Methylated Polycyclic Aromatic Hydrocarbons Research Articles

Establishing Criteria of Relative Abundance of Alkyl Polycyclic Aromatic Hydrocarbons (PAHs) for Differentiation of Pyrogenic and Petrogenic PAHs: An Application to Indian Sediment

Various source materials such as crude oil, automobile exhaust, coal, and wood combustion products were analyzed for polycyclic aromatic hydrocarbons (PAHs) and hopanes. Hopanes were detected in coal combustion-derived soot, along with petrogenic sources. Based on the correlation between the alkylated PAHs/parental PAHs ratio with their respective C30 hopane/PAHs ratio in these source materials, the criteria for differentiating the petrogenic and pyrogenic sources of PAHs were established. This study concluded that sediments contained exclusively pyrogenic PAHs when the following conditions were met: MP/P (methylphenanthrenes to phenanthrene) = 0.5, MPy/Py [(methylpyrenes + methylfluoranthenes) to (pyrene + fluoranthene)] = 0.15, and MC/C [(methylchrysenes + methylbenz[a]anthracenes) to (chrysene + benz[a]anthracene)] = 0.2. By contrast, the sediments contained exclusively petrogenic PAHs when the ratios of MP/P, MPy/Py, MC/C, and methyl PAHs (MPAHs)/PAHs exceeded 3.5, 1.5, 2.0, and 2.2, respectively. Based on these criteria, an intensive source apportionment of PAHs to sediments from three Indian metro cities, Chennai (n = 6), Mumbai (n = 6) and Kolkata (n = 12) (2003–2007), was determined. This investigation found that sedimentary PAHs from India were depleted in alkylated PAHs with low MP/P ≈ 0.7, MPy/Py ≈ 0.14, and MC/C ≈ 0.3, which contrasted with other tropical Asian countries, where elevated values of these ratios were detected. Among the three cities, Kolkata waters were more strongly influenced by pyrogenic sources of PAHs with very low alkylated PAHs ratios than Chennai and Mumbai sediments. The compositional differences were attributed to the dominant coal and biomass in Kolkata compared to the dominant usage of petrogenic cooking fuel (e.g., kerosene) and automobile-derived sources in Chennai and Mumbai.

The use of limpets as monitor of PAHs pollution in the Cantabrian coast

The concentrations of 24 parental and methylated polycyclic aromatic hydrocarbon (PAH) levels in common limpet ( Patella vulgata) were measured in 15 coastal localities of the southern Bay of Biscay and 2 sites per locality for 2 years (spring and autumn of 2004 and 2005, respectively). Although the average concentrations shown a wide variability among the sites and the season, several sites showed a remarkable high concentration (PAH tot>900 ng g −1 d.w.) or medium (300–800 ng g −1 d.w) levels, but in most of the cases the initial high concentrations decreased sharply from one campaign to the following. Moreover, based on ratios of individual isomers concentration, petrogenic and pyrolitic sources were distinguished suggesting more than one source. Finally, based on the results of this work and comparing with previously reported data, it can concluded that limpets can be used as sentinel organisms in coastal environmental monitoring, even when oil spills are involved.

Characterization of PM 2.5-bound polycyclic aromatic hydrocarbons in Atlanta—Seasonal variations at urban, suburban, and rural ambient air monitoring sites

Twenty-eight polycyclic aromatic hydrocarbons (PAH) and methylated PAHs (Me-PAH) were measured in daily PM 2.5 samples collected at an urban site, a suburban site, and a rural site in and near Atlanta during 2004 (5 samples/month/site). The suburban site, located near a major highway, had higher PM 2.5-bound PAH concentrations than did the urban site, and the rural site had the lowest PAH levels. Monthly variations are described for concentrations of total PAHs (∑PAHs) and individual PAHs. PAH concentrations were much higher in cold months than in warm months, with average monthly ∑PAH concentrations at the urban and suburban-highway monitoring sites ranging from 2.12 to 6.85 ng m −3 during January–February and November–December 2004, compared to 0.38–0.98 ng m −3 during May–September 2004. ∑PAH concentrations were found to be well correlated with PM 2.5 and organic carbon (OC) within seasons, and the fractions of PAHs in PM 2.5 and OC were higher in winter than in summer. Methyl phenanthrenes were present at higher levels than their un-substituted homologue (phenanthrene), suggesting a petrogenic (unburned petroleum products) input. Retene, a proposed tracer for biomass burning, peaked in March, the month with the highest acreage and frequency of prescribed burning and unplanned fires, and in December, during the high residential wood-burning season, indicating that retene might be a good marker for burning of all biomass materials. In contrast, potassium peaked only in December, indicating that it might be a more specific tracer for wood-burning.

