Trace organic contaminants, including toxaphene and trifluralin, in cotton field soils from Georgia and South Carolina, USA.

Residues of organic contaminants--including toxaphene, DDT, trifluralin, hexachlorocyclohexanes, polychlorinated biphenyls, polycyclic aromatic hydrocarbons (PAHs) and nonylphenol--were measured in 32 cotton field soils collected from South Carolina and Georgia in 1999. Toxaphene, trifluralin, DDT and PAHs were the major contaminants found in these soils. The maximum concentration of toxaphene measured was 2,500 ng/g dry weight. Trifluralin was detected in all the soils at concentrations ranging from 1 to 548 ng/g dry weight. Pesticide residues were not proportional to soil organic carbon content, indicating that their concentrations were a reflection of application history and dissipation rates rather than air-soil equilibrium. Soil extracts were also subjected to in vitro bioassays to assess dioxinlike, estrogenic, and androgenic/glucocorticoid potencies. Relatively more polar fractions of the soils elicited estrogenic and androgenic/glucocorticoid activities, but the magnitude of response was much less than those found in coastal marine sediments from industrialized locations.

CYP1B1 expression in human lung.

Cytochrome P450 1B1 is a recently recognized phase I bioactivating enzyme with high affinity for both inhaled tobacco carcinogens and 17beta-estradiol. We evaluated the human lung expression of this multifunctional member of the P450 superfamily across 16 individuals. Expression of CYP1B1 was evaluated by qualitative reverse transcription-polymerase chain reaction and Western immunoblots performed on human tumor and nontumor lung tissue. Expression at both mRNA and protein levels was then correlated with smoking history, plasma biomarkers of tobacco exposure (nicotine and cotinine), gender, and tumor histology. CYP1B1 mRNA and protein were detected in 94 and 100% of individuals, respectively. Multivariate analysis confirmed that there were more subjects displaying CYP1B1 mRNA expression in tumor than nontumor tissue (p = 0.0003). Correlation of CYP1B1 protein with plasma cotinine levels was statistically marginal (p = 0.027). Self-reported smoking history, gender, and tumor histology did not correlate with gene expression in the multivariate model. After multivariate modeling for confounding factors, the expression patterns of 5 of 16 individuals appeared to differ from the group as a whole for mRNA and/or protein. We conclude that CYP1B1 is commonly expressed in human lung and hypothesize that it may be an important phase I enzyme with respect to human lung carcinogen metabolism, warranting an understanding of regulatory control and coding region polymorphisms.

SULT1A1 catalyzes 2-methoxyestradiol sulfonation in MCF-7 breast cancer cells.

In a previous study of nine human breast-derived cell lines, rates of metabolism of 17beta-estradiol (E(2)) were greatly enhanced when cultures were exposed to the aromatic hydrocarbon receptor agonist, 2,3,7,8-tetrachlorodibenzo-p-dioxin. Elevated rates of E(2) hydroxylation at the C-2, -4, -6alpha and -15alpha positions were observed concomitant with the induction of cytochromes P450 1A1 and 1B1. In each cell line, 2- and 4-hydroxyestradiol (2- and 4-OHE(2)) were converted to 2- and 4-methoxyestradiol (2- and 4-MeOE(2)) by the action of catechol O:-methyltransferase. In this study, conjugation of these estrogen metabolites was investigated. A comparison of the levels of metabolites determined with and without prior treatment of the media with a crude beta-glucuronidase/sulfatase preparation showed that most of the 2-MeOE(2) present was in conjugated form, whereas 4-MeOE(2), 6alpha-OHE(2) and 15alpha-OHE(2) were minimally conjugated. Inhibitor studies suggested that it was the sulfatase activity of the preparation that hydrolyzed the 2-MeOE(2) conjugates in MCF-7 cell media; the presence of 2-MeOE(2)-3-sulfate in MCF-7 culture media was confirmed by electrospray ion-trap mass spectrometry. To identify the enzyme catalyzing this conjugation, the expression of mRNAs encoding five sulfotransferases (SULT1A1, SULT1A2, SULT1A3, SULT1E1 and SULT2A1) was evaluated in the nine cell lines by use of the reverse transcription-polymerase chain reaction. Only expression of SULT1A1 mRNA correlated with the observed conjugation of nanomolar levels of 2-MeOE(2) in these cell lines. Cloning and sequencing of SULT1A1 cDNA from MCF-7 cells revealed that mRNAs encoding two previously identified allelic variants, SULT1A1*1 ((213)Arg) and SULT1A1*2 ((213)His), were expressed in these cells. Heterologous cDNA-directed expression of either variant in MDA-MB-231 cells, which do not normally express SULT1A1, conferred 2-MeOE(2) sulfonation activity. The SULT1A1 allelic variants were also expressed in SF:9 insect cells, from which post-microsomal supernatants were used to determine K:(m) values of 0.90 +/- 0.12 and 0.81 +/- 0.06 microM for SULT1A1*1 and SULT1A1*2, respectively, with 2-MeOE(2) as substrate. These results show that SULT1A1 is an efficient and selective catalyst of 2-MeOE(2) sulfonation and, as such, may be important in modulating the anticarcinogenic effects of 2-MeOE(2) that have been described recently.

