Soil mutagens are airborne mutagens: variation of mutagenic activities induced in Salmonella typhimurium TA 98 and TA 100 by organic extracts of agricultural and forest soils in dependence on location and season.

As our hypothesis was that soil mutagens are airborne mutagens, possibly modified by soil microorganisms, we checked solvent extracts from agricultural and forest soils collected during late summer in the environment of Mainz, a region highly charged by anthropogenic air pollution, or near Bayreuth, a rural low charged region of Germany, or in a remote region of western Corsica without anthropogenic air pollution for the presence of mutagenicity in Salmonella typhimurium. Levels of mutagenic activities were quantified by calculation of revertants/g from the initial slope of dose-response curves applying tester strains S. typhimurium TA 98 and TA 100 in the absence and presence of an activat…

Protective Effects of Fruits and Vegetables against In Vivo Clastogenicity of Cyclophosphamide or Benzo[a]pyrene in Mice

Seven fruits and 10 vegetables commonly consumed in Germany were investigated for their anticlastogenic potencies against cyclophosphamide (CP) and benzo[a]pyrene (BaP) in the in vivo mouse bone marrow micronucleus assay. We detected protective effects in 76.5% and 70.6% of the samples, respectively, and more or less distinct quantitative differences between the various plant materials and the two clastogens investigated. With respect to CP, moderate activities were exerted by sweet cherries, strawberries, cucumber, radish and tomatoes, average activities by bananas, oranges, peaches, asparagus and red beets and strong activities by yellow red peppers and especially spinach. Apples (cultiva…

Isolation and Characterization of Structurally Novel Antimutagenic Flavonoids from Spinach (Spinacia oleracea)

Thirteen compounds, isolated from spinach (Spinacia oleracea), acted as antimutagens against the dietary carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline in Salmonella typhimurium TA 98. The antimutagens were purified by preparative and micropreparative HPLC from a methanol/water (70:30, v/v) extract of dry spinach (commercial product) after removal of lipophilic compounds such as chlorophylls and carotenoids by solid-phase extraction (SPE). Pure active compounds were identified by instrumental analysis including FT-IR, (1)H and (13)C NMR, UV-vis spectroscopy, and mass spectrometry. All of these compounds were flavonoids and related compounds that could be attributed to five groups: (A, m…

Fruits and vegetables protect against the genotoxicity of heterocyclic aromatic amines activated by human xenobiotic-metabolizing enzymes expressed in immortal mammalian cells

Heterocyclic aromatic amines (HAAs) can be formed during the cooking of meat and fish at elevated temperatures and are associated with an increased risk for cancer. On the other hand, epidemiological findings suggest that foods rich in fruits and vegetables can protect against cancer. In the present study three teas, two wines, and the juices of 15 fruits and 11 vegetables were investigated for their protective effect against the genotoxic effects of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). To closely mimic the enzymatic activation of these HAAs in humans, genetically engineered V79 Chinese hamster fibroblasts were employed tha…

Structure-Activity Relations of Flavonoids as Antimutagens Against Cooked Food Mutagens and Nitroarenes in the Salmonella Reversion Assay

When about 60 flavonoids were tested for their antimutagenic potencies against 2-amino-3-methylimidazo [4,5-f]quinoline (IQ) and the nitroarenes 2-nitrofluorene (2-NF), 1-nitropyrene (1-NP), and 3-nitrofluoranthene (3-NFA) in the Salmonella reversion assay, distinct structure-activity relations were detected. With both groups of compounds, flavonoid glycosides were inactive or at best weakly active. Catechins and anthocyanidins, which lack the carbonyl function at C-4 of the flavane nucleus, were inactive. In contrast, chalcones exerted strong antimutagenicity against IQ and the nitroarenes. With respect to IQ, flavonoids of low polarity were the most active compounds: The parent compounds …

Antimutagenic effects and possible mechanisms of action of vitamins and related compounds against genotoxic heterocyclic amines from cooked food.