QUANTITATIVE STRUCTURE–ACTIVITY RELATIONSHIPS FOR KINETIC PARAMETERS OF POLYCYCLIC AROMATIC HYDROCARBON BIOTRANSFORMATION

Quantitative structure-activity relationships (QSARs) were developed for three Monod-type parameters--qmax, Ks, and qmax/Ks--that express the kinetics of polycyclic aromatic hydrocarbon (PAH) biotransformation by Sphingomonas paucimobilis strain EPA505. The training sets contained high-quality experimental values of the kinetic parameters for 20 unsubstituted and methylated PAHs as well as values of 41 meaningful molecular descriptors. A genetic function approximation algorithm was used to develop the QSARs. Statistical evaluation of the developed QSARs showed that the relationships are statistically significant and satisfy the assumptions of linear-regression analysis. The Organization for Economic Co-operation and Development principles for (Q)SAR validation were followed to evaluate the developed QSARs, which showed that the QSARs are valid. The QSARs contain spatial, spatial and electronic, topological, and thermodynamic molecular descriptors. Whereas spatial descriptors were essential in explaining biotransformation kinetics, electronic descriptors were not. Mechanistic interpretation of the QSARs resulted in evidence that is consistent with the hypothesis of membrane transport as being the rate-limiting process in PAH biotransformation by strain EPA505. The present study demonstrates the value of QSAR not only as a predictive tool but also as a framework for understanding the mechanisms governing biodegradation at the molecular level.

SENCAR Mouse Skin Papillomas Induced by 6-Methylbenzo[a]pyrene Carry H-ras Codon 52 (CTALeu → CCAPro) and Codon 13 (GGCGly → GTCVal) Mutations

Organic smoke contains various methylated polycyclic aromatic hydrocarbon (PAH) carcinogens. Among these, 6-methylbenzo[a]pyrene (6-CH3BP) is a moderate carcinogen. In vitro, it can form both stable and depurinating adducts. Of these, only two depurinating adducts [BP-6-CH2-N7G and 6-CH2BP-(1&3)-N7G] have been detected in mouse skin. We examined seven 6-CH3BP-induced SENCAR mouse skin papillomas for the presence of oncogenic H-ras mutations. Two papillomas contained the codon 52 (CTALeu → CCAPro) and a third contained the codon 13 (GGCGly → GTCVal) mutation. These results suggest that the H-ras codon 52 mutation is an oncogenic mutation.

Concentrations of methylated naphthalenes, anthracenes, and phenanthrenes occurring in Czech river sediments and their effects on toxic events associated with carcinogenesis in rat liver cell lines

Alkylated polycyclic aromatic hydrocarbons (PAHs) are important environmental pollutants. In the present study, we determined levels of monomethylated naphthalenes (MeNap), phenanthrenes (MePhe), and anthracenes (MeAnt) in Czech river sediments. The levels of MePhe generally were lower than the concentrations of phenanthrene. In contrast, both MeNap and MeAnt were found at levels higher than their respective parent compounds in the majority of sampling sites. We then investigated their aryl hydrocarbon receptor (AhR)-mediated activity, accumulation of phosphorylated p53 protein, induction of expression of cytochrome P450 1A1 (CYP1A1), inhibition of gap junctional intercellular communication (GJIC), and effects on cell proliferation in rat liver cell models to evaluate the relative importance of these toxicity mechanisms of low-molecular-weight methylated PAHs. Methylated phenanthrene and anthracene compounds were weak inducers of AhR-mediated activity as determined both in a reporter gene assay system and by detection of the endogenous gene (Cyp1a1) induction. 2-Methylphenanthrene was the most potent AhR ligand. Contribution of MeAnt and MePhe to overall AhR-inducing potencies should be taken into account in PAH-contaminated environments. Nevertheless, their effects on AhR were not sufficient to modulate cell proliferation in a normal rat liver progenitor cell model system. These PAHs only had a marginal effect on p53 phosphorylation at high doses of 1-, 3-, and 9-MePhe as well as 1 MeAnt. On the other hand, both 2- and 9-MeAnt as well as all the MePhe under study were efficient inhibitors of GJIC, suggesting that these compounds might act as tumor promoters. In summary, inhibition of GJIC and partial activation of AhR seem to be the most prominent toxic effects of the methylated PAHs in the present study.

