Direct determination of estrogenic and antiestrogenic activities using an enhanced plant two-hybrid system.

This paper reports a simple, low-cost, and extremely sensitive reporter-gene assay system for comprehensive analysis of estrogenic activity using transgenic Arabidopsis thaliana: the EPTH system. It had the capability to detect 17beta-estradiol at a concentration of 10 pM. The system was rendered 5 times more sensitive than a previous system [Tojo, T.; Tsuda, K.; Wada, T.; Yamazaki, K. Ecotoxicol. Environ. Saf. 2006, 64, 106-114) (1)] by increasing the copy number of the transactivation domain fused to a nuclear receptor co-activator. The system can efficiently detect other estrogenic and antiestrogenic substances. Estrogenic activities were determined in treated sewage samples from four distinct sewage farms using the system. Results showed that the system can detect estrogenic activity directly and more efficiently than a yeast two-hybrid system without any manipulation for extraction and condensation of hydrophobic compounds and aseptic treatment. Furthermore, the system also is useful as a powerful tool for discovery of a new category of natural estrogenic substances that are undetectable by previous plant and yeast systems.

Characterization of polychlorinated biphenyls, pentachlorobenzene, hexachlorobenzene, polychlorinated dibenzo-p-dioxins, and dibenzofurans in surface sediments of Muroran Port, Japan.

We determined the distribution of polychlorinated biphenyls (PCBs), pentachlorobenzene (PeCBz), hexachlorobenzene (HxCBz), and polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in surface sediments at 21 sites inside and outside Muroran Port, Japan. The concentration ranges and geometric means of PCBs (Σ209PCB), PeCBz, HxCBz, and PCDD/Fs and toxicity equivalence quantity (total TEQ) of dioxins inside Muroran Port were 1,100-65,000 (mean, 17,000) pg/g dw, 37-220 (100) pg/g dw, 31-810 (84) pg/g dw, 69-410 (170) pg/g dw, and 0.51-6.2 (2.3) pg TEQ/g dw, respectively. Their corresponding inventories inside Muroran Port were estimated to be 76, 0.31, 0.32, 0.55, and 7.8 g TEQ, respectively. The amounts of these pollutants were higher inside the port than outside the port and especially large in the inner part of the port. Most PCBs were homologues and congeners of penta- to hepta-chlorinated compounds, and the PCBs around Muroran Port were derived from technical PCBs, especially KC500 and KC600. As for PCDD/Fs, the influence of pentachlorophenol was significant, although pollution due to chloronitrofen and combustion was detected. The congeners of PCDD/Fs predominantly contributed to total TEQ. The concentration distributions of PeCBz, HxCBz, and PCDD/Fs and total TEQ were highly correlated with one another. This indicates that they are derived from the same combustion process.