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1.
Effect of acute doses of technical grade of dichloro diphenyl trichloro ethane (DDT) and piperonyl butoxide (PB) on hepatic microsomal cytochrome P450 (Cyt. P450) and cytosolic glutathione-S-transferase (GST) activity in pigeon were studied after 24 hours of treatment. A completely reverse trend of changes in Cyt. P450 and GST activity were found as increase in Cyt. P450 paralleled with decrease in GST activity following exposure to DDT. However, intensity of changes in Cyt. P450 was greater than that of GST. A dose dependent decrease in Cyt. P450 and GST activity was observed after PB treatment. The study may, therefore, throw some light on metabolic alterations in wild birds resulting from environmental pollution by DDT and allied chemicals.  相似文献   

2.
Abstract

Effect of acute doses of technical grade of dichloro diphenyl trichloro ethane (DDT) and piperonyl butoxide (PB) on hepatic micro‐somal cytochrome P450 (Cyt. P450) and cytosolic glutathione‐S‐transferase (GST) activity in pigeon were studied after 24 hours of treatment. A completely reverse trend of changes in Cyt. P450 and GST activity were found as increase in Cyt. ?450 paralleled with decrease in GST activity following exposure to DDT. However, intensity of changes in Cyt. P450 was greater than that of GST. A dose dependent decrease in Cyt. ?450 and GST activity was observed after PB treatment. The study may, therefore, throw some light on metabolic alterations in wild birds resulting from environmental pollution by DDT and allied chemicals.  相似文献   

3.
The effect of bisphenol A (BPA) on the kinetics of cytochrome P450 (P450)-dependent monooxygenases in rat liver microsomes was studied. Testosterone 16beta-hydroxylase (TS16BH) and testosterone 2alpha-hydroxylase (TS2AH) activities were extensively inhibited by BPA at 100 microM (69% and 74%, respectively). The inhibition type was mixed for both P450-dependent monooxyganases. The Ki of TS16BH and TS2AH from Lineweaver-Burk plots were 25.9 and 24.9 microM, respectively. The activities of acetanilide 4-hydroxylase (AA4H), 7-ethoxycoumarin O-deethylase (ECOD), bufuralol 1'-hydroxylase (BF1'H), chlorzoxazone 6-hydroxylase (CZ6H) and testosterone 6beta-hydroxylase (TS6BH) were also effectively inhibited by BPA at 100 microM (43-52%). The inhibition type of these P450-dependent monooxygenases was mixed or uncompetitive, and the K(i)s (50.5-88.5 microM) were higher than those of TS16BH and TS2AH. By contrast, the values of IC50 and Ki of testosterone 7alpha-hydroxylase (TS7AH) and lauric acid omega-hydroxylase (LAOH) for BPA were >1000 microM. These results suggest that BPA interacts with rat hepatic CYP1A2, CYP2A2, CYP2B2, CYP2C11, CYP2D1, CYP2E1 and CYP3A2 in vitro.  相似文献   

4.
Bisphenol A is an endocrine disrupting compound, which is ubiquitous in the environment due to its wide use in plastic and resin production. Seven day old cultures of the litter-decomposing fungus Stropharia coronilla removed the estrogenic activity of bisphenol A (BPA) rapidly and enduringly. Treatment of BPA with purified neutral manganese peroxidase (MnP) from this fungus also resulted in 100% reduction of estrogenic activity, as analyzed using a bioluminescent yeast assay, and in the formation of polymeric compounds. In cultures of Stropharia rugosoannulata, estrogenic activity also quickly disappeared but temporarily re-emerged in the further course of cultivation. LC-MS analysis of the extracted estrogenic culture liquid revealed [M−H] ions with m/z values of 219 and 235. We hypothesize that these compounds are ring fission products of BPA, which still exhibit one intact hydroxyphenyl group to interact with estrogen receptors displayed by the yeast.  相似文献   

