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P XXI.6 Screening for genotoxicity with SOS chromotest
S206
S-XXI: SOS chromotest
Ir XXI.61
Screenln2 for 2enotoxlclty with 50S chromatest
R. Vachkova-PetrdVa, E. Tyagunenko. National Center of Hygiene, Medical Ecology and Nutrition, Sofia, Bulgaria
In the present study SOS chromotest was performed by the use of the Bioscreen Automated Analyser System, Labsystems, Finland. The method adapted to Bioscreen is based on the colorimetric assay, described by P. Quillardet and M. Hofnung, 1985, but makes use of kinetic assay pnnciple instead of a fixed time assay to quantify the enzymatic activities measured in the test. Usually the conditions of protocol B were used: +/- S9; 10 dikns of sample, max 3 samples; chromogen - yellow 420 nm; incubation time 2 h; positive controls - 4-nilroquinoline-oxide, highest dose 2.63 nmoVassay and 2-arninoanlltracene, 2.59 nmoVassay. Resultsof the screening for genotoxicityof differentcompounds are presented. They include environmental chemicals, food additives, drugs: chloracetophane, alachlor, tetrapol A 159. propylenethiourea, acesulfame K, sodium nitrite. nalidixic acid, ciprofloxacin, cadmium and lead, complex mixtures (biotechnologyproducts for food industry). The correlation between results by SOS chromotest ans Ames test are discussed also.
Ir XXI.71
The SOS chromotest: An analysis from published data on 950 chemicals
P. Quillardet, E. Touati, M. Hofnung. Unite de Programmation Moleculaire et Toxlcologi« Genetique, CNRS URA1444, Bdtiment des Biotechnologies. Institut Pasteur; 25 rue du Dr Raux. 75724 Porn Cedex, Prance In 1982. we have made use of an E. coli strain carrying a fusion of gene lacZ to gene s./iA, one of the SOS gene, to devise a rapid assay for genotoxins: the SOS ChlOmotest. The assay i. performed in few hout! and involves simple enzymatic assays. It allows to classify compounds according to their SOS inducing potency (SOSIP), defined as their ability to induce the expression of the s./iA::lacZ fusion. To day. work from a number of laboratories using the SOS Chromotest have been published. We have reviewed data obtained with the SOS Chromotest on more than 950 chemicals issued from 130 publications arising from about 40 different laboratories. This led us to evaluate further the potential of the SOS Chromotest to detect carcinogens and to compare its response to that of the Salmanellafmicrosome assay. Although far from to be perfect, the results confirm that the SOS Chromotest is a satisfaying method to detect and evaluate genotoxic agents. However, we think that the utility of the SOS chromotcst in a battery of tests of genetic toxicology is mainly due to its simplicity. It constitute an unrivalledtool in the rapid screening of genotoxiccompounds to a large scale and in the monitoring of genotoxie substances in the environment itself. KeywOrd(s): SOS Chromotest; Genotoxicity; Evaluation
Ir XXI.81
Inhibition of SOS response In E. coli PQ37 by heated turmeric and curcumln
Kalpagam Polasa, A. Nadamuni Naidu, R. Krishnaswamy. Food & Drug Toxicology Research Centre, National Institute of Nutrition. Indian Council ofMedical Research. Hyderabad - 500 007, A.P., India Turmeric and its active principle curcumin, commonly used oriental spice has been proved to possess antimutagenic potential. These have been tested in their raw fornu . It is nOC known if antimutagenicityexistsafter exposureto cooking conditions. So heat treatment of these two substances were carried out to simulate cooking conditions namely boiling and frying and tested for antimutagenic property using SOS chromotest. Various concentrations of boiled/fried turmeric and cureumin were tested against 3. $ and 10 ng of 4NQO for inhibition of SOS response. Pooled analysis of the data indicated that both turmeric and curcumin are capable of inhibiting SOS response in E. coli PQ37 even after heat treatment suggesting that cooking processes are unlikely to destroy its antimutagenic property. Keyword(s): SOS test; turmeric; curcumin
