A Biomimetic Approach to the Detection and Identification of Estrogen Receptor Agonists in Surface Waters Using Semipermeable Membrane Devices (SPMDs) and Bioassay-Directed Chemical Analysis (12 pp)

Goal, Scope and Background Some anthropogenic pollutants posses the capacity to disrupt endogenous control of developmental and reproductive processes in aquatic biota by activating estrogen receptors. Many anthropogenic estrogen receptor agonists (ERAs) are hydrophobic and will therefore readily partition into the abiotic organic carbon phases present in natural waters. This partitioning process effectively reduces the proportion of ERAs readily available for bioconcentration by aquatic biota. Results from some studies have suggested that for many aquatic species, bioconcentration of the freely-dissolved fraction may be the principal route of uptake for hydrophobic pollutants with logarithm n-octanol/water partition coefficient (log Kow) values less than approximately 6.0, which includes the majority of known anthropogenic ERAs. The detection and identification of freely-dissolved readily bioconcentratable ERAs is therefore an important aspect of exposure and risk assessment. However, most studies use conventional techniques to sample total ERA concentrations and in doing so frequently fail to account for bioconcentration of the freely-dissolved fraction. The aim of the current study was to couple the biomimetic sampling properties of semipermeable membrane devices (SPMDs) to a bioassay-directed chemical analysis (BDCA) scheme for the detection and identification of readily bioconcentratable ERAs in surface waters. Methods SPMDs were constructed and deployed at a number of sites in Germany and the UK. Following the dialytic recovery of target compounds and size exclusion chromatographic clean-up, SPMD samples were fractionated using a reverse-phase HPLC method calibrated to provide an estimation of target analyte log Kow. A portion of each HPLC fraction was then subjected to the yeast estrogen screen (YES) to determine estrogenic potential. Results were plotted in the form of 'estrograms' which displayed profiles of estrogenic potential as a function of HPLC retention time (i.e. hydrophobicity) for each of the samples. Where significant activity was elicited in the YES, the remaining portion of the respective active fraction was subjected to GC-MS analysis in an attempt to identify the ERAs present. Results and Discussion Estrograms from each of the field samples showed that readily bioconcentratable ERAs were present at each of the sampling sites. Estimated log Kow values for the various active fractions ranged from 1.92 to 8.63. For some samples, estrogenic potential was associated with a relatively narrow range of log Kow values whilst in others estrogenic potential was more widely distributed across the respective estrograms. ERAs identified in active fractions included some benzophenones, various nonylphenol isomers, benzyl butyl phthalate, dehydroabietic acid, sitosterol, 3-(4-methylbenzylidine)camphor (4-MBC) and 6-acetyl-1,1,2,4,4,7-hexamethyltetralin (AHTN). Other tentatively identified compounds which may have contributed to the observed YES activity included various polycyclic aromatic hydrocarbons (PAHs) and their alkylated derivatives, methylated benzylphenols, various alkylphenols and dialkylphenols. However, potential ERAs present in some active fractions remain unidentified. Conclusions and Outlook Our results show that SPMD-YES-based BDCA can be used to detect and identify readily bioconcentratable ERAs in surface waters. As such, this biomimetic approach can be employed as an alternative to conventional methodologies to provide investigators with a more environmentally relevant insight into the distribution and identity of ERAs in surface waters. The use of alternative bioassays also has the potential to expand SPMD-based BDCA to include a wide range of toxicological endpoints. Improvements to the analytical methodology used to identify ERAs or other target compounds in active fractions in the current study could greatly enhance the applicability of the methodology to risk assessment and monitoring programmes.     

Bisphenol A exposure alters release of immune and developmental modulators and expression of estrogen receptors in human fetal lung fibroblasts

