Age-related settlement success by cyprids of the barnacleBalanus amphitrite,with special reference to consumption of cyprid storage protein

Newly molted (0-d-old) cyprids of the barnacleBalanus amphitrite Darwin were prevented from settling for 0 to 14 d at four different temperatures (25, 20, 15 and 5°C treatments). The effect on settlement success of prolonging the cyprid lifetime was evaluated using a nitrilocellulose membrane assay. In addition, protein extract prepared from these cyprids was analyzed using gel electrophoresis to characterize the effect of age on protein content and composition. Settlement success was significantly affected for larvae aged at 25 (P < 0.001), 20 (P < 0.001) and 15°C (P < 0.05), while differences in settlement success between age groups was negligible at 5°C (P = 0.09). Settlement success of cyprids increased with time for up to 3 d (P < 0.001, Phase 1), following which settlement success significantly declined (P < 0.001, Phase 11). Temperature had no significant effect on settlement in Phase I (P = 0.17), but did enhance the decline in settlement success with age during Phase II (P < 0.001). Gel electrophoresis revealed a significant decline in the quantity of the cyprid storage protein CMP (Cyprid Major Protein) with increasing age at 25, 20 and 15°C, but CMP levels remained constant at 5°C. These results suggest that, upon molting to the cyprid stage, larvae may still require a settlement-competence attainment period. This may be achieved by CMP utilization during Phase I, depletion of which during Phase II may be responsible for reduction in settlement success with cyprid age such that remaining CMP stores can no longer support the production of adult structures following settlement.     

Identification of the dimethylbenzyl mercapturic acid in urine of rats administered with 1,2,4-trimethylbenzene

A study was undertaken of the mercapturic acid metabolism of 1,2,4-trimethylbenzene in the rat. Of three regioisomeric dimethylbenzyl mercapturic acids, i.e. 2,4-, 2,5- and 3,4-dimethylbenzyl isomers, the third isomer was not found in the urinary mercapturic acid isolated by preparative HPLC, from the comparison of NMR spectrum of the isolate with those of authentic specimens. The urinary mercapturate was then assigned to 2,4- and/or 2,5-dimethylbenzyl isomers. The excretion rate of the mercapturic acid was 14-20% of dose as 2,4-dimethylbenzyl isomer.     

Simulation of dioxin accumulation in human tissues and analysis of reproductive risk

Polychlorinated dibenzo-p-dioxin, polychlorinated dibenzofuran and dioxin-like polychlorinated biphenyl concentrations in human liver, kidney, fat, blood, muscle, richly perfused tissue (brain, lung etc.) and skin were simulated to assess the health risk for Japanese fetuses. A 40-year time course of dioxin accumulation via food ingestion was simulated using a physiologically based pharmacokinetic (PBPK) model. In richly perfused tissue, the concentration estimated by the PBPK model showed better agreement with measured concentrations than that calculated by the one-compartment model. Fetal dioxin concentration was simulated based on the assumption that the fetal concentration was almost equal to the concentration in the mother's richly perfused tissue. To assess the reproductive risk, the estimated concentration in human fetus was compared with that in rat fetus in which reproductive function showed signs of alteration by 2,3,7,8-TCDD in previous reports [Toxicol. Appl. Pharmacol. 114 (1992) 118; 146 (1997) 11; Toxicol. Sci. 53 (2000) 411; 57 (2000) 275]. The present daily intake of 2,3,7,8-TCDD is approximately 1/50 of the amount that leads to possible reproductive toxicity in the next generation. However, when 29 kinds of dioxin congeners are considered, the present level is 1/5 of the hazardous levels. For species extrapolation of dioxin risk, further study on tissue concentration versus toxicity is required.     