Ir XXI.91
Tbe SOS Inducln2 potency offour textile dyes
R. C. Sobti • Meenu Kaushal2 , P.Quillarder', I DepartmentofBiotechnology, Punjab University. Chandigarh - /60014. India: 2Deparrment of Zoology. Panjab Unive~ity. Chandigarh - /60014. India: J Pasteur tnsutute; Paris. France l
SOS responses induced by four textile dyes viz.• Sulphur Red Brown 360 (SRB), lade Green 2G (JG), Reactofix Turquoise Blue 5 GFL (RTB) and Direct Scarlet 4BS (OS) were measured in Escherichia coli PQ37 tester strain, both in the absence and presence of metabolic activating mixture. SOS inducing potencies for SRB, IG, RTB, OS were 0.05, 0.0127, 1.1473, 0.035 in absence and 1.19, 0.022, 0.146, 0.33 in presence of metabolic activatingmixturerespectively. All dyes were toxic to bacteria altheir higher concentrations. IG induced SOS responses at very high concentrations but dye SRB triggered SOS responses at its lower concentrations only. lesions induced by dye RTBappear to be repairable in the presence of S9 mix, while activation mixture appeared to activate dye IG slightly and OS significantly. SRB inhibited general protein synthesis drastically in the presence of S9 mix. Keyword(s): SOS response; Genotoxicity; Textile dyes; Sulpbur dye; Vat dye; Reactivedye; Direct dye
Ir XXI.tOI
A study on tbe mutagenic elTectsof some plant growth
hormones by salmonella/microsome and SOS chromotest systems 6ksilzo~lu Emine, Diril Nuran, Durusoy Milbeccel. Haceuepe University, Faculty of Science, Department of Biology, Molecular Biology, Ankara;
1lvkey In this study the mutagenic effect of some plant growth hormones have been investigated by short term bacterial mutagenity test systems namely Salmonella/microsome and SOS chromotest. In the Salmonella/microsome test system the mutant strains used are S. typhimurium TA98 and TAI00. The tests have been performed in the presence and absence of rat liver S9 fraction. The mutant strain E. coli PQ 37 have been used for the SOS chromotest. Among the plant growth hormones tested only banzyladenine was found to be weaklymutagenic for the S. typhimuriumTA 98 strain and in the presence of S9 fraction. The other plant growth hormones tested were nonmutagenic in both test systems. Keyword(s): Plant growth hormones; SalmonellafMicrosome test system; SOS chromotest system
Ir
XXI.ttl
Tbe SOS lUlltest for the detection of gcnotollc agents
Petra Rettberg', Leonid R. Ptitsyn2 , Olga Komova1 , Stanislav Kozubek", Eugene Krasavin1, Marin BoncvS, Gerda Horncckl . I DLR. Institute of Aerospace Medicine. D - 5/170 Kiiln, Germany; 1Slate Scientific Center of RussianFederation. GNII Genetica, 1/3545 Moscow. Russia: J JINR, 141980 Dubna, Russia; 'Institute of Biophysics, ASCR. Brno. Czech Republic; sInstitute of Nuclear Researchand Nuclear Energy. Sofia. Bulgaria A SOS controlled luminescent bacterial reporter assay, the SOS lux test, was developed for the rapid detection of environmental genotoxins. The bioassayis based on the recombinant plasmid pPLS-I, which was constructed as derivative of pBR322, carrying the promoterless Iw:CDABFE genes of Photobacterium leiognathi downstream of a truncated cda gene from ColD with a strong SOS promoter. E. coli recA' strains containing this plasmid are inducibleto high levels of light production in the presence of substances or agents thal cause DNA damage. The light signal, reflecting the SOS inducing potency, is recorded from the growing culture and the results are availablewithin 1-2 h. The induction of biolummescence was demonstrated by treatment of E. coli C600 (pPLS. I) with 6 genotoxic chemicals and with UV and y radiation, The sensitivity of the SOS lux test is similar to that of other bioassays for genotoxicity, such as the SOS chromotest, IImll test and Ames mutaltst. These rcsull& indicate that the SOS lux test is potentially useful for the in situ and continuous detection of genotoxins,