Possible antimutagenic activity of 26 vitamins and related compounds - ascorbic acid, beta-carotene, cyanocobalamin, folic acid, nicotinic acid, nicotinamide, pantothenic acid, pyridoxale, pyridoxamine, pyridoxine, retinal, retinol, retinoic acid, retinyl acetate, retinyl palmitate, riboflavin, riboflavin 5'-phosphate, flavin adenine dinucleotide (FAD), alpha-tocopherol, alpha-tocopherol acetate, vitamins K(1), K(3), K(4), 1, 4-naphthoquinone, and coenzyme Q(10) - was tested against six heterocyclic amine (HCA) mutagens, i.e., 2-amino-3-methyl-imidazo[4, 5-f]quinoline (IQ), 2-amino-3,4-dimethyl-imidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethyl-imidazo[4,5-f]quinoxaline (MeIQx), 2-amino-1…

Effects of β-Carotene, Retinal, Riboflavin, α-Tocopherol and Vitamins C and K1 on Sister-chromatid Exchanges Induced by 3-Amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2) and Cyclophosphamide in Human Lymphocyte Cultures

The vitamins and related compounds cited in the title were investigated for their abilities to modulate sister-chromatid exchanges (SCEs) induced by Trp-P-2 or cyclophosphamide (CP) in human peripheral lymphocyte cultures in the presence of an exogenous metabolizing system from rat liver. When inducer and test substances were given simultaneously, beta-carotene, retinal and alpha-tocopherol caused a dose-dependent decrease of SCE frequencies induced by Trp-P-2 and CP. Vitamin K1, however, brought about an identical effect with Trp-P-2 only, while with CP an initial decrease of SCEs was followed by a statistically significant re-increase at higher concentrations. Vitamin C was ineffective ag…

Inhibition of clastogenicity of benzo[a]pyrene and of its trans-7,8-dihydrodiol in mice in vivo by fruits, vegetables, and flavonoids.

In the in vivo mouse bone marrow micronucleus assay, homogenates of spinach, artichoke, peaches, and blue grapes as well as commercial concentrates of these vegetables and fruits reduced induction of micronuclei by benzo[a]pyrene (BaP) by 43-50%. Concentrates of strawberries (31% reduction) and of cauliflower (20% reduction) were less potent. Inhibition of genotoxicity by spinach and peaches was not caused by any delay in maturation of micronucleated erythrocytes as shown by experiments with sampling times of 24, 48, and 72 h after dosing of BaP. Pre-treatment of the mice with spinach 48, 24, and 12h before application of BaP resulted in a 44% reduction of micronuclei while peaches generate…

Inhibition of the mutagenicity of 2-nitrofluorene, 3-nitrofluoranthene and 1-nitropyrene by vitamins, porphyrins and related compounds, and vegetable and fruit juices and solvent extracts

When 21 vitamins including related compounds haemin, chlorophyllin, chlorophyll, biliverdin and bilirubin, as well as juices from five fruits and 25 vegetables and solvent extracts from the residues of fruits and vegetables were tested for their antimutagenic potencies with respect to mutagenicity induced by 2-nitrofluorene (2-NF), 3-nitrofluoranthene (3-NFA) and 1-nitropyrene(1-NP) in Salmonella typhimurium TA98 the following results were obtained. The tetracyclic nitroarenes 3-NFA and 1-NP were in general more effectively antagonized by potent antimutagenic compounds than the tricyclic 2-NF. beta-Carotene, retinol, retinal, retinoic acid, retinol palmitate, riboflavin 5'-phosphate, alpha-…

Development and application of test methods for the detection of dietary constituents which protect against heterocyclic aromatic amines

This article describes the development and use of assay models in vitro (genotoxicity assay with genetically engineered cells and human hepatoma (HepG2) cells) and in vivo (genotoxicity and short-term carcinogenicity assays with rodents) for the identification of dietary constituents which protect against the genotoxic and carcinogenic effects of heterocyclic aromatic amines (HAs). The use of genetically engineered cells expressing enzymes responsible for the bioactivation of HAs enables the detection of dietary factors that inhibit the metabolic activation of HAs. Human derived hepatoma (HepG2) cells are sensitive towards HAs and express several enzymes [glutathione S-transferase (GST), N-…

Ernährung und Colonkrebsätiologie: Von der deskriptiven Epidemiologie zur diätetischen Prävention