In Situ Direct Sampling Mass Spectrometric Study on Formation of Polycyclic Aromatic Hydrocarbons in Toluene Pyrolysis

The gas-phase reaction products of toluene pyrolysis with and without acetylene addition produced in a flow tube reactor at pressures of 8.15-15.11 Torr and temperatures of 1136-1507 K with constant residence time (0.56 s) have been detected in an in situ direct sampling mass spectrometric study by using a vacuum ultraviolet single-photon ionization time-of-flight mass spectrometry technique. Those products range from methyl radical to large polycyclic aromatic hydrocarbons (PAHs) of mass 522 amu (C(42)H(18)) including smaller species, radicals, polyynes, and PAHs, together with ethynyl, methyl, and phenyl PAHs. On the basis of observed mass spectra, the chemical kinetic mechanisms of the formation of products are discussed. Especially, acetylene is mixed with toluene to understand the effect of the hydrogen abstraction and acetylene addition (HACA) mechanism on the formation pathways of products in toluene pyrolysis. The most prominent outputs of this work are the direct detection of large PAHs and new reaction pathways for the formation of PAHs with the major role of cyclopenta-fused radicals. The basis of this new reaction route is the appearance of different sequences of mass spectra that well explain the major role of aromatic radicals mainly cyclopenta fused radicals of PAHs resulting from their corresponding methyl PAHs, with active participation of c-C(5)H(5), C(6)H(5), C(6)H(5)CH(2) ,and C(9)H(7) in the formation of large PAHs. The role of the HACA only seemed important for the formation of stable condensed PAHs from unstable primary PAHs with zigzag structure (having triple fusing sites) in one step by ring growth with two carbon atoms.

Measurement of biodegradability parameters for single unsubstituted and methylated polycyclic aromatic hydrocarbons in liquid bacterial suspensions

Substrate depletion experiments were conducted to characterize aerobic biodegradation of 20 single polycyclic aromatic hydrocarbons (PAHs) by induced Sphingomonas paucimobilis strain EPA505 in liquid suspensions. PAHs consisted of low molecular weight, unsubstituted, and methyl-substituted homologs. A material balance equation containing the Andrews kinetic model, an extension of the Monod model accounting for substrate inhibition, was numerically fitted to batch depletion data to estimate extant kinetic parameters including the maximal specific uptake rates, q(max), the affinity coefficients, K(S), and the substrate inhibition coefficients, K(I). Strain EPA505 degraded all PAHs tested. Applied kinetic models adequately simulated experimental data. A cell proliferation assay involving reduction of the tetrazolium dye WST-1 was used to evaluate the ability of strain EPA505 to utilize individual PAHs as sole energy and carbon sources. Of the 22 PAHs tested, 9 supported bacterial growth. Evaluation of the biokinetic data showed that q(max) correlated highly with transmembrane flux as theoretically estimated by a diffusion model, pointing to transmembrane transport as a potential rate-determining process. The biodegradability data generated in this study is essential for the development of quantitative structure-activity relationships (QSARs) for biodegradability and for modeling biodegradation of simple PAH mixtures.

Stabilities and Spin Distributions of Benzannulated Benzyl Radicals.

