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Pyrene and chrysene fate in surface soil and sand microcosms.

Polycyclic aromatic hydrocarbons (PAHs) are major components of wastes from municipal gas plants and many wood preservatives. Soil contaminated with these wastes is a potential threat to human health because of the carcinogenicity of many PAHs. This study follows the fate of two four-ring PAHs, pyrene and chrysene, in three matrices: an adapted soil (obtained from a site contaminated with PAHs for more than 75 years), an uncontaminated soil (with and without an inoculum of adapted soil), and sand mixed with an inoculum of adapted soil. Radiolabeled pyrene, chrysene, and salicylic acid (a metabolite of PAH biodegradation) were used to trace the mineralization, transformation, extractability, and formation of an unextractable residual over time. Linear approximations of the rates of these processes were made. High-performance liquid chromatography (HPLC) analysis of extracts from inoculated soil showed the transient formation of two known metabolites: 1-hydroxypyrene (from pyrene) and 1-hydroxy-2-naphthoic acid (from chrysene). The amount of extractable label diminished steadily over the course of the study in systems that were not inhibited with sodium azide, whereas the amount of extractable label remained relatively constant in inhibited systems. Correspondingly, the amount of nonextractable residual label generally increased during each incubation in uninhibited systems, whereas the amount of this residual label remained relatively constant in inhibited systems. In contrast, the rate and extent of mineralization varied widely across matrix types. This suggests that alterations of the PAH that impact extractability and residual formation are common, in contrast to mineralization, which was apparently limited to adapted communities.

Biodegradation, Environmental↗

Water solubility enhancements of pyrene by single and mixed surfactant solutions.

Water solubility enhancements of pyrene by both single-surfactant and mixed-surfactant solutions were compared and evaluated. The solubility of pyrene in water was greatly enhanced by each of Triton X-100 (TX100), Triton X-405 (TX405), Brij 35 and SDS, in which the water solubility enhancements increased with increasing surfactant concentrations. The extent of solubility enhancements at surfactant concentrations below the CMC is the order of TX100 > Brij 35 > TX405 > SDS; the sequence at surfactant concentrations above the CMC is TX100 > Brij 35 > SDS > TX405. Pyrene was solubilized synergistically by anionic-nonionic mixed surfactant solutions, especially at low surfactant concentrations. The synergistic power of the mixed surfactants is SDS-TX405 > SDS-Brij 35 > SDS-TX100. The synergism as noted is attributed to increasing Kmc is and/or decreasing the CMC of the mixed surfactant solution. For SDS-TX405 and SDS-Brij 35 mixed surfactant solutions, an increase in Kmc is coupled with a decrease in the CMC; for SDS-TX100, only a decreased in the CMC value is noted. Mixed-surfactant solutions may improve the performance of the surfactant-enhanced remediation (SER) of soils by increasing the bioavailability and biodegradation of non-aqueous-phase organic pollutants and reducing the level of surfactant pollution and remediation expenses.

Biological Availability↗

Effects of two barrier creams on the diffusion of benzo[a]pyrene across human skin.

Barrier hand creams, often containing antiseptic agents, may provide a form of protection not only for health care professionals, but also for workers in the chemical and pharmaceutical industries. To evaluate the efficacy of two such barrier creams available on the South African market, the in vitro diffusion of a model compound, benzo[a]pyrene, through human skin at 20 degrees and 37 degrees C was studied. Treated (10 min) and untreated human skin disks (4 mm in diameter) were mounted in flow cells of a continuous flow-through diffusion apparatus. Buffer/tritiated benzo[a]pyrene was collected from the acceptor chambers at 2-hour intervals for a total of 24 hours and counted in a liquid scintillation counter. At 20 degrees C no significant differences could be detected between the flux rates of benzo[a]pyrene across barrier cream treated and untreated skin. However, at 37 degrees C Skinguard significantly increased flux rates of this carcinogen. Skin barrier creams therefore need to be carefully scrutinised with respect to their protective effects because the latter may vary for molecules with different chemical properties.

Analysis of Variance↗

[Determination of benzo[a]pyrene in mainstream smoke by solid-phase extraction and gas chromatography-mass spectrometry].

A novel method is suggested for the determination of benzo[a]pyrene in mainstream smoke. Extraction of Cambridge pads was cleaned up with methanol/n-heptane extraction system and silica solid-phase extraction (SPE). The eluted solution containing benzo[a]pyrene was evaporated gently to complete dryness with N2 flow, then reconstituted in 200 microL ethyl acetate for determination with gas chromatography-mass spectrometry selected ion monitoring (SIM). The proposed method had good results and precision for the separation of pyrene in complex matrices.