5.
Organic solvents, such as dimethylsulfoxide (DMSO) and methanol are widely used as vehicles to solubilise lipophilic test compounds in toxicity testing. However, the effects of such solvents upon innate detoxification processes in aquatic organisms are poorly understood. This study assessed the effect of solvent exposure upon cytochrome P450 (CYP)-mediated xenobiotic metabolism in Daphnia magna and zebrafish larvae (4 d post fertilisation). Adult D. magna were demonstrated to have a low, but detectable, metabolism of ethoxyresorufin in vivo and this activity was not modulated by pre-exposure to DMSO or methanol (24 h, up to 0.1% and 0.05% v/v, respectively). In contrast, the metabolism of ethoxyresorufin in zebrafish larvae was significantly reduced by both solvents (0.1% and 0.05% v/v, respectively) after 24 h of exposure. In zebrafish, these observed decreases in activity towards ethoxyresorufin were accompanied by decreased expression of a variety of genes coding for drug metabolising enzymes (corresponding to CYP1, CYP2, CYP3 and UDP-glucuronyl transferase [UGT] family enzymes), measured by quantitative PCR. Reduction of gene expression and CYP1 enzyme activities by methanol (0.05% v/v) in zebrafish larvae was partially reversed by co-exposure with Aroclor 1254 (100 μg L−1). Overall this study suggests that relatively low concentrations of organic solvents can impact upon the biotransformation of certain xenobiotics in zebrafish larvae, and that this warrants consideration when assessing compounds for metabolism and toxicity in this species.  相似文献   

6.
7.
In Kyrgyzstan, many former storehouses and dump sites for obsolete pesticides exist. In 2009/2010, an inventory and assessment of these sites including risks of environmental hazard has been conducted by FAO and the World Bank. Monitoring revealed high concentration of pesticides listed as persistent organic pollutants (POPs). The purpose of this research was to study the microbial structural complexes of the pesticide-contaminated soils in these dumping zones, and to search for and select microorganism’s destructors with cytochrome P450 genes for pesticide degradation. Culture-dependent and culture-independent approaches were used to determine the taxonomic composition of these bacterial communities. The universal primer set for the 16S ribosomal RNA (rRNA) gene and the specific primer set P450R were used to amplify the cytochrome P450 hydroxylase gene. In soils from Suzak A and B and soils from Balykchy dumping sites, the bacteria from the Actinobacteria phylum (Micrococcus genus) were dominant. These bacteria made up 32–47% of the indigenous local microflora; bacteria species from the Pseudomonas genus (Gammaproteobacteria phylum) made up 23% in Suzak, 12% in Balykchy soils. Bacillus species from the Firmicutes phylum were found only in Suzak soils. The 16S rRNA analyses and the specific primer set P450R have revealed bacteria with cytochrome genes which are directly involved in the degradation process of organic carbon compounds. Experiments were carried out to help select active degraders from the bacterial populations isolated and used to degrade Aldrin in laboratory. Active bacterial strains from the Pseudomonas fluorescens and Bacillus polymyxa population were selected which demonstrated high rates of degradation activity on Aldrin.  相似文献   

8.
Cytochrome P450-dependent microsomal monooxygenase (P450) activity was measured in control and atrazine-exposed third instar midge larvae, Chironomus tentans. Significantly elevated O-demethylase activity was observed in gut homogenates taken from midges exposed to atrazine concentrations from 1 to 10 ppm for 90 h. No significant induction was observed at atrazine concentrations below 1 ppm. A region of a cytochrome P450 family 4 gene was amplified and sequenced from C. tentans larvae. Alignments of inferred amino acid sequences with other insect CYP4 gene homologues indicate a high degree of similarity. Northern blot analysis employing the CYP4 gene fragment as a probe showed an over-expression in C. tentans exposed to atrazine. The results support the previously identified inducibility of cytochrome P450-dependent activity and provide insight into the potential consequences of atrazine exposure to aquatic organisms.  相似文献   