A simple, reliable method for the determination of chlorinated volatile organics in human breath and air using glass sampling tubes.

A method for determining chloroform, 1,1,1-trichloroethane, carbon tetrachloride, and trichloroethene in breath samples was developed. It consisted of collecting samples in 40-ml. glass-silanized tubes that were 16-in. long and had a 0.64-in. diameter. The ends tapered, resulting in a tube with a 1/4-in. diameter that was 1 3/4-in. long; each end had a shutoff valve attached. One end had a strip of rubber tube attached to the shutoff valve for collecting the breath sample, and the other end contained a 1/4-in. Swagelok nut with a rubber septum for withdrawing the sample. Samples were withdrawn using a pressure-lock, gastight syringe, and they were injected onto a gas chromatograph fitted with an electron-capture detector. The analytes were stable for at least 22 days in these tubes. The method detection limit was determined to be 0.03, 0.08, 0.04, and 0.04 pg/ml. for chloroform, 1,1,1-trichloroethane, carbon tetrachloride, and trichloroethene, respectively. Precision, based on 13 injections, was determined to be 13% for 0.09 pg chloroform, 13% for 0.21 pg 1,1,1-trichloroethane, 8% for 0.16 pg carbon tetrachloride, and 14% for 0.1 pg trichloroethene. In all, the proposed method is a sensitive and reliable one for determining volatile organic compounds in breath and a method that can also be applied to air sampling.

Human exposure to volatile organic compounds: a comparison of organic vapor monitoring badge levels with blood levels.

We undertook a study in Albany, New York, to investigate whether volatile organic compounds (VOCs) were measurable in the blood and in the breathing-zone air of people exposed to gasoline fumes and automotive exhaust. We sampled blood of 40 subjects, placed organic vapor badges on 40 subjects, and obtained personal breathing-zone samples from 24 subjects. We limited this analysis to 19 subjects who wore the organic vapor badges for at least 5 h. VOC levels, as determined by the organic vapor badges, were highly correlated with blood levels of these same compounds. Using detection in blood as the gold standard, we found the badges to be more sensitive than conventional charcoal tube samples in detecting low levels of methyl tert-butyl ether (0.60 vs 0.08), toluene (0.95 vs 0.64), and o-xylene (0.85 vs 0.64). In this study, organic vapor badges provided data on VOC exposure that correlated with blood assay results. These organic vapor badges might provide a convenient means of determining human exposure to VOCs in epidemiologic studies.

Stimulation of 17 beta-estradiol metabolism in MCF-7 cells by bromochloro- and chloromethyl-substituted dibenzo-p-dioxins and dibenzofurans: correlations with antiestrogenic activity.

