Characterization of Trp(+) reversions in Escherichia coli strain WP2uvrA. (1/15)

The Escherichia coli strain WP2uvrA is widely used in general mutagenicity screening tests because of its high sensitivity to many kinds of mutagens and it serves as a supplement to the standard Salmonella typhimurium tester strains. In contrast to Salmonella His(+) revertants, E.coli Trp(+) revertants have not been characterized at the molecular level. In this study we found that in the trpE65 allele of WP2uvrA the triplet that codes for the fourth amino acid from the N-terminus of anthranilate synthetase was an ochre stop codon (TAA) instead of a glutamine codon (CAA). In spontaneous Trp(+) revertants the ochre codon had been changed to glutamine (CAA), lysine (AAA), glutamic acid (GAA), leucine (TTA), serine (TCA) or tyrosine (TAC, TAT). Since tryptophan prototrophy could also be restored by ochre suppressor mutations at the anticodon sites in the genes for tRNA(Glu) (glnU), tRNA(Lys) (lysT) and tRNA(Tyr) (tyrT, tyrU), the Trp(+) reversion system with E.coli WP2uvrA detected five types of base substitutions, A.T-->T.A, A.T-->C.G, A.T-->G.C, G.C-->A.T and G.C-->T.A. About 30-50% of Trp(+) revertants induced by N-ethyl-N'-nitro-N-nitrosoguanidine, captan and angelicin plus UVA irradiation were attributable to reversion at the trpE65 ochre locus; the others were attributable to suppressor mutations. In contrast, almost all revertants induced by N-methyl-N'-nitro-N-nitrosoguanidine, 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone and furylfuramide were caused by suppressor mutations. Thus, the high mutagen sensitivity of WP2uvrA is due to several target sites consisting of A.T base pairs (trpE65, lysT) and G.C base pairs (glnU, tyrT, tyrU).  (+info)

Induction of hsp70 in transgenic Drosophila: biomarker of exposure against phthalimide group of chemicals. (2/15)

The expression of stress genes is suggested to be a potentially sensitive indicator of any chemical or physical assault. This led us to explore the possibility of using expression of one of the major stress genes, hsp70, in Drosophila as a biomarker against phthalimide group of chemicals, which may accordingly provide an early indication of exposure to these hazardous chemicals. We exposed third instar larvae of transgenic Drosophila melanogaster (hsp70-lacZ) Bg(9) to different concentrations of the test chemicals (Captan, Captafol and Folpet) for various time intervals (2-48 h) to evaluate expression of hsp70 by X-gal staining, ONPG assay and whole organ in situ immunohistochemistry. The study was further extended to examine the effect of the said chemicals on development of the organism and tissue damage occurring in them, thus raising the possibility of evaluating comparative deleterious effect inducing potential of the test chemicals. Our results showed a strong hsp70 expression in the Captafol-exposed larvae followed by weaker expression in Captan- and Folpet-treated larvae. The effect was further reflected on development as revealed by a delay in emergence of the flies by 3 days in 200 ppm Captafol-exposed group. Hsp70 was found not to be induced at 0.0002 ppm Captafol and at 0.002 ppm Captan and Folpet. The present study suggests that (a). hsp70 induction is sensitive enough to be used as a biomarker against phthalimide group of chemicals, (b). amongst the three test chemicals, Captafol is the most deleterious compound followed by Captan and Folpet, (c). 0.0002 ppm for Captafol and 0.002 ppm for Captan and Folpet, respectively, can be regarded as no observed adverse effect level (NOAEL).  (+info)

Urine mutagenicity and lymphocyte DNA damage in fruit growers occupationally exposed to the fungicide captan. (3/15)

AIMS: To determine haematological parameters, urine mutagenicity (on three Salmonella typhimurium strains), and DNA damage (using the comet assay) in mononuclear leucocytes of farmers before and after a one-day spraying period of pear and apple trees with the fungicide captan in usual conditions. METHODS: Fruit growers were exposed to captan during the 1998 (n = 12) and/or the 2000 spraying seasons (n = 17). Biological samples were collected on the morning of the day of spraying (S1), the evening after spraying (S2), and the morning of the day after (S3). The UK Predictive Operator Exposure Model (UK-POEM) was used to quantify pesticide exposure intensity. RESULTS: No effect was observed on haematological parameters for these two spraying seasons. Proportions of mutagenic urine samples did not significantly differ between S1 and S2/S3 sampling points. In contrast with strains TA97a and YG1041 mainly sensitive to frameshift mutations, a positive trend was observed between the difference (S3-S1) of mutagenic power on strain TA102 detecting base-pair mutations and the exposure predicted value given by UK-POEM, mainly due to parameters related to protective clothing. No significant variations in DNA damage levels were observed between S1 and S3, nor were correlations observed with parameters of pesticide exposure. CONCLUSIONS: A one-day spraying period with captan and other pesticides does not significantly induce DNA damages in mononuclear leucocytes. In contrast, an inefficient protective clothing could correlate with an increase in urine mutagenicity as assessed by the TA102 tester strain.  (+info)

Captan exposure and evaluation of a pesticide exposure algorithm among orchard pesticide applicators in the Agricultural Health Study. (4/15)

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Rhinitis associated with pesticide use among private pesticide applicators in the agricultural health study. (5/15)

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Tetrabutylammonium bromide media aza-Michael addition of 1,2,3,6-tetrahydrophthalimide to symmetrical fumaric esters and acrylic esters under solvent-free conditions. (6/15)

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Determinants of captan air and dermal exposures among orchard pesticide applicators in the Agricultural Health Study. (7/15)

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Biological monitoring of pesticide exposures in residents living near agricultural land. (8/15)

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