Isomer distribution of nitrotriphenylenes in airborne particles, diesel exhaust particles, and the products of gas-phase radical-initiated nitration of triphenylene

The formation of mutagenic nitro-polycyclic aromatic hydrocarbons (NPAHs) 1- and 2-nitrotriphenylene (1- and 2-NTP) via gas-phase OH or NO₃ radical-initiated reactions of triphenylene was demonstrated for the first time using a flow reaction system. In contrast with the results of conventional electrophilic nitration, 2-NTP was formed in larger yield than 1-NTP, but this is consistent with the mechanism proposed for gas-phase radical-initiated nitration of PAH. In diesel exhaust particle (DEP) samples, both 1- and 2-NTP were identified and their concentrations determined, as well as 1-nitropyrene (1-NP), which is a representative combustion-derived NPAH: the mean concentrations of 1-NTP, 2-NTP, and 1-NP were 4.7, 1.9, and 32 pmol mg_DEP^–1, respectively. The mean 2-NTP/1-NTP, 1-NTP/1-NP, and 2-NTP/1-NP ratios in samples of airborne particles collected in a residential area in Osaka, Japan, were>1.55,<0.25, and 0.37, respectively; these values are much higher than those of the DEP samples. This finding indicates that there is another source for airborne NTPs, especially 2-NTP, apart from diesel exhaust. These results strongly suggest that airborne NTPs originate from atmospheric processes such as radical-initiated reactions of triphenylene, and this has a significant influence on the atmospheric occurrence of NTPs.     

Determination of tissue-blood partition coefficients for a physiological model for humans, and estimation of dioxin concentration in tissues

The tissue-blood partition coefficients for a physiologically based pharmacokinetic (PBPK) model were determined, and the concentrations of 17 congeners of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs) in tissues in Japanese people were estimated using the model. According to the PBPK model established by Lawrence and Gobas [Chemosphere 35 (1997) 427-452], we assumed a steady-state fugacity model for Japanese people in general, and set the route of PCDD/Fs exposure only from food intake. The required partition coefficients for liver, kidney, adipose, muscle, skin, bile, gut and viscera (richly perfused tissue) were calculated using available autopsy data from eight Japanese men and women who were not accidentally exposed to PCDD/Fs. For validation of the partition coefficients, estimated PCDD/F concentrations in liver, kidney, fat, blood and muscle using the model were compared to other two sets of measured concentration data in Japanese tissues. Good agreement was obtained between estimated data and measured data, and most of the measured data were within the simulated concentration range in liver, kidney, blood and muscle. From these results, our model and calculated partition coefficients seem applicable for the estimation of congener-specific concentrations in human tissues.     

Formation of mutagens by photolysis of amino acids in neutral aqueous solution containing nitrite or nitrate ion

Nine amino acids, i.e., alanine, threonine, cysteine, glutamic acid, arginine, proline, tryptophan, phenylalanine and tyrosine, were irradiated with UV light in water containing nitrite or nitrate ion under neutral conditions. The mutagenicities of the ether extracts and the residual water layers of the reaction mixtures were assayed with and without S-9 mix using $\text{Salmonella typhimurium}$ TA98 and TA100. Three aromatic amino acids, tryptophan, phenylalanine and tyrosine, were found to give direct-acting and frameshift mutagens by irradiation in aqueous nitrite solution. Among them, the ether extract of tryptophan exhibited the strongest mutagenicity toward TA98. In the case of irradiation in aqueous nitrate solution, only the ether extract of tryptophan exhibited weak mutagenicity toward TA98 without S-9 mix. The effects of nitrite concentration, irradiation time and pH on mutagen formation from tryptophan and some characteristics of the produced mutagens were examined.     