Benzo(a)pyrene↗

[Antisense ERCC1 RNA decreases the repair capability of damaged DNA in lung cancer cells induced by benzo[a]pyrene].

OBJECTIVE: To investigate the effect of ERCC1 gene on the repair capability of damaged DNA in lung cancer A549 cells induced by benzo[a]pyrene. METHODS: Recombinant plasmid expressing ERCC1 antisense RNA was constructed and transfected into A549 cells by Lipofectin reagent. The stable-transfected cell colonies were selected by hygromycin. Cell viability was determined by the MTT assay. The level of ERCC1 mRNA was measured by Northern Blot analysis. Single cell gel electrophoresis assay was applied to determine the cellular DNA damage and fifty cells for each group were counted. RESULTS: Seven positive colonies expressing ERCC1 antisense RNA were screened. There was no growth rate difference between the antisense-transfected cells and the parental cells. The endogenous mRNA level in transfected colonies decreased in varied degrees, i.e. 12% approximately 86% of that of the parental cells in Northern Blot assay. After 24 h treatment of 10 micro mol/l benzo[a]pyrene, the repair capability for DNA damage in transfected colonies was reduced to 29% approximately 71% of that of the parental cells. Also, a statistically significant correlation was observed between expression of ERCC1 mRNA and repair capability (r = 0.84). CONCLUSION: Antisense ERCC1 RNA decreased the repair capability for damaged DNA in lung cancer cells induced by benzo[a]pyrene.

Benzo(a)pyrene↗

Oxygen radical-dependent epoxidation of (7S,8S)-dihydroxy-7,8-dihydrobenzo[a]pyrene in mouse skin in vivo. Stimulation by phorbol esters and inhibition by antiinflammatory steroids.

(7S,8S)--Dihydroxy--7,8--dihydrobenzo[a]pyrene ((+)-BP-7,8-diol) is epoxidized to (7S,8R)-dihydroxy-(9S,10R)-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene ((+)-syn-BPDE) by cytochrome P-450 isoenzymes and to (7S,8R)-dihydroxy-(9R,10S)-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene ((-)-anti-BPDE) by peroxyl free radicals. 32P postlabeling analysis of the diastereomeric BPDE-DNA adducts was used to investigate the pathways of (+)-BP-7,8-diol oxidation in mouse skin in vivo. The pattern of deoxynucleoside 3',5'-bisphosphate adducts in epidermal scrapings from female CD-1 mice indicated that cytochrome P-450 was the major oxidant. Similar results were obtained when the tumor-promoting phorbol ester tetradecanoylphorbolacetate (TPA) was coadministered with (+)-BP-7,8-diol. However, when animals were pretreated with TPA 24 h before coadministration of TPA and (+)-BP-7,8-diol, the pattern of BPDE-DNA adducts indicated that peroxyl radicals made a major contribution to (+)-BP-7,8-diol epoxidation. Peroxyl radical-dependent epoxidation was maximal when the time between the two TPA administrations was 24-72 h. No increase in (-)-anti-BPDE-DNA was observed when the non-tumor-promoting phorbol ester 4-O-methyl-TPA was substituted for TPA. The calcium ionophore A23187 stimulated peroxyl radical generation when substituted for the first, but not the second, TPA treatment. The antiinflammatory steroid fluocinolone acetonide inhibited (-)-anti-BPDE-DNA adduct formation when coadministered with the first but not the second TPA treatment. These findings demonstrate the existence of two independent pathways of metabolic activation of (+)-BP-7,8-diol in mouse epidermis, one dependent on cytochrome P-450 and the other dependent on peroxyl free radicals. The results also suggest that repetitive topical administration of tumor-promoting phorbol esters remodels epidermal metabolism leading to a significant increase in free radical generation.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

Mono- and diglucuronide formation from chrysene and benzo(a)pyrene phenols by 3-methylcholanthrene-inducible phenol UDP-glucuronosyltransferase (UGT1A1).

