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Purpose: Crude coal tar and its derivatives have been used in modern medicine for the treatment of psoriasis since at least 1925 as part of the Goeckerman regimen. To this day, coal tar remains a safe and highly effective option for the treatment of psoriasis vulgaris. However, the mechanism by which coal tar has its therapeutic effect is unknown. This review summarizes current knowledge of the mechanism by which coal tar has its therapeutic effect in the treatment of psoriasis vulgaris., Material and Methods: A Pubmed search was conducted on March 13, 2017 for relevant English language journal articles on the subject and were relevant journal articles were included in this review., Results: Crude coal tar consists of thousands of ingredients, many of which are unidentified. Of these ingredients, the most research has gone into analyzing polycyclic aryl hydrocarbons. These hydrocarbons are thought to be the most likely component of crude coal tar that leads to its effects in psoriasis. Of the aryl hydrocarbons, carbazole has been the most well studied in psoriasis and is hypothesized as being responsible for the treatment efficacy of crude coal tar., Conclusions: Polycyclic aryl hydrocarbons, and specifically carbazole, are thought to be the mechanism by which crude coal tar has its effect in psoriasis. However, further research is warranted to fully characterize the mechanism of action of crude coal tar, with the potential to create new therapies for psoriasis.

Objective: To evaluate the inner or extra-exposure of workers in carbon and coal tar process industries. Methods: In May 2015, 271 exposure and 75 non-exposure workers in two of carbon and coal tar process industries in Shandong province were selected. Coal tar pitch (CTP) was detected according to the methods of GB17054-1997. The 1-OH-Nap、2-OH-Nap、1-OH-Pyr in urine were detected by high performance liquid Chromatography (HPLC) . Results: The concentration of 1-OH-Nap、2-OH-Nap、1-OH-Pyr in urine of exposure group were significant higher than that of control group ( P <0.05) , respectively. The urine concentration of 1-OH-Nap、2-OH-Nap、1-OH-Py in contact group tar and asphalt were 12.20, 12.55, 7.08 and 10.62, 8.73, 3.07 μg/gCr, and the concentration of them were 7.25, 8.54, 3.00 μg/gCr in container handle workers. Significant positive correlations were found between high or median CTP exposure and the urine concentration of 1-OH-Nap、2-OH-Nap、1-OH-Pyr in exposure groups ( P <0.05) . Conclusions: The inner or extra exposure level is relative and different for different type of work in these carbon and coal tar process industries.

Goeckerman therapy (GT) for psoriasis combines the therapeutic effect of crude coal tar (CCT) and ultraviolet radiation (UVR). CCT contains polycyclic aromatic hydrocarbons, some of which can form DNA adducts that may induce mutations and contribute to carcinogenesis. The aim of our work was to evaluate the relationship between concentrations of benzo[a]pyrene-7,8-diol-9,10-epoxide-DNA adducts (BPDE-DNA adducts) and rs4646903 (CYP1A1 gene), rs1048943 (CYP1A1), rs1056836 (CYP1B1), rs1051740 (EPHX1), rs2234922 (EPHX1) and rs8175347 (UGT1A1) polymorphic sites, and GSTM1 null polymorphism in 46 patients with chronic stable plaque psoriasis who underwent GT. The level of BPDE-DNA adducts was determined using the OxiSelect BPDE-DNA Adduct ELISA Kit. Polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis (rs4646903, rs1048943, rs1051740, and rs2234922), fragment analysis (rs8175347), real-time PCR (rs1056836), and digital droplet PCR polymorphism (GSTM1) were used. CYP1B1*1/*1 wild-type subjects and CYP1B1*3/*1 heterozygotes for rs1056836 formed significantly higher amounts of BPDE-DNA adducts than CYP1B1*3/*3 homozygotes (p=0.031 and p=0.005, respectively). Regarding rs1051740, individuals with EPHX1*3/*1 heterozygosity revealed fewer adducts than EPHX1*1/*1 wild-type subjects (p=0.026). Our data suggest that CYP1B1/EPHX1 genotyping could help to predict the risk of DNA damage and to optimize doses of coal tar and UVR exposure in psoriatic patients in whom GT was applied., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Polaromonas naphthalenivorans strain CJ2 is a Gram-negative betaproteobacterium that was identified, using stable isotope probing in 2003, as a dominant in situ degrader of naphthalene in coal tar-contaminated sediments. The sequenced genome of strain CJ2 revealed several genes conferring nitrogen fixation within a 65.6 kb region of strain CJ2's chromosome that is absent in the genome of its closest sequenced relative Polaromonas sp. strain JS666. Laboratory growth and nitrogenase assays verified that these genes are functional, providing an alternative source of nitrogen in N-free media when using naphthalene or pyruvate as carbon sources. Knowing this, we investigated if nitrogen-fixation activity could be detected in microcosms containing sediments from the field site where strain CJ2 was isolated. Inducing nitrogen limitation with the addition of glucose or naphthalene stimulated nitrogenase activity in amended sediments, as detected using the acetylene reduction assay. With the use of fluorescence microscopy, we screened the microcosm sediments for the presence of active strain CJ2 cells using a dual-labelling approach. When we examined the carbon-amended microcosm sediments stained with both a strain CJ2-specific fluorescent in situ hybridization probe and a polyclonal fluorescently tagged antibody, we were able to detect dual-labelled active cells. In contrast, in sediments that received no carbon addition (showing no nitrogenase activity), no dual-labelled cells were detected. Furthermore, the naphthalene amendment enhanced the proportion of active strain CJ2 cells in the sediment relative to a glucose amendment. Field experiments performed in sediments where strain CJ2 was isolated showed nitrogenase activity in response to dosing with naphthalene. Dual-label fluorescence staining of these sediments showed a fivefold increase in active strain CJ2 in the sediments dosed with naphthalene over those dosed with deionized water. These experiments show that nitrogen fixation may play an important role in naphthalene biodegradation by strain CJ2 and contribute to its ecological success., (© 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.)

Bioaccumulation of polycyclic aromatic hydrocarbons (PAHs) from coal tar pitch polluted sediments was predicted by (1) a generic approach based on organic carbon-water partitioning and Gibbs linear free energy relationship (between K(OW) and K(OC)), and (2) measurements of freely dissolved concentrations of PAHs in the sediment pore water, using passive samplers and solid phase extraction. Results from these predictions were compared with those from in vivo bioaccumulation experiments using Nereis diversicolor (Polychaeta), Hinia reticulata (Gastropoda), and Nuculoma tenuis (Bivalvia). Measured sediment/water partition coefficients were higher than predicted by the generic approach. Furthermore, predicted biota-to-sediment accumulation factors (BSAFs) derived from measured pore water concentrations were more in agreement with the bioaccumulation observed for two of the three species. Discrepancies associated with the third species (N. tenuis) were likely a result of particles remaining in the intestine (as shown by microscopic evaluation). These results indicate the importance of conducting site-specific evaluations of pore water concentrations and/or bioaccumulation studies by direct measurements to accurately provide a basis for risk assessment and remediation plans. The importance of knowledge regarding specific characteristics of model organisms is emphasized.

