Enantioselective recognition of mono-demethylated methoxychlor metabolites by the estrogen receptor

Metabolites of methoxychlor such as 2-(p-hydroxyphenyl)-2-(p-methoxyphenyl)-1,1,1-trichloroethane (mono-OH-MXC) and 2,2-bis(p-hydroxyphenyl)-1,1,1-trichloroethane (bis-OH-MXC), have estrogenic activity. Mono-OH-MXC is a chiral compound in which the carbon atom bridging two benzene rings is the chiral centre. In previous studies the estrogenic activity of racemic mono-OH-MXC has been measured, and the activity of each enantiomer of this compound has not yet been elucidated. In this study, we evaluated the estrogen receptor-binding activity of each enantiomer of mono-OH-MXC to clarify the enantioselective recognition by the estrogen receptor. (S)-mono-OH-MXC showed 3-fold higher binding activity than that of the (R) enantiomer. The activity of bis-OH-MXC was only 1.7-fold higher than that of (S)-mono-OH-MXC. This result suggests that the one hydroxy group and the orientation of the CCl3 group of mono- and bis-OH-MXCs are important for the interaction with the estrogen receptor. The result also points out the estrogenic activity of methoxychlor after metabolic activation in vivo, which predominantly produces the (S)-mono-OH-MXC, may be higher than estimated from the in vitro activity of racemic mixtures.     

Enzymatic Degradation and Aminolysis of Microbial Poly(3-hydroxybutyrate-co-4-hydroxybutyrate) Single Crystals

Solution-grown single crystals of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] were hydrolyzed by polyhydroxybutyrate (PHB) depolymerase from Ralstonia pickettii T1. Enzymatic degradation proceeded from the edges of lamellar crystals, yielding serrated contour and small crystal fragments. Gel permeation chromatography analysis revealed that the molecular weights of the crystals decreased during enzymatic degradation, suggesting that the enzymatic hydrolysis of chain-folding regions at the crystal surfaces occurred in addition to the enzymatic degradation at crystal laterals or edges. After P(3HB-co-4HB) single crystals were aminolysed in 20% aqueous methylamine solution to remove the folded-chain regions and enzymatic degradation by lipase from Rhizopus oryzae to remove 4HB components at crystal surfaces of single crystal aminolyzed, it was found that a small amount (up to ca. 2 mol%) of 4HB component can be incorporated into the P(3HB) mother crystal lattice irrespective of the 4HB content.     

Combination of adsorption and biodegradation processes for textile e uent treatment using a granular activated carbon-biofilm configured packed column system

The objective of this study was to investigate the feasibility of using a granular activated carbon-biofilm configured packed column system in the deeolodzation of azo dye Acid Orange 7-containing wastewater.The Acid Orange 7-degrading microbial from anaerobic sequencing batch reactor which treating the azo dye-containing wastewater for more than 200 d was immobilized on spent granular activated carbon(GAC)through attachment.The GAC-biofilm configured packed column system showed the ability to decolorize...     

Restoration of cadmium-contaminated paddy soils by washing with ferric chloride: Cd extraction mechanism and bench-scale verification

The ability of FeCl3 to extract Cd from three paddy soils was compared with that of various irons, manganese, and zinc salts to elucidate the extraction mechanism. Manganese, zinc and iron salts (including FeCl3) extracted 4-41%, 8-44% and 24-66% of total Cd, respectively. This difference reflected the pH of the extraction solution, indicating that the primary mechanism of Cd extraction by FeCl3 is proton release coupled with hydroxide generation, as iron hydroxides are insoluble. Washing with FeCl3 led to the formation of Cd-chloride complexes, enhancing Cd extraction from the soils. FeCl3 effectively extracted Cd from all of the three soils compared to HCl that is a conventional washing chemical, when the concentrations of the two washing chemicals were between 15 and 60mM(c) (at above extraction pH 2.4), while the corresponding extraction pH of FeCl3 was slightly higher than HCl. As HCl is the strong acid of complete dissociation, if excess amount of HCl was added to soil, it is possible to give the dissolution of clay minerals in soils. In contrast, proton release from FeCl3 is controlled by the chemical equilibrium of hydroxide formation. While soil fertility properties were affected by a bench-scale soil washing with 45mM(c) FeCl3, adverse effects were not crucial and could be corrected. The bench-scale test confirmed the effectiveness of FeCl3 for removal of soil Cd. The washing had no negative effect on rice yield and lowered the Cd concentration of rice grain and rice straw in a pot experiment.     

