Efficiency of butyl rubber sorbent to remove the PAH toxicity.

Large amounts of polycyclic aromatic hydrocarbons (PAHs) have been released to the marine environment as a result of oil spills and from other sources including wastewaters, surface runoff, industrial processes, atmospheric deposition, biosynthesis, and natural events such as forest fires. PAHs have been known to affect a variety of biological processes and can be potent cell mutagens/carcinogens and toxic. In this study, PAH toxicity removal was investigated by using a novel macroporous butyl rubber (BR) sorbent. To find out the toxicity removal efficiency of the sorbents, the toxicity tests with Vibrio fisheri (luminescence bacteria) and Phaeodactylum tricornutum (marine algae) were applied to the acenaphthene (Ace) and phenanthrene (Phen) solutions in seawater (Ace: 500- 1000 µg/L; Phen; 100-1000 µg/L) before and after sorbent applications. Additionally, lysosomal stability and filtration rate biomarker techniques were applied to the mussels (Mytilus galloprovincialis) exposed to 1000 µg/L Phen solution and bioaccumulation was measured. The results showed that the toxicity of the PAH solutions decreased 50-100 percent depending on the concentration of the solutions and organisms. Phaeodactylum was found as the most sensitive organism to Phen and Ace. Since the application of BR sorbent removed the Phen from the solution, the bioaccumulated Phen amount in the mussels decreased accordingly.

Acenaphtho[1,2-b]pyrrole-based selective fibroblast growth factor receptors 1 (FGFR1) inhibitors: design, synthesis, and biological activity.

A novel series of acenaphtho[1,2-b]pyrrole derivatives as potent and selective inhibitors of fibroblast growth factor receptor 1 (FGFR1) were designed and synthesized. In silico target prediction revealed that tyrosine kinases might be the potential targets of the representative compound 2, which was subsequently validated by enzyme-linked immunosorbent assay (ELISA) for its selective and active FGFR1 inhibition of various tyrosine kinases. The structure-activity relationship (SAR) analysis aided by molecular docking simulation in the ATP-binding site demonstrated that acenaphtho[1,2-b]pyrrole carboxylic acid esters (2-5) are potent inhibitors of FGFR1 with IC(50) values ranging from 19 to 77 nM. Furthermore, these compounds exhibited favorable growth inhibition property against FGFR-expressing cancer cell lines with IC(50) values ranging from micromolar to submicromolar. Western blotting analysis showed that compounds 2, 3, and 2b inhibited activation of FGFR1 and extracellular-signal regulated kinase 1/2 (Erk1/2).

Polycyclic aromatic hydrocarbons (PAHs) in surface sediments of Liaodong Bay, Bohai Sea, China.

MATERIALS AND METHODS: The levels and possible sources of 16 polycyclic aromatic hydrocarbons (PAHs) were investigated in the surface sediments of Liaodong Bay, Bohai Sea, China. RESULTS: The sum of 16 PAHs (∑PAH(16)) concentrations varied from 144.5 to 291.7 ng/g, with a mean value of 184.7 ng/g, indicating low PAH levels compared with reported values for other bays and rivers in China and developed countries. High concentrations of PAHs were observed in the Luan River Estuary and in the vicinity of Qinhuangdao, implying that sewage from the Luan River and shipping activities are important sources of PAHs in Liaodong Bay. CONCLUSION: An ecological risk assessment of PAHs, based on the effect range-low quotients, indicated that adverse biological effects caused by acenaphthene occasionally may take place in the sediments of Liaodong Bay. PAH source identification suggested that PAHs in most sediments were mainly from incomplete combustion of grass, wood, and coal. At other stations near the Luan River Estuary, both petrogenic and pyrogenic inputs were significant, and the petroleum-derived PAHs were mainly from shipping activities and discharge of pollutants via rivers.

Toxicity of xenobiotics during sulfate, iron, and nitrate reduction in primary sewage sludge suspensions.