9.
Agradi E  Baga R  Cillo F  Ceradini S  Heltai D 《Chemosphere》2000,41(10):1555-1562
Groups of eels (Anguilla anguilla) were exposed to Po river water under control conditions at "la Casella" Fluvial Hydrobiology Station for 30-day periods between November and May. At the end of the exposure period, fish were sacrificed and cytochrome P4501A was evaluated in liver microsomes using both catalytic (EROD) and semiquantitative immunodetection (ELISA) assays. At the same time, water samples were taken for chemical analyses of PCBs, PAHs and pesticides. Eel was chosen as bio-indicator on the bases of experiments on cytochrome P450 expression and activity under basal or induced conditions and in consideration of the ecological and economic relevance and ease of handling. Low levels of cytochrome P450 during Winter were followed by a step increase during Spring, associated with a substantial concentration increase of agrochemicals. A good correlation was found between measurements of cytochrome P4501A by EROD and ELISA in dose-effect experiments, however, ELISA showed a higher sensitivity. Immunochemical techniques may be used in addition to enzyme activity measurements both to detect lower levels of cytochrome P450 induction (where they proved more sensitive) and as quality control. These results suggest that measurement of cytochrome P4501A under controlled conditions, using eel as bio-indicator, can be a useful tool in monitoring Po river ecosystems.  相似文献   

10.
Natural steroidal hormone estrone (E1) was treated with the white rot fungus Phanerochaete sordida YK-624 under ligninolytic condition with low-nitrogen and high-carbon culture medium. E1 decreased by 98% after 5 d of treatment and the activities of ligninolytic enzymes, manganese peroxidase (MnP) and laccase, were detected during treatment, which suggested that the disappearance of E1 is related to ligninolytic enzymes produced extracellularly by white rot fungus. Therefore, E1 was treated with MnP and laccase prepared from the culture of white rot fungi. HPLC analysis demonstrated that E1 disappeared completely in the reaction mixture after 1 h of treatment with either MnP or laccase. Using the yeast two-hybrid assay system, it was also confirmed that both enzymatic treatments completely removed the estrogenic activity of E1 after 2 h. These results strongly suggest that ligninolytic enzymes are effective in removing the estrogenic activity of E1.  相似文献   

11.
Drugs inducing different forms of P-450 cytochrome isoenzymes and binding the Ah receptor or not were investigated for their ability to modify 2,3,7,8, tetrachlorodibenzo-p-dioxin (TCDD) immunotoxicity in mice. 3-Methyl-cholanthrene (3MC) and β-naphthoflavone (BNF) administered to TCDD-treated mice caused additive inhibition of humoral antibody production and of responsiveness to concanavallin A (ConA) but not to phytohemagglutinin (PHA) and lipopolisaccharide (LPS) while phenobarbital (PB) never modified TCDD immunosuppression. Natural killer (NK) cell activity was not reduced by single drug treatment or by combined treatments. Hepatic aryl hydrocarbon hydroxylase (AHH) induction by TCDD was not significantly modified by PB, 3MC or BNF.  相似文献   

12.
Abstract

The present study was under taken to demonstrate the effect of some commonly used insecticides on the activity of cytochrome P450 system including cytochrome b5, aryl hydrocarbon [benzo(a)pyrene] hydroxylase (AHH), N-nirosodimethylamine N-demethylase I [NDMA-dI] and NADPH-cytochrome c reductase as phase I of drug oxidation. In addition, the activity of glutathione S-transferase (GST), glutathione reductase (GR), and the level of glutathione (GSH) were determined in the liver of male mice after oral administration of sumithion, dursban, chlordane, methoxychlor, heptachlor epoxide, and lindane as single (24 h) or as repeated doses for six consecutive days. Oral administration of sumithion, dursban, chlordane, methoxychlore, and heptachlor epoxide as repeated doses decreased: (i) the hepatic content of cytochrome P450 by 36, 37, 47, 37, and 67%, respectively, (ii) AHH activity by 28, 29, 70, 31, and 79%, respectively, (iii) NDMA-dI activity by 43, 44, 32, 27, and 31, respectively. On the other hand, sumithion, chlordane, and methoxychlore induced the activity of NADPH-cytochrome c reductase by 45, 62, and 43 respectively after repeated dose treatments. In addition, single and repeated-dose treatments of mice with lindane induced; (i) cytochrome P450 by 23 and 65%, respectively, (ii) cytochrome b5 by 49 and 131%, respectively, (iii) AHH activity by 64 and 50%, respectively. Repeated-dose treatments of mice with chlordane, methoxychlore, and heptachlor epoxide decreased the GSH level by 42, 38, and 68%, respectively and GST activity by 44, 44, and 55% respectively. Moreover, single- and repeated-dose treatments of mice with lindane decreased the GSH levels by 40 and 54%, respectively, and induced GST activity by 25 and 41%, respectively. Interestingly, single-dose treatments with chlordane, methoxychlore, and heptachlor epoxide decreased the activity of GR by 32, 38, and 31, respectively whereas repeated doses of these compounds induced such activity by 83, 50, and 64%, respectively. It is concluded that modifications in cytochrome P450 system by pesticides could potentiate the toxicity and carcinogenicity of environmental carcinogens such as polycyclic aromatic hydrocarbon and N-nirosodimethylamine (NDMA).  相似文献   

