The OECD Validation Program of the H295R Steroidogenesis Assay for the Identification of In Vitro Inhibitors and Inducers of Testosterone and Estradiol Production. Phase 2: Inter-Laboratory Pre-Validation Studies (8 pp)

Background, Goals and Scope In response to concerns that have been raised about chemical substances that may alter the function of endocrine systems and result in adverse effects on human health, an OECD initiative was undertaken to develop and validate in vitro and in vivo assays to identify chemicals that may interfere with endocrine systems of vertebrates. Here we report on studies that were conducted to develop and standardize a cell-based screening assay using the H295R cell line to prioritize chemicals that may act on steroidogenic processes in humans and wildlife. These studies are currently ongoing as part of the ‘Special Activity on the Testing and Assessment of Endocrine Disruptors’ within the OECD Test Guidelines Program to review, develop, standardize, and validate a number of in vitro and in vivo toxicological assays for testing and assessment of chemicals concerning their potential to interact with the endocrine system of vertebrates. Study Design Six laboratories from five countries participated in the pre-validation studies. Each laboratory tested the effects of three model chemicals on the production of testosterone (T) and estradiol (E2) using the H295R Steroidogenesis Assay. Chemicals tested were well described inducers or inhibitors of steroidogenic pathways (forskolin, prochloraz and fadrozole). All experiments were conducted in 24 well plates following standard protocols. Six different doses per compound were analyzed in triplicate per plate. A quality control (QC) plate was run in conjunction with the chemical exposure plate to account for inter-assay variation. Each chemical exposure was conducted two or three times. Results All laboratories successfully detected increases and/or decreases in hormone production by H295R cells after exposure to the different model compounds and there was good agreement in the pattern of response for all groups. Forskolin increased both T and E2 while fadrozole and prochloraz decreased production of both hormones. All chemicals affected hormone production in a dose-dependent manner with the exception of fadrozole which caused maximum inhibition of E2 at the two least concentrations tested. Some inter-laboratory differences were noted in the alteration of hormone production measured in chemically exposed cells. However, with the exception of the production of T measured at one laboratory in cells exposed to forskolin, the EC₅₀s calculated were comparable (coefficients of variation 34–49%) for all hormones. Discussion and Perspectives The results indicated that the H295R Steroidogenesis Assay protocol was robust, transferable and reproducible among all laboratories. However, in several instances that were primarily related to one laboratory there were unexplained minor uncertainties related to the inter-laboratory hormone production variation. Based on the findings from this Phase 2 prevalidation study, the H295R Steroidogenesis Assay protocol is currently being refined. The next phase of the OECD validation program will test the refined protocol among the same group of laboratories using an extended set of chemicals (∼30) that will include positive and negative chemical controls as well as a broad spectrum of different potential inducers and inhibitors of steroidogenic pathways. Submission Editor: Dr. Carsten Brühl (bruehl@uni-landau.de)     

Improved quartz furnace method for chlorine and sulfur determination in municipal solid waste

 A method of determining the chlorine (Cl) and sulfur (S) in municipal solid waste (MSW) was studied. The quartz furnace method was improved in two ways: recovery from ash by hot extraction with dilute nitric acid, and avoidance of the volatilization of alkali (earth) metal chlorides by setting the sample combustion temperature at 600°C. In a comparison with the bomb method, using nine sets of kitchen garbage and waste plastics, the bomb method yielded a 15%–25% lower value than the improved quartz furnace method. Combustion in the bomb was frequently incomplete, resulting in recovery losses of Cl and S. The average kitchen garbage involved 5.2 mg Cl/g, of which at least 24.1% would be converted to HCl. Plastics contained 23 mg Cl/g generating 88.1% HCl on average. In the same way, kitchen garbage contained 3.0 mg S/g, generating 52.3% SO _x , whereas plastics contained 1.1 mg S/g with 55.1% SO _x formation. Received: March 20, 2002 / Accepted: October 13, 2002     

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...     