Der Colonkrebs ist eine der haufigsten Krebserkrankungen in der Bundesrepublik Deutschland und in den meisten westlichen Landern, in Afrika, Asien und Lateinamerika jedoch gewohnlich selten. Aufgrund epidemiologischer Untersuchungen sind verschiedenartige Ernahrungsgewohnheiten die Hauptursache fur diese Unterschiede. Ein hoher Fett- und Eiweiskonsum wurde als Risikofaktor erkannt, wahrend ein hoher Ballaststoffgehalt der Nahrung sich als protektiv erwies. Die existierenden Hypothesen zur Atiologie des Colonkrebses, die vom „Zwei-Stufen-Initiations-Promotionsmodell“ ausgehen, betrachten das Wechselspiel der Einwirkung von Initiatoren, Promotoren und Inhibitoren als entscheidend fur die Real…

Gnotobiotische untersuchungen an SPF-Mäusen im rahmen einer studie zur tumorentwicklung im kolon nach cholezystektomie

Summary The relation between cholecystectomy and colon carcinogenesis has not been fully elucidated. As bacteria may be involved in the carcinogenic process, we investigated the effect of cholecystectomy and dimethylhydrazine (DMH) administration to SPF NMRI mice with regard to tumour genesis and bacterial colonisation of the intestine. It results from this study that cholecystectomy does not influence tumour genesis and that 6–7 months post operationem and DMH administration tumours and bacteria originally not found in the animals develop: Clostridia, eubacteria spec, which cannot be differentiated and E. lentum. Theses changes appear in group II of mice (laparotomy and DMH) and group III …

Free radical scavenging abilities of flavonoids as mechanism of protection against mutagenicity induced by tert-butyl hydroperoxide or cumene hydroperoxide in Salmonella typhimurium TA102

Mutagenicity induced by tert-butyl hydroperoxide (BHP) or cumene hydroperoxide (CHP) in Salmonella typhimurium TA102 was effectively reduced by flavonols with 3',4'-hydroxyl groups such as fisetin, quercetin, rutin, isoquercitrin, hyperoxide, myricetin, myricitrin, robinetin, and to a lesser extent also by morin and kaempferol (ID50=0.25-1.05 micromol per plate). With the exception of isorhamnetin, rhamnetin, morin, and kaempferol, closely similar results were obtained with both peroxides. Hydrogenation of the double bond between carbons 2 and 3 (dihydroquercetin, dihydrorobinetin) as well as the additional elimination of the carbonyl function at carbon 4 (catechins) resulted in a loss of a…

Epimerization, oxidation and reduction of bile acids by Eubacterium lentum

Summary Twenty-eight strains of Eubacterium lentum were divided into five groups on the basis of their ability to metabolize cholic acid and dehydrocholic acid: (I) Strains with 3α- and 3/β-dehydrogenating activities (1 strain), (II) strains with 12α-dehydrogenating activity (5 strains), (III) strains with 3α-, 3β-, 7α-, and 12α-dehydrogenating activities (8 isolates), (IV) transition stages between group II and III (1 strain), (V) strains devoid of any measurable bile acid metabolizing activities or with only trace activities (11 isolates). Using appropriate bile acids it was demonstrated that hydroxyl or keto groups at positions C 3 , C 7 , and C 12 did not influence bile acid transformat…

Inhibition of the mutagenicity of 2-nitrofluorene, 3-nitrofluoranthene and 1-nitropyrene by flavonoids, coumarins, quinones and other phenolic compounds

When 56 flavonoids, 32 coumarins, five naphthoquinones, 12 anthraquinones and five structurally-related compounds were tested for their antimutagenic potencies with respect to mutagenicities induced by 2-nitrofluorene (2-NF), 3-nitrofluoranthene (3-NFA) and 1-nitropyrene (1-NP) in Salmonella typhimurium TA98 distinct structure-activity relationships were detected. First, the tetracyclic nitroarenes 3-NFA and 1-NP were in general more effectively antagonized by potent antimutagenic flavonoids and coumarins than the tricyclic 2-NF, while there were only minor differences with quinones. Secondly, antimutagenicity of natural compounds of plant origin correlated with the aglyconic nature 10 of a…

The inhibition by naphthoquinones and anthraquinones of 2-amino-3-methylimidazo[4,5- f  ]quinoline metabolic activation to a mutagen: a structure-activity relationship study