The effect and persistence of six organic xenobiotics was tested under sulfate-, iron-, and nitrate-reducing conditions in primary sewage sludge suspensions. The xenobiotics tested were acenaphthene, phenanthrene, di(2-ethylhexyl)phthalate (DEHP), 4-nonylphenol (4-NP), linear alkylbenzene sulfonate (LAS), and 1,2,4-trichlorobenzene (1,2,4-TCB) added to initial analytical concentrations of 54-117 mgL(-1). The suspensions were incubated at 30 degrees C for 15 weeks and rates of sulfate, iron, and nitrate reduction were estimated from the time course of hydrogen sulfide accumulation, Fe(II) accumulation, and nitrate depletion, respectively. Chemical analysis showed that the xenobiotics were persistent under the different electron acceptor regimes for the duration of the experiment. This was partly attributed to low bioavailability and microbial toxicity of the xenobiotics. Rates of anaerobic respiration in control suspensions (without added xenobiotics) showed a weekly reduction potential of 0.84 mM SO(4)(2-), 0.92 mM Fe(III), and 9.25 mM NO(3)(-). All three processes were completely inhibited by 1,2,4-TCB (54 mgL(-1)) whereas there was no significant (P<0.05) toxicity of phenanthrene (109 mgL(-1)) and DEHP (105 mgL(-1)). Sulfate reduction was inhibited completely by LAS (105 mgL(-1)), 76% by acenaphthene (54 mgL(-1)) and 57% by 4-NP (117 mgL(-1)), and likewise iron reduction was inhibited 62% by LAS and 55% by 4-NP (the latter though at P<0.10). Nitrate reduction was not significantly inhibited by acenaphthene and 4-NP and furthermore was resistant to LAS toxicity (105 mgL(-1)). Nitrate reduction also had the highest potential for mineralization of organic matter and thus was the most robust of the tested anaerobic processes in the sewage sludge suspensions.

Novel acenaphtho[1,2-b]pyrrole-carboxylic acid family: synthesis, cytotoxicity, DNA-binding and cell cycle evaluation.

A family of 8-oxo-8H-acenaphtho[1,2-b]pyrrole-9-carboxylic acid derivatives were synthesized as a result of our efforts to modify a series of acenaphthopyrrole aromatic-heterocycle compounds that proved to be potent anticancer drugs. Among the derivatives, 3d (3-(dimethylamino-propylamino)-8-oxo-8H-acenaphtho-[1,2-b]pyrrole-9-carboxylic acid) and 3g (3-piperidine-8-oxo-8H-acenaphtho-[1,2-b]pyrrole-9-carboxylic acid) showed potential anticancer activity and different action mechanism from our previously reported compounds. UV-vis absorption, circular dichroism and viscosity measurement indicated that effect of both compounds on the advanced DNA conformation was different, although they could bind to DNA in the same way. Cell cycle analysis showed that 3d could induce S-phase arrest followed by apoptosis, while 3g induced apoptosis. The results seem to imply that different action mechanism could contribute to the dissimilitude of biological activities toward 3d and 3g.

DNA double helix unwinding triggers transcription block-dependent apoptosis: a semiquantitative probe of the response of ATM, RNAPII, and p53 to two DNA intercalators.

We have previously shown the binding modes of two DNA interacting analogues (1)a {3-(4-methyl-piperazin)-8-oxo-8H-acenaphtho[1,2-b]pyrrole-9-carbonitrile} and (3)a {3-(3-dimethylamino-propylamino)-8-oxo-8H-acenaphtho[1,2-b]pyrrole-9-carbonitrile} with the DNA double helix. In this study, we have determined the notably different DNA damage signal pathway elicited by (1)a and (3)a due to the different extents to which they unwind the DNA double helix. First, we have identified that ataxia-telangiectasia-mutated (ATM) protein kinase can respond to DNA double helix unwinding caused by both (1)a and (3)a. In addition, the amount of ATM activation is consistent with the degree to which the DNA double helix was unwound. Consequently, we used (1)a and (3)a to semiquantitatively probe the response of RNA polymerase II (RNAPII) and p53 toward DNA double helix unwinding in vivo. By means of flow cytometry, immunocytochemistry, ChIP, quantitative real-time polymerase chain reaction, and Western blot analyses, we measured the level of p53 and RNAPII phosphorylation, in addition to the dynamics of the RNAPII distribution along the c-Myc gene. These results provided novel evidence for the impact of subtle DNA structural changes on the activity of RNAPII and p53. Moreover, DNA double helix conformational damage-dependent apoptosis was studied for the first time. These results indicated that (1)a can induce transcriptional blockage following a shift of the unphosphorylated IIa form of RNAPII to the phosphorylated IIo form, while (3)a is unable to induce the same effect. Subsequently, p53 accumulation and phosphorylation events occur that lead to apoptosis in the case of (1)a exposure. This suggests that the transcriptional blockage is also correlated to the degree of double helix unwinding. Furthermore, we found that the degree of DNA conformational damage determines whether or not apoptosis occurs through transcriptional blockage. Under our experimental conditions, ATM does not participate in the downstream events even when it has been activated. Thus, p53-mediated apoptosis may be independently triggered by transcriptional blockage.