The monitoring of a windrow treatment applied to soil contaminated by mostly 2-, 3- and 4-ring PAHs produced by coal tar distillation was performed by following the evolution of both PAH concentration and the bacterial community. Total and PAH-degrading bacterial community structures were followed by 16S rRNA PCR-DGGE in parallel with quantification by bacterial counts and 16 PAH measurements. Six months of biological treatment led to a strong decrease in 2-, 3- and 4-ring PAH concentrations (98, 97 and 82% respectively). This result was associated with the activity of bacterial PAH-degraders belonging mainly to the Gamma-proteobacteria, in particular, the Enterobacteria and Pseudomonas genera, which were detected over the course of the treatment. This group was considered to be a good bioindicator to determine the potential PAH biodegradation of contaminated soil. Conversely, other species, like the Beta-proteobacteria, were detected after 3months, when 2-, 3- and 4-ring PAHs were almost completely degraded. Thus, presence of the Beta-proteobacteria group could be considered a good candidate indicator to estimate the endpoint of biotreatment of this type of PAH-contaminated soil., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Background: Goeckerman therapy (GT) for psoriasis is based on cutaneous application of crude coal tar [polycyclic aromatic hydrocarbons (PAH)] and exposure to ultraviolet radiation (UVR). PAH and UVR are mutagenic, carcinogenic and immunotoxic agents that promote apoptosis., Methods: We evaluated dermal absorption of PAH as well as the genotoxic and apoptotic effects of GT in 20 patients with psoriasis, by determining numbers of chromosomal abnormalities in peripheral lymphocytes, and levels of 1-hydroxypyrene (1-OHP), p53 protein and soluble FasL (sFasL) in urine and/or blood, before and after GT. Psoriasis Area and Severity Index (PASI) score was used to evaluate clinical efficacy of GT., Results: Compared with pre-treatment levels, there was a significant increase in urine 1-OHP, indicating a high degree of dermal absorption of PAH (P < 0.01). We also found a significant increase in the number of chromosomal abnormalities in peripheral blood lymphocytes (P < 0.001), suggesting that GT is genotoxic; significantly increased p53 protein in plasma (P < 0.05), an indicator of cell response to DNA damage; and significantly increased sFasL in serum (P < 0.01), an indicator of apoptosis. The PASI score was significantly decreased after GT (P < 0.001), confirming clinical benefit of this treatment., Conclusion: Our results demonstrate high dermal absorption of PAH during GT and provide evidence that GT promotes genotoxicity and apoptosis.

The propensity for groundwater ecosystems to recover from contamination by organic chemicals (in this case, coal-tar waste) is of vital concern for scientists and engineers who manage polluted sites. The microbially mediated cleanup processes are also of interest to ecologists because they are an important mechanism for the resilience of ecosystems. In this study we establish the long-term dynamic nature of a coal-tar waste-contaminated site and its microbial community. We present 16 years of chemical monitoring data, tracking responses of a groundwater ecosystem to organic contamination (naphthalene, xylenes, toluene, 2-methyl naphthalene and acenaphthylene) associated with coal-tar waste. In addition, we analyzed small-subunit (SSU) ribosomal RNA (rRNA) genes from two contaminated wells at multiple time points over a 2-year period. Principle component analysis of community rRNA fingerprints (terminal-restriction fragment length polymorphism (T-RFLP)) showed that the composition of native microbial communities varied temporally, yet remained distinctive from well to well. After screening and analysis of 1178 cloned SSU rRNA genes from Bacteria, Archaea and Eukarya, we discovered that the site supports a robust variety of eukaryotes (for example, alveolates (especially anaerobic and predatory ciliates), stramenopiles, fungi, even the small metazoan flatworm, Suomina) that are absent from an uncontaminated control well. This study links the dynamic microbial composition of a contaminated site with the long-term attenuation of its subsurface contaminants.

Expedited site characterization and groundwater monitoring using direct-push technology and conventional monitoring wells were conducted at a former manufactured gas plant site. Biogeochemical data and heterotrophic plate counts support the presence of microbially mediated remediation. By superimposing solutions of a two-dimensional reactive transport analytical model, first-order degradation rate coefficients ((day-1) ) of various compounds for the dissolved-phase plume were estimated (i.e., benzene [0.0084], naphthalene [0.0058], and acenaphthene [0.0011]). The total mass transformed by aerobic respiration, nitrate reduction, and sulfate reduction around the free-phase coal-tar dense-nonaqueous-phase-liquid region and in the plume was estimated to be approximately 4.5 kg/y using a biogeochemical mass-balance approach. The total mass transformed using the degradation rate coefficients was estimated to be approximately 3.6 kg/y. Results showed that a simple two-dimensional analytical model and a biochemical mass balance with geochemical data from expedited site characterization can be useful for rapid estimation of mass-transformation rates.

The biodegradation of 16 USEPA-listed PAHs (SigmaPAHs) during simulated in-vessel composting-bioremediation of an aged coal-tar-contaminated soil amended with fresh green waste compost (FGWC) collected from two landfill sites in the United Kingdom (UK) were studied over 56 days. The experimental design compared three constant temperature profiles (TC=38, 55 and 70 degrees C) with one variable temperature profile including treatment at 70 degrees C to comply with regulatory requirements (TP1). The highest disappearance of SigmaPAHs was observed in the soil amended with FGWC (53.2% and 48.1% SigmaPAHs disappearance in soil amended with FGWC-Site 1 and FGWC-Site 2, respectively) containing lower initial organic mater (TOM) (Initial TOM(FGWC-Site 1)=25.6+/-0.6%

In-vessel composting of an aged coal-tar contaminated soil from a manufactured gas plant site was investigated over 98days using laboratory-scale in-vessel composting reactors. The composting reactors were operated at 18 different operational conditions using a logistic three-factor factorial design with three temperatures (T=38, 55 and 70 degrees C), four soil to green waste ratios (S:GW; 0.6:1, 0.7:1, 0.8:1 and 0.9:1 on a dry weight basis) and three moisture contents (MC; 40%, 60% and 80%). Excitation-emission matrix (EEM) fluorescence spectroscopy was used to investigate organic matter dynamics in the composting mixture. The results of this investigation indicated that formation of humic substances can be monitored by fluorescence excitation-emission matrix, and provided evidence of progressive mineralization or humification of the composting mixture. Peak excitation wavelength shifts and peak fluorescence intensity can both be used as indicators to monitor the humification or maturation of compost. Finally, the fluorescence index can be applied to investigate the origin of humic substances and fulvic acids, and the humification or maturation of compost.