13.
The purpose of this study was to examine neuroendocrine-disrupting effects of two domestic wastewater aeration lagoons on freshwater mussels. Mussels were caged and placed in two final aeration lagoons for treating domestic wastewaters for 60 days, at a site 1km downstream of the dispersion plume on the eastern shores of the Richelieu River; the western shore served as the reference site. The mussels were analysed for gonad activity, oxidative metabolism of xenobiotics, stress biomarkers and neuroendocrine status (monoamine and arachidonic acid metabolism). The domestic wastewaters produced many different effects at all levels examined. The gonado-somatic index and vitellogenin-like proteins were significantly induced in both aeration lagoons and gonad pyrimidine synthesis (aspartate transcarbamylase activity) was significantly reduced, indicating that vitellogenin-like proteins were produced while DNA synthesis in gametes remained constant. Biomarkers of oxidative metabolism revealed that global heme oxidase (HO), glutathione S-transferase and xanthine (caffeine) oxydoreductase (XOR) activities were significantly induced in at least one of the aeration lagoons, but not downstream of the dispersion plume. The activities of 7-ethoxyresorufin (cytochrome P4501A1), dibenzoylfluorescein (cytochrome P450 3A4 and 3A5) and benzoyloxyresorurufin (cytochrome P450 3A4 and 2B6) dealkylases were readily induced by substances sharing structural similarities with coplanar polyaromatic hydrocarbons and hydroxylated or aminated aromatic or cyclic hydrocarbon compounds such as pharmaceuticals or steroids in the domestic wastewaters. Biomarkers of toxic stress revealed that exposure to aeration lagoons led to increased production of metallothioneins, lipid peroxidation and DNA strand breaks, with decreased heme oxygenase activity. LPO was significantly correlated with XOR, HO and cytochrome P4501A1 activities. Neuroendocrine effects included significant increases in dopamine and serotonin levels and in monoamine oxidase (MAO). Dopamine transport in synaptosome was significantly increased while serotonin transport activity was significantly decreased, suggesting the mussels were in a state of serotonergicity. Moreover, arachidonic acid cyclooxygenase (COX) activity was also readily increased in one aeration lagoon. Aeration lagoons for the treatment of domestic wastewaters are toxic, estrogenic and disrupt the metabolism of monoamines and COX in freshwater mussels.  相似文献   

14.
Endocrine disrupting compounds (EDCs) are exogenous environmental chemicals that can interfere with normal hormone function and present a potential threat to both environmental and human health. The fate, distribution and degradation of EDCs is a subject of considerable investigation. To date, several studies have demonstrated that conventional water treatment processes are ineffective for removal of most EDCs and in some instances produce multiple unknown transformation products. In this study we have investigated the use of direct photolysis with low-pressure (LP) Hg UV lamps and UV+hydrogen peroxide (H(2)O(2)) advanced oxidation process (AOP) for the degradation of a prototypic endocrine disrupter, bisphenol A (BPA), in laboratory water. Removal rates of BPA and formation of degradation products were determined by high performance liquid chromatography (HPLC) analysis. Changes in estrogenic activity were evaluated using both in vitro yeast estrogen screen (YES) and in vivo vitellogenin (VTG) assays with Japanese medaka fish (Oryzias latipes). Our results demonstrate that UV alone did not effectively degrade BPA. However, UV in combination with H(2)O(2) significantly removed BPA parent compound and aqueous estrogenic activity in vitro and in vivo. Removal rates of in vivo estrogenic activity were significantly lower than those observed in vitro, demonstrating differential sensitivities of these bioassays and that certain UV/AOP metabolites may retain estrogenic activity. Furthermore, the UV/H(2)O(2) AOP was effective for reducing larval lethality in treated BPA solutions, suggesting BPA degradation occurred and that the degradation process did not result in the production of acutely toxic intermediates.  相似文献   

15.
The present study was under taken to demonstrate the effect of some commonly used insecticides on the activity of cytochrome P450 system including cytochrome b5, aryl hydrocarbon [benzo(a)pyrene] hydroxylase (AHH), N-nirosodimethylamine N-demethylase I [NDMA-dI] and NADPH-cytochrome c reductase as phase I of drug oxidation. In addition, the activity of glutathione S-transferase (GST), glutathione reductase (GR), and the level of glutathione (GSH) were determined in the liver of male mice after oral administration of sumithion, dursban, chlordane, methoxychlor, heptachlor epoxide, and lindane as single (24h) or as repeated doses for six consecutive days. Oral administration of sumithion, dursban, chlordane, methoxychlore, and heptachlor epoxide as repeated doses decreased: (i) the hepatic content of cytochrome P450 by 36, 37, 47, 37, and 67%, respectively, (ii) AHH activity by 28, 29, 70, 31, and 79%, respectively, (iii) NDMA-dI activity by 43, 44, 32, 27, and 31, respectively. On the other hand, sumithion, chlordane, and methoxychlore induced the activity of NADPH-cytochrome c reductase by 45, 62, and 43 respectively after repeated dose treatments. In addition, single and repeated-dose treatments of mice with lindane induced: (i) cytochrome P450 by 23 and 65%, respectively, (ii) cytochrome b5 by 49 and 131%, respectively, (iii) AHH activity by 64 and 50%, respectively. Repeated-dose treatments of mice with chlordane, methoxychlore, and heptachlor epoxide decreased the GSH level by 42, 38, and 68%, respectively and GST activity by 44, 44, and 55% respectively. Moreover, single- and repeated-dose treatments of mice with lindane decreased the GSH levels by 40 and 54%, respectively, and induced GST activity by 25 and 41%, respectively. Interestingly, single-dose treatments with chlordane, methoxychlore, and heptachlor epoxide decreased the activity of GR by 32, 38, and 31, respectively whereas repeated doses of these compounds induced such activity by 83, 50, and 64%, respectively. It is concluded that modifications in cytochrome P450 system by pesticides could potentiate the toxicity and carcinogenicity of environmental carcinogens such as polycyclic aromatic hydrocarbon and N-nirosodimethylamine (NDMA).  相似文献   

16.
Kanaly RA  Hur HG 《Chemosphere》2006,63(2):202-211
Generally, the white-rot fungus Phanerochaete chrysosporium performs its biodegradative activities in liquid culture while growing on easily utilized carbon sources such as malt- or potato-extract. However, less is known about the potential of this organism to grow directly on environmental pollutants without regard to special conditions. Growth of P. chrysosporium on a middle fraction (MF) of diesel fuel at neutral pH in mineral medium under non-ligninolytic conditions was explored. After 14 d, the GC-analyzable n-alkanes of 1000 mg l(-1)MF were reduced to background, with most biodegradation occurring by day 7 when quantified relative to the biodegradation of the internal fuel biodegradation marker, pristane. Investigations with n-hexadecane and unmodified diesel fuel further confirmed these biodegradation results. Biomass production was monitored and indicated that fungal biomass was more than 10 times less than positive controls (potato dextrose broth, PDB) but that biomass increased relative to negative controls. When P. chrysosporium was incubated with diesel fuel and PDB, fuel biodegradation was delayed for at least 4d and inhibited overall through 14 d. Experiments with P. chrysosporium growing on n-hexadecane in the presence of 1 mM 1-aminobenzotriazole (ABT), an inhibitor of the cytochrome P-450 enzyme system, resulted in inhibition of biomass production relative to positive controls implicating the utilization of this enzyme system in n-alkane metabolism. Finally, when P. chrysosporium was incubated in a non-aqueous phase liquid (NAPL) mixture of polycyclic aromatic hydrocarbons (PAHs) and MF, n-alkanes and phenanthrene were degraded in 2 weeks while anthracene, chrysene and benzo[a]pyrene were not.  相似文献   