Measurements of short-lived cosmic-ray-produced radionuclides in rainwater

Cosmic-ray-produced (CP) nuclides with half-lives shorter than 21h were measured in rainwater by ultra-low-background gamma spectrometry at Ogoya Underground Laboratory. As levels of CP nuclides are extremely low and their half-lives are very short, quick sampling methods for a large volume of rainwater and rapid chemical separations by ion exchange method were developed. The nuclides measured were short-lived (24)Na, (28)Mg, (38)S, (38)Cl, (39)Cl, as well as nuclides with longer half-lives (7)Be and (22)Na. The number of atoms of CP nuclides in rainwater were evaluated to range from 30 to 1500L(-1) for (24)Na (n=16, mean; 520 [6.7mBqL(-1)]), 80 to 600L(-1) for (28)Mg (n=13, mean; 260 [2.4mBqL(-1)]), 400 to 1900L(-1) for (39)Cl (n=6, mean; 1200 [250mBqL(-1)]), 1x10(6) to 4x10(7)L(-1) for (7)Be (n=16, mean; 7x10(6) [1.05BqL(-1)]) and 2x10(3) to 1x10(5)L(-1) for (22)Na (n=9, mean; 2x10(4) [0.2mBqL(-1)]).     

Bacterial community succession during the enrichment of chemolithoautotrophic arsenite oxidizing bacteria at high arsenic concentrations

To generate cost-effective technologies for the removal of arsenic from water, we developed an enrichment culture of chemolithoautotrophic arsenite oxidizing bacteria (CAOs) that could effectively oxidize widely ranging concentrations of As(III) to As(V). In addition, we attempted to elucidate the enrichment process and characterize the microbial composition of the enrichment culture. A CAOs enrichment culture capable of stably oxidizing As(III) to As(V) was successfully constructed through repeated batch cultivation for more than 700 days, during which time the initial As(III) concentrations were increased in a stepwise manner from 1 to 10-12 mmol/L. As(III) oxidation activity of the enrichment culture gradually improved, and 10-12 mmol/L As(III) was almost completely oxidized within four days. Terminal restriction fragment length polymorphism analysis showed that the dominant bacteria in the enrichment culture varied drastically during the enrichment process depending on the As(III) concentration. Isolation and characterization of bacteria in the enrichment culture revealed that the presence of multiple CAOs with various As(III) oxidation abilities enabled the culture to adapt to a wide range of As(III) concentrations. The CAOs enrichment culture constructed here may be useful for pretreatment of water from which arsenic is being removed.     

关于资源技术与工程学科建设若干问题的讨论

资源技术与工程是现阶段资源科学学科建设的主要方向之一,是研究资源直接利用、间接利用、循环利用的理论、方法和实践的学科,其研究对象为物质实体。目前国内外资源技术与工程学科建设的进展仍处于纵向分异的阶段,单项资源(水、土、气、生物、矿产、能源等)利用的技术与工程研究仍是其主要特征。资源技术与工程的基本学科定位应为工学,其学科体系可分为3个二级学科和12个三级学科。在开展资源技术与工程学科建设时,理科院校应侧重于新型资源、替代资源的开发利用技术与工程研究,而工科、农科院校则应注重单项资源以及多项资源之间的再生利用、循环利用、保护利用、协同利用和综合利用技术与工程研究。     

Why did the project fail to achieve its objectives in some villages? The experience of the Upland Settlement Project (USP) in Bangladesh

By sampling an upland settlement project (USP) village, this study examined why the project failed to achieve the planned objectives in some villages, and what livelihood strategies villagers (planters) have adopted to sustain their livelihoods. The paper uses data obtained by qualitative and quantitative methods. The USP was formulated with the objectives of settling 50 landless and marginal tribal families in a project village and introducing agroforestry and rubber plantations for short- and long-term socioeconomic development of planters, in a participatory manner. There are now only 15 planters in the project village and, although agroforestry has progressed well, only 30 to 40% of poor quality rubber trees remain. Planter participation in project activities was limited to wage labour, indicating that the USP failed to adequately achieve all its proposed objectives. In the face of project failure, planters have adopted diversified strategies to maintain their livelihoods. Several shortcomings caused failure of the project, such as the inability of project staff members to organize planters, failure to create awareness among planters about project benefits, lack of regular project money disbursement, sociocultural ignorance, and lack of social capital among planters. Recommendations are made for securing effective participation of local people to assure successful and sustainable project outcomes. Lessons learned will be helpful in formulating future development programmes in this country and elsewhere.     

