Development of a testing method for asbestos fibers in treated materials of asbestos containing wastes by transmission electron microscopy.

Appropriate treatment of asbestos-containing wastes is a significant problem. In Japan, the inertization of asbestos-containing wastes based on new treatment processes approved by the Minister of the Environment is promoted. A highly sensitive method for testing asbestos fibers in inertized materials is required so that these processes can be approved. We developed a method in which fibers from milled treated materials are extracted in water by shaking, and are counted and identified by transmission electron microscopy. Evaluation of this method by using asbestos standards and simulated slag samples confirmed that the quantitation limits are a few million fibers per gram and a few µg/g in a sample of 50mg per filter. We used this method to assay asbestos fibers in slag samples produced by high-temperature melting of asbestos-containing wastes. Fiber concentrations were below the quantitation limit in all samples, and total fiber concentrations were determined as 47-170×10(-6) f/g. Because the evaluation of treated materials by TEM is difficult owing to the limited amount of sample observable, this testing method should be used in conjunction with bulk analytical methods for sure evaluation of treated materials.

An automated system for high-throughput single cell-based breeding.

When establishing the most appropriate cells from the huge numbers of a cell library for practical use of cells in regenerative medicine and production of various biopharmaceuticals, cell heterogeneity often found in an isogenic cell population limits the refinement of clonal cell culture. Here, we demonstrated high-throughput screening of the most suitable cells in a cell library by an automated undisruptive single-cell analysis and isolation system, followed by expansion of isolated single cells. This system enabled establishment of the most suitable cells, such as embryonic stem cells with the highest expression of the pluripotency marker Rex1 and hybridomas with the highest antibody secretion, which could not be achieved by conventional high-throughput cell screening systems (e.g., a fluorescence-activated cell sorter). This single cell-based breeding system may be a powerful tool to analyze stochastic fluctuations and delineate their molecular mechanisms.

Cell surface-fluorescence immunosorbent assay for real-time detection of hybridomas with efficient antibody secretion at the single-cell level.

For establishing cells that secrete antibodies most efficiently (e.g., hybridomas, CHO (Chinese hamster ovary) cells), the screening and subsequent breeding of promising cells have been performed at the single-colony level, which requires several weeks to propagate a substantial number of cells by forming colonies from single cells for evaluation by the conventional assays. However, this screening process lacks high-throughput performance in time and colony numbers. Therefore, development of novel methods is expected to identify single cells secreting higher amounts of antibodies in real-time and in a nondestructive manner without colony formation. In this study, we prepared lipid-labeled antimouse IgG Fc antibodies (capture molecules) that were uniformly displayed on the surface of candidate cells. Secreted nascent antibodies were subsequently sandwiched between capture molecules and fluorescence-labeled antimouse IgG F(ab')(2) F(ab')(2) (detection molecules). This newly developed method is hereinafter referred to as a cell surface-fluorescence immunosorbent assay (CS-FIA). The fluorescence intensity of each cell was found to correlate well with the amount of sandwiched antibodies (from 6.25 fg/cell to 6.40 pg/cell). When about 4 × 10(3) cells of mouse hybridomas were subjected to CS-FIA, we isolated 28 hybridomas showing the highest fluorescence intensity within a day. Furthermore, after propagation of single cells to about 10(5) cells (after 2 weeks), 20 hybridomas were still able to secrete higher amounts (up to 7-fold) of antibodies than parental hybridomas. Our results demonstrate that CS-FIA is a powerful method for the single-cell-based establishment of cells that secrete most efficiently not only antibodies but also various biomolecules.

Atmospheric mercury emissions from waste combustions measured by continuous monitoring devices.

Atmospheric mercury emissions have attracted great attention owing to adverse impact of mercury on human health and the ecosystem. Although waste combustion is one of major anthropogenic sources, estimated emission might have large uncertainty due to great heterogeneity of wastes. This study investigated atmospheric emissions of speciated mercury from the combustions of municipal solid wastes (MSW), sewage treatment sludge (STS), STS with waste plastics, industrial waste mixtures (IWM), waste plastics from construction demolition, and woody wastes using continuous monitoring devices. Reactive gaseous mercury was the major form at the inlet side of air pollution control devices in all combustion cases. Its concentration was 2.0-70.6 times larger than elemental mercury concentration. In particular, MSW, STS, and IWM combustions emitted higher concentration of reactive gaseous mercury. Concentrations of both gaseous mercury species varied greatly for all waste combustions excluding woody waste. Variation coefficients of measured data were nearly equal to or more than 1.0. Emission factors of gaseous elemental mercury, reactive gaseous mercury, and total mercury were calculated using continuous monitoring data. Total mercury emission factors are 0.30 g-Hg/Mg for MSW combustion, 0.21 g-Hg/Mg for STS combustion, 0.077 g-Hg/Mg for STS with waste plastics, 0.724 g-Hg/Mg for industrial waste mixtures, 0.028 g-Hg/Mg for waste plastic combustion, and 0.0026 g-Hg/Mg for woody waste combustion. All emission factors evaluated in this study were comparable or lower than other reported data. Emission inventory using old emission factors likely causes an overestimation.