Nine naphthoquinones, 19 anthraquinones, and nine structurally related monoketonic compounds such as anthrone, xanthone, etc., inhibited mutagenicity induced by 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in Salmonella typhimurium TA 98 in the presence of rat liver S9 with distinct structure-activity relationships. A carbonyl function was a prerequisite for antimutagenicity while, in general, anthraquinones (IC50 values: 2.3–>213 nmol/ml top agar) were more potent antimutagens than structurally related monoketonic compounds (IC50 values: 25.3–94.9 nmol/ml top agar) and naphthoquinones (IC50 values: 3.7–90.7 nmol/ml top agar). The parent compounds and methyl substituted derivatives were alr…

In vitro antimutagenic and in vivo anticlastogenic effects of carotenoids and solvent extracts from fruits and vegetables rich in carotenoids.

The water insoluble residues of some carotenoid-rich fruits and vegetables, such as apricots, oranges, brussels sprouts, carrots, yellow-red peppers, and tomatoes, were sequentially extracted with n-hexane, dichloromethane, acetone, and 2-propanol, and solvent extracted materials were tested for inhibition of mutagenicities induced by aflatoxin B1 (AFB1), benzo[a]pyrene (BaP), 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), and cyclophosphamide (CP) in histidine-deficient strains of Salmonella typhimurium. Antimutagenic activities were found in many extracts, but especially in the n-hexane extracts. For example, in the case of oranges, 100 microg of this extract reduced the bacterial mutageni…

Genotoxicity of selected pesticides in the mouse bone-marrow micronucleus test and in the sister-chromatid exchange test with human lymphocytes in vitro

Selected pesticides (aldicarb, 1,3-dichloropropene, methidathion, parathion, triadimefon, vinclozolin) were tested for their clastogenic and aneugenic activities in the mouse bone-marrow micronucleus (MN) test in vivo and for their sister-chromatid exchange-inducing activities in human lymphocytes in vitro in the presence and absence of an exogenous metabolizing system from rat-liver S9. 1,3-Dichloropropene significantly increased the frequencies of micronucleated polychromatic erythrocytes (PCE) in bone-marrow cells of female mice from 3.3 MN/1000 PCE to 15.3 MN/1000 PCE (187 mg per kg body weight). 1,3-Dichloropropene (100 microM) induced 16.0 SCE/metaphase after 24 h of incubation as com…

Antimutagenic effects of flavonoids, chalcones and structurally related compounds on the activity of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and other heterocyclic amine mutagens from cooked food.

Abstract Sixty-four flavonoids were tested for their antimutagenic potencies with respect to IQ in Salmonella typhimurium TA98 and in part also towards MeIQ, MeIQx, Trp-P-2, and Glu-P-1 and in S. typhimurium TA100. Antimutagenic potencies were quantified by the inhibitory dose for 50% reduction of mutagenic activity (ID 50 ). A carbonyl function at C-4 of the flavane nucleus seems to be essential for antimutagenicity: two flavanols and four anthocyanidines were inactive. Again, five isoflavons, except biochanin A, were inactive. Within the other groups of 21 flavones, 16 flavonols and 16 flavanones the parent compounds flavone, flavonol, and flavonone possessed the highest antimutagenic pot…

In vivo genotoxicity of selected herbicides in the mouse bone-marrow micronucleus test

The herbicides alachlor, atrazine, terbuthylazine, gluphosinate-ammonium, isoproturon, pendimethaline and trifluralin were tested for genotoxicity in the mouse bone-marrow micronucleus test (MNT). Both atrazine and trifluraline caused a significant increase in the number of micronuclei at doses of 1,400 mg/kg body weight in female mice only. Alachlor, terbuthylazine, gluphosinate-ammonium, isoproturon and pendimethaline did not have any genotoxic effect in the mouse bone-marrow micronucleus test in either female or male animals.

