Formation of estrogenic products from environmental phthalate esters under light exposure

Phthalate esters (PEs) have been suspected to be environmental endocrine disruptors and the detailed mechanism remains unclear. The activities of these chemicals can be enhanced through chemical modification under the environmental conditions. We demonstrate that PEs acquire unequivocal estrogenic activity by light exposure. Through UV exposure of an aqueous PE solution, one active photoproduct, identified as 4-hydroxyPE (PE-4OH) based on its characteristic UV and mass spectra, was detected in an estrogen receptor alpha-dependent transactivation assay. PE-4OH was effectively generated by UV 290 nm. The PE-4OH production accompanied H2O2 generation in a UV dose-dependent manner. Both PE and UV irradiation were indispensable in the generation of H2O2. Addition of H2O2 to the PE solution increased PE-4OH production under UV irradiation. The PE-4OH production was also observed in the PE reaction with the Fenton reagent generating hydroxyl radical without UV irradiation. The proposed mechanism for PE-4OH production based on these results is such that by PE-mediated photosensitization H2O2 is generated from O2 and H+ and decomposed to hydroxyl radical, thus oxidizing the PE benzene ring. The PEs-4OH are remarkably active estrogenic products of PEs and would be involved in ER-mediated endocrine disruption.     

Induction of larval settlement and metamorphosis of the sea urchin Strongylocentrotus intermedius by glycoglycerolipids from the green alga Ulvella lens

In aquaculture centers of the northern region of Japan, "Nami-ita" (waved polycarbonate plates), on which the green alga Ulvella lens Crouan frat. (Chaetophoraceae: Chaetophorales) was cultured, are used to promote larval settlement and metamorphosis of the sea urchin species Strongylocentrotus intermedius (A. Agassiz) and S. nudus (A. Agassiz). We investigated chemical inducer(s) for larval settlement and metamorphosis of these sea urchins with extracts of U. lens. Bioassay-guided separation of the methanol extract using a combination of column and thin-layer chromatography led to the isolation of several active compounds, the chemical structures of which were determined by spectral and chemical methods. These active compounds were identified as glycoglycerolipids, all comprising several molecular species: sulfoquinovosyl monoacylglycerols (SQMGs), sulfoquinovosyl diacylglycerols (SQDGs), monogalactosyl monoacylglycerols (MGMGs), monogalactosyl diacylglycerols (MGDGs), digalactosyl monoacylglycerols (DGMGs) and digalactosyl diacylglycerols (DGDGs). Among these glycolipids, SQMGs, MGMGs, MGDGs and DGMGs induced larval metamorphosis of the sea urchin S. intermedius. SQMGs and MGDGs induced larval metamorphosis at a concentration of 5 µg ml^-1, whereas SQDGs and DGDGs only induced larval settlement. These glycoglycerolipids are new congeners of chemical inducers to settlement and metamorphosis of planktonic larvae of sea urchins. The findings would provide a better understanding of larval settlement and metamorphosis in sea urchins.     

Depth profiles of volatile halogenated hydrocarbons in seawater in the Bay of Bengal

Measurements were made of bromocarbons (CHBr3 and CH2Br2), iodocarbons (CH2I2 and CH2ClI), and dimethylsulfide (DMS, CH3SCH3) in seawater collected from the Bay of Bengal under tropical stratified conditions. These compounds showed different depth profiles, characteristic of each group. CH2I2 and CH2ClI showed very similar depth profiles to chlorophyll-a, suggesting their production by phytoplankton followed by rapid decay in seawater. The CH2I2 maximum at a depth a little below the CH2ClI maximum was consistent with its more significant photolytic decay. The bromocarbons were less localized in their distributions than were the iodocarbons, suggesting their longer residence time in seawater after their release from phytoplankton. Both of these profiles were different from the pattern of DMS, which had its maxima above the chlorophyll-a maximum layer near the surface.     

Annual life cycle of Chattonella spp., causative flagellates of noxious red tides in the Inland Sea of Japan

Effects of storage temperature on the dormancy and maturation of dormant cells of Chattonella spp. were investigated by the extinction dilution method using sediment samples collected in 1984 after the blooming in Harima-Nada. Seasonality of germinability in fresh sediments was also studied in 1985. A storage period of more than 4 months with a low temperature (11°C) was essential for the maturation to induce the germination capacity of dormant cells. The optimal temperature for germination (22°C) had no effects on the maturation. Using fresh Suo-Nada sediments, a marked seasonality of germinability was confirmed. It was weak from autumn to early winter, then strengthened gradually up to a high level, which was maintained between spring and early summer, and again decreased rapidly during summer. The annual life cycle of Chattonella spp., including vetetative and dormant phases, was summarized as follows: (1) vegetative cells in early summer originate from germination of dormant cells in sediments; (2) they form overwintering dormant cells during the summer season; (3) dormant cells spend a period of spontaneous dormancy until next spring; (4) the duration of post dormancy, an enforced one due to low temperatures, follows until early summer. The life cycle of Chattonella spp. is therefore well adapted to the temperature regime in temperate seas such as the Seto Inland Sea of Japan.     

The effect of female wings on male courtship behavior in the cricket Gryllus bimaculatus

Sexual dimorphism exists in the shape and the structure of the forewings of the cricket (Gryllus bimaculatus). However, the functional significance of the wings in the G. bimaculatus female has been unclear. In common blue butterflies (Polyommatus icarus), wings in females have been suggested as being important for attracting males. To test whether female crickets need wings for conspecific males to recognize them and initiate mating behavior, we removed all wings from females and observed the behavior of males towards them. Most males (87.5%) showed mating behavior towards the wingless females: they produced courtship song and transferred spermatophores to the wingless females. Similarly, 88.5% of the males showed mating behavior towards intact females. When males were placed with both a wingless female and an intact female, no significant difference was detected in male mate choice. The findings demonstrate that the wing of the G. bimaculatus female is not necessary for female recognition by conspecific males and the initiation of male mating behavior, and that it is not important in male mate choice.     

Screening of agonistic activities against four nuclear receptors in wastewater treatment plants in Japan using a yeast two-hybrid assay

To assess the potential endocrine disruptive effects through multiple nuclear receptors(NRs),especially non-steroidal NRs,in municipal wastewater,we examined the agonistic activities on four NRs(estrogen receptor α,thyroid hormone receptor α,retinoic acid receptor α and retinoid X receptor α) of untreated and treated wastewater from municipal wastewater treatment plants(WWTPs) in Japan using a yeast two-hybrid assay.Investigation of the influent and effluent of seven WWTPs revealed that agonistic activities...     

Patchiness and bistability in the comprehensive cyanobacterial model (CCM)

We analyzed spatiotemporal dynamics of cyanobacteria using a four-component mathematical model with nutrients, unicellular algae, colonial algae and herbivorous zooplankton. One of the astonishing abilities of cyanobacteria is the morphological change from the unicellular type to the colonial or filamentous type in response to grazing activities of zooplankton, the phenomenon known as phenotypic plasticity. Our model, referred to as comprehensive cyanobacterial model (CCM), includes the effects of phenotypic plasticity. Depending on parameter values, CCM exhibits not only bistability but also limit cycle oscillations without showing the paradox of enrichment, which has been a controversy among mathematical ecologists. Further, CCM is modified to reaction–advection–diffusion equations, the simulation results of which indicate that the ratio of the lateral diffusivity to the turbulent velocity significantly affects the appearance of patchiness patterns.     

Efficacy of forming biofilms by naphthalene degrading Pseudomonas stutzeri T102 toward bioremediation technology and its molecular mechanisms

In natural environments, bacteria often exist in close association with surfaces and interfaces. There they form "biofilms", multicellular aggregates held together by an extracellular matrix. The biofilms confer on the constituent cells high resistance to environmental stresses and diverse microenvironments that help generate cellular heterogeneity. Here we report on the ability of Pseudomonas stutzeri T102 biofilm-associated cells, as compared with that of planktonic cells, to degrade naphthalene and survive in petroleum-contaminated soils. In liquid culture system, T102 biofilm-associated cells did not degrade naphthalene during initial hours of incubation but then degraded it faster than planktonic cells, which degraded naphthalene at a nearly constant rate. This delayed but high degradation activity of the biofilms could be attributed to super-activated cells that were detached from the biofilms. When the fitness of T102 biofilm-associated cells was tested in natural petroleum-contaminated soils, they were capable of surviving for 10 wk; by then T102 planktonic cells were mostly extinct. Naphthalene degradation activity in the soils that had been inoculated with T102 biofilms was indeed higher than that observed in soils inoculated with T102 planktonic cells. These results suggest that inoculation of contaminated soils with P. stutzeri T102 biofilms should enable bioaugmentation to be a more durable and effective bioremediation technology than inoculation with planktonic cells.     

Evaluation of pressurized liquid extraction for the analysis of four pesticides in unpolished rice

The aim of to evaluate efficiency of this study was extraction pressurized liquid extraction (PLE) for the analysis of four pesticides, fthalide, etofenprox, fenitrothion, and isoprothiolane, in unpolished rice by comparing with homogenization as a reference technique. The concentrations of four selected pesticides obtained by PLE with acetonitrile at 130°C for 10 min × 2 cycles were comparable to those by homogenization with water-soaking. The repeatability of the analysis, represented as relative standard deviations (RSDs), were 1.4–3.6% (n = 3) for PLE at 130°C and 1.2–3.8% (n = 3) for homogenization with water-soaking. Recovery yields of surrogates were 75–88% and 87–109% for PLE at 130°C and homogenization with water-soaking, respectively, and these were satisfactory according to the method of positive list. This study suggested that PLE can be applied for the analysis of selected four pesticides in unpolished rice as well as homogenization with water-soaking.     

Two-stage thermal gasification of polyolefins

Plastic pellets of polyethylene (PE), polypropylene (PP), and polystyrene (PS) were gasified in a two-stage thermal degradation process. The first stage is the conversion of polyolefins to distilled oils using a melting vessel. In the second stage, the oils from the first stage are gasified using a tubular reactor. The distilled oil yields of PE, PP, and PS in the first stage were 84, 89, 92 wt%, respectively, each at 470°C. The total gas yields of PE, PP, and PS in the second stage were 80, 74, and 6.2 wt%, respectively, each at 800°C. The main components of the product gas for PE and PP were methane and olefins such as ethene and propene. Some aromatic oils, including benzene, toluene, and xylene, were also produced as by-products. The amount of carbonaceous residue, or coke, was very low (less than 1 wt%). By dividing the process into two stages, the coking rate was considerably reduced compared with direct gasification of the polyolefins. Received: July 19, 2000 / Accepted: September 17, 2000