The effects have been studied of mono- and dibenzannulation of a benzyl radical with hybrid density functional theory (B3LYP) and quadratic configuration interaction theory (QCISD). Bond dissociation energies and enthalpies are reported that were determined at the common level QCISD/6-311G**//B3LYP/6-31G* for the benzylic C-H bonds of toluene 1H, the monobenzannulated polycyclic aromatic hydrocarbons (PAH) 1- and 2-methylnaphthalene 2H and 3H, the dibenzannulated PAHs 9-methylanthracene 4H and 9-methylphenanthrene 5H, and the model hydrocarbons 1-phenylpropene 6H and propene 7H. The conformational preferences and the symmetries of 1H-7H and of their corresponding radicals 1-7 have been determined. The analysis of the electron and spin density distributions of radicals 1-7 at the QCI level are reported, and these high-level data are discussed in comparison to results obtained with density functional theory and with an awareness of a general perception shaped by Hückel molecular orbital theory. The results show in a compelling fashion that electron and spin delocalization onto an annulated arene is not the decisive principle for stabilization of the benzyl radicals formed by homolysis of the methylated PAHs C10H7-CH3 and C14H9-CH3, and instead, the analysis of QCI spin density distributions suggests that spin delocalization onto annulated arenes is avoided as much as possible while spin polarization does occur to a significant extent.

Dermal Exposure to Polycyclic Aromatic Hydrocarbons among Road Pavers

Dermal exposure to polycyclic aromatic hydrocarbons (PAHs) and the role of an industrial by-product, coal fly ash, on workers' PAH exposure were investigated during stone mastic asphalt (SMA) paving and remixing. PAH exposure was measured at eight sites during the laying of SMA containing coal fly ash or limestone (conventional SMA) as the filler. Six of the surveys were carried out during SMA paving and two during remixing of SMA (hot recycling at the paving site). Dermal PAH exposure was measured by hand washing (using sunflower oil and wiping with Kleenex tissues) before and after the work shift, and by placing exposure pads on the workers' wrists during the work shift. The analyses included 15 native PAHs from the hand-washing samples determined using high-performance liquid chromatography equipped with a two-channel fluorescence detector and 16 native PAHs and four methylated PAHs from the exposure pads using gas chromatography with mass-selective detection. The PAH results obtained using the pad and hand-washing methods (concentrations after the work shift) were equivalent and showed a strong correlation (r = 0.757, P < 0.001, N = 23 for total PAHs). There was a statistically significant difference between pre- and post-shift samples as measured by hand washing. The skin contamination by PAHs was significantly higher (P < 0.01) during remixing than during SMA paving. The variation in PAH contamination on the skin explained more of the variation in the excretion of urinary 1-hydroxypyrene and phenanthrols than the variation in the respiratory PAH concentrations. The industrial by-product investigated in asphalt, coal fly ash, had no statistically significant effect on the workers' dermal PAH exposure. The dermal exposure of paving workers to PAHs was higher during remixing than during SMA paving.

Mass spectrometric analysis of 7‐sulfoxymethyl‐12‐methylbenz[a]anthracene and related electrophilic polycyclic aromatic hydrocarbon metabolites

The Meso-region theory of polycyclic aromatic hydrocarbon (PAH) carcinogenesis predicts that the development of pronounced carcinogenicity depends on the introduction of a good leaving group on alkyl side-chains attached to the exceptionally reactive meso-anthracenic or L-region positions of PAHs. Thus, the first step in carcinogenesis by methylated PAHs such as 7,12-dimethylbenz[a]anthracene (DMBA) would be the hydroxylation of the L-region methyl groups, particularly the 7-methyl group. The second would be the formation of a metabolite, e.g. a sulfate ester, which is expected to be a good leaving group capable of generating a highly reactive benzylic carbocation. 7-Hydroxymethyl-12-methylbenz[a]anthracene (7-HMBA) is a metabolite of DMBA, and sulfation of 7-HMBA to a 7-sulfoxymethyl metabolite (7-SMBA) is a known Phase II metabolic process designed to facilitate excretion, but actually enabling more destructive side-reactions. These side-reactions occur with generation of an electrophilic 7-methylene carbonium ion, and/or by in vivo halide exchange to provide neutral side-products more capable of entering cells, especially those of DMBA target tissues. Electrospray ionization mass spectrometry (MS) enabled us to visualize 7-SMBA as an intact m/z 351 conjugate anion by negative mode, and as a released m/z 255 carbonium ion by positive mode. Upon prolonged refrigeration, 7-SMBA accumulated an m/z 383 photooxide, which appeared capable of re-evolving the starting material as visualized by tandem quadrupole MS, or MS/MS. The 7-SMBA carbonium ion provided interpretable fragments when studied by fragment ion MS/MS, including those representing the loss of up to several protons. Subtle differences in this property were encountered upon perturbing 7-SMBA, either by warming it at 37 degrees C for 2 h or by substituting the initial sulfoxy group with an iodo group. Side-reactions accounting for such proton losses are proposed, and are of interest whether they occur in the mass spectrometer, in solution or both; these proposals include acidity at the 12-methyl position and cyclization between the 12-methyl group and the adjacent C-1 position. It is also suggested that such side-reactions may comprise one route to relieving steric strain arising between the 12-methyl group and the angular benzo ring of 7-SMBA.