17.
An OECD initiative for the development of mollusc-based toxicity tests for endocrine disrupters and other chemicals has recommended three test species with respective test designs for further standardisation. Preparing a subsequent pre-validation study we performed a reproduction test with Potamopyrgus antipodarum, determining the concentration range of the selected test substances, bisphenol A (BPA) and cadmium (Cd). At 16 °C, the recommended test temperature, the number of embryos in the brood pouch was increased by BPA and decreased by Cd (NOEC: 20 μg BPA/L and 1 μg Cd/L). Coinstantaneous BPA tests at 7 °C and 25 °C demonstrated a temperature dependency of the response, resulting in lower NOECs (5 μg/L respectively). As expected, reproduction in control groups significantly varied depending on temperature. Additional observations of the brood stock showed seasonal fluctuations in reproduction under constant laboratory conditions. The recommended temperature range and test conditions have to be further investigated.  相似文献   

18.
Biodegradation of the polychlorinated naphthalenes (PCNs) 1,4-dichloronaphthalene (1,4-DCN), 2,7-dichloronaphthalene (2,7-DCN), and 1,2,3,4-tetrachloronaphthalene (1,2,3,4-TCN), by the white-rot fungus Phlebia lindtneri was investigated. 1,4-DCN was metabolized to form six metabolites by the fungus. It was estimated from GC–MS fragment patterns that the metabolites were four putative hydroxylated and two dihydrodihydroxylated compounds. One of the hydroxylated products was identified as 2,4-dichloro-1-naphthol by GC–MS analysis using an authentic standard. This intermediate indicated chlorine migration in a biological system of P. lindtneri. 2,7-DCN was metabolized to five hydroxylated metabolites and a dihydrodihydroxylated metabolite. Significant inhibition of the degradation of DCNs and formation of their metabolic products was observed in incubation with the cytochrome P-450 monooxygenase inhibitor piperonyl butoxide. The formation of the dihydrodiol-like metabolites, chlorine migration and the experiment with P-450 inhibitor suggested that P. lindtneri provides hydroxyl metabolites via benzene oxide intermediates of DCNs by a cytochrome P450 monooxygenase. In addition, P. lindtneri degraded 1,2,3,4-TCN; two hydroxylated compounds and a dihydrodihydroxylated compound were formed.  相似文献   

19.
Studies were carried out to investigate the metabolism of herbicide chlortoluron in the microsomal fractions and whole cells of Saccharomyces cerevisiae expressing human cytochrome P450 3A4. Both whole cells and microsomal fractions of yeast expressing human cytochrome P450 3A4 exhibited a typical dithionite-reduced, CO-difference absorbance spectrum with maximum absorbance at 448 nm. Chlortoluron produced a type I binding spectrum with cytochrome P450 3A4 with a Ks value of 200 μM. Chlortoluron was metabolised into four metabolites; hydroxylated-N-monodemethylated, hydroxylated ring methylated, N-didemethylated and N-monodemethylated products. Chlortoluron metabolism was absolutely dependent on NADPH and no metabolism was observed in control transformants.  相似文献   

20.
Bisphenol A (BPA) is a well-known environmental toxic substance, which exerts unfavorable effects through endocrine disruptor (ER)-dependent and ER-independent mechanisms to threaten ecological systems seriously. BPA may also interact with other environmental factors, such as light and heavy metals, to have a synergetic effect in plants. However, there is little data concerning the toxic effect of BPA on the primary producers-plants and its possible interaction with light-dependent response. Here, the effects of BPA on germination, fresh weight, tap root length, and leaf differentiation were studied in Arabidopsis thaliana under different parts of light spectrum (dark, red, yellow, green, blue, and white light). Our results showed that low-dose BPA (1.0, 5.0 μM) caused an increase in the fresh weight, the tap root length and the lateral root formation of A. thaliana seedlings, while high-dose BPA (10.0, 25.0 μM) show an inhibition effect in a dose-dependent manner. Unlike karrikins, the effects of BPA on germination fresh weight and tap roots length under various light conditions are similar, which imply that BPA has no notable role in priming light response in germination and early seedling growth in A. thaliana. Meanwhile, BPA exposure influences the differentiation of A. thaliana leaf blade significantly in a light-dependent manner with little to no effect in dark and clear effect under red illumination.  相似文献   

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