Mixed halo- and haloalkyl-substituted dibenzo-p-dioxins (DD) and dibenzofurans (DF) are known environmental contaminants, although there is limited information on the toxic effects of these compounds in human cells. In this study antiestrogenicity, a property of 2,3,7,8-Cl4-DD, was investigated with a series of bromochloro- and chloromethyl-substituted DDs and DFs. The effects of these compounds on the metabolism of 17 beta-estradiol (E2) and on the estrogen-dependent formation of multicellular foci in cultures of MCF-7 human breast cells were examined. Pretreatment of MCF-7 cells with 2,3,7,8-Cl4-DD induced pathways of E2 metabolism involving cytochrome P-450-catalyzed hydroxylation, methylation of the catechol estrogens, and conjugation. Several Br-Cl3-DD and Br2-Cl2-DD congeners with halogen substitution at the 2, 3, 7, and 8 positions also stimulated E2 metabolism with similar potency to that of 2,3,7,8-Cl4-DD; however, compounds with substitution of a methyl group for a halogen at any of these positions did not stimulate the metabolism of E2. For the series of compounds tested in MCF-7 cultures, a close correlation was observed between the antiestrogenicity as measured by the inhibition of estrogen-dependent postconfluent growth that results in focus formation and the efficacy with which the compounds stimulated the metabolism of E2. 2,3,7,8-TetrahaloDDs with one or two bromine atoms at these positions were highly antiestrogenic as determined by their inhibition of estrogen-dependent focus formation, whereas the methyl-substituted polychlorinated DDs and DFs investigated did not inhibit focus formation. These results indicate that the 2,3,7,8-substituted mixed halo-substituted DDs and DFs are of importance when the biologic effects of halogenated DD and DF congeners are considered, and provide additional evidence for the role of increased metabolism of E2 in the antiestrogenic effects of halogenated DDs and DFs.

Analysis of oxidative warfarin metabolites by thermospray high-performance liquid chromatography/mass spectrometry.

Oxidative metabolites of the anticoagulant, warfarin [4-hydroxy-3-(3-oxo-1-phenylbutyl)-2H-1-benzopyran-2-one], produced by the actions of cytochromes P450 were analyzed by thermospray high-performance liquid chromatography/mass spectrometry. Warfarin, dehydrowarfarin, and the 6-, 7-, 8-, and 4'-hydroxy derivatives of warfarin were found to ionize well by the thermospray process in the presence of ammonium acetate. Thermospray mass spectra of these compounds were generally dominated by the protonated molecule, (M + H)+, and ions formed by the loss of water from the protonated molecule, (M + H - H2O)+. Fragment ions arising from the hydroxycoumarin, benzylhydroxycoumarin, and phenylbutanone portions of the molecules were observed, and the relative intensity of these fragment ions was greatly increased with filament ionization and application of a high repeller potential (100-130 V). Selected-ion monitoring of the (M + H)+ and (M + H - H2O)+ ions provided sensitivities for these compounds in the 2 to 10 ng range. A method employing thermospray HPLC/MS with selected-ion monitoring and internal standard quantitation for the analysis of the oxidative metabolites of warfarin is described.

Thermally induced formation of polychlorinated dibenzofurans from Aroclor 1254-contaminated mineral oil.

Numerous laboratory simulations and real-world events have demonstrated the thermal conversion of neat or high concentration of PCBs into the much more toxic PCDFs. Since millions of mineral oil transformers currently in service contain PCB concentrations in the 50 to 5000 ppm range, the thermal behavior of dilute PCB solutions is of practical and regulatory significance. In this work, neat Aroclor 1254 and 5000 ppm Aroclor 1254 in mineral oil were subjected to pyrolysis and combustion under a range of experimental conditions to define parameters resulting in maximal PCDF yields. The dependence of PCDF yield on Aroclor 1254 concentrations was then investigated in the 5000 to 50 ppm range. Combustion experiments demonstrated that PCDF yields expressed as micrograms PCDF/gram PCB were independent of concentration range, confirming that the process is kinetically first order in PCB. Much lower yields of PCDF were observed in the open tube pyrolysis experiments, as compared to combustion experiments and to earlier and concurrent sealed tube experiments. Slightly improved yields were observed in the pyrolysis experiments at lower concentrations, suggesting the existence of a PCB or PCDF destruction process of higher than first order kinetics. In all cases, yields expressed as micrograms PCDF/gram mixture were sharply and monotonically lower as concentrations decreased between neat or 5000 ppm Aroclor 1254 and 50 ppm Aroclor 1254.