Association of the mutagenicity of airborne particles with the direct emission from combustion processes investigated in Osaka, Japan

The association of the direct-acting mutagenicity of soluble organic fraction of airborne particles toward Salmonella typhimurium YG1024 strain with the direct emission was investigated at a roadside and at a residential area in Osaka, Japan. The direct-acting mutagenicity was evaluated as mutagenic activity per unit volume of ambient air (rev m⁻³) and/or that per airborne particulate weight collected on a filter (rev mg⁻¹). The annual or diurnal changes of the mutagenicity of airborne particles at the residential site showed similar patterns to those of some gaseous pollutants such as NO₂ and SO₂, which were emitted from combustion processes. This result indicates that the mutagenicity is mainly attributable to the primary emissions. From the analysis of the relationship between the wind sector and the mutagenic intensity, rev m⁻³ and rev mg⁻¹ values were strongly affected by the emissions from the fixed sources and from the mobile sources, respectively. The rev m⁻³ value and concentration of 1-nitropyrene (1-NP) in unit per m³ at the roadside were a factor of 2.6 and 2.8 higher than those at the residential site, respectively, but the rev mg⁻¹ value and concentration of 1-NP in unit per mg at the roadside were substantially comparable to those at the residential area. These observations suggest that the characteristics of the airborne particles can be attributed to the automotive emissions even at the suburban area.     

Oyster Contamination with Human Noroviruses Impacted by Urban Drainage and Seasonal Flooding in Vietnam

This study investigated the level of norovirus contamination in oysters collected at a lagoon receiving urban drainage from Hue City for 17 months (August 2015–December 2016). We also investigated the genetic diversity of norovirus GI and GII in oyster and wastewater samples by using pyrosequencing to evaluate the effect of urban drainage on norovirus contamination of oysters. A total of 34 oyster samples were collected at two sampling sites (stations A and B) in a lagoon. Norovirus GI was more frequently detected than GII (positive rate 79 vs. 41%). Maximum concentrations of GI and GII were 2.4 × 10⁵ and 2.3 × 10⁴ copies/g, respectively. Co-contamination with GI and GII was observed in 35% of samples. Norovirus GII concentration was higher at station A in the flood season than in the dry season (P = 0.04, Wilcoxon signed-rank test). Six genotypes (GI.2, GI.3, GI.5, GII.2, GII.3, and GII.4) were identified in both wastewater and oyster samples, and genetically similar or identical sequences were obtained from the two types of samples. These observations suggest that urban drainage and seasonal flooding contribute to norovirus contamination of oysters in the study area.     

Microcystin accumulation and biochemical responses in the edible clam Corbiculaleana P. exposed to cyanobacterial crude extract

We investigated the accumulation and effects of cyanobacterial crude extract(CCE)containing microcystins(MCs) on the edible clam Corbicula leana P. Toxic effects were evaluated through the activity of antioxidant and detoxification enzymes: catalase(CAT),superoxide dismutase(SOD), and glutathione-S-transferases(GSTs) from gills, foot, mantle and remaining soft tissues. Clams were exposed to CCE containing 400 μg MC-LReq/L for10 days and were then kept in toxin-free water for 5 days. Clam accumulated MCs(up to 3.41 ± 0.63 μg/g dry weight(DW) of unbound MC and 0.31 ± 0.013 μg/g DW of covalently bound MC). Detoxification and antioxidant enzymes in different organs responded differently to CCE during the experiment. The activity of SOD, CAT, and GST in the gills and mantle increased in MC-treated clams. In contrast, CAT and GST activity was significantly inhibited in the foot and mostly only slightly changed in the remaining tissues. The responses of biotransformation, antioxidant enzyme activity to CCE and the fast elimination of MCs during depuration help to explain how the clam can survive for long periods(over a week) during the decay of toxic cyanobacterial blooms in nature.     

Indigo carmine removal by charcoal from rice bran as an agricultural by-product

On the utilization of agricultural by-product, charcoal from rice bran was produced. The adsorption characteristics of indigo carmine onto charcoals from rice bran were investigated by the batch method. The yield and surface area of charcoal were decreased and increased with increase of carbonization temperature, respectively. The removal ratio of indigo carmine was high with the charcoal carbonized at higher temperature. In the relationship between the amount of indigo carmine adsorbed and the square root of elapsed time, a good linearity was recognized. The kinetic constant of adsorption removal for indigo carmine was rapid in the charcoal carbonized at higher temperature. The adsorption characteristics for indigo carmine removal by charcoal from rice bran were dominated by the value of their surface area.