Several distinct naphthalene dioxygenases have been characterized to date, which provides the opportunity to investigate the ecological significance, relative distribution, and transmission modes of the different analogs. In this study, we showed that a group of naphthalene-degrading isolates from a polycyclic aromatic hydrocarbon (PAH)-contaminated hillside soil were phenotypically and genotypically distinct from naphthalene-degrading organisms isolated from adjacent, more highly contaminated seep sediments. Mineralization of (14)C-labeled naphthalene by soil slurries suggested that the in situ seep community was more acclimated to PAHs than was the in situ hillside community. phnAc-like genes were present in diverse naphthalene-degrading isolates cultured from the hillside soil, while nahAc-like genes were found only among isolates cultured from the seep sediments. The presence of a highly conserved nahAc allele among gram-negative isolates from the coal tar-contaminated seep area provided evidence for in situ horizontal gene transfer and was reported previously (J. B. Herrick, K. G. Stuart-Keil, W. C. Ghiorse, and E. L. Madsen, Appl. Environ. Microbiol. 63:2330-2337, 1997). Natural horizontal transfer of the phnAc sequence was also suggested by a comparison of the phnAc and 16S ribosomal DNA sequences of the hillside isolates. Analysis of metabolites produced by cell suspensions and patterns of amplicons produced by PCR analysis suggested both genetic and metabolic diversity among the naphthalene-degrading isolates of the contaminated hillside. These results provide new insights into the distribution, diversity, and transfer of phnAc alleles and increase our understanding of the acclimation of microbial communities to pollutants.

The influence of the white rot fungus Pleurotus ostreatus on the degradation of selected poly- and heterocyclic aromatic hydrocarbons (referred to as polycyclic aromatic hydrocarbons [PAHs]) in soil was investigated under field conditions representing the Northern temperate zone. Pleurotus ostreatus was added to two contaminated soils in the form of homogenized refuse from the commercial production of fungus. The soils were collected from a former shipyard (the B&W soil) and underneath a former coal tar storage at an old asphalt factory in Denmark (the Ringe soil). Treatments (control, soil mixed with autoclaved sawdust medium, and soil mixed with P. ostreatus refuse) were set up in triplicate in concrete cylinders (height, 50 cm; diameter, 60 cm). The activity of P. ostreatus was measured as laccase activity and phenanthrene (PHE)- and pyrene (PYR)-degrading bacteria were enumerated. Twenty-one different PAHs were quantified. After nine weeks the concentrations of the 3-, 4-, 5-, and 6-ring PAHs in the Ringe soil were reduced by 78, 41, and 4%, respectively. These reductions corresponded with high initial laccase activity, a decrease in pH caused by the fungus, and an increase in the number of PHE- and PYR-degrading bacteria. No significant PAH degradation was observed in the B&W soil. Reasons for the difference in performance of P. ostreatus in the two soils are discussed in terms of soil histories and bioavailability. The use of P. ostreatus refuse holds promising potential for bioremediation purposes.

Coal tars in soil at a gasworks site in South Eastern Australia led to groundwater contamination with polycyclic aromatic hydrocarbons (PAHs), mono-aromatic compounds (BTEX) and phenols. The scope of the study included testwork in laboratory scale bioreactors and evaluation of available commercial groundwater treatment units. Two bioreactor configurations, a submerged fixed film reactor (SFFR) and a fluidized bed bioreactor (FBR) were effective, with high efficiencies of contaminant removal (typically >90%) over a range of hydraulic retention times (HRT) (3-29 h). Specifically, concentrations of total PAH, naphthalene, pyrene and total phenols in the feedstock and effluent of the SFFR were 123, 60, 51, 1.38 and 0.004, 0.001, 0.004, 0.1mg/l, respectively. The FBR was only marginally less effective than the SFFR for the same groundwater contaminants. Discharge to sewer was the most appropriate end use for the effluent. SFFRs are regarded as being simpler in design and operation, and a commercially available unit has been identified which would be suitable for treating small volumes (<10 m(3) per day) of contaminated water collected at an interception trench at the site.

The aim of this study was to investigate the genotoxic potential of DNA adducts and to compare DNA adduct levels and patterns in petroleum vacuum distillates, coal tar distillate, bitumen fume condensates, and related substances that have a wide range of boiling temperatures. An in vitro assay was used for DNA adduct analysis with human and rat S-9 liver extract metabolic activation followed by 32P-postlabeling and 32P-high-performance liquid chromatography (32p-HPLC). For petroleum distillates originating from one crude oil there was a correlation between in vitro DNA adduct formation and mutagenic index, which showed an increase with a distillation temperature of 250 degrees C and a peak around a distillation point of approximately 400 degrees C. At higher temperatures, the genotoxicity (DNA adducts and mutagenicity) rapidly declined to very low levels. Different petroleum products showed a more than 100-fold range in DNA adduct formation, with severely hydrotreated base oil and bitumen fume condensates being lowest. Coal tar distillates showed ten times higher levels of DNA adduct formation than the most potent petroleum distillate. A clustered DNA adduct pattern was seen over a wide distillation range after metabolic activation with liver extracts of rat or human origin. These clusters were eluted in a region where alkylated aromatic hydrocarbons could be expected. The DNA adduct patterns were similar for base oil and bitumen fume condensates, whereas coal tar distillates had a wider retention time range of the DNA adducts formed. Reference substances were tested in the same in vitro assay. Two- and three-ringed nonalkylated aromatics were rather low in genotoxicity, but some of the three- to four-ringed alkylated aromatics were very potent inducers of DNA adducts. Compounds with an amino functional group showed a 270-fold higher level of DNA adduct formation than the same structures with a nitro functional group. The most potent DNA adduct inducers of the 16 substances tested were, in increasing order, 9,10-dimethylanthracene, 7,12-dimethylbenz[a]anthracene and 9-vinylanthracene. Metabolic activation with human and rat liver extracts gave rise to the same DNA adduct clusters. When bioactivation with material from different human individuals was used, there was a significant correlation between the CYP 1A1 activity and the capacity to form DNA adducts. This pattern was also confirmed using the CYP 1A1 inhibitor ellipticine. The 32P-HPLC method was shown to be sensitive and reproducible, and it had the capacity to separate DNA adduct-forming substances when applied to a great variety of petroleum products.

Four white-rot fungi (Phanerochaete chrysosporium IMI 232175, Pleurotus ostreatus from the University of Alberta Microfungus Collection IMI 341687, Coriolus versicolor IMI 210866 and Wye isolate #7) and all possible combinations of two or more of these fungi, were incubated in microcosms containing wheat straw and non-sterile coal-tar contaminated soil to determine their potential to degrade polycyclic aromatic hydrocarbons (PAHs). Biotic and abiotic controls were prepared similarly and PAH concentrations remaining in each microcosm were determined after 8, 16 and 32 weeks by GC-MS following extraction with dichloromethane. The greatest PAH losses were in the biotic control, compared to small or negligible differences in microcosms inoculated with one or more fungi. These results suggest that in the biotic control native microorganisms colonised the straw added as organic substrate and degraded PAH as an indirect consequence of their metabolism. By contrast, in other microcosms, colonisation of straw by the natural microflora was inhibited because the straw was previously inoculated with fungi. Soil cultures prepared at the end of the experiment showed that though introduced fungi were still alive, they were unable to thrive and degrade PAH in such a highly contaminated soil and remained in a metabolically inactive form.