북극 스발바드 군도 스피츠베르겐섬 콩스피요르드에서의 다환 방향족 탄화수소화합물의 분포 특성

In order to elucidate the polycyclic aromatic hydrocarbon concentration and its origin in arctic area, four arctic brown algae (Laminaria saccharina, L. digita, Alaria esculenta, Desmarestia aculeata), one marine invertebrate (Echinoidea) and sediments were collected from Kongsfjorden in Spitsbergen from the late July to early August, 2003. In case of macroalgae, the young blade part above growth point and the old stipes and blades beneath growth point were separated and analyzed for polycyclic aromatic hydrocarbons (PAHs) in an attempt to check the mechanism of uptake in macroalgae to accumulate PAH. There was no difference in PAH concentrations between sampling sites (Stations B and C), species, and blades beneath and above growth point. PAH concentrations in all samples collected in this study were relatively higher than those reported in other areas of arctic. Especially, station C, which is known as an unpolluted area, showed 10 times higher PAH concentration (8,765 ng/g) in sediment than station A (694 ng/g) around harbor. In addition high PAH concentration, station C had very higher proportion of methylated PAH to parent PAH in sediment than station A. Source analysis using PAH isomer pair ratios as indicators showed that Kongsfjorden area seemed to be relatively contaminated with PAH derived from direct petroleum input.

Polycyclic aromatic hydrocarbons, aliphatic hydrocarbons, trace elements, and monooxygenase activity in birds nesting on the North Platte River, Casper, Wyoming, USA

Tree swallow (Tachycineta bicolor) and house wren (Troglodytes aedon) eggs and chicks were collected near a refinery site on the North Platte River, Casper. Wyoming, USA and at a reference site 10 km upstream. Total polycyclic aromatic hydrocarbon (PAH) concentrations in swallow and wren chicks were higher at the refinery site than at the reference site. Polycyclic aromatic hydrocarbon concentrations in sediment and chick dietary samples were consistent with these findings. The general lack of methylated PAHs in sediment, diet, and bird carcasses suggested that the PAHs were derived from combustion and not from petroleum. The predominance of odd-numbered aliphatic hydrocarbons and the low ratios (< or =0.25) of pristane:n-C17 and phytane:n-C18 in chick and diet samples also suggested that swallow and wren chicks were not being chronically exposed to petroleum. Mean ethoxyresorufin-O-dealkylase and benzyloxyresorufin-O-dealkylase activities in tree swallow livers averaged nine times higher at the refinery site than at the reference site and were probably induced by exposure to PAHs. Trace element concentrations in eggs and livers of swallows and wrens were similar or greater at the reference site than at the refinery site. Selenium, strontium, and boron concentrations were elevated in eggs and livers of swallows and wrens at both the refinery and reference sites.

