Improved application of recombinant yeast assays on environmental samples by size exclusion chromatography

The utility of HPLC with a size exclusion chromatography (SEC) column for the fractionation of environmental samples was examined to expand the applicability of recombinant yeast assays (RYAs) for the measurement of estrogenic activity in environmental samples. The elution of steroidal hormone standards through SEC was tested, and river water and sediment samples were subjected to SEC fractionation. The retention times of all estrogen standards were longer than those of the major components with absorbance at 230 nm of the environmental sample matrix. The estrogenic activity of SEC fractions of environmental samples from highly polluted sites was measured using RYA and revealed the existence of estrogenic compounds that could not be detected or quantified in extracted samples before fractionation. The fractions from environmental samples that corresponded to the retention time of estrone, 17[small beta]-estradiol, and estriol were analyzed with LC/MS/MS. These three estrogens were separated into three different fractions, and the concentration of estrone coincided with the estrogenic activity of the fraction in which it was detected. The profile of estrogenic activity of SEC fractions indicated the molecular size of the estrogenic compounds and the yeast growth inhibitors in samples.     

Concentrations of polychlorinated dibenzo-p-dioxins, dibenzofurans, non-ortho polychlorinated biphenyls, and mono-ortho polychlorinated biphenyls in Japanese flounder, with reference to the relationship between body length and concentration

The concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), non-ortho polychlorinated biphenyls (non-ortho PCBs), and mono-ortho polychlorinated biphenyls (mono-ortho PCBs) in Japanese flounder (Paralichthys olivaceus, body length 10.4-36.6 cm) collected from Sendai Bay, Japan, were determined using high-resolution gas chromatography-mass spectrometry. The relationship between the concentrations of these compounds (dioxins) and the body length of the Japanese flounder was examined. The total PCDD and total PCDF concentrations did not correlate with body length (both r(2) < 0.1, both p > 0.05), whereas the total non-ortho PCB and total mono-ortho PCB (coplanar polychlorinated biphenyls, Co-PCBs) concentrations were significantly correlated (r(2)= 0.8, p < 0.05 and r(2)= 0.63, p < 0.05, respectively). The bioaccumulation properties of PCDD/Fs in Japanese flounder differed from those of Co-PCBs. Toxicity equivalency quotient (TEQ) values derived from the Co-PCBs made up 46.3%-63.7% of the total TEQ value for all the dioxins. Although the concentrations of non-ortho PCBs were lower than those of mono-ortho PCBs, the TEQ value for non-ortho PCBs was higher than that for mono-ortho PCBs. The TEQ value for non-ortho PCBs increased more with increasing body length than did the values for PCDDs, PCDFs, and mono-ortho PCBs. These results show that from the standpoint of risk management, non-ortho Co-PCBs are the most important of the dioxins in Japanese flounder.     

Carbon Molecular Sieving Membranes Derived from Lignin-Based Materials

Carbon molecular sieving membranes were prepared by pyrolysis of lignocresol derived from lignin by the phase-separation method. Lignocresol membranes formed by a dip process on a porous -alumina tubing were carbonized at 400–800°C under nitrogen atmosphere. The thickness of the membrane formed on the outer surface of the substrate was about 400 nm judging from SEM observation. Gas-evolving behavior of lignocresol was measured using thermogravimetry-mass spectrometry (TG-MS). The gaseous products evolved from lignocresol included a number of fragments with higher molecular weights; whereas those from phenolic resin are mainly due to phenol and methylphenol. These evolved pyrolysis fragments effectively contribute to micropore formation of carbonized lignocresol membranes. Gas permeation rates through the membrane decreased in the order of increasing kinetic molecular diameter of the penetrant gas, and the membrane behaved like a molecular sieve. The permeation properties were dependent on heating conditions, and a pyrolysis temperature of 600°C gave the best membrane performance. Gas selectivities of the membrane prepared at 600°C were 50, 8, 290, and 87 for CO₂/N₂, O₂/N₂, H₂/CH₄, and CO₂/CH₄ at 35°C, respectively.     

Empirical research on diffusion behavior of leaked gas in the ground

Chemical plants and gas utilities own large underground pipelines to transport material such as combustible gas. For example, city gas utilities in Japan have about 230,000 km of pipelines, even if only the pipelines to deliver gas to their customers are considered. Any accidents involving such pipelines can lead to enormous human and physical damage, and their security is therefore a top-priority issue for utilities. For the safety management of underground pipelines, in addition to assessment of the long-term reliability of pipeline materials, it is extremely important to understand the diffusion behavior of gases in the ground, such as the diffusion range and time, in the case of leakage, and the impact on the surrounding area is a fundamental factor to be considered in the design and maintenance of safe facilities and for emergency response. Although many papers introduce the situations of gas diffusion in the atmosphere such as indoor and outdoor conditions, only fundamental surveys have been conducted on gas diffusion in the ground, and there have been few full-scale empirical studies. This study reports the results of the verification of the diffusion behavior with full-scale gas leakage experiments simulating real underground pipelines, as well as the outcomes of the applicability test of a numerical simulation model investigated and proposed based on the results. This technical knowledge regarding security will contribute to further improvement of safety in the industry.     