High-resolution inventory of Japanese anthropogenic mercury emissions.

Heavy metals like mercury that are emitted into the environment remain there indefinitely, posing a long-term threat to both the environment and human health. Elemental mercury is volatile and is in gaseous form, and because of the long residence time, transported over long distances. Comprehensive control of mercury emissions therefore remains an important international issue. The crucial steps for designing effective approaches for such control include the quantification of mercury emissions by sources and the identification of geographical characteristics of the emissions. In this study a detailed, high-resolution inventory of Japanese mercury emissions in 2005 was developed to improve understanding of their geographical distribution. Proceeding from a national emissions inventory per source category, emissions were spatially allocated with increasing geographical resolution in a stepwise procedure using statistics from geographic information resources, yielding mercury emissions per prefecture, per municipality and per grid cell of approximately 1 × 1 km. The five prefectures with the highest emissions were Fukuoka, Yamaguchi, Hyogo, Oita, and Hokkaido, accounting for 35.2% of all emissions. In each prefecture a small number of municipalities account for a major share of emissions. Distribution by grid cell is characterized by a concentration of 50% of all emissions in a mere 32 of the 255 954 grid cells over which emissions are distributed in this study. It was also quantitatively confirmed that use of larger grid cells leads to greater uncertainty in emissions distribution. Problems with data collection are clarified and measures to improve the accuracy of future estimation are proposed.

Size-dependent enrichment of waste slag aggregate fragments abraded from asphalt concrete.

Authors consider the environmental prospects of using melted waste slag as the aggregate for asphalt pavement. In particular, the enrichment of slag-derived fragments in fine abrasion dust particles originated from slag asphalt concrete and its size dependency were concerned. A series of surface abrasion tests for asphalt concrete specimens, containing only natural aggregates as reference or 30 wt% of substituted slag aggregates, were performed. Although two of three slag-asphalt concretes generated 1.5-3.0 times larger amount of abrasion dust than the reference asphalt concrete did, it could not be explained only by abrasion resistance of slag. The enrichment of slag-derived fragments in abrasion dust, estimated on the basis of the peak intensity of quartz and heavy metal concentrations, had size dependency for all slag-asphalt concretes. Slag-derived fragments were enriched in abrasion dust particles with diameters of 150-1000 µm. Enrichment factors were 1.4-2.1. In contrast, there was no enrichment in abrasion dust particles with diameter less than 75 µm. This suggests that prior airborne-size fragmentation of substituted slag aggregates does not need to be considered for tested slag aggregates when environmental risks of abrasion dust of slag-asphalt pavement are assessed.

A preliminary categorization of end-of-life electrical and electronic equipment as secondary metal resources.

End-of-life electrical and electronic equipment (EEE) has recently received attention as a secondary source of metals. This study examined characteristics of end-of-life EEE as secondary metal resources to consider efficient collection and metal recovery systems according to the specific metals and types of EEE. We constructed an analogy between natural resource development and metal recovery from end-of-life EEE and found that metal content and total annual amount of metal contained in each type of end-of-life EEE should be considered in secondary resource development, as well as the collectability of the end-of-life products. We then categorized 21 EEE types into five groups and discussed their potential as secondary metal resources. Refrigerators, washing machines, air conditioners, and CRT TVs were evaluated as the most important sources of common metals, and personal computers, mobile phones, and video games were evaluated as the most important sources of precious metals. Several types of small digital equipment were also identified as important sources of precious metals; however, mid-size information and communication technology (ICT) equipment (e.g., printers and fax machines) and audio/video equipment were shown to be more important as a source of a variety of less common metals. The physical collectability of each type of EEE was roughly characterized by unit size and number of end-of-life products generated annually. Current collection systems in Japan were examined and potentially appropriate collection methods were suggested for equipment types that currently have no specific collection systems in Japan, particularly for video games, notebook computers, and mid-size ICT and audio/video equipment.

Statistical estimate of mercury removal efficiencies for air pollution control devices of municipal solid waste incinerators.

Although representative removal efficiencies of gaseous mercury for air pollution control devices (APCDs) are important to prepare more reliable atmospheric emission inventories of mercury, they have been still uncertain because they depend sensitively on many factors like the type of APCDs, gas temperature, and mercury speciation. In this study, representative removal efficiