Application of the buccal micronucleus cytome assay and analysis of PON1Gln192Arg and CYP2A6*9(−48T>G) polymorphisms in tobacco farmers

Tobacco is a major Brazilian cash crop. Tobacco farmers apply large amounts of pesticides to control insect growth. Workers come into contact with green tobacco leaves during the tobacco harvest and absorb nicotine through the skin. In the present study, micronucleus frequency, cell death, and the frequency of basal cells were measured in tobacco farmers using the buccal micronucleus cytome assay (BMCyt), in parallel with measurement of blood butyrylcholinesterase (BChE) and nicotine levels. Polymorphisms in PONIGln192Arg and CYP2A6*9(?48T>G) were evaluated to verify the relationship between genetic susceptibility and the measured biomarkers. Peripheral blood and buccal cell samples were collected from 106 agricultural workers, at two different crop times (during pesticide application and leaf harvest), as well as 53 unexposed controls. BMCyt showed statistically significant increases in micronuclei, nuclear buds, and binucleated cells among exposed subjects in differentiated cells, and in micronuclei in basal cells. In addition, the exposed group showed higher values for condensed chromatin, karyorrhectic, pyknotic, and karyolitic cells, indicative of cell death, and an increase in the frequency of basal cells compared to the unexposed control group. A slight difference in mutagenicity using the BMCyt assay was found between the two different sampling times (pesticide application and leaf harvest), with higher micronucleus frequencies during pesticide application. Elevated cotinine levels were observed during the leaf harvest compared to the unexposed controls, while BChE level was similar among the farmers and controls. PONIGln192Arg and CYP2A6*9(?48T>G) polymorphisms were associated with DNA damage induced by pesticides and cell death. Environ. Mol. Mutagen., 2012. © 2012 Wiley Periodicals, Inc.     

Interaction of microorganisms with enterocytes after oral administration of pesticides

Intragastral administration of the pesticides Sumi-alpha and Omait to rats significantly increases the number of parietal microorganisms in the jejunum, ileum, and particularly in the cecum. Electron microscopy shows that parietal microorganisms invade goblet cells during secretion and then enter prismatic cells via the lateral plasma membrane. The number of parietal microorganisms entering enterocytes after Sumi-alpha is higher than after less toxic Omait, reaching the maximum 5 h after administration. Translated fromByulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 123, No. 4, pp. 467–470, April, 1997     

有机磷对作业工人红细胞乙酰胆碱酯酶和血清对氧磷酶活力的影响

目的探讨接触有机磷对作业工人红细胞乙酰胆碱酯酶(eAChE)和血清对氧磷酶(sPON)活力的影响。方法采用GC-NPD(气相色谱-氮磷检测器)监测作业场所空气中有机磷农药浓度,试剂盒和标准曲线法测定作业工人和对照者eAChE和sPON活力。结果对硫磷包装车间呼吸带的浓度为(0.022±0.012)mg/m3,敌敌畏包装车间呼吸带的浓度为(1.960±1.180)mg/m3;有机磷暴露使作业工人的eAChE及sPON活力降低并与接触工龄呈负相关;对硫磷暴露组中≥35岁作业工人的eAChE活力高于<35岁的作业工人,饮酒可以降低eAChE活力而升高sPON活力。结论eAChE及sPON活力可以作为评价有机磷农药中毒程度的生物标志物。     

Dermal in vitro penetration of methiocarb, paclobutrazol, and pirimicarb

OBJECTIVES—The dominant route of occupational exposure to pesticides in horticulture is dermal. However, preventive measures are seldom used when handling plant cultures recently treated with pesticides, thus causing significant dermal exposure and potential absorption. Assessment of exposure often depends on biological monitoring of blood or urine samples. The skin often acts as a temporary reservoir for chemicals before absorption. Failure to consider the lag time between dermal exposure and appearance of pesticide or metabolites in the general circulation may lead to false conclusions about assessment of exposure.
METHODS—In an experimental model in which in vitro static diffusion cells were mounted with human skin, dermal penetration of three extensively used pesticides (methiocarb, paclobutrazol, pirimicarb) was evaluated.
RESULTS—Pirimicarb and paclobutrazol had comparable rates of dermal penetration and lag times of around 18 hours. Methiocarb had a considerably shorter lag time. Dermal penetration continued for extended periods after exposure had ended.
CONCLUSIONS—With lag times sometimes considerably longer than a normal working day, biological monitoring at the end of exposure may seriously underestimate the actual exposure. There may be implications for regulatory guidelines, which often require only 24 hour observation periods.