Mono- and diphenols of chrysene and benzo(a)pyrene are suspected substrates of a 3-methylcholanthrene (MC)-inducible phenol UDP-glucuronosyltransferase (UGT1A1). Mono- and diglucuronide formation from these compounds was studied in two systems, (a) livers of MC-treated rats (homologous expression) and (b) a Chinese hamster lung fibroblast cell line (V79) containing rat UGT1A1 cDNA and stably expressing this isozyme (heterologous expression). In liver microsomes of MC-treated rats, glucuronidation of 6-hydroxychrysene was stimulated 11-fold by MC treatment. With 3,6-dihydroxychrysene, formation of its 3-hydroxy-6-monoglucuronide and of the diglucuronide was increased 24- and 310-fold, respectively. Induction factors obtained for monoglucuronide formation (but not for diglucuronide formation) were in line with published data on the increase of immunodetectable UGT1A1 protein and of its mRNA. It is suggested that the high induction factors for diglucuronide formation are the result of a combination of the induction of UGT1A1 and facilitated interaction of neighboring UGT1A1 molecules in endoplasmic reticulum membranes. Glucuronidation of 6-hydroxychrysene, 3-hydroxybenzo(a)pyrene, and 3,6-dihydroxybenzo(a)pyrene to their mono- and diglucuronides was clearly detectable in V79 cells expressing UGT1A1. However, conjugation of 3,6-dihydroxychrysene to its monoglucuronides was low and diglucuronide formation was not detectable, suggesting that UGT isozymes other than UGT1A1 are responsible for these reactions.

Animals↗

Inhibition of benzo[a]pyrene-induced mouse forestomach neoplasia by a principal flavor component of Japanese-style fermented soy sauce.

A refined diet supplemented with Japanese-style fermented soy sauce (shoyu) inhibits benzo[a]pyrene-induced forestomach neoplasia in mice (Cancer Res., 51:2940-2942, 1991). In the present study, soy sauce was extracted with ethyl acetate. The soluble fraction contained flavor/aroma compounds and antioxidants, whereas amino-carbonyl compounds that impart color were concentrated in the ethyl acetate insoluble fraction. Both fractions inhibited benzo[a]pyrene-induced forestomach neoplasia in a protocol in which the test material was fed following exposure to the carcinogen. A principal flavor/aroma component of soy sauce, 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone, was fed to mice following benzo[a]pyrene administration and found to inhibit the subsequent development of forestomach neoplasia. 4-Hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone was effective when fed at 4 mg/kg body weight/day, indicating that it is a potent anticarcinogen.

Animals↗

Sorption of pyrene on two paddy soils and their particle-size fractions.

In the present study, the sorption of pyrene on two kinds of bulk paddy soils, Gleyic Stagnic Anthrosols, and Ferric accumulic Stagnic Anthrosols as well as their particle-size fractions was investigated. The sorption isotherms fitted well with Freundlich equation. For both soils, the clay fraction( < 2 microm) and coarse sand fraction(2000-250 microm) had higher sorption capacity than fine sand fraction(250-20 microm) and silt fraction(20-2 microm). The log Koc values obtained of each soil and its particle-size fractions were similar, proving that SOM content was a key factor affecting pyrene sorption. The Kd values showed a significant correlation with contents of dithionite-extractable Fe in both paddy soils and a good relationship with CEC in Gleyic Stagnic Anthrosols, indicating possible effects of surface properties of particle-size fractions on the sorption of pyrene.

Adsorption↗

Chemical analysis of smoke of Indian cigarettes, bidis and other indigenous forms of smoking--levels of steam-volatile phenol, hydrogen cyanide and benzo(a)pyrene.

Levels of steam-volatile phenol, hydrogen cyanide and benzo(a)pyrene in various types of tobacco smoking products marketed in the country have been determined for the first time. Steam-volatile phenol levels in six popular brands of Indian cigarettes varied from 118 to 226 micrograms, and in six popular brands of bidis, from 129 to 273 micrograms. Cheroot and cigarillos yielded 400 micrograms and 333 micrograms steam-volatile phenol respectively. The hydrogen cyanide levels in the mainstream smoke of five popular brands of Indian cigarettes varied from 366 to 638 micrograms and in the mainstream smoke of four popular brands of bidis from 688 to 904 micrograms. Cheroot and cigarillos yielded 588 micrograms and 1119 micrograms hydrogen cyanide respectively. The values of benzo(a)pyrene content in Indian cigarettes varied from 85 to 114 ng and in bidis from 108 to 144 ng. Herbal bidi and cheroot had 1315 ng and 2519 ng benzo(a)pyrene respectively. Cigarettes were smoked as per international standard smoking conditions and the levels of noxious agent were found to be higher than in currently marketed western cigarettes. However, these levels in all the indigenous products including bidis cannot be directly compared with those of cigarettes as they were smoked under modified conditions.

