts equivalent to 50 mg tea leaves/plate (mgEq) and potent inhibition was frequently achieved even with 10 mgEq/plate. Decaffeinated tea produced the same effect as observed for 'regular' teas. However, lower concentrations of some tea extracts enhanced mutagenic activity of 2-amino-3,4,7,8-tetramethyl-3H-imidazo[4,5-f]quinoxaline (4,7,8-TriMeIQx) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). Herbal tea extracts displayed variable effects on the mutagenicity of different HAA. While some extracts had no effect, others exhibited a moderate inhibitory effect on the mutagenicity of IQ-type HAA. In contrast to common tea, herbal teas showed substantial potentiating effects on the mutagenicity of several HAA, especially Trp-P-2 and 4,7,8-TriMeIQx.

Amines↗
PubMed1995

Evaluation of hamburgers and hot dogs for the presence of mutagens.

Food products derived from heat-treated (fried, broiled, baked) meat may contain traces of mutagenic heterocyclic amine contaminants, some of which are proven carcinogens in rodents. To confirm their presence and range in Canadian foods, and estimate the average human intake of these types of mutagens from frequently consumed heat-processed foods, several commercially prepared fried-beef patties (hamburgers) and hot dogs (weiners) were analysed for their mutagenic capacity. The mutagenicity of the extracts was tested in the Salmonella/microsome assay using strain TA98 with metabolic activation. 16 samples of hamburgers and 14 samples of hot dogs, randomly obtained from 'fast food' commercial establishments or street vendors, were used in this survey. The mutagenic activity of these samples ranged from very low to 1042 revertants/g equivalent for the hamburgers and from non-detectable to 4875 revertants/g equivalent for the hot dogs. The average values were 199 and 424 revertants/g for the hamburgers and hot dogs, respectively. The wide range in mutagenicity was found even for the same type of product obtained from the same outlet at different times. This indicates possible inconsistency in cooking procedures during the preparation of these products. It also shows the difficulty in accurately assessing the intake of mutagenic heterocyclic amines from hamburgers and hot dogs prepared in 'fast food' outlets.

Amines↗
PubMed1993

Cytotoxic and genotoxic properties of tert.-butyl-p-quinone (TBQ) in an in vitro assay system with Chinese hamster V79 cells and in strain D7 of Saccharomyces cerevisiae.

tert.-Butyl-p-quinone (TBQ), a major metabolite of the phenolic antioxidant tert.-butyl-4-hydroxyanisole (BHA), was examined for cytotoxic and genotoxic properties in an in vitro assay system with Chinese hamster V79 cells and in diploid strain D7 of Saccharomyces cerevisiae. TBQ was prepared from BHA by oxidative demethylation with sodium nitrite at low pH. Spectroscopic analyses identified the crystalline reaction product as TBQ with a purity close to 100%. Cytotoxicity of TBQ was determined by cloning efficiency in the absence of hepatocyte activation. TBQ reduced colony size of V79 cells at 0.4 micrograms/ml, prevented growth of 50% of the cells at 0.6 micrograms/ml, and was lethal to 100% of the cells at concentrations above 1.0 micrograms/ml. TBQ was 6-7 times more cytotoxic to V79 cells than TBHQ, a related BHA metabolite, and 100 times more cytotoxic than BHA. At dose levels of 0.2, 0.4 and 0.6 micrograms/ml of medium, TBQ did not increase significantly the frequency of sister-chromatid exchanges (SCE) in V79 cells and did not consistently increase the frequency of mutation to thioguanine resistance (TGR) at the hgprt gene locus either alone or with activation by rat hepatocytes. Incubation with TBQ for 4 h at pH 3.6 without activation resulted in only small increases in the frequency of gene conversion and reverse mutation in strain D7 of Saccharomyces cerevisiae. However, exposure to TBQ alone in growth medium for 24 h, produced inconsistent results. From these studies it was concluded that TBQ was cytotoxic but not genotoxic to V79 cells; and may be weakly genotoxic to strain D7 of S. cerevisiae.

Analysis of Variance↗
PubMed1993

Analysis of commercial bouillons for trace levels of mutagens.

A new method, developed specifically for the extraction of heterocyclic aromatic amine (HAA) type mutagens from different food matrices, was applied to various forms of commercially available bouillons. This procedure is based on liquid-liquid extraction of the sample at different pH values. Recovery and reproducibility of the procedure was determined by processing spiked samples using a mutagenicity bioassay technique as an endpoint. The mutagenicity was tested in the Salmonella/microsome assay using strain TA98 with metabolic activation. 22 bouillon samples in liquid, cube or powder forms from seven manufacturers were extracted and tested for potential mutagenicity. The mutagenic activity of these samples varied and ranged from non-detectable to about 1200 induced revertants per gram of solid material, with a median value of approximately 250 revertants/g. The mutagenic response appeared to be dependent on the source rather than the type or form of the product tested. A negative response was obtained from only one chicken bouillon, and the highest positive response was obtained from a beef bouillon in cube form. It appears that the average beef sample, regardless of form, has a higher mutagenic potency than chicken or chicken and turkey samples. Overall, the intake of mutagens from commercial bouillons (obtained as cubes, concentrates or dry mixes) to prepare one serving (as bouillon, soup, casseroles, etc.) is considerably less than that reported in the literature for one serving of fried beef or pork. The extractability and mutagenic characteristics of these samples indicate the presence of HAA-type mutagens. Work is in progress to identify the mutagenic factors in bouillons.