S-XXI: SOS chromotest
Ir XXI.61
Screenln2 for 2enotoxlclty with 50S chromatest
R. Vachkova-PetrdVa, E. Tyagunenko. National Center of Hygiene, Medical Ecology and Nutrition, Sofia, Bulgaria
In the present study SOS chromotest was performed by the use of the Bioscreen Automated Analyser System, Labsystems, Finland. The method adapted to Bioscreen is based on the colorimetric assay, described by P. Quillardet and M. Hofnung, 1985, but makes use of kinetic assay pnnciple instead of a fixed time assay to quantify the enzymatic activities measured in the test. Usually the conditions of protocol B were used: +/- S9; 10 dikns of sample, max 3 samples; chromogen - yellow 420 nm; incubation time 2 h; positive controls - 4-nilroquinoline-oxide, highest dose 2.63 nmoVassay and 2-arninoanlltracene, 2.59 nmoVassay. Resultsof the screening for genotoxicityof differentcompounds are presented. They include environmental chemicals, food additives, drugs: chloracetophane, alachlor, tetrapol A 159. propylenethiourea, acesulfame K, sodium nitrite. nalidixic acid, ciprofloxacin, cadmium and lead, complex mixtures (biotechnologyproducts for food industry). The correlation between results by SOS chromotest ans Ames test are discussed also.
Ir XXI.71
The SOS chromotest: An analysis from published data on 950 chemicals
P. Quillardet, E. Touati, M. Hofnung. Unite de Programmation Moleculaire et Toxlcologi« Genetique, CNRS URA1444, Bdtiment des Biotechnologies. Institut Pasteur; 25 rue du Dr Raux. 75724 Porn Cedex, Prance In 1982. we have made use of an E. coli strain carrying a fusion of gene lacZ to gene s./iA, one of the SOS gene, to devise a rapid assay for genotoxins: the SOS ChlOmotest. The assay i. performed in few hout! and involves simple enzymatic assays. It allows to classify compounds according to their SOS inducing potency (SOSIP), defined as their ability to induce the expression of the s./iA::lacZ fusion. To day. work from a number of laboratories using the SOS Chromotest have been published. We have reviewed data obtained with the SOS Chromotest on more than 950 chemicals issued from 130 publications arising from about 40 different laboratories. This led us to evaluate further the potential of the SOS Chromotest to detect carcinogens and to compare its response to that of the Salmanellafmicrosome assay. Although far from to be perfect, the results confirm that the SOS Chromotest is a satisfaying method to detect and evaluate genotoxic agents. However, we think that the utility of the SOS chromotcst in a battery of tests of genetic toxicology is mainly due to its simplicity. It constitute an unrivalledtool in the rapid screening of genotoxiccompounds to a large scale and in the monitoring of genotoxie substances in the environment itself. KeywOrd(s): SOS Chromotest; Genotoxicity; Evaluation
Ir XXI.81
Inhibition of SOS response In E. coli PQ37 by heated turmeric and curcumln
Kalpagam Polasa, A. Nadamuni Naidu, R. Krishnaswamy. Food & Drug Toxicology Research Centre, National Institute of Nutrition. Indian Council ofMedical Research. Hyderabad - 500 007, A.P., India Turmeric and its active principle curcumin, commonly used oriental spice has been proved to possess antimutagenic potential. These have been tested in their raw fornu . It is nOC known if antimutagenicityexistsafter exposureto cooking conditions. So heat treatment of these two substances were carried out to simulate cooking conditions namely boiling and frying and tested for antimutagenic property using SOS chromotest. Various concentrations of boiled/fried turmeric and cureumin were tested against 3. $ and 10 ng of 4NQO for inhibition of SOS response. Pooled analysis of the data indicated that both turmeric and curcumin are capable of inhibiting SOS response in E. coli PQ37 even after heat treatment suggesting that cooking processes are unlikely to destroy its antimutagenic property. Keyword(s): SOS test; turmeric; curcumin