Coal tar is a complex mixture containing hundreds of compounds, at least 30 of which are polycyclic aromatic hydrocarbons, including the carcinogen benzo[a]pyrene (BaP). Although humans are exposed to complex mixtures on a daily basis, the synergistic or individual effects of components within a mixture on the carcinogenic process remain unclear. We have compared DNA adduct formation and cell proliferation in mice fed coal tar or BaP for 4 weeks with tumor formation in a 2 year chronic feeding study. Additionally, we have analyzed tumor DNA for mutations in the K-ras, H-ras and p53 genes. In the forestomach of mice fed either coal tar or BaP an adduct indicative of BaP was detected, with adduct levels increasing in a dose-responsive manner. K-ras mutations were detected in the forestomach tumors, with the incidence being similar in mice fed coal tar or BaP. These results suggest that the BaP within coal tar is associated with forestomach tumor induction in coal tar-fed mice. DNA adduct levels in the small intestine were not predictive of tumor incidence in this tissue; instead, the tumors appeared to result from compound-induced cell proliferation at high doses of coal tar. K-ras mutations were detected in lung tumors. Since lung tumors were not increased by BaP, coal tar components other than BaP appear to be responsible for the tumors induced in this tissue. H-ras mutations, primarily occurring at codon 61, were the most common mutation observed in liver tumors induced by coal tar. Since this mutation profile is observed in spontaneous hepatic tumors, components in the coal tar may be promoting the expansion of pre-existing lesions.

Twelve bacterial strains isolated from tar-contaminated soil were subjected to a polyphasic taxonomic study. The strains possessed meso-diaminopimelic acid as the diagnostic diamino acid of the peptidoglycan, MK-9(H2) as the predominant menaquinone, long-chain mycolic acids of the Gordonia-type, straight-chain saturated and monounsaturated fatty acids, and considerable amounts of tuberculostearic acid. The G + C content of the DNA was 68 mol%. Chemotaxonomic and physiological properties and 16S rDNA sequence comparison results indicated that these strains represent a new species of the genus Gordonia. Because of the ability of these strains to use alkanes as a carbon source, the name Gordonia alkanivorans is proposed. The type strain of Gordonia alkanivorans sp. nov. is strain HKI 0136T (= DSM 44369T).

The biological degradation of complex mixtures of recalcitrant substances is still a major challenge in environmental biotechnology and the remediation of coal-tar constitutes one such problem area. Biofilm bioreactors offer many advantages and may be successfully used for this purpose. Two stirred-tank reactors and one packed-bed reactor were tested in a continuous mode. Continuous cultivation allows microbial selection to take place whilst adhesive growth provides a high degradation capacity and process stability. The reactors were inoculated with mixed microbial populations to favour complete metabolism and to prevent metabolite accumulation and substrate inhibition effects. Phenol, o-cresol, quinoline, dibenzofuran, acenaphthene and phenanthrene were used as model contaminants and constituted the sole energy and carbon sources. The hydraulic retention time (HRT) was initially set to 2.5 days for a period of several months to allow the establishment of a stable biofilm and was then gradually decreased. All the compounds were found to be degraded by more than 90% at HRT of 3 h or more. Neither substrate inhibition nor metabolite accumulation effects were observed. The stirred-tank configuration was found to be the most efficient for use with high loads. No improvement in the degradation capacity could be achieved by increasing the biofilm surface in these reactors, illustrating that the limiting factor may be the mass transfer limitations rather than the availability of the biofilm surface. Finally, anaerobic treatment was successfully achieved, confirming the potential for remediation of contaminated sites under anaerobic conditions, providing that alternative electron acceptors are present.

Fifteen hospitalized, non-smoking, dermatological patients were treated with ointment containing 2% coal tar (CT) in order to assess the influence of metabolic genotype GSTM1 on urinary mutagen levels. Urinary 1-pyrenol, the main metabolite of pyrene, was used to check the high exposure to PAH of this population. The mean levels of urinary 1-pyrenol found in the 24-h urine of our patients were 467. 8+/-211.0 nmoles-24 h (range 94.6-890.1 nmoles-24 h). Mutagenicity was assessed on urine samples collected over a period of 24 h, after three consecutive days of topical application, using the bacterial mutagenesis test on Salmonella typhimurium strains TA98 and YG1024 in the presence of microsomal enzymes. The latter strain turned out to be more sensitive than the former in revealing urinary mutagens in these patients (42 693+/-30 867 vs. 6877+/-6040 net revertants-24 h). The mutagenicity on YG1024 strain and 1-pyrenol levels of urine samples were correlated (Spearman's rank correlation coefficient=0. 6678, P<0.01, z=2.795). The influence of genotype GSTM1 on urinary mutagen levels was assessed on strain YG1024. The values of urinary mutagenicity of subjects with genotype GSTM1-null (n=6) were on average higher than those of GSTM1-positive subjects (n=9) (55 498+/-45 957 vs. 34 156+/-11 933 net rev.-24 h), a non-significant statistical difference. The mean total excretion of mutagens corrected for PAH exposure (net rev./nmoles of urinary 1-pyrenol) in GSTM1-null patients was double that of GSTM1-positive ones (136. 8+/-34.7 vs. 70.8+/-23.3 net rev./nmoles of urinary 1-pyrenol; one-tailed Mann-Whitney U-test, U=11.5, P<0.05). These results indicate a greater body burden of promutagens, resulting from skin application of CT, in GSTM1-null subjects., (Copyright 1999 Elsevier Science B.V.)