Amines↗
PubMed1993

DNA binding and mutagenicity of aflatoxin B1 catalyzed by isolated rabbit lung cells.

The abilities of different rabbit lung cell types to bioactivate aflatoxin B1 (AFB1) to a DNA-binding and mutagenic metabolite have been examined. Microsomes were prepared from centrifugal elutriation-enriched preparations of isolated rabbit lung cell types. The activation of [3H]AFB1 (5.0 or 200 microM), measured indirectly as covalent binding to calf thymus DNA, was concentrated in microsomes from the non-ciliated bronchiolar epithelial (Clara) cell-rich fractions (13-22 times the activity of whole lung microsomes). Microsomes from type II cell-rich fractions had minimal activity. Significant correlations were detected between the rates of microsomal DNA binding and the percentages of Clara cells in the fractions. Prior treatment of rabbits with the cytochrome P450 class 1A inducer beta-naphthoflavone had no significant effect on the microsomal activation of AFB1. In other experiments, intact, enriched isolated rabbit lung cells were incubated with AFB1 (0-1.5 microM) in a modification of the Ames mutagenicity assay, using Salmonella typhimurium strain TA100. The ability to activate AFB1 to mutagenic metabolite(s) in this system was localized in Clara cell-rich fractions, with no significant activity being detected in other fractions. The results of these studies indicate that the biotransformation of AFB1 to DNA-binding and mutagenic metabolite(s) in rabbit lung is heterogeneous, and that the Clara cell is specifically implicated in this ability.

Aflatoxin B1↗
PubMed1992

Evaluation of genotoxicity of tert.-butylhydroquinone in an hepatocyte-mediated assay with V79 Chinese hamster lung cells and in strain D7 of Saccharomyces cerevisiae.

tert.-Butylhydroquinone (TBHQ) has been reported to be genotoxic in some short-term assays but non-genotoxic in others. We have examined cytotoxicity and genotoxicity of TBHQ, a principal metabolite of the phenolic antioxidant 2(3)-tert.-butyl-4-hydroxyanisole (BHA), in an hepatocyte-mediated assay with V79 Chinese hamster lung cells including both sister-chromatid exchange (SCE) and thioguanine-resistance (TGR) endpoints. The ability of BHA and of TBHQ to elicit a genotoxic response in Saccharomyces cerevisiae strain D7 was also investigated. In V79 cytotoxicity tests, TBHQ without hepatocytes produced a 50% reduction in colony formation at 4.2 micrograms/ml and was lethal to 100% of the cells at concentrations above 5 micrograms/ml. At partially cytotoxic dose levels, (0.17-3.4 micrograms/ml of medium), TBHQ sometimes increased significantly the frequency of SCE. TBHQ also produced sporadic statistically significant increases in the mutation frequency at the HGPRTase (TGR) gene locus when tested alone or with activation by rat or hamster hepatocytes. Mitotic gene conversion and reverse mutation were not induced in strain D7 of Saccharomyces cerevisiae by exposure to BHA or to TBHQ for 4 h at concentrations as high as 200 micrograms/ml for BHA or 500 micrograms/ml for TBHQ, either alone or with activation by rat-liver S9. Incubation of the yeast cells with BHA or TBHQ for 24 h in growth medium without activation also did not induce genotoxic activity. The slight and sporadic response to TBHQ in the V79 test system may indicate weak genotoxicity which is sensitive to slight differences in test conditions. The classification and test strategies adopted for compounds such as TBHQ could have important implications for regulatory decisions and for the validation of short-term tests.

Animals↗
PubMed1990

Evaluation of genotoxicity of N-nitrosodibenzylamine in Chinese hamster V79 cells and in Salmonella.

Health concerns have arisen due to the formation of N-nitrosodibenzylamine (NDBzA; CAS No. 5336-53-8) in pork processed in a new type of rubber netting. In view of the potent carcinogenicity of related nitrosamines (e.g. N-nitroso-n-dibutylamine and N-nitrosodiethylamine), NDBzA was evaluated for genotoxicity in vitro in both Chinese hamster V79 cells and in Salmonella. In V79 cells, concentrations up to 25 micrograms/ml were tested with and without activation by rat or hamster hepatocytes. Significant elevation of SCE frequency was seen only at 25 micrograms/ml in the presence of uninduced hamster hepatocytes. Mutation to 6-thioguanine resistance was observed at 25 micrograms/ml, in the absence of hepatocytes and in the presence of induced (Aroclor 1254) or uninduced hamster hepatocytes, but not with rat hepatocytes. With uninduced rat hepatocytes, a small but significant (p less than 0.05) increase in the mutation frequency was seen with 10 micrograms/ml NDBzA. In the Salmonella assay, using a pre-incubation protocol and concentrations up to 1000 micrograms/ml, NDBzA was negative in strain TA98, and in TA100 with rat S9, but was positive at the highest dose in TA100 with hamster S9, and more strongly with Aroclor 1254-induced hamster S9. When activated by uninduced rat or hamster hepatocytes, as opposed to S9, NDBzA was negative with all tester strains. Hamster hepatocytes activated more than rat in the V79 studies, and hamster S9 was more strongly activating in the Salmonella assay. These results indicate that NDBzA is weakly mutagenic to both Salmonella and V79 cells.