Chemical fingerprinting applied to the evaluation of marine oil pollution in the coasts of Canary Islands (Spain)

Chemical fingerprinting approach to environmental assessment is illustrated in the evaluation of marine oil pollution in the coasts using two limpet species as bioindicator organisms, and based on profiles and concentrations of n-alkanes and aromatic hydrocarbons in their tissues. Accidental and chronic releases of hydrocarbons can contaminate the marine environment of the Canary Islands not only because of their geographical situation but also because of the very dense tanker traffic around. This situation affects coastal areas, fishing activities, tourism resort, etc. Concentrations of n-alkanes and aromatic hydrocarbons (polycyclic aromatic hydrocarbons and methyl-polycyclic aromatic hydrocarbons) in the soft tissues of the marine intertidal and subtidal limpets, Patella crenata and Patella ullysiponensis aspera, were evaluated. Limpet samples were collected at monthly intervals, at three locations on the southeast coast of Tenerife over a 3-year period (1991–93). Levels of hydrocarbons found in limpets are similar to concentrations found in unpolluted areas around the world. From application of principal component analysis, the interpretation of variable loading plots gives information on variable correlation and can be used to distinguish among potential sources of pollution and the ability of studied molluscs to be used as bioindicator organisms.

Identifying carcinogenic activity of methylated and non-methylated polycyclic aromatic hydrocarbons (PAHs) through electronic and topological indices

Polycyclic aromatic hydrocarbons (PAHs) are a class of planar molecules, abundant in urban environment, which can induce chemical carcinogenesis. Their carcinogenic power varies in a large range, from very strong carcinogens to inactive ones. In a previous study, we proposed a methodology to identify the PAHs carcinogenic activity exploring electronic and topological indices. In the present work, we show that it is possible to simplify that methodology and expand its applicability to include methylated PAHs compounds. Using very simple rules, we can predict their carcinogenic activity with high accuracy (~ 89%).

Identifying carcinogenic activity of methylated polycyclic aromatic hydrocarbons (PAHs)

Polycyclic aromatic hydrocarbons (PAHs) are a class of planar molecules that can induce chemical carcinogenesis. Their carcinogenic power vary in a large range, from the very strong carcinogens to the inactive ones. Recently a new methodology was proposed (using the topological simple Hückel theory) to group and identify PAHs carcinogenic activity in terms of very simple rules based on the concept of electronic local density of states over specific molecular regions. In the present work we have extended this study to include methylated compounds. Our results show that the previous methodology can also be simplified and successfully used to predict the carcinogenic activity of methylated PAHs.

Reproductive toxicology of hexachlorocyclohexane isomers: Induction of eggshell protein synthesis in atlantic salmon

Immunoassays based on monoclonal antibodies (mAbs) are highly sensitive for the detection of polycyclic aromatic hydrocarbons (PAHs) and can be employed to determine concentrations in near real-time. A sensitive generic mAb against PAHs, named as 2G8, was developed by a three-step screening procedure. It exhibited nearly uniformly high sensitivity against 3-ring to 5-ring unsubstituted PAHs and their common environmental methylated PAHs, with IC50 values between 1.68 and 31 μg/L (ppb). 2G8 has been successfully applied on the KinExA Inline Biosensor system for quantifying 3–5 ring PAHs in aqueous environmental samples. PAHs were detected at a concentration as low as 0.2 μg/L. Furthermore, the analyses only required 10 min for each sample. To evaluate the accuracy of the 2G8-based biosensor, the total PAH concentrations in a series of environmental samples analyzed by biosensor and GC–MS were compared. In most cases, the results yielded a good correlation between methods. This indicates that generic antibody 2G8 based biosensor possesses significant promise for a low cost, rapid method for PAH determination in aqueous samples.

Nongenotoxic Effects of Polycyclic Aromatic Hydrocarbons and Their Ozonation By-Products on the Intercellular Communication of Rat Liver Epithelial Cells