A group of eczema patients topically treated with coal tar (CT) ointments was used as a model population to examine the applicability of DNA adducts in WBC subpopulations as a measure of dermal exposure to polycyclic aromatic hydrocarbons (PAHs). Aromatic DNA adducts were examined by 32P-postlabeling in exposed skin and WBC subsets, and urinary excretion of PAH metabolites was determined to assess the whole-body burden. The median urinary excretion of 1-hydroxypyrene and 3-hydroxybenzo(a)pyrene was 0.39 (range, 0.12-1.57 micromol/mol creatinine) and 0.01 micromol/mol creatinine (range, <0.01-0.04 micromol/mol creatinine), respectively, before the dermal application of CT ointments. After treatment for 1 week, these levels increased to 139.7 (range, 26.0-510.5 micromol/mol creatinine) and 1.18 micromol/mol creatinine (range, <0.01-2.14 micromol/mol creatinine), respectively, indicating that considerable amounts of PAHs were absorbed. Median aromatic DNA adduct levels were significantly increased in skin from 2.9 adducts/10(8) nucleotides (nt; range, 0.7-10.0 adducts/10(8) nt) before treatment to 63.3 adducts/10(8) nt (range, 10.9-276.2 adducts/10(8) nt) after treatment with CT, in monocytes from 0.28 (range, 0.25-0.81 adducts/10(8) nt) to 0.86 adducts/10(8) nt (range, 0.56-1.90 adducts/10(8) nt), in lymphocytes from 0.33 (range, 0.25-0.89 adducts/10(8) nt) to 0.89 adducts/10(8) nt (range, 0.25-3.01 adducts/10(8) nt), and in granulocytes from 0.28 (range, 0.25-0.67 adducts/10(8) nt) to 0.54 adducts/10(8) nt (range, 0.25-1.58 adducts/10(8) nt). A week after stopping the CT treatment, the DNA adduct levels in monocytes and granulocytes were reduced to 0.38 (range, 0.25-0.71 adducts/10(8) nt) and 0.38 adducts/10(8) nt (range, 0.25-1.01 adducts/10(8) nt), respectively, whereas the adduct levels in lymphocytes remained enhanced [1.59 adducts/10(8) nt (range, 0.25-2.40 adducts/10(8) nt)]. Although the adduct profiles in skin and WBC subsets were not identical, and the adduct levels in WBCs were significantly lower as compared with those in skin, the total DNA adduct levels in skin correlated significantly with the adduct levels in monocytes and lymphocytes, but not with those in granulocytes. Excretion of urinary metabolites during the first week of treatment was correlated with the percentage of the skin surface treated with CT ointment and decreased to background levels within a week after the cessation of treatment. 3-Hydroxybenzo(a)pyrene excretion, but not that of 1-hydroxypyrene, correlated significantly with the levels of DNA adducts in skin that comigrated with benzo(a)pyrene-diol-epoxide-DNA. This study indicates that the DNA adduct levels in mononuclear WBCs can possibly be used as a surrogate for skin DNA after dermal exposure to PAHs.

The importance of chemical conditions and mass transfer effects to in situ bioremediation of PAHs is presented using a case study. In situ bioremediation is being evaluated as a means for remediating a coal-tar contaminated aquifer at the site of a former manufactured gas plant. Two objectives of this work have been to evaluate the potential for the indigenous bacteria to biodegrade coal tar constituents and to identify factors controlling biodegradation rates. Aquifer sediments collected from a variety of locations across the site contain bacteria capable of aerobically mineralizing some of the principal aromatic compounds in the groundwater plume (benzene, naphthalene, and phenanthrene). Parallel mineralization assays incubated under aerobic and anaerobic conditions strongly suggest that O2 availability is a primary factor controlling the rate and extent of biodegradation. Data indicate that sorption may have also significantly affected biodegradation rates by limiting the bioavailability of the aromatic compounds. A mass transfer-limited numerical model was developed to explore the effect of sorption and bioavailability on biodegradation rates. In this model biodegradation rates are proportional to aqueous concentration, which is directly reduced by sorption. Both biotransformation and bacterial growth are described as being controlled by the rate of desorptive mass transfer. The influence of sorption on biodegradation is quantified by defining a Bioavailability Factor, Bf. A Thiele Modulus which indicates the ratio of characteristic times for sorption and biodegradation is helpful for determining the extent of mass transfer control during biodegradation of the aromatic compounds. This approach is preferred to equilibrium partitioning models, which may overestimate biodegradation rates by failing to consider the effect of rate-limited desorption on bioavailability.

An in vitro system was utilized to measure DNA adduct-forming ability of petroleum oils and oil coal tar mixtures to define correlations between DNA adduct levels and their mutagenic potencies. The system consisted of reaction of dimethyl sulfoxide extracts of oils with calf thymus DNA in the presence of Aroclor-induced hamster liver microsomes for 30 min. Following DNA extraction, DNA adducts were measured by the nuclease P1-enhanced postlabeling assay coupled with two-dimensional polyethyleneimine (PEI)-cellulose TLC. Thin layer plates showed putative aromatic DNA adducts, with levels ranging from 60 to 1400 adducts per 10(9) DNA nucleotides. TLC mobilities suggested adducts to be aromatic compounds containing 4 or more rings. A good correlation (coefficient of correlation = 0.91) was observed between DNA adduct levels and Salmonella mutagenicity for 19 oils. All 19 samples tested produced DNA adducts. To expedite the TLC procedure, adducts were resolved by one-dimensional TLC and the radioactivity measured using a mechanical scanner. Results were comparable to those obtained by two-dimensional TLC and quantification after scraping. Our data show that the in vitro incubation system coupled with the postlabeling adduct assay is a useful screening method to identify mutagenic and potentially carcinogenic oils.

Horizontal transfer of genes responsible for pollutant biodegradation may play a key role in the evolution of bacterial populations and the adaptation of microbial communities to environmental contaminants. However, field evidence for horizontal gene transfer between microorganisms has traditionally been very difficult to obtain. In this study, the sequences of the 16S rRNA and naphthalene dioxygenase iron-sulfur protein (nahAc) genes of nine naphthalene-degrading bacteria isolated from a coal tar waste-contaminated site, as well as a naphthalene-degrading bacterium from a contaminated site in Washington state and two archetypal naphthalene-degrading strains, were compared. Seven strains from the study site had a single nahAc allele, whereas the 16S rRNA gene sequences of the strains differed by as much as 7.9%. No nahAc alleles from the site were identical to those of the archetypal strains, although the predominant allele was closely related to that of Pseudomonas putida NCIB 9816-4, isolated in the British Isles. However, one site-derived nahAc allele was identical to that of the Washington state strain. Lack of phylogenetic congruence of the nahAc and 16S rRNA genes indicates that relatively recent in situ horizontal transfer of the nahAc gene has occurred, possibly as a direct or indirect consequence of pollutant contamination. Alkaline lysis plasmid preparations and pulsed-field gel electrophoresis have revealed the presence of plasmids ranging in size from 70 to 88 kb in all site isolates. Southern hybridizations with a 407-bp nahAc probe have suggested that the nahAc gene is plasmid borne in all the site isolates but one, a strain isolated from subsurface sediment 400 m upstream from the source of the other site isolates. In this strain and in the naphthalene-degrading strain from Washington state, nahAc appears to be chromosomally located. In addition, one site isolate may carry nahAc on both chromosome and plasmid. Within the group of bacteria with identical nahAc sequences the Southern hybridizations showed that the gene was distributed between plasmids of different sizes and a chromosome. This suggests that plasmid modification after transfer may have been effected by transposons. Horizontal transfer of catabolic genes may play a significant role in the acclimation of microbial communities to pollutants.