An environmental quinoid polycyclic aromatic hydrocarbon, acenaphthenequinone, modulates cyclooxygenase-2 expression through reactive oxygen species generation and nuclear factor kappa B activation in A549 cells.

Diesel exhaust particles (DEPs) contain oxygen-containing polycyclic aromatic hydrocarbons (PAHs) called quinoid PAHs. Some quinoid PAHs generate free radicals as they undergo enzymatic and nonenzymatic redox cycling with their corresponding semiquinone radicals. Reactive oxygen species (ROS) produced by these reactions can cause severe oxidative stress connected with inflammatory processing. Although humans and animals are continuously exposed to these chemicals in the environment, little is known about which quinoid PAHs are active. In this study, we estimated the intracellular ROS production and nuclear factor kappa B (NF-kappaB) translocation in A549 cells exposed to isomers of quinoid PAHs having two to four rings. We found that both acenaphthenequinone (AcQ) and 9,10-phenanthrenequinone (PQ) enhanced ROS generation and that AcQ translocated NF-kappaB from the cytosol to the nucleus. However, PQ, which has been reported to induce apoptosis, did not influence NF-kappaB activation. In addition, AcQ induced cyclooxygenase-2 (COX-2) expression which is a key enzyme in the inflammatory processing involved in the activation of NF-kappaB. Upregulation of NF-kappaB and COX-2 expression by AcQ treatment was suppressed by the antioxidant N-acetylcysteine (NAC). These results provide that AcQ might play an important role in human lung inflammatory diseases as an air pollutant.

Acute and chronic life cycle toxicity of acenaphthene and 2,4,6-trichlorophenol to the midge Paratanytarsus parthenogeneticus (Diptera: Chironomidae).

The acute and life cycle toxicities of acenaphthene and 2,4,6-trichlorophenol (2,4,6-TCP) were evaluated using the parthenogenic dipteran larva, Paratanytarsus parthenogeneticus, of the family Chironomidae. Static 48 h acute toxicity tests employed third instar larvae, and the flow-through 20-day life cycle tests were initiated with eggs. No acute lethal effect was observed in the saturated concentration (2.1 mg/l) of acenaphthene in water, and the median lethal concentration of 2,4,6-TCP was approximately 40 mg/l. In the life cycle toxicity test with acenaphthene, hatchability of the midge was affected by 50% at an exposure concentration of approximately 0.17 mg/l. The median effective concentration for larval, pupal and adult development was found between 0.06 and 0.07 mg/l, respectively, and these were significantly lower than the effective concentration for hatching success. With regard to 2,4,6-TCP toxicity, the median effective concentration for midge hatchability was 4.23 mg/l. The larval development, pupal formation and adult emergence, however were impeded at 1.60, 1.46 and 1.33 mg/l, respectively. Each level was noticeably lower than the median effective concentration for hatchability. The concentrations for each compound that interfered with the development to fourth instar, pupa and adult were not significantly different. These results indicate that larval developments after hatching were the most sensitive stages and affected the success of midge growth.

Acetaminophen produces cataract in DBA2 mice by Ah receptor-independent induction of CYP1A2.

The metabolic transformation of acetaminophen to N-acetyl-p-benzoquinone imine by cytochrome P450 enzymes (e.g., cytochrome P450 1A2) is a prerequisite for acetaminophen-induced cataract formation in mice. Aromatic hydrocarbons, such as beta-naphthoflavone, induce cytochrome P450 1A2 in C57BL6 mice via the mediation of the aromatic hydrocarbon receptor and render the animals susceptible to cataract formation by acetaminophen administration but not in DBA2 mice which do not respond to cytochrome P450 1A2 induction by these compounds. Polycyclic hydrocarbons, such as acenaphthylene, were recently found to induce cytochrome P450 1A2 gene expression in young DBA2 mice by aromatic hydrocarbon receptor-independent pathways. In this work, we investigated whether enhanced metabolism of acetaminophen to N-acetyl-p-benzoquinone by cytochrome P450 1A2 induction by acenaphthylene could produce cataract in young DBA2 mice. Fifteen-day-old DBA2 mice were pretreated with two intraperitoneal injections of acenaphthylene and, 24 hr later, with one injection of acetaminophen. In most mice, cataract developed 18-24 hr after acenaphthylene injection. Acenaphthylene treatment of young DBA2 mice resulted in a 2-fold increase in cytochrome P450 1A2-dependent methoxyresorufin O-demethylase activity in the liver. These results support the hypothesis that the aromatic hydrocarbon receptor-independent induction of cytochrome P450 1A2 enzyme leads to accumulation of sufficient N-acetyl-p-benzoquinone in the liver and cataract development in the eye.