Keywords: dermal penetration; pesticides; in vitro     

PXR-dependent induction of human CYP3A4 gene expression by organochlorine pesticides

OCP are xenobiotics which display various toxic effects on animal and human health. One of their effects is to bind and activate estrogen receptor alpha (ERalpha). We have previously studied the down-regulation of induced CYP1A1 (cytochrome P450) expression by this class of molecules in mammary carcinoma cells and shown the importance of ERalpha in this process. However, an alternative mechanism was suggested by those experiments in hepatoma cells. In this study, we have performed Northern blot and transient transfection assays in various cell lines and shown that OCP activate human pregnane X receptor (PXR) and subsequent CYP3A4 mRNA expression. This effect is mediated by the distal xenobiotic responsive element modulator of the promoter. The induction of CYP3A4 by OCP was dose-dependent within the 1-10 microM range. The data suggest that chronic exposure to OCP could alter a major metabolite pathway in human liver and putatively modify the pharmacokinetics of drugs and pollutants.     

Effects of a mixture of two insecticides in freshwater microcosms: I. Fate of chlorpyrifos and lindane and responses of macroinvertebrates

Effects of chronic application of a mixture of the insecticides chlorpyrifos and lindane were studied in indoor freshwater microcosms. The exposure concentrations (based on 0, 0.005, 0.01, 0.05, 0.1 and 0.5 times the LC50 of the most sensitive standard test organism for each compound) were kept at a constant level for four weeks. The calculated mean concentrations for chlorpyrifos were found to be almost at their corresponding nominal level during the treatment period. The mean calculated lindane concentrations, however, were found to be 15–40% higher than intended. In the post treatment period both insecticides dissipated fast (t _1/2: chlorpyrifos 9 days, lindane 22 days) from the water phase. The concentrations of the mixture at the highest treatment level corresponded to 0.53 toxic units (TU) for Daphnia magna and 0.61 TU for the most sensitive fish. The decomposition of Populus leaves in litter bags was significantly lower at the three highest insecticide concentrations. The macroinvertebrate community was seriously affected at the three highest treatment levels, with Crustacea and the Chironomidae Corynoneura proving to be the most sensitive groups. Gastropoda and Oligochaeta were relatively insensitive and some taxa (e.g. Valvata piscinalis, juvenile Physa fontinalis, Nemertea and Stylaria lacustris) increased in numbers. The observed effects could be explained from the individual toxicity of the insecticides to the invertebrates, and did not indicate synergistic effects. A second paper (Van den Brink et al., 2002) addresses the effects on other endpoints, as well as the overall risk assessment of the insecticide mixture.     

Effects of a mixture of two insecticides in freshwater microcosms: II. Responses of plankton and ecological risk assessment

This paper reports on the chronic effects of a mixture of the insecticides chlorpyrifos and lindane in freshwater microcosms. Chronic treatment levels corresponding to concentrations of 0, 0.005, 0.01, 0.05, 0.1 and 0.5 times the LC50 of the most sensitive standard test organism were evaluated. The zooplankton community structure was altered from the 0.05*LC50 treatment level upwards. Cladocerans were the most susceptible group, followed by Copepoda and Ostracoda. Rotifera increased in abundance at the higher treatment levels. Increased abundance of some phytoplankton taxa and increased chlorophyll-a levels were found at the two highest treatment levels, most probably a consequence of decreased grazing pressure. Threshold levels for the mixture, both at population and community/ecosystem level, corresponded well with those reported in the literature for the individual compounds. The overall risk assessment indicates no antagonistic or synergistic effects of the mixture at ecosystem level. It was found that the safety factors set by the Uniform Principles for individual compounds also ensure protection against chronic exposure to a mixture of insecticides at community level, though not always at species level.     

The organochlorine pesticide residues and antioxidant enzyme activities in human breast tumors: is there any association?

The levels of some organochlorine pesticides (OCP)s (hexachlorobenzene, HCB, -hexachlorocyclohexane, -HCH, -HCH, -HCH, heptachlorepoxide, HE, bis (4-chlorophenyl)-1,1-dichloroethene, p.pDDE, bis (4-chlorophenyl)-1,1,1-trichloroethane, p.p DDT and total DDT (-DDT) and antioxidant enzyme activities namely Cu, Zn superoxide dismutase (SOD), catalase (CAT), selenium-dependent glutathione peroxidase (Se-GSH-Px), total glutathione peroxidase (T-GSH-Px), selenium independent glutathione peroxidase (GSH-Px II), glutathione reductase (GRd), level of reduced glutathione (GSH) and lipid peroxidation (LP), glutathione S-transferase (GST) activity toward several substrates including 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), ethacrynic acid (EAA), 1,2-epoxy-3-(p-nitrophenoxy)-propane (ENPP) were measured in tumor and surrounding tumor free tissues of 24 female breast cancer patients and was evaluated whether there exist any association between the levels of OCPs and antioxidants. The mean levels of GSH, -BHC, -BHC and HE, and activities of SOD, Se-GSH-Px, T-GSH-Px, GSH-Px II,GRd, GST CDNB, and GST DCNB were significantly higher in tumors than in controls. In tumors, significant correlations were noted between: SOD and -BHC; Se-GSH-Px and -BHC; T-GSH-Px and -BHC; GSH-Px II and -BHC, -BHC; GSH and -BHC, -BHC, HE; GRd and -BHC; CDNB GST and -BHC, -BHC. These results show that free-radical mediated oxidative stress is, at least partly, associated with some of these OCP residues in human breast tumors.