The present study determined tumorigenicity, tumor classification and DNA damage induced in infant mice by benzo[a]pyrene (B[a]P) or Manufactured Gas Plant (MGP) residues after a single exposure. Male and female B6C3F1 mice were exposed to B[a]P or MGP residue from a single environmental site (MGP-4) and males were also exposed to MGP residue composite from seven different sites (MGP-M7). At 26, 39 and 52 weeks after exposure tumorigenesis was assessed in lung, forestomach and liver. Formation and persistence of DNA adducts were quantified by 32P-postlabeling. Exposure of males to B[a]P induced liver tumors in a dose and time dependent manner. MGP induced more advanced tumors than B[a]P. Only a single liver tumor was found in MGP-4 treated females. No forestomach and few pulmonary adenomas were induced in males or females. MGP-4, MGP-M7 or B[a]P induced DNA adducts in males and females. Adducts in liver, lung and forestomach peaked on different days and decreased at different rates. At 24 h post-exposure, no significant differences in initial DNA adduct levels occurred in males and females exposed to MGP-4 or B[a]P. Lack of DNA damage (adducted DNA) did not account for non-responsiveness of lung and forestomach in B6C3F1 genders as well as in liver in females. MGP tumorigenicity could not be accounted for solely by B[a]P content nor did it reflect additivity of B[a]P and other carcinogenic polycyclic aromatic hydrocarbons (PAHs) in MGP. Synergy among MGP-PAHs, presence of unidentified carcinogens and/or promoters in MGP may account for MGP potency. The B6C3F1 infant male model is a convenient and rapid assay for assessing MGP liver tumorigenicity and potency.

A method for the simultaneous determination of urinary phenanthrene, fluoranthene, pyrene, chrysene and benzo[a]pyrene metabolites has been developed for individual risk assessment at polycyclic aromatic hydrocarbon (PAH)-burdened workplaces. The concentration of urinary metabolites as a measure for individual PAH exposure takes account not only of PAH masses resorbed by the respiratory tract but also those incorporated percutaneously. The method allows the determination of 25 different components with a low margin of error; the individual metabolite profiles thereby allow conclusions on the individual characteristics of PAH-oxidizing enzymes (monooxygenases). The coefficients of variation are lower than 10%. After enzymatic treatment of the urine with glucuronidase and arylsulfatase one part of the benzene or toluene extract is treated with diazomethane to convert phenols into methylethers, while another part is used to convert dihydrodiols into phenols. After further purification the metabolites are determined by means of a combination of gas chromatography and mass spectrometry. The PAH exposure of cock plant workers during several consecutive days resulted in fairly constant individual urinary metabolite profiles which, however, exhibited significant inter-individual variability. This held true also for Wistar rats exposed to tar pitch aerosol on 5 days during a period of 10 days. It was also demonstrated that in the case of coke plant workers there is a correlation between inhaled PAH and metabolites excreted. Mass relationships between inhaled PAH and metabolites excreted were found to differ from one individual to another.

Weathered coal tar collected from the Connecticut River near Holyoke, Massachusetts, was toxic to shortnose sturgeon embryos and larvae in whole sediment flow-through and elutriate static-renewal laboratory exposures. Sterile laboratory sand and clean Connecticut River sand, collected upstream from the coal tar deposits, produced no significant difference in toxicity to sturgeon embryos-larvae, while coal tar-contaminated sediment produced over 95% embryo-larval mortality. Hydrocarbon transfer and subsequent toxicity appeared to be via direct contact of the embryos with contaminated sediment, rather than via exposure to soluble hydrocarbons. This conclusion was supported by exposure of embryos and larvae to elutriates (e.g., water soluble extract) of coal-tar sediments, that resulted in embryo and larval mortality at low molecular weight PAH concentrations-0.47 mg/L, higher than would occur naturally. No decrease in petroleum hydrocarbon concentration was observed in sediments exposed to flowing water for 14 d, supporting the contention that soluble hydrocarbons were not responsible for the observed toxicity in whole sediment exposures under the conditions employed in this study.

The present study characterized the DNA adducts induced by manufactured gas plant residue (MGP) and benzo[alpha]pyrene (B[alpha]P) in mouse lung and forestomach. The dose levels used in the present study were comparable to the levels used in a previous animal bioassay. Adduct formation was evaluated in female A/J mice (7 weeks old) fed MGP (0.25%) or B[alpha]P (16 and 98 ppm) for 14 days. In addition, adduct formation was also evaluated in mice 24 h after the ip administration of 1.8 mg of B[alpha]P in 0.5 mL of tricaprylin. 32P-Postlabeling combined with multidimensional TLC and reverse phase HPLC was used to evaluate hydrocarbon-DNA adducts. HPLC separation of chemical-DNA adducts formed in lung following MGP ingestion resulted in three distinct peaks of radioactivity eluting at 22, 32.4, and 33.5 min. These peaks accounted for 13, 10, and 41% of the total adducts detected. The adducts isolated from forestomach eluted as a series of minor peaks with two more distinct peaks of radioactivity at 32.4 and 33.5 min. These peaks accounted for 47 and 32% of the total adducts detected in forestomach, respectively. Ingestion of B[alpha]P (16 or 98 ppm) and the ip administration of B[alpha]P resulted in a single major adduct with a retention time of 32.4 min. The DNA adducts formed from MGP administration were further characterized by comparison with adducts formed following the administration of individual hydrocarbons and a mixture of hydrocarbons.(ABSTRACT TRUNCATED AT 250 WORDS)

Coal tar treated psoriasis patients were used as a model population to evaluate a panel of immunoassays for monitoring exposure to benzo[a]pyrene (BP) and related polycyclic aromatic hydrocarbons (PAH). The assays included measurement of PAH diol epoxide-DNA adducts in white blood cells by competitive enzyme-linked immunosorbent assay (ELISA) with fluorescence endpoint detection, PAH-albumin adducts by competitive ELISA with color endpoint detection and serum levels of antibodies recognizing BP diol epoxide-DNA adducts by noncompetitive color ELISA. PAH-DNA adducts by ELISA were elevated in patients (mean 6.77 +/- 12.05/10(8)) compared to controls (4.90 +/- 8.81/10(8), p = 0.12). There was no difference in PAH-albumin adducts between patients (mean 0.61 +/- 0.31 fmol/micrograms) and controls (0.63 +/- 0.30 fmol/micrograms). Glutathione S-transferase M1 genotype was also determined but no relationship was found between presence of the gene and either DNA or protein adduct levels. About 30% of both patients and controls had measurable titer of antibodies recognizing BPDE-I-DNA adducts. Measurement of white blood cell DNA adducts by ELISA was the most sensitive method for detecting PAH exposure in coal tar-treated psoriasis patients.