Decolorization of Orange II using an anaerobic sequencing batch reactor with and without co-substrates

We investigated the decolorization of Orange II with and without the addition of co-substrates and nutrients under an anaerobic sequencing batch reactor (ASBR). The increase in COD concentrations from 900 to 1750 to 3730 mg/L in the system treating 100 mg/L of Orange II-containing wastewater enhanced color removal from 27% to 81% to 89%, respectively. In the absence of co-substrates and nutrients, more than 95% of decolorization was achieved by the acclimatized anaerobic microbes in the bioreactor treating 600 mg/L of Orange II. The decrease in mixed liquor suspended solids concentration by endogenous lysis of biomass preserved a high reducing environment in the ASBR, which was important for the reduction of the Orange II azo bond that caused decolorization. The maximum decolorization rate in the ASBR was approximately 0.17 g/hr in the absence of co-substrates and nutrients.     

Accelerated degradation of a variety of aromatic compounds by Spirodela polyrrhiza-bacterial associations and contribution of root exudates released from S. polyrrhiza

Removal experiments of phenol,aniline,2,4-dichlorophenol,nonylphenol and bisphenol A(BPA) using Spirodela polyrrhizabacterial associations revealed that all compounds but BPA underwent accelerated removal.The mechanisms differed depending on the substrates.It was found that S.polyrrhiza has a great ability to release phenolic compound-rich root exudates,and the exudates seem to stimulate bacterial degradation of a variety of aromatic compounds.     

Variation in estrogenic activity among fractions of a commercial nonylphenol by high performance liquid chromatography

Estrogenic activity by recombinant yeast screen assay of the commercial NP was considerably higher when compared with that of n-nonylphenol (n-NP). Fractionation of the commercial NP by high performance liquid chromatography (HPLC) afforded seven isomers: 4-(1,3-dimethyl-1-propyl-butyl)-phenol, 4-(1,1,3-trimethyl-hexyl)-phenol, 4-(1,1-dimethyl-3-ethyl-pentyl)-phenol, 4-(1,1,4-trimethyl-hexyl)-phenol, 4-(1-methyl-1-propyl-pentyl)-phenol, 4-(1,1,2-trimethyl-hexyl)-phenol and 4-(1-ethyl-1-methyl-hexyl)-phenol. The structures of these isomers were determined by GC-MS and nuclear magnetic resonance spectroscopy (NMR). All of these isomers possessed tertiary alpha-carbon in their chemical structures. Another tertiary NP, 4-(1,1-dimethyl-heptyl)-phenol was synthesized in the present study and this synthetic NP also exhibited the estrogenic activity. One fractionated compound was identified as one of decylphenol, 4-(1-ethyl-1,4,4-trimethyl-pentyl)-phenol. The isomer, 4-(1,1,4-trimethyl-hexyl)-phenol exhibited the highest estrogenic activity corresponding to 1/10000 that of 17beta-estradiol (E2). The activity of n-NP was the least. This suggests that it may be possible to develop a technical NP mixture with relatively low estrogenic activity.     

The Effect of Estrogens,River Water,and Treated Wastewater on Vitellogenin Induction of Japanese Medaka1

Abstract: The objective of this study is to evaluate the effect of estrogens on fish endocrine disruption in river water and treated wastewater. Endogenous estrogen estrone (E1), 17β‐estradiol (E2), river water, and treated wastewater were used for exposure tests on male Japanese medaka. Vitellogenin induction in male medaka was regarded as the endpoint of endocrine disruption. The effective concentrations of E1 and E2 on vitellogenin induction in medaka were evaluated by breeding medaka for 14 days in tanks with various concentrations of E1 or E2. Vitellogenin induction increased with elapse of time during exposure, with higher estrogen concentrations causing greater vitellogenin induction. According to the test results, the lowest observed effect concentrations (LOECs) of E1 and E2 were estimated to be 31.6 and 5.0 ng/l, respectively; and the E2 equivalent (EEQ) LOEC value of E1 was 5.2 ng‐E2/l, derived by multiplying the relative potency of E1 by that of E2 as estimated in this study. In continuous exposure tests using river water or treated wastewater, the estrogenic activity, an index of total estrogenic potential measured by yeast screen assay and expressed as EEQ, varied widely during the exposure tests, and significant vitellogenin induction was observed after several days of high levels of estrogenic activity in water. Vitellogenin concentration tended to significantly increase if EEQ exceeded the level of 5 ng‐E2/l. The threshold value was substantially the same as the results for E1 and E2 exposure tests of medaka. Consequently, EEQ over 5 ng‐E2/l was revealed to have the potential to cause endocrine disruption of male medaka. As estrogenic activity exceeding 5 ng‐E2/l was observed in some rivers in Japan, total estrogenic potential needs to be further decreased at wastewater treatment plants to prevent fish endocrine disruption.