Modulation of steroidogenesis by coastal waters and sewage effluents of Hong Kong, China, using the H295R assay

BACKGROUND, AIM, AND SCOPE: The presence of a variety of pollutants in the aquatic environment that can potentially interfere with the production of sex steroid hormones in wildlife and humans has been of increasing concern. The aim of the present study was to investigate the effects of extracts from Hong Kong marine waters, and influents and effluents from wastewater treatment plants on steroidogenesis using the H295R cell bioassay. After exposing H295R cells to extracts of water, the expression of four steroidogenic genes and the production of three steroid hormones were measured. MATERIALS AND METHODS: Water samples were collected during the summer of 2005 from 24 coastal marine areas and from the influents and effluents of two major waste water treatment plants (WWTPs) in Hong Kong, China. Samples were extracted by solid phase extraction (SPE). H295R cells were exposed for 48 h to dilutions of these extracts. Modulations of the expression of the steroidogenic genes CYP19, CYP17, 3betaHSD2, and CYP11beta2 were determined by measuring mRNA concentrations by real-time polymerase chain reaction (Q-RT-PCR). Production of the hormones progesterone (P), estradiol (E2), and testosterone (T) was quantified using enzyme linked immunosorbent assays (ELISA). RESULTS: Extracts from samples collected in two fish culture areas inhibited growth and proliferation of H295R cells at concentrations greater or equal to 10(5) L equivalents. The cells were exposed to the equivalent concentration of active substances in 10,000 L of water. Thus, to observe the same level of effect as observed in vitro on aquatic organisms would require a bioaccumulation factor of this same magnitude. None of the other 22 marine samples affected growth of the cells at any dilution tested. Twelve of the marine water samples completely inhibited the expression of CYP19 without affecting E2 production; inhibition of CYP17 expression was observed only in one of the samples while expression of CYP11beta2 was induced as much as five- and ninefold after exposure of cells to extracts from two locations. The expression of the progesterone gene 3betaHSD2 was not affected by any of the samples; only one sample induced approximately fourfold the production of E2. Although more than twofold inductions were observed for P and T production, none of these values were statistically significant to conclude effects on the production of these two hormones. While influents from WWTPs did not affect gene expression, an approximately 30% inhibition in the production of E2 and a 40% increase in P occurred for the exposure with influents from the Sha Tin and Stonecutters WWTPs, respectively. Effluents from WWTPs did not affect the production of any of the studied hormones, but a decrement in the expression of the aldosterone gene CYP11beta2 was observed for the Sha Tin WWTP exposure. No direct correlation could be established between gene expression and hormone production. DISCUSSION: Observed cytotoxicity in the two samples from fish culture areas suggest the presence of toxic compounds; chemical analysis is required for their full identification. Although effluents from WWTPs did not affect hormone production, other types of endocrine activity such as receptor-mediated effects cannot be ruled out. Interactions due to the complexity of the samples and alternative steroidogenic pathways might explain the lack of correlation between gene expression and hormone production results. CONCLUSIONS: Changes observed in gene expression and hormone production suggest the presence in Hong Kong coastal waters of pollutants with endocrine disruption potential and others of significant toxic effects. The aromatase and aldosterone genes seem to be the most affected by the exposures, while E2 and P are the hormones with more significant changes observed. Results also suggest effectiveness in the removing of compounds with endocrine activity by the WWTPs studied, as effluent samples did not significantly affect hormone production. The H295R cell showed to be a valuable toll in the battery required for the analysis of endocrine disrupting activities of complex environmental samples. RECOMMENDATIONS AND PERSPECTIVES: Due to the intrinsic complexity of environmental samples, a combination of analytical tools is required to realistically assess environmental conditions, especially in aquatic systems. In the evaluation of endocrine disrupting activities, the H295R cell bioassay should be used in combination with other genomic, biological, chemical, and hydrological tests to establish viable modes for endocrine disruption and identify compounds responsible for the observed effects.     