Bisphenol A(BPA) has been shown to exert biological effects through estrogen receptor(ER)-dependent and ER-independent mechanisms. Recent studies suggest that prenatal exposure to BPA may increase the risk of childhood asthma. To investigate the underlying mechanisms in the actions of BPA, human fetal lung fibroblasts(h FLFs) were exposed to varying doses of BPA in culture for 24 hr. Effects of BPA on localization and uptake of BPA,cell viability, release of immune and developmental modulators, cellular localization and expression of ERα, ERβ and G-protein coupled estrogen receptor 30(GPR30), and effects of ERs antagonists on BPA-induced changes in endothelin-1(ET-1) release were examined.BPA at 0.01–100 μmol/L caused no changes in cell viability after 24 hr of exposure. h FLFs expresses all three ERs. BPA had no effects on either cellular distribution or protein expression of ERα, however, at 100 μmol/L(or 23 μmol/L intracellular BPA) increased ERβprotein levels in the cytoplasmic fractions and GPR 30 protein levels in the nuclear fractions.These paralleled with increased release of growth differentiation factor-15, decreased phosphorylation of nuclear factor kappa B p65 at serine 536, and decreased release of ET-1,interleukin-6, and interferon gamma-induced protein 10. ERs antagonists had no effects on BPA-induced decrease in ET-1 release. These data suggest that BPA at 100 μmol/L altered the release of immune and developmental modulators in h FLFs, which may negatively influence fetal lung development, maturation, and susceptibility to environmental stressors, although the role of BPA in childhood asthma remains to be confirmed in in vivo studies.     

Study of the effects of bisphenol A using human fetal lung fibroblasts

正Bisphenol A(BPA,2,2-bis(4-hydroxyphenyl)propane;CAS#80-05-7)is a highly produced industrial chemical,with an estimated four billion kilograms produced worldwide each year(reviewed in Vandenberg et al.,2010).BPA serves as a monomer in the manufacture of polycarbonate,a hard,clear plastic.Polycarbonate plastics are used in many consumer products,such as reusable water bottles.BPA is also widely used to make epoxy resins,which are used as coating on the inside of some     

Microfluidic photomechanic infrared receptors in a pyrophilous flat bug

Infrared (IR) receptors are so far known only in boid and crotalid snakes and in three genera of pyrophilous beetles that seek out forest fires. Pyrophilous insects can also be found in other orders, however, so it can be hypothesised that IR receptors also occur in some of these species. We investigated the pyrophilous Australian flat bug Aradus albicornis and found a small number of dome-shaped sensilla (diameter 13 microm) on the prothorax, which have previously not been described. Ultrastructural investigations revealed that the sensilla are characterised by a fluid-filled inner compartment enclosed in a round cuticular shell. The cuticular apparatus is innervated by the dendrite of a ciliary mechanoreceptor, which is fluidically coupled to the inner compartment. Electrophysiological recordings demonstrated that the sensilla respond to brief warming by red laser light or to broadband IR radiation. Depending on the radiation intensity (4.4-549 mW/cm(2) tested, threshold measured as 11.3 mW/cm(2)), first spike latencies varied between 3.4 and 7.5 ms. Thus, our findings demonstrate that A. albicornis most probably possesses photomechanic IR sensilla resembling the metathoracic IR sensilla of buprestid beetles of the genus Melanophila. In the Melanophila sensillum, IR radiation causes thermal expansion of a fluid, which rapidly deforms the dendritic membrane of a mechanosensory cell. The existence of photomechanic IR receptors in both beetles and bugs demonstrates a remarkable convergent evolution towards this particular biophysical transduction mechanism and suggests that it provides selective advantages over other possible solutions.     

Alkaloid tolerance in Manduca sexta and phylogenetically related sphingids (Lepidoptera: Sphingidae)

Summary. Nicotine tolerance is well known for Manduca sexta. It also occurs in several other sphingids of the subfamilies Macroglossinae and Sphinginae. Only members of the subfamily Smerinthinae appear to be more susceptible to nicotine intoxication. Phylogenetic trees have been reconstructed from mitochondrial 16S rDNA and nuclear DNA to map nicotine tolerance.?The nicotine binding site of both α-subunits of nicotinic acetylcholine receptors (nAChR) have been amplified and sequenced. No apparent amino acid substitution can be seen in the putative nicotine binding site of the α-subunits of nAChR from nicotine tolerant and nicotine sensitive sphingids. Thus, a simple target-site modification can be ruled out as a cause for nicotine tolerance. This finding agrees with feeding experiments: larvae of M. sexta and other sphingids of the Macroglossinae and Sphinginae not only tolerated nicotine, but also many other alkaloids that affect neuroreceptors other than acetylcholine receptors (nAChR, mAChR).?Only 10 to 20% of nicotine injected into larvae of nicotine-tolerant taxa could be recovered later as free nicotine, nicotine N-oxide or cotinine, i.e., 80 to 90% must have been converted to polar conjugates or degradation products which are not detectable with the methods applied. Usually more than 98% of the recoverable alkaloids were found in the faeces. Excretion reached a maximum 6 h after injection in tolerant taxa. Larvae of Manduca sexta, which were reared on a nicotine-rich diet, showed higher nicotine degradation and faster nicotine elimination than na?ve larvae. Application of the cytochrome P450 inhibitor SKF 525A (proadifen) reduced the formation of nicotine N-oxide and the rate of alkaloid degradation. Thus, an inducible detoxification mechanism, coupled with a rapid and inducible excretion, appear to be a strategy in Sphingidae that helps them to live on host plants rich in otherwise toxic secondary metabolites. Received 23 March 2001; accepted 4 August 2001.     