Pathway of 17��-estradiol degradation by Nitrosomonas europaea and reduction in 17��-estradiol-derived estrogenic activity

In recent years, natural and synthetic estrogens have been recognized as endocrine disruptors in aquatic organisms. Although natural and synthetic estrogens are known to be degraded by microbes, only limited information about their degradation pathways is available. Here, we studied the degradation pathways of a natural estrogen, 17β-estradiol, by the nitrifying microorganism Nitrosomonas europaea, and we determined whether the degradation products of 17β-estradiol had estrogenic activity. To identify the degradation products, we subjected the culture solution to solid-phase extraction, and the extract was analyzed by gas chromatography–mass spectrometry. The potential estrogenic activity of the degradation products was investigated by means of a yeast two-hybrid assay. 1,3,5(10),16-Estratetraen-3-ol (estratetraenol) was newly identified as a degradation intermediate produced by dehydration of 17β-estradiol. Estratetraenol was also degraded by N. europaea, and its degradation rate was faster than that of 17β-estradiol. The two-hybrid assay confirmed that estratetraenol acted as a ligand for the estrogen receptor; estratetraenol thus has potential estrogenic activity. N. europaea eliminated the estrogenic activity derived from 17β-estradiol. This paper is the first to report dehydration as a mechanism of microbial estrogen degradation.     

Transformations of selected pesticides in flooded rice-field soil — A review

Rice is mostly cultivated on soil held under flooded conditions during the growing season, where the plow layer differentiates into a thin oxidized surface layer and an underlying reduced layer. Under these conditions, certain pesticides undergo reductive transformations which are characteristic to rice fields and other anaerobic systems. Thus, lindane and pentachlorophenol are reductively dechlorinated into less toxic products, whereas, in highly reduced soils, thiobencarb is dechlorinated into a substance phytotoxic to rice plants. Nitrophenyl compounds, such as parathion and chlornitrofen, are converted into respective amino analogs, which are apt to remain as soil-bound residues. Transformation processes of these and several other environmentally important pesticides used in rice fields in the past and/or at present are reviewed.     

Estrogen content and relative performance of Japanese and British sewage treatment plants and their potential impact on endocrine disruption.

Both the UK and Japan are densely populated islands with relatively short rivers. Therefore, both countries are likely to be highly exposed to contaminants emanating from their human populations. This review considered how effective the different sewage treatment facilities of the two countries are at removing steroid estrogens from the effluent. The methods of estrogen analysis in sewage effluent, the number and importance of different sewage treatment types, and their apparent effectiveness at removing estrogens were all considered. In both countries, the activated sludge treatment was dominant in terms of people served and water discharged. The analytical techniques used by those studying estrogen concentrations in effluents in both countries were broadly similar. Activated sludge plant (ASP) effluent in the UK typically contained around 2 ng/L estradiol (E2) and 8 ng/L estrone (E1), while Japanese ASPs typically reported E2 as below detection, and 10 ng/L E1 in their effluents. When estrogenic bioassays were used in Japan, they typically record an estrogenic potency of 10 ng/L E2 equivalents. Even taking into account ethinylestradiol (EE2) (not found in Japanese effluents), the overall estrogenicity of British sewage effluents would appear to be the same as that of Japanese sewage effluents (around 10 ng/L E2 equivalents). This suggests that the ASPs serving the large urban communities in Japan and the UK would have effluent of similar estrogenic potencies. Less information is available about the more numerous biological (trickling) filter plants (BFP) in the UK and oxygen ditches (OD) in Japan which tend to serve smaller, more rural communities. The available data would suggest that the BFPs are significantly less efficient than the ODs at removing E1. This would suggest that in similar circumstances, British headwaters (where this sewage treatment plant (STP) type is often found) might be more at risk from endocrine disruption than their Japanese counterparts. Overall, the apparently higher incidence of endocrine disruption in British wild fish than in Japanese wild fish cannot be attributed to differences in the efficiency of their respective STPs.     

Studies on automated survey system of environmental chemicals by gas chromatography-mass spectrometry-computer III. Construction of an automated survey system and its application

An automated survey system on the chemicals in environments by a gas chromatography-mass spectrometry-computer system has been constructed of Three Dimensional Mass Chromatography, Modified Probability Based Matching method, and Self Training Interpretive and Retrieval method. And it was applied to analyze environmental materials, with interesting results.     

High (210)Po atmospheric deposition flux in the subtropical coastal area of Japan

Bulk atmospheric deposition fluxes of (210)Po and (210)Pb were measured at three coastal regions of Japan, the Pacific Ocean coastal area of the Japanese mainland (Odawa Bay), the Chinese continental side of Japanese coastal area (Tsuyazaki), and an isolated island near Okinawa (Akajima). Wet and dry fallout collectors were continuously deployed from September 1997 through August 1998 for periods of 3 to 31 days depending on the frequency of precipitation events. Annual (210)Pb deposition fluxes at Odawa Bay (35 degrees N 139 degrees E), Tsuyazaki (33 degrees N 130 degrees E) and Akajima (26 degrees N 127 degrees E) were 73.3+/-8.0, 197+/-35 and 78.5+/-8.0 Bqm(-2)y(-1), respectively. Higher (210)Pb deposition was observed at the Chinese continental side of Japanese coast than at the Pacific Ocean coastal site. The high (210)Pb atmospheric flux at the Chinese continental side coast was thought to be attributable to (222)Rn-rich air-mass transport from the Chinese continent during the winter monsoon. In contrast, the annual (210)Po deposition fluxes at the three study sites were 13.0+/-2.3 (Odawa Bay), 21.9+/-4.4 (Tsuyazaki) and 58.4+/-7.7 (Akajima)Bqm(-2)y(-1), respectively, indicating unusual high (210)Po deposition at Akajima during winter. Anomalous unsupported (210)Po input was observed during summer 1997, suggesting unknown source of (210)Po at this area.