Benzo(a)pyrene↗

Effects of dietary benzo(a) pyrene on intestinal phase I and phase II drug metabolizing systems in normal and vitamin A-deficient rats.

Feeding of vitamin A-deficient diet to male weanling rats for 10 weeks caused significant increase in the activities of Phase I enzyme system, i.e., cytochrome P-450, cytochrome b5 and arylhydrocarbon hydroxylase in the proximal, middle and distal segments of the intestine. Of the Phase II enzymes studied, UDP-glucuronyltransferase showed significant decrease whereas glutathione S-transferase showed significant increase. Treatment with benzo(a)pyrene caused greater induction in the levels of Phase I enzymes in deficient animals as compared to controls. In contrast to this, benzo(a)pyrene treatment induced the level of UDP-glucuronyltransferase in control rats more than in deficient rats. Intestinal NADPH cytochrome C-reductase and glutathione S-transferase remained insensitive to benzo(a)pyrene induction.

Animals↗

[Metabolic activation of benzo(a)pyrene by human amniotic fluid].

The in vitro activation of benzo(a)pyrene was studied in amniotic fluid from ten 4-month pregnant women. Benzo(a)pyrene monooxygenase and epoxide hydrolase activities were in the same range in amniotic fluid as in human liver. Glutathione epoxide transferase activity was markedly lower than in hepatocytes. Human amniotic fluid also catalyzed the formation of hydrocarbon metabolites mutagenic to Salmonella typhimurium TA98 (Ames system). Profiles of amniotic fluid aromatic hydrocarbons from non smokers exhibited low benzo(a)pyrene concentration (less than 0.1 ng/ml).

Amniotic Fluid↗

Regioselectivity of rat liver microsomal UDP-glucuronosyltransferase activities toward phenols of benzo(a)pyrene and dibenz(a,h)anthracene.

Inducibility of rat liver microsomal UDP-glucuronosyltransferase was investigated with regard to the substrate structure using 3-, 6-, 7-, 8-, and 9-hydroxybenzo(a)pyrene, all seven phenols of dibenz(a,h)anthracene, 3-hydroxybenz(a)anthracene, and 3-hydroxyfluoranthene as substrates. Glucuronide formation of the majority of the planar phenols was preferentially inducible by 3-methylcholanthrene (4- to 8-fold, group 1). However, glucuronidation of 8-hydroxybenzo(a)pyrene, 3-hydroxybenz(a)anthracene, and 3-hydroxydibenz(a,h)anthracene was markedly inducible by phenobarbital (3- to 8-fold, group 2). Glucuronidation of 9-hydroxybenzo(a)pyrene and 3-hydroxyfluoranthene was only moderately induced by the two inducers (less than 2-fold, group 3). Glucuronidation was also determined with purified phenol UDP-glucuronosyltransferase from 3-methylcholanthrene-treated rats. A close correlation (r = 0.95) was observed between purification factors (ratio of enzyme activities in purified enzyme and microsomes) and induction factors (ratio of microsomal enzyme activities from 3-methylcholanthrene-treated rats and untreated controls). Interestingly, differences in size and shape of group 1 and 2 substrates could be recognized. Group 1 substrates were shorter (less than 1.3 nm) and broader (greater than 1.1 nm) than group 2 substrates when viewed from the hydroxy group, along the axis of the C-O bond, to the plane of the polycyclic aromatic hydrocarbon, suggesting a distinct geometry of the binding site of the 3-methylcholanthrene-inducible phenol UDP-glucuronosyltransferase.

Animals↗

Inhibitory effect of nicotine on benzo(a)pyrene elimination and marked pulmonary metabolism of nicotine in isolated perfused rat lung.

Concomitant administration of nicotine to the perfusion medium of an isolated perfused rat lung decreased the pulmonary elimination of benzo(a)pyrene entering the lung by vascular access. This inhibitory effect of nicotine was observed at a concentration of 0.5 microM nicotine in the perfusion medium. The elimination half-life of benzo(a)pyrene given as a bolus to establish an initial concentration of 1 microM in the perfusate was increased by a factor of about 3 after the addition of nicotine at 60 min of perfusion. A comparably high increase of the benzo(a)pyrene steady-state concentration was observed during constant infusion experiments when nicotine as the interfering substrate was infused parallelly after a control period of 60 min. In perfused rat lungs the rate of metabolism of nicotine given as a bolus (initial concentration of 0.5 microM) was comparable to the rate in isolated rat livers for 5,6-benzoflavone, oil or saline pretreated animals. Induction of cytochrome P-450 by pretreatment of the rats with phenobarbital caused an eightfold increase of the nicotine clearance in isolated livers, whereas the pulmonary nicotine clearance was almost doubled. Overall, expressed per g organ weight, the isolated perfused rat lung exhibited a marked capacity to metabolize nicotine while even the maximally induced phenobarbital-stimulated rat liver reached only 60% of the corresponding clearance values (per g organ) of the isolated rat lung. For the 5,6-benzoflavone and control treatments, values of 14%-21% were estimated for the liver compared with lung.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Preferential modification of replicating DNA by benz(a)pyrene)].