Cenosis of microorganisms, coal resin destructors, is selected by the percolation method. A microbiological analysis of bacterial associations able to destruction of aromatic hydrocarbons composing the coal resin is carried out. Pure cultures are isolated as the most active destructors of these substrates. Destructive cultures are identified as Pseudomonas fluorescens 1-D biovar II and Bacillus subtilis 2-D. Efficiency of the microbiological refinement of coal-resin-contaminated soil by the isolated pure cultures and by their mixture is compared with self-refinement of soil. It is shown that complete refinement of soil from coal resin with contamination of 1 g per 1 kg of soil was attained after 160 days using the mixture of isolated destructive cultures P. fluorescens 1-D biovar II and B. subtilis 2-D. Usage of only P. fluorescens 1-D biovar II provided destruction of 87% of contaminating substances for the same period, whereas self-purification of soil by the natural cenosis of microorganisms made the purification level of 42%.

The relationships between increase of urinary 1-hydroxypyrene over the workweek and the airborne concentrations of benzo(a)pyrene and coal tar pitch volatiles (CTPVs) were studied among groups of workers in a vertical-stud Söderberg potroom of an aluminum smelter. There was a strong correlation between the natural logarithm of the pyrene concentration and the natural logarithm of the total polycyclic aromatic hydrocarbon (PAHs) concentration in personal air samples (r = 0.94). 1-Hydroxypyrene as the major metabolite of pyrene, a polycyclic aromatic hydrocarbon, was used as a marker for exposure to PAHs. A strong positive correlation was found between the natural logarithm of increase of urinary 1-hydroxypyrene and the natural logarithm of the estimated airborne PAH exposure (R2 = 0.84 with CTPV) when the use of facial protective clothing was taken into account. The relationship between increase of urinary 1-hydroxypyrene and PAHs differed for workers who used facial protective clothing under their respirators compared to those who did not. A contradictory fact was found: the use of facial protection seemed to lead to an elevated increase of 1-hydroxypyrene over the workweek. The regression model for the relationship between increase of urinary 1-hydroxypyrene over the workweek and airborne CTPV was: LN(change in 1-hydroxypyrene) = -4.7 + 1.2 LN(CTPV) -0.44 LN(CTPV)*(use of facial protection) + 3.5 (use of facial protection).

Relatively few reported attempts have been made to substitute HPLC for the thin-layer ion-exchange chromatography (TLC) conventionally used in the 32P-postlabelling assay. Using a reverse-phase phenyl-modified silica gel column and a gradient of methanol in 0.5 M sodium phosphate buffer (pH 2.0), we were able to improve the resolution of very similar adducts. Combined with on-line detection of Cerenkov radiation, this method allows separation of sub-femtomole quantities of 32P-labelled nucleoside 3',5'-bisphosphates modified by bulky carcinogens. Using this method, we were able to separate nine of the ten major adducts formed by reaction of the diol-epoxides of ten polycyclic aromatic hydrocarbons with DNA, and resolve different adducts formed by a single carcinogen. The major adducts formed by benzo[b]fluoranthene (BbF) or dibenz[a,h]anthracene in mouse skin in vivo have been shown to be distinct from the adducts formed directly by the bay-region diol-epoxides. The heterocyclic amines IQ and MeIQ have each been shown to form one major DNA adduct in several in vitro and in vivo systems; using HPLC we were able to resolve the two adducts formed by these food mutagens. HPLC is especially useful for the identification of adducts by means of chromatographic comparisons and in the analysis of the multiple adducts formed by complex mixtures of environmental carcinogens. The major adducts formed by benzo[a]pyrene (BaP) and BbF in mouse skin in vivo that were not resolved on TLC were well separated by HPLC and thus a major DNA adduct formed in the skin of mice treated topically with coal tar was found to be derived from BaP rather than BbF.

Treatment of mouse skin with coal tar is known to initiate tumour formation, with the carcinogenic activity associated mainly with polycyclic aromatic hydrocarbons (PAHs). A sample of pharmaceutical coal tar was analysed by gas chromatography and 19 major PAHs were identified. 32P-postlabelling analysis was used to characterize those PAHs that are responsible for the DNA binding of coal tar and, by inference, its biological activity. PAHs were grouped according to their reported carcinogenic activities and applied as mixtures to mouse skin. Group A contained all of the 19 PAHs, group B seven PAHs for which there is sufficient evidence for carcinogenicity and group C 12 PAHs with only limited or inadequate evidence of carcinogenicity in experimental animals. 32P-Labelled DNA adducts formed by coal tar were resolved on TLC into a pattern of three discrete spots (2, 4 and 6) and four areas of diffuse radioactivity (1, 3, 5 and 7). By comparison of the pattern of adducts formed by coal tar with those formed by the synthetic mixtures it appeared that PAHs in group B formed coal tar-DNA adduct spots 4 and 6, and that adduct spot 2 was formed by PAHs in group C. Attempts to identify those PAHs responsible for the formation of coal tar-DNA adducts 4 and 6 were made by comparing the chromatographic mobilities of 32P-labelled coal tar-derived DNA adducts formed in mouse skin, using TLC and HPLC, with those formed by PAHs in group B. As benzo[ghi]perylene (B[ghi]P), a component of group C, has been demonstrated to exhibit significant DNA binding ability previously, the chromatographic mobility of coal tar-DNA adduct spot 2 was compared to that of the major DNA adducts formed by B[ghi]P in vivo and in vitro. It appeared that coal tar adduct spot 2 was the major adduct formed by B[ghi]P in vitro and that benzo[a]pyrene, benzo[b]fluoranthene, benzo [j]fluoranthene and benzo[k]fluoranthene contributed to the formation of adduct spot 6. None of the PAHs examined appeared to be responsible for the formation of adduct spot 4.

Mutagenicity and toxicity of urine samples of 30 coke oven workers (group I) and 26 individuals composing a control group (group II) was estimated using Ames test with Salmonella typhimurium strains TA 98 and TA 100. Urine mutagenicity and toxicity, tested with strain TA 98 and metabolic activation, was significantly higher in group I than in group II. Using the strain TA 100, urine samples of coke oven workers showed only direct toxicity. Mutagenic and toxic activity was observed mostly in workers exposed to the highest BaP concentrations. No correlation was found between obtained results and tobacco smoking. Increased urinary mutagenic activity in group I resulted from exposure to very high BaP concentrations. It is concluded that urine samples of exposed coke oven workers should be periodically tested for mutagenic activity. Workers with mutagenic urine ought to be shifted to work posts with lower exposure to mutagens.