化学质量平衡受体模型新技术的应用──TEDA大气颗粒物来源解析实例

本文结合实例介绍了大气颗粒物来源解析最常用的模型－化学质量平衡（ＣＭＢ）的原理、有效方差最小二乘解法及诊断检验技术，并用ＭＰＩＮ矩阵检验了拟合元素对拟合源的灵敏度，奇异值分解法（ＳＶＤ）鉴别了共线源，得出天津市经济技术开发区（ＴＥＤＡ）的ＴＳＰ主要来源为风沙、扬尘，其次为燃煤飞灰和木灰。     

Receptor modeling for analyzing PCDD/F and dioxin-like PCB sources in Hong Kong

A total of 96 ambient air samples were collected from January 2004 to December 2004 for the investigation of dioxins (polychlorinated dibenzo-para-dioxins and polychlorinated dibenzofurans [PCDD/Fs]) and dioxin-like profile for polychlorinated biphenyls (PCBs) in the atmosphere of Hong Kong. Two-day back-trajectories were obtained using the Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) model from National Oceanic and Atmospheric Administration (NOAA) Air Resources Laboratory. Receptor models – potential source contribution function (PSCF) and concentration-weighted trajectory (CWT) – were built based on the measured PCDD/Fs and dioxin-like PCBs total toxic equivalent (TEQ) concentrations. Both models were modified in this study so as to integrate the information obtained from three sampling sites in Hong Kong. The PSCF modeling resolved two main PCDD/F and dioxin-like PCB polluting paths from Mainland China. In line with previous studies, the northerly path around the Pearl River Delta might be one of the pollution sources of Hong Kong air. On the other hand, the receptor models also indicated another potential northeasterly source from Yangtze River Delta, which was seldom mentioned as the source region of Hong Kong air. Results from the CWT modeling suggested that the most toxic air mass affecting Hong Kong is probably attributable to the northeast path rather than the north path.     

Myasthenia gravis with polyhydramnios in the fetus of an asymptomatic mother

Transient myasthenia gravis was observed in an infant whose mother had no signs of neuro-muscular disease. The case presented with severe polyhydramnios at 31 weeks. Sonographic and invasive work-up showed only an absence of fetal swallowing. At birth, the infant had severe muscle weakness and respiratory distress. He had high titres of anti-acetylcholine receptor (anti-AChR) antibodies of maternal origin. Anti-AChR antibodies were also found in stored samples of fetal serum and amniotic fluid. To our knowledge, this is the first case of neonatal myasthenia gravis for which neither past nor present evidence of disease could be obtained in the mother. Anti-AChR antibody testing should be considered in cases of unexplained polyhydramnios.     

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...     

Estrogen-related receptor disruption of source water and drinking water treatment processes extracts

Environmental chemicals in drinking water can impact human health through nuclear receptors. Additionally, estrogen-related receptors (ERRs) are vulnerable to endocrine-disrupting e ects. To date, however, ERR disruption of drinking water potency has not been reported. We used ERR two-hybrid yeast assay to screen ERR disrupting activities in a drinking water treatment plant (DWTP) located in north China and in source water from a reservoir, focusing on agonistic, antagonistic, and inverse agonistic activity to 4-hydroxytamoxifen (4-OHT). Water treatment processes in the DWTP consisted of pre-chlorination, coagulation, coal and sand filtration, activated carbon filtration, and secondary chlorination processes. Samples were extracted by solid phase extraction. Results showed that ERR antagonistic activities were found in all sample extracts, but agonistic and inverse agonistic activity to 4-OHT was not found. When calibrated with the toxic equivalent of 4-OHT, antagonistic e uent e ects ranged from 3.4 to 33.1 g/L. In the treatment processes, secondary chlorination was e ective in removing ERR antagonists, but the coagulation process led to significantly increased ERR antagonistic activity. The drinking water treatment processes removed 73.5% of ERR antagonists. To our knowledge, the occurrence of ERR disruption activities on source and drinking water in vitro had not been reported previously. It is vital, therefore, to increase our understanding of ERR disrupting activities in drinking water.