Nematode community composition in hydrothermal vent and adjacent non-vent fields around Myojin Knoll,a seamount on the Izu-Ogasawara Arc in the western North Pacific Ocean

In contrast to specific large benthic invertebrates in chemosynthetic ecosystems such as hydrothermal vents, meiofaunal communities in such habitats have been reported to have strong taxonomic overlap with meiofauna in the adjacent “normal” environments. However, meiofauna have only recently been included in studies of those environments and detailed information on these communities is still rare. This is especially true in the Northwest Pacific Ocean, even though there are many seamounts with active vents in the calderas of the region. Nematode community composition at the genus level in sediments from a hydrothermal vent field in the caldera of Myojin Knoll (32°06′N, 139°52′E, depth 1,300 m), a seamount on the Izu-Ogasawara Arc, Japan, was investigated for the first time and was compared with adjacent non-vent areas inside and outside the caldera. Multivariate analyses showed that the composition of nematodes in the hydrothermal field was significantly different from that in the non-hydrothermal fields around the caldera. However, the common genera, such as Oxystomina, Pareudesmoscolex, Desmoscolex, and Microlaimus were found in two, or all three vent fields while their rank contributions differed among the three fields. When the data from Myojin Knoll were compared with those from other deep-sea vent environments in different regions (e.g., North Fiji Basin, East Pacific Rise, Mid-Atlantic Ridge), the nematode composition in the vent field of the Myojin caldera was more similar to that of the non-vent fields around the caldera than the composition in vent fields of other regions. These data from the Northwest Pacific Ocean also suggest the absence of long-range transport systems and local adaptations for meiofauna in hydrothermal vent fields.     

Chlorine promotes antimony volatilization in municipal waste incineration

Antimony volatilization in municipal waste incineration was studied. Two municipal waste samples and antimony(III) oxide (Sb₄O₆) were heated to 500°C and 700°C in an air stream in a quartz furnace. The volatilization of Sb₄O₆ occurred more at 700°C that at 500°C. Conversely, antimony volatilization form municipal waste was stronger at 500°C than at 700°C. This implies that antimony from municipal waste is volatilized as chloride instead of oxide. The chlorine sources for antimony chlorination, a gas-phase reaction involving hydrochloric acid and a solid-phase reaction of inorganic chlorine, e.g., CaCl₂, were compared. Only the solid-phase reaction could offer enough active chlorine to induce chlorination of antimony oxide. Received: July 2, 1998 / Accepted: January 28, 1999     

Driver exposure to particulate matter in Bangkok

The aims of this study were to determine the particulate matter with aerodynamic diameters > or = 2.5 microm (PM2.5) and 2.5-10 microm (PM10-2.5) exposure levels of drivers and to analyze the proportion of elemental carbon (EC) and organic carbon (OC) in PM2.5 in Bangkok, Thailand. Four bus routes were selected. Measurements were conducted over 10 days in August (rainy season) 2008 and 8 days in January (dry season) 2009. The mean PM2.5 exposure level of the Tuk-tuk drivers was 86 microg/m3 in August and 198 microg/m3 in January. The mean for the non-air-conditioned bus drivers was 63 microg/m3 in August and 125 microg/m3 in January. The PM2.5 and PM10-2.5 exposure levels of the drivers in January were approximately twice as high as those in August. The proportion of total carbon (TC) in PM2.5 to the PM2.5 level in August (0.97 +/- 0.28 microg/m3) was higher than in January (0.65 +/- 0.13 microg/m3). The proportion of OC in the TC of the PM2.5 in August (0.51 +/- 0.08 microg/m3) was similar to that in January (0.65 +/- 0.07 microg/m3). The TC exposure by PM25 in January (81 +/- 30 microg/m3) remained higher than in August (56-21 microg/m3). The mean level of OC in the PM2.5 was 29 +/- 13 microg/m3 in August and 50 +/- 24 microg/m3 in January. In conclusion, the PM exposure level in Bangkok drivers was higher than that in the general environment, which was already high, and it varied with the seasons and vehicle type. This study also demonstrated that the major component of the PM was carbon, likely derived from vehicles.