Stability of Antimutagenic Activities in Fruit and Vegetables during High Pressure Processing

The influence of high pressure processing on antimutagenic activities of fruit and vegetable juices

Abstract In model experiments using the Salmonella/reversion assay (Ames test) and the carcinogen 2-amino-3-methyl-imidazo[4,5-f]quinoline (IQ), antimutagenic activities against IQ had been detected in about 70% of 62 fruit and vegetable species [Edenharder, Kurz, John, Burgard and Seeger (1994) Fd Chem. Toxic. 32, 443–459]. However, thermal stability of the antimutagenic factors varied considerably. As hydrostatic pressure is an alternative to heat in food processing and preservation, homogenates of 14 fruit and vegetable species were exposed to different pressures and temperatures. According to the response of the antimutagenic potential, three groups of produce could be identified: (a) m…

Deoxycholic Acid Methyl Ester — a Novel Bacterial Metabolite of Cholic Acid

Summary Methyl deoxycholate was identified as a novel bacterial metabolite of cholate, produced anaerobically by strains of most saccharolytic Bacteroides species ( B. distasonis, B. eggerthii, B. fragilis, B. incommunis, B. thetaiotaomicron, B. uniformis, B. variabilis, B. vulgatus , and unnamed species). It was also detected with a few strains of Eubacterium and Lactobacillus species. Among 2476 freshly isolated human fecal cultures the frequency of carboxyl group esterification was comparable with that of 7α-dehydroxylation (92 versus 102 cultures). Both activities were, however, lost for unknown reasons after serial transfers. Two of ten mixed fecal cultures tested esterified 3α,7α- or …

The inhibition by flavonoids of 2-amino-3-methylimidazo[4,5-f]quinoline metabolic activation to a mutagen: a structure-activity relationship study.

The mutagenicity of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) in Salmonella typhimurium TA98 is inhibited by flavonoids with distinct structure-antimutagenicity relationships (Edenharder, R., I. von Petersdorff I. and R. Rauscher (1993). Antimutagenic effects of flavonoids, chalcones and structurally related compounds on the activity of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and other heterocyclic amine mutagens from cooked food, Mutation Res., 287, 261-274). With respect to the mechanism(s) of antimutagenicity, the following results were obtained here. (1) 7-Methoxy- and 7-ethoxyresorufin-O-dealkylase activities in rat liver microsomes, linked to cytochrome P-450-dependent 1A1 and…

Changes in functional properties of vegetables induced by high pressure treatment

Abstract Ultrahigh-pressure (UHP) hydrostatic exposure in excess of 100 MPa is being used to inactivate micro-organisms as well as to induce texture changes to foods without thermal degradation. The UHP process is said not only to preserve flavour and food product taste but also natural properties of biological products. The influence of high pressure on functional properties of a choice of vegetables was evaluated. The treatments were at pressures around 600 MPa and also in combination with elevated temperatures and in comparison with thermal treatment. Carrots, tomatoes and broccoli were investigated (crushed or liquid extracts). The following characteristics were assessed: content of hea…

Gas chromatographic and mass spectrometric analysis of bile acids as trifluoroacetyl-hexafluoroisopropyl and heptafluorobutyryl derivatives

Abstract The gas chromatographic retention times on QF-1 of 38 bile acids in the form of their trifluoroacetyl-hexafluoroisopropyl (TFA-HFIP), trifluoroacetyl-methyl, and heptafluorobutyryl derivatives are given. In general hexafluoroisopropyl ester trifluoroacetates proved superior with regard to simplicity of preparation, absence of artifacts, and resolution on QF-1. The main disadvantages of heptafluorobutyrates were the production of artifacts with some ketonic bile acids and the impossibility of separating any of the dihydroxy bile acids with substituents in the 3,6- and 3,7-positions. Mass spectra of TFA-HFIP derivatives were recorded with both direct and gas chromatographic inlet sys…

The influence of automobile exhausts on mutagenicity of soils: contamination with, fractionation, separation, and preliminary identification of mutagens in the Salmonella/reversion assay and effects of solvent fractions on the sister-chromatid exchanges in human lymphocyte cultures and in the in vivo mouse bone marrow micronucleus assay.

To test the assumption that automobile exhausts contribute to soil mutagenicity, two soils with low levels of mutagenic activities were exposed to traffic exhausts at a heavily charged junction of German motorways (Autobahnen) for 3, 7, 10, 13, 17, 21, and 26 weeks. Indeed, in the presence of a metabolic activation system from rat liver (S9), an average increase of 8 and 9 (4 and 12) revertants per gram per week was found in Salmonella typhimurium TA 98 (TA 100). In the absence of S9, meaningful measurements were impossible on account of a concurrent dose dependent increase of toxicity. No correlation between the increase of mutagenicity and the contents of polycyclic aromatic hydrocarbons …