Improvement of an efficient separation method for chemicals in diesel exhaust particles: analysis for nitrophenols

GOAL, SCOPE, AND BACKGROUND: Diesel exhaust is believed to consist of thousands of organic constituents and is a major cause of urban pollution. We recently reported that a systematic separation procedure involving successive solvent extractions, followed by repeated column chromatography, resulted in the isolation of vasodilatory active nitrophenols. These findings indicated that the estimation of the amount of nitrophenols in the environment is important to evaluate their effect on human health. The isolation procedure, however, involved successive solvent extractions followed by tedious, repeated chromatography, resulting in poor fractionation and in a significant loss of accuracy and reliability. Therefore, it was crucial to develop an alternative, efficient, and reliable analytical method. Here, we describe a facile and efficient acid-base extraction procedure for the analysis of nitrophenols. MATERIALS AND METHODS: Diesel exhaust particles (DEP) were collected from the exhaust of a 4JB1-type engine (ISUZU Automobile Co., Tokyo, Japan). Gas chromatography-mass spectrometry (GC-MS) analysis was performed with a GCMS-QP2010 instrument (Shimadzu, Kyoto, Japan). RESULTS: A solution of DEP in 1-butanol was extracted with aqueous NaOH to afford a nitrophenol-rich oily extract. The resulting oil was methylated with trimethylsilyldiazomethane and subsequently subjected to GC-MS analysis, revealing that 4-nitrophenol, 3-methyl-4-nitrophenol, 2-methyl-4-nitrophenol, and 4-nitro-3-phenylphenol were present in significantly higher concentrations than those reported previously. DISCUSSION: Simple acid-base extraction followed by the direct analysis of the resulting extract by GC-MS gave only broad peaks of nitrophenols with a poor detection limit, while the GC-MS analysis of the sample pretreated with (trimethylsilyl)diazomethane gave satisfactorily clear chromatograms with sharp peaks and with a significantly lowered detection limit (0.5 ng/ml, approximately 100 times). CONCLUSION: The present method involving an acid-base extraction, in situ derivatization, and GC-MS analysis has shown to be a simple, efficient, and reliable method for the isolation and identification of the chemical substances in DEP.     

The effects of coal-smoke pollutants on the growth, yield and leaf epidermal features of Abelmoschus esculentus moench

In Abelmoschus esculentus var. Pusa Sawani, coal-smoke pollutants, experienced close to a thermal power plant, led to decreases of plant height and stem diameter, jeopardised the production of leaves and flower buds and stimulated leaf and flower fall, leading to a decrease in the number of fruits set per plant. The length of fruits was affected a little, and the circumference and fresh weight suffered significantly, causing a decrease in plant yield. The net productivity of stems, roots and leaves incurred losses of 22.5%, 24.0% and 37.5%, respectively, and resulted in a 28.3% loss in the total net productivity of polluted plants. A decrease in stomatal density, aperture and index, and a lower density of epidermal cells of greater dimensions were observed on both the leaf surfaces. Abnormalities of the stomatal complexes were also observed on the upper surface of polluted leaves.     

世界铅、锌矿产资源的开发与展望

只要我们回顾1990～1991年全世界经济增长速度减退的状况,就不难发现有许多因素影响了世界金属的消费量;但是随着东欧经济的复苏,南亚经济的持续增长,拉丁美洲贸易壁垒的打破、投资增多,以及工业化国家基本建设对金属需求量增多,促使全世界对包括铅、锌在内的金属矿产资源的消费量持续增长。1990年西欧和前苏联的经济衰退,外费     

21世纪世界水资源的开发前景

过去数年,发展中国家特别是位于干旱、半干旱地区的发展中国家日益认识到合理开发水资源的重要性,越来越多的规划工作者和决策者也已认识到有效刺用水资源是非常重要的。目前,世界上新开发的水源与原开发的水源相比,新开发的水源水量显著减少,且新水源的开发费用越来越高;因此,需要对新水源的开发进行规划、设计和管理,需要有关方面的知识和技术。当今,决策者已认识到水资源并不     

Heavy metals alter photosynthetic pigment profiles as well as activities of chlorophyllase and 5-aminolevulinic acid dehydratase (ALAD) in Amaranthus lividus seedlings

Varied concentrations of PbCl2 and CdCl2 in the germinating media reduced the total chlorophyll and carotenoid contents in primary leaves of Amaranthus lividus seedlings (168 h old). When chlorophyll a and chlorophyll b contents were measured separately, greater loss of chl b than chl a under the identical conditions of heavy metal treatment was observed In addition, the loss of total chlorophyll was more than carotenoids under the same magnitude of heavy metal treatment. The effect of heavy metal treatment at germination stage was further studied on chlorophyll accumulation in primary leaves in relation to the activities of 5-aminolevulinic acid dehydratase (ALAD) and chlorophyllase. The activities of ALAD gradually diminished in response to both the heavy metals in a concentration-guided manner, while the activities of chlorophyllase did not exhibit any significant change.     