Since polycyclic aromatic hydrocarbons (PAHs) are known to have epigenetic effects, we evaluated the effect of the parent chemical and the ozonated products on in vitro cell to cell communication bioassays which measures a nongenotoxic event. The scrape loading/dye transfer (SL/DT) technique was used to determine the effect of the following PAHs on gap-junction intercellular communication (GJIC): flourene. I-methyl-fluorene, fluoranthene, anthracene, 9-methyl-anthracene, phenanthrene, pyrene, benzo(a)pyrene, and benzo(e)pyrene. The methylated PAHs were more inhibitory to GJIC than the unmethylated counterparts. Fluoranthene, which has an additional ring added to fluorene, was more effective in inhibiting GJIC than flourene. The three-ringed PAHs were also more inhibitory than the four- and five-ringed PAHs. A time-course study of fluoranthene and of pyrene resulted in maximal inhibition occurring within 30 min of incubation with the cells. The cells recovered from the inhibition within 1 hr after fluoranthene and pyrene were removed front the cell culture medium. Pyrene, benzo(a)pyrene, fluorene, and fluoranthene were ozonated until the parent compound was completely eliminated as determined by reverse-phase high-pressure liquid chromatography (RP-HPLC). An increased level of inhibition of GJIC was observed for the ozonated mixtures of by-products of pyrene, fluoranthene, and benzo(a)pyrene, but not for fluorene, as monitored with the SL/DT technique. The products of the ozonated pyrene mixture were fractionated and collected by RP-HPLC. Each fraction was found to be inhibitory to GJIC as monitored by fluorescence recovery after photobleaching. In conclusion, current treatment technologies, such as ozonation or biologically based oxidations and methylations, do not necessarily eliminate toxicity. Therefore, it is imperative that toxicological studies be used to complement traditional chemical detection techniques used to monitor the fate of a pollutant in environmental treatment systems.

Persistence of Calcium Elevation in the HPB-ALL Human T Cell Line Correlates with Immunosuppressive Properties of Polycyclic Aromatic Hydrocarbons

The immunosuppressive synthetic methylated polycyclic aromatic hydrocarbon (PAH), 7,12-dimethylbenz[a]anthracene (DMBA), has been shown to cause both an immediate and a sustained elevation of free intracellular calcium (Ca2+) in human T cells. In the present studies, a series of anthracene- and pyrene-based PAHs were tested for rapid (3 min) and sustained (4 hr) Ca2+ mobilization in the HPB-ALL human T cell line measured by flow cytometry using Fluo-3 as a Ca2+ indicator. Immunosuppressive PAHs produced a sustained Ca2+ elevation for at least 4 hr, while weakly immunosuppressive PAHs caused only a transient increase in Ca2+. The immunosuppressive PAHs, DMBA, benzo[a]pyrene, dibenz[a,h]anthracene, and 9,10-dimethylanthracene, produced a sustained increase in intracellular Ca2+ in HPB-ALL cells. Those PAHs with moderate to minimal immunosuppressive properties (i.e., dibenz[a,c]anthracene, benz[a]anthracene, benzo[e]pyrene, and anthracene) produced small and transient Ca2+ mobilization responses in HPB-ALL cells. It appeared that methylation of anthracene at the 9,10-positions increased the duration of Ca2+ mobilization, whereas the addition of a benzene group in the "a" position was associated with a transient increase in Ca2+ levels. Genistein, a protein tyrosine kinase (PTK) inhibitor, partially inhibited the rapid and sustained PAH-induced Ca2+ mobilization responses, while the protein kinase C (PKC) inhibitors, staurosporine and calphostin C, had essentially no effect on PAH-induced Ca2+ elevation. It appears that the action of PAHs on PTKs is important in the rapid Ca2+ response of human T cells. However, additional biochemical mechanisms appear to be responsible for the sustained elevation of Ca2+ produced by PAHs in T cells. The results of these studies demonstrate that persistent elevation of intracellular Ca2+ by PAHs correlates with their known immunosuppressive properties.

Absence of mutagenicity of benzo[ c]phenanthridine alkaloids in somatic cells of Drosophila melanogaster: Comparison with 7,12-dimethylbenz[ a]anthracene and chrysene

The wing somatic mutation and recombination test (SMART) of Drosophila melanogaster was used to study the mutagenic potential of three benzo[ c]phenanthridines with antileukemic properties, fagaronine, nitidine and O-methylfagaronine, as compared with that of two structurally related aromatic polycyclic hydrocarbons: 7,12-dimethylbenz[ a]anthracene and chrysene. Although toxic to larvae, the benzo[ c]phenanthridines and chrysene gave negative or inconclusive results while 7,12-dimethylbenz[ a]anthracene was found to be highly mutagenic and recombinogenic as previously reported. These results suggest that the alkoxy groups and the quaternary nitrogen of the benzo[ c]phenanthridines may reduce or eliminate their mutagenicity in spite of their similarity to methylated polycyclic aromatic hydrocarbons.