The nuclei of cells from regenerating rat liver were incubated with benzo(a)pyrene and the concentrations of the metabolites that covalently bound to DNA of different nuclear fractions were compared. It appeared that DNA associated with nuclear matrix (containing replicating DNA) is modified most intensively. The synchronized mouse embryo cells were incubated with benzo(a)pyrene during S phase and the levels of modifications in short and long single-stranded DNA fragments were compared. It has been observed that replicating DNA is represented in short fragments. These short DNA fragments were found to be modified by benzo(a)pyrene 4-9 times more intensively than total DNA. The possible mechanisms of both the increase in the number of DNA modifications in proliferating cells and the reason for the enhancement of carcinogenic effect on dividing cells are being discussed.

Animals↗

Fatty acid modification of C3H 10T 1/2 fibroblast cells: changes in benzo(a)pyrene metabolism and phorbol ester binding.

The mouse embryo fibroblast cell line, C3H 10T 1/2 Cl8, was studied as an in vitro experimental model to investigate the mechanism and specificity behind the modulation of carcinogenesis by dietary lipid. The cells were grown in medium supplemented with 95 microM stearate, linoleate, or palmitate as fatty acid/albumin complexes, during which time they maintained normal growth and morphology characteristics. After 5 days of supplementation total cellular lipid fatty acid was enriched in the supplemented fatty acid. By Day 40, however, fatty acid profiles of all groups were the same. Cellular uptake and utilization of 14C-radiolabeled fatty acids were measured. Within 24 h of supplementation, label was incorporated into cholesterol and diglycerides, cholesterol ester, alkyldiacylglycerols, and phospholipids. Approximately half of the radiolabel was found in phosphatidylcholine. Supplementation significantly increased the rate of benzo(a)pyrene metabolism, but did not affect DNA modification by benzo(a)pyrene. Phorbol dibutyrate binding to C3H 10T 1/2 cells at 4 degrees C was modified by lipid supplementation. At 37 degrees C and 23 degrees C, phorbol dibutyrate binding was characterized by both high- and low-affinity sites for linoleate- and stearate-supplemented cells. At 4 degrees C high-affinity binding was absent in stearate- and palmitate-supplemented cells, but was maintained in linoleate-supplemented cells. These studies suggest that the unsaturated fatty acid content of the diet may not significantly affect the initiation stage of benzo(a)pyrene carcinogenesis, but may instead affect promotion. One possible mechanism for this could involve changes in the lipid microenvironment of membrane receptors involved in tumor promotion.

Animals↗

[Benzo(a)pyrene and the carcinogenic effect of coal tar].

The carcinogenic effects of coal tar and benzo(a)pyrene were compared. The agents were applied to murine skin, with coal tar dose being adjusted for benzo(a)pyrene. The carcinogenic effect of coal tar appeared higher than that of benzo(a)pyrene, the relative carcinogenic risk of the former being 3.2 times that of the latter. The coefficient of relative carcinogenic risk of coal tar calculated on the basis of TD56 regression coefficients for dose-effect relationship and mean latent period ratios was averagely 1.5.

Animals↗

Oxidation of benzo(a)pyrene by extracellular ligninases of Phanerochaete chrysosporium. Veratryl alcohol and stability of ligninase.

Benzo(a)pyrene was oxidized with crude and purified extracellular ligninase preparations from Phanerochaete chrysosporium. Both the crude enzyme and the purified fractions oxidized the substrate to three organic soluble products, namely benzo(a)pyrene 1,6-, 3,6-, and 6,12-quinones. These findings support the recent proposition that lignin-degrading enzymes are peroxidases, mediating oxidation of aromatic compounds via aryl cation radicals. The ligninase which was unstable in the presence of hydrogen peroxide could be stabilized by addition of 3,4-dimethoxy benzyl alcohol to the reaction mixture. The oxidation of benzo(a)pyrene was enhanced in the presence of this alcohol.

Basidiomycota↗