Calculation of 2,3,7,8-TCDD equivalent concentrations of complex environmental contaminant mixtures.

Sufficient toxicological data are now available to permit use of conventional risk assessment techniques to estimate the hazards associated with human exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD). However, many real-world exposures involve complex mixtures of dibenzodioxins, dibenzofurans, and related compounds. Historical approaches to risk assessment on such mixtures have ranged from ignoring all compounds except 2,3,7,8-TCDD itself to assuming that all compounds have potencies equal to 2,3,7,8-TCDD. An alternative approach which uses existing literature data and analytical results to calculate the "2,3,7,8-TCDD equivalent" concentration of a mixture in order to "predict" its biological potency relative to 2,3,7,8-TCDD itself is advanced here. Previously reported in vivo acute and subchronic studies and some recently obtained analytical chemistry data are integrated here to clarify the utility of this important approach and to assess the uncertainties associated with its use. This predictive approach, and various conceptually similar ones, have now found wide applicability to the risk assessment process associated with exposure to complex mixtures of dioxins, dibenzofurans, and related compounds.

Chemical and biological investigations of a transformer accident at Binghamton, NY.

A transformer fire occurred in a state office building in Binghamton, NY on February 5, 1981. Particulates from inside surfaces of ceiling panels on 16 of the 17 floors had concentrations of polychlorinated dibenzofurans (PCDFs) ranging from less than 1 part per million (ppm) to 1200 ppm while polychlorinated biphenyl (PCB) concentrations varied from 28 ppm to 23,000 ppm. In spite of the wide variations in contaminant concentrations, complete analytical data from 11 floors showed that there was a consistent PCDF/PCB ratio (0.067 +/- 0.026) and also consistent PCDF isomer group distributions (tetra-CDFs, 33 +/- 5%; penta-CDFs, 40 +/- 3%; hexa-CDFs, 18 +/- 7%; hepta-CDFs, 6 +/- 3%). It was found that the particulate samples could be successfully ranked in order of their degree of chemical contamination by an in vitro bioassay. The bioassay was based on induction of keratinization or changes in morphology in mouse epithelial cells. Animal toxicology experiments were carried out with a soot sample containing a PCDF concentration which approximated the mean value found on the ceiling particulates. The single dose oral LD values of the soot and its benzene extract equivalent, each administered to female guinea pigs in 0.75% methyl cellulose, were 410 and 327 mg/kg, respectively. These results demonstrated that the soot matrix had virtually no effect on the toxicity of the chemical contaminants in the soot. Morphological alterations in liver tissues from animals receiving the soot were found after examination by electron and light microscopy.(ABSTRACT TRUNCATED AT 250 WORDS)

Lead content of foodstuffs.

The lead content of a number of foodstuffs, particularly baby fruit juices and milk, is reported. Samples were analyzed in quadruplicate by using an automated Delves cup atomic absorption procedure. A large proportion of the products examined contained significant amounts of lead. Of 256 metal can examined, the contents of 62% contained a lead level of 100 mug/l. or more, 37% contained 200 mug/l. or more and 12% contained 400 mug/l. lead or more. Of products in glass and aluminum containers, only 1% had lead levels in excess of 200 mug/l. Lead levels of contents also correlate with the seam length/volume ratio of the leaded seam can. A survey of bulk milk showed a mean lead level of 40 mug/l. for 270 samples; for canned evaporated milk the mean level was 202 mug/l. These data indicate a potential health hazard.

Some optical studies with the air-acetylene flame.

The application of Schlieren and shadow techniques for the study of flame processes is discussed in relation to analytically useful premixed flames. The information obtained, particularly with shadowgraphs, may be correlated with the measured signals and signal noise in atomic-absorption and flame-emission spectrometry.