Animals↗
PubMed1990

Validity of in vitro testing.

In summary, the extent of toxicity studies which can be carried out with bound residues of nitroimidazole drugs will be dictated by the availability and the amount of residues that can be produced. For evaluating their toxicity the use of the Salmonella assay, which has been proven to be sensitive for the parent drugs, is proposed. Furthermore, it is suggested that the toxicity of bound residues for in vivo effects should be investigated in cells of target tissues, such as the epithelial cells of the gastrointestinal tract. Nuclear aberration and sister chromatid exchange assays in this tissue would be good candidates for evaluation. It should be pointed out also that an examination of the literature for genotoxic effects of nitroimidazole drugs reveals an apparent contradiction, especially in the context of genotoxicity testing strategy discussed above. It appears first that these drugs are potent mutagens in vitro in microbial systems (Salmonella and yeast). However, they are not active in mammalian cells in vitro, in both mutation and chromosomal aberration assays, as well as in vivo, in assays such as the dominant lethal test and the micronucleus assay. Thus, it may be of interest to speculate that, although these drugs are in vitro mutagens, they may not be in vivo mutagens in mammals. Their tumorigenic effects, which have been detected at high doses in rodents, may, therefore, be due to other than genotoxic activity. To resolve this conflict, reassessment of genotoxicity of these drugs in vivo would be a worthwhile pursuit. In addition, such seeming discrepancies would also argue strongly for toxicity screening to be conducted in a battery of complementary short-term in vitro and in vivo tests. This would help insure the likelihood of detection of at least some meaningful biological activity, which would serve to flag chemicals warranting further testing.

Animals↗
PubMed1989

The classification of chemical carcinogens.

Various approaches to classification of chemical carcinogens developed by international and national agencies are described. Special features of each classification scheme with their strengths and weaknesses and their usefulness in regulation are discussed. Some recent examples of regulatory decisions on chemical hazards in Canada are presented, illustrating the many factors that need to be considered for risk assessment, in addition to merely categorizing carcinogens.

Animals↗
PubMed1986

Evaluation of the cytotoxicity and genotoxicity of the spermicides nonoxynol-9 and octoxynol-9.

The cytotoxic and genotoxic potentials of the spermicidal agents, nonoxynol-9 (N-9) and octoxynol-9 (0-9), were evaluated in in vitro test systems using rat liver cells. N-9 was also tested for the induction of sperm abnormalities in mice. Dose-related cytocidal effects were seen after the addition of N-9 and O-9 to the culture medium for 24 h. The mean concentrations of N-9 and O-9 necessary to decrease the number of viable cells by 50% (LC50) were 24 and 43 micrograms/ml of media, respectively. The spermicides neither induced DNA repair in freshly isolated hepatocytes, nor caused any mutations at HGPRT locus in the T51B rat liver cell line. There was also a lack of malignant transformation response in the low-calcium assay. Further, the germinal cells of mice remained unaffected by N-9.

Animals↗
PubMed1980

Mutagenicity of constituents identified in pulp and paper mill effluents using the Salmonella/mammalian-microsome assay.

About 300 compounds have been reported in the literature as constituents of pulp-mill effluent. Previously, in our screening program, 10 resin acids identified in effluent were examined for potential mutagenicity in the Salmonella/mammalian-microsome assay. Neoabietic acid was the only resin acid which was found to be mutagenic. Now, a program to screen for mutagenicity of 48 additional compounds, belonging to chemical classes of chlorinated aliphatic and aromatic hydrocarbons, phenols, aldehydes, quinones, and carboxylic acids, has been completed. Only 2 of these compounds, tetrachloropropene and pentachloropropene, were found to be mutagenic, showing dose-related increases in His+ reversion mutations, in the standard Salmonella test. Metabolic activation with a preparation of Aroclor 1254-induced liver homogenate (S9) greatly reduced the mutagenic responses of these 2 compounds. Modifications of the Salmonella test for volatile mutagens enabled the detection of the mutagenicity of 3 additional chlorinated aliphatic hydrocarbons dichloromethane, dichloroethane and trichloroethane.

Animals↗
PubMed1980

Short-lived mutagen in Salmonella produced by reaction of trichloroacetic acid and dimethyl sulphoxide.

A chemical reaction occurs when trichloroacetic acid (TCA) is dissolved in dimethyl sulphoxide (DMSO), with the production of a short-lived mutagenic derivative which was detected using the Salmonella/mammalian-microsome plate overlay assay. Other interactions between test chemicals and solvents are discussed. Choice of proper solvents in mutagenicity testing is emphasized.

Dimethyl Sulfoxide↗