Urinary excretion of polycyclic aromatic hydrocarbon (PAH) metabolites and DNA binding of coal tar components in male mice were investigated following the ingestion of a coal tar adulterated diet. Male B6C3F1 mice were able to tolerate an F0927 basal gel diet which contained from 0.1 to 1% coal tar (tar weight/dry food weight) for 15 days. Mice maintained on a 0.1 and 0.2% coal tar diet had body weight gains similar to those of control animals. However, mice maintained on the 0.5 and 1.0% diet had body weight gains considerably lower than control values. Chemical-DNA adduct formation was detected and quantified in lung and forestomach tissue of animals on 0.1, 0.2, 0.5, and 1% coal tar containing diets. A dose-related effect was observed in lung DNA adduct formation while no dose effect was observed in forestomach tissue. In addition, overall adduct levels in lung tissue were considerably higher than forestomach levels for animals on the 0.5 or 1% diet. In contrast, DNA adduct levels were highest in the forestomach of animals on diets lower in coal tar content (0.1 or 0.2%). Chemical-DNA adducts of coal tar components were also evaluated for four other coal tar samples which varied in chemical composition. Mice were maintained on diets containing 0.25% of each coal tar for 15 days. Chemical-DNA adducts were detected in lung, liver, and spleen for all animals receiving these coal tar diets. DNA adduct patterns were similar while quantitative differences were observed between coal tar samples and tissue sites. Highest adduct levels were detected in lung DNA. Benzo[a]pyrene content in coal tar samples could not account for the DNA adduct levels observed with coal tar ingestion. The urinary excretion of select PAH metabolites following coal tar ingestion was evaluated by using urine collected on days 1 and 14 of diet administration. The levels of 1-hydroxypyrene in urine, the major PAH metabolite detected, correlated with the pyrene content of these coal tars. These data demonstrate that coal tar components are readily bioavailable following ingestion.

Adult and foetal human skin samples maintained in short-term organ culture were treated topically with solutions of coal-tar, creosote or bitumen, then DNA was isolated and analysed by 32P-post-labelling for the presence of aromatic DNA adducts. Autoradiographs of the 32P-labelled adducts resolved on polyethyleneimine-cellulose tlc sheets revealed a band of radioactivity indicative of the formation of adducts by a large number of components in these complex carcinogenic mixtures. Single doses of the materials, similar to those used to initiate tumours in experimental animals, resulted in the formation of approximately 0.3 fmol total adducts/micrograms DNA. The levels of adducts formed in human skin are thus similar to those formed in mouse skin after administration of doses known to be carcinogenic to the latter. The results provide direct evidence of DNA damage in human skin by materials strongly suspected of being carcinogenic to humans, and point to a method for evaluating other complex mixtures of aromatic chemicals for their potential carcinogenic hazard.

The urinary mutagenicity and the excretion of polycyclic aromatic hydrocarbons (PAH) in three non-smoking male patients, treated for psoriasis with cutaneous applications of crude coal tar, were analysed. Mutagenicity of the urinary extracts was measured by the plate incorporation assay using Salmonella typhimurium strains TA 98 and TA 100 in the presence of liver S9 fraction from Aroclor-induced rats with or without beta-glucuronidase. After concentration, hydrolysis and reduction of the urine sample, PAH levels were measured by high resolution gas chromatography/mass spectrometry. Following cutaneous treatment with coal tar, the urine of all three subjects showed noticeable levels of PAH and/or metabolites and marked mutagenicity both on strain TA 98 and TA 100 in the presence of S9 fraction. The addition of beta-glucuronidase increased the mutagenicity of the urinary extracts, the maximum values being attained on strain TA 100 in the presence of both microsomal fraction and deconjugating enzymes. The mutagenicity of urinary extracts from subjects treated therapeutically with crude coal tar was correlated (r = 0.788, P less than 0.01) with the total PAH levels in their urine. The PAH excreted in urine were mainly low molecular weight compounds, while benzo[a]anthracene was present in scarce amounts and the excretion of benzo[a]pyrene did not increase following the cutaneous exposure to the crude coal tar.

Biological monitoring of exposure to coal-tar products has been carried out using the nonselective urinary mutagenicity and thioether assays and a selective method for the detection of 1-hydroxypyrene in urine. The sensitivities of the three methods have been compared. In several work environments, no increase that could be related to exposure to coal-tar products was found with the nonselective methods, whereas the levels of 1-hydroxypyrene in urine were enhanced. The applicability of the method for the detection of 1-hydroxypyrene in urine is further demonstrated.

The toxicity of xanthene dyes were studied by various interaction between the dyes and the components in vital body. (1) An increase in the amount of Rose Bengale adsorbed on the gill of fish was followed by the increase in red corpuscle number, and it was assumed that the death of fish in xanthene dye solution was due to anoxemia. (2) Binding capacity of xanthene dyes with bovine serum albumin decreased in the following; Rose Bengale, Phloxine, Erythrosine, Eosine and Uranine. This order was quite coincident with the toxicity compared by TLm values. (3) From the results of rec-assay test by use of Bacillus subtilis, it was confirmed that Phloxine and Rose Bengale had DNA-damaging capacity related to the mutagenecity.

Liver S9 fractions were prepared from male Wistar rats, either non-induced or induced with Aroclor 1254 and from 5 human kidney transplant donors. The preparations were compared for their ability to metabolize the premutagens present in coal tar to mutagenic metabolites in the Salmonella mutagenicity assay towards strain TA98. Low levels of mutagenicity of coal tar were seen with human S9 preparations. The differences between the S9 mix of the 5 donors in capacity to activate premutagens were approximately 6-fold. The activation of coal tar by rat liver S9 preparations was higher than by the human S9 preparations. The metabolic conversion of pyrene in coal tar to 1-hydroxypyrene by the same human S9 preparations was determined in a parallel assay. 3 human preparations showed a high correlation between the formation of 1-hydroxypyrene and bioactivation of coal tar to mutagenic metabolites. The slope values of the individual regression lines were equal, suggesting that 1-hydroxypyrene is a good indicator for the activation of premutagens present in coal tar.

Three methods for the biological monitoring of human exposure to coal tar were compared. Levels of 1-hydroxypyrene(1-OH PYR), polycyclic aromatic hydrocarbons (PAH) and mutagens (Ames plate incorporation assay using Salmonella typhimurium strain TA98 in the presence of S9 and beta-glucuronidase) were determined in urinary samples from psoriatic patients undergoing topical treatment with mineral coal tar. A single sample of urine with a high content of PAH was diluted with urine of nonexposed, non-smoking subjects in order to obtain nine samples with a decreasing content of PAh metabolites. Mutagenicity of the extracts was detectable down to the dilution corresponding to a content in 1-OH PYR of about 50 micrograms/g creatinine and total PAH of 7 micrograms/g creatinine. In a second phase the three indicators of exposure to PAH were compared in 16 urinary samples from four psoriatic patients. The total PAH levels determined by the acidic deconjugation/reduction method were confirmed to be nearly always lower than the corresponding levels of 1-OH PYR alone. Most of the extracts were mutagenic, however, some of the samples with a high content in PAh metabolites were not mutagenic. In all the urinary samples analyzed the excretion of 1-OH PYR was markedly greater than in control subjects. 1-OH PYR and urinary mutagenicity levels were well correlated. The present data suggest that both the determination of mutagenicity and 1-OH PYR in urine may be used to monitor occupational exposure to PAH, the latter method being cheaper and of greater specificity and sensitivity.