Ultra-structural observations on the lymphoid organs of the freshwater catfish, Clarias batrachus (Linnaeus)

Light microscopic and ultra-structural studies of the lymphoid tissues such as blood immunocytes, spleen and pronephros of the freshwater catfish, Clarias batrachus, were carried out. The peripheral blood showed nucleated erythrocytes, total leucocytic count (TLC) more than that observed in mammalian blood and leucocytes with morphological appearance similar to the mammalian white blood cells (WBCs). The spleen and pronephros showed presence of numerous lymphocytes, monocytes and nucleated red blood cells (RBCs) along with hemosiderin-containing macrophages. The morphology of lymphoid organs of the catfish has been discussed in light of the evolution of the immune system in this class of vertebrates.     

Development and performance of an alternative biofilter system

Step tracer tests were carried out on lab-scale biofilters to determine the residence time distributions (RTDs) of gases passing through two types of biofilters: a standard biofilter with vertical gas flow and a modified biofilter with horizontal gas flow. Results were used to define the flow patterns in the reactors. "Non-ideal flow" indicates that the flow reactors did not behave like either type of ideal reactor: the perfectly stirred reactor [often called a "continuously stirred tank reactor" (CSTR)] or the plug-flow reactor. The horizontal biofilter with back-mixing was able to accommodate a shorter residence time without the usual requirement of greater biofilter surface area for increased biofiltration efficiency. Experimental results indicated that the first bed of the modified biofilter behaved like two CSTRs in series, while the second bed may be represented by two or three CSTRs in series. Because of the flow baffles used in the horizontal biofilter system, its performance was more similar to completely mixed systems, and hence, it could not be modeled as a plug-flow reactor. For the standard biofilter, the number of CSTRs was found to be between 2 and 9 depending on the airflow rate. In terms of NH3 removal efficiency and elimination capacity, the standard biofilter was not as good as the modified system; moreover, the second bed of the modified biofilter exhibited greater removal efficiency than the first bed. The elimination rate increased as biofilter load increased. An opposite trend was exhibited with respect to removal efficiency.     

Assessment of imidacloprid in Brassica environment

Imidacloprid was applied as seed treatment (Gaucho 70 WS, 5 and 10 g ai kg(-1) seed) and foliar spray (Confidor 200 SL, 20 and 40 g ai ha(-1)) at 50% pod formation stage on mustard (Brassica campestris Linn.) to control mustard aphid, Lipaphis erysimi Kalt. It was detectable upto 82 and 96 days in plants after sowing from lower and higher doses of seed treatment. However, it dissipated faster and became nondetectable after 7 and 15 days of foliar treatments from lower and higher rates of application, respectively. The dissipation models yielded the rate constants of 0.0209 and 0.0230 and 0.0736 and 0.0779 day(-1) from seed and foliar treatment. The corresponding half-lives of 14.40 and 13.07 and 4.09 and 3.86 days were recorded. This suggested that the dissipation was independent of initial doses and followed a first order rate kinetics. The projected TMRC of imidacloprid from seed (0.136 and 0.225 mg person(-1) day(-1)) and foliar (0.069 and 0.1497 mg person(-1) day(-1)) treatments were found lower than the MPI (3.135 mg person(-1) day(-1)). At harvest mustard grains did not contain imidacloprid residues. The absence of imidacloprid in 0-10 and 10-20 cm soil layers indicated no leaching of insecticide. Therefore, imidacloprid treatments could be taken as safe for crop protection, consumption of leaves and environmental contamination point of view.     

Social perceptions of the impacts of Colombo water supply projects

The complexity of development activities, in which the interactions between various forces often mean that outcomes are unpredictable and unanticipated, highlights the importance of objective and comprehensive evaluations. In the specific case of the evaluation of towns east and south of Colombo water supply projects, the findings have major implications for the implementation of the Millennium Development Goals, if the water-related objectives are to be fulfilled. The study found that 30% of the households of these towns that are receiving clean, piped water are not using it for drinking. Instead, they are continuing to drink contaminated well water. Whereas it is considered that the main beneficiaries of water supply projects have been women and people who are sick, elderly, or disabled, the general perception is that the health conditions of the families have not improved with the provision of clean water (in fact, many claim that their health conditions have deteriorated). In spite of the higher cost of the supplied water, people are willing to pay for it as long as the supply is reliable.