Ir XXI.91
Tbe SOS Inducln2 potency offour textile dyes
R. C. Sobti • Meenu Kaushal2 , P.Quillarder', I DepartmentofBiotechnology, Punjab University. Chandigarh - /60014. India: 2Deparrment of Zoology. Panjab Unive~ity. Chandigarh - /60014. India: J Pasteur tnsutute; Paris. France l
SOS responses induced by four textile dyes viz.• Sulphur Red Brown 360 (SRB), lade Green 2G (JG), Reactofix Turquoise Blue 5 GFL (RTB) and Direct Scarlet 4BS (OS) were measured in Escherichia coli PQ37 tester strain, both in the absence and presence of metabolic activating mixture. SOS inducing potencies for SRB, IG, RTB, OS were 0.05, 0.0127, 1.1473, 0.035 in absence and 1.19, 0.022, 0.146, 0.33 in presence of metabolic activatingmixturerespectively. All dyes were toxic to bacteria altheir higher concentrations. IG induced SOS responses at very high concentrations but dye SRB triggered SOS responses at its lower concentrations only. lesions induced by dye RTBappear to be repairable in the presence of S9 mix, while activation mixture appeared to activate dye IG slightly and OS significantly. SRB inhibited general protein synthesis drastically in the presence of S9 mix. Keyword(s): SOS response; Genotoxicity; Textile dyes; Sulpbur dye; Vat dye; Reactivedye; Direct dye
Ir XXI.tOI
A study on tbe mutagenic elTectsof some plant growth
hormones by salmonella/microsome and SOS chromotest systems 6ksilzo~lu Emine, Diril Nuran, Durusoy Milbeccel. Haceuepe University, Faculty of Science, Department of Biology, Molecular Biology, Ankara;
1lvkey In this study the mutagenic effect of some plant growth hormones have been investigated by short term bacterial mutagenity test systems namely Salmonella/microsome and SOS chromotest. In the Salmonella/microsome test system the mutant strains used are S. typhimurium TA98 and TAI00. The tests have been performed in the presence and absence of rat liver S9 fraction. The mutant strain E. coli PQ 37 have been used for the SOS chromotest. Among the plant growth hormones tested only banzyladenine was found to be weaklymutagenic for the S. typhimuriumTA 98 strain and in the presence of S9 fraction. The other plant growth hormones tested were nonmutagenic in both test systems. Keyword(s): Plant growth hormones; SalmonellafMicrosome test system; SOS chromotest system
Ir
XXI.ttl
Tbe SOS lUlltest for the detection of gcnotollc agents
Petra Rettberg', Leonid R. Ptitsyn2 , Olga Komova1 , Stanislav Kozubek", Eugene Krasavin1, Marin BoncvS, Gerda Horncckl . I DLR. Institute of Aerospace Medicine. D - 5/170 Kiiln, Germany; 1Slate Scientific Center of RussianFederation. GNII Genetica, 1/3545 Moscow. Russia: J JINR, 141980 Dubna, Russia; 'Institute of Biophysics, ASCR. Brno. Czech Republic; sInstitute of Nuclear Researchand Nuclear Energy. Sofia. Bulgaria A SOS controlled luminescent bacterial reporter assay, the SOS lux test, was developed for the rapid detection of environmental genotoxins. The bioassayis based on the recombinant plasmid pPLS-I, which was constructed as derivative of pBR322, carrying the promoterless Iw:CDABFE genes of Photobacterium leiognathi downstream of a truncated cda gene from ColD with a strong SOS promoter. E. coli recA' strains containing this plasmid are inducibleto high levels of light production in the presence of substances or agents thal cause DNA damage. The light signal, reflecting the SOS inducing potency, is recorded from the growing culture and the results are availablewithin 1-2 h. The induction of biolummescence was demonstrated by treatment of E. coli C600 (pPLS. I) with 6 genotoxic chemicals and with UV and y radiation, The sensitivity of the SOS lux test is similar to that of other bioassays for genotoxicity, such as the SOS chromotest, IImll test and Ames mutaltst. These rcsull& indicate that the SOS lux test is potentially useful for the in situ and continuous detection of genotoxins,