Ability of 16 priority PAHs to be directly cytotoxic to a cell line from the rainbow trout gill.

Sixteen polycyclic aromatic hydrocarbons (PAHs) were screened for their ability to be directly cytotoxic to a cell line from the rainbow trout gill, RTgill-W1. Exposure times of 2 h or less were sufficient for direct cytotoxicity to be detected, which appeared to be caused by a common mechanism, the general perturbation of membranes. This was judged by the similarity of results obtained for three fluorescent indicator dyes, alamar Blue, 5-carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM) and neutral red. Among the 16 PAHs tested, just two- and three-ring PAHs were found to be directly cytotoxic. These were naphthalene approximately = acenaphthylene approximately = acenaphthene > fluorene approximately = phenanthrene. The results suggest that water solubility and lipophilicity are the critical properties determining the direct cytotoxicity of PAHs and that they do so by influencing PAH accumulation in membranes. Only naphthalene was effective at concentrations well below its water solubility limit. Therefore, direct cytotoxicity is likely to be most environmentally relevant only with naphthalene.

Polycyclic aromatic hydrocarbons-induced vasorelaxation through activation of nitric oxide synthase in endothelium of rat aorta.

In the present study, the effect of polycyclic aromatic hydrocarbons (PAHs) on isolated rat aorta was investigated. Acenaphthylene and naphthalene dose-dependently relaxed the phenylephrine-induced contraction of rat aorta with 50% vasorelaxation at 40.8+/-19.83 and 118.75+/-9.83 microM, respectively. The vasorelaxation effect was diminished in the denuded (endothelium removed) aorta suggesting that the relaxation effect of PAHs was endothelium dependent. By comparing PAHs with different ring structures, we have found that acenaphthylene has the highest potency to induce vasorelaxation. Pretreatment with the nitric oxide synthase inhibitor, L-N(G)-nitroarginine methyl ester, and the guanylate cyclase inhibitor, methylene blue, prevents the vasorelaxation induced by PAHs. These results indicate that the vasorelaxation effect of PAHs is mediated by activation of nitric oxide synthase of endothelium.

Assessment of the mutagenic activity of some acenaphtho(1,2-b)quinolines.

The mutagenic activity of four substituted acenaphtho(1,2-b)quinolines were evaluated using the Ames test (Salmonella assay). The compounds tested were acenaphtho(1,2-b)quinoline (1) and its 10-methoxy (2), 10-methyl (3) and 10-nitro (4) derivatives. Compounds 1 and 3 were found to be non-mutagenic, but compound 2 was found to be mutagenic with metabolic activation only. However, compound 4 was found to be very active with or without activation.

Early life stage toxicity tests with a saltwater fish: effects of eight chemicals on survival growth, and development of sheepshead minnows (Cyprinodon variegatus).

Flow-through, acute (96-h), and early life stage (28-d after hatch) toxicity tests were performed with eight chemical on a saltwater fish, sheepshead minnows (Cyprinodon variegatus). Chemical effects on survival, growth, and development were determined. Maximum acceptable toxicant concentrations (MATCs) were greater than 3.2 less than 7.7 mg/l for toluene, greater than 0.52 greater than 0.97 mg/l for acenaphthene, greater than 80 less than 156 mg/l for isophorone, greater than 10 less than 16 mg/l for 4-nitrophenol, greater than 4.8 less than 8.5 mg/l for bromoform, greater than 0.39 less than 0.79 mg/l for 1-chloronaphthalene, greater than 0.09 less than 0.18 mg/l for 1, 2, 4, 5-tetrachlorobenzene, and less than 0.36 mg/l for 2, 4-dichloro-6-methylphenol; application factors were 0.25-0.59, 0.17-0.31, greater than or equal to 0.57, 0.31-0.50, greater than or equal to 0.68, greater than or equal to 0.56, 0.27-0.54, and less than 0.10. respectively. Test results reported here were compared with results of static, acute toxicity tests conducted previously with six species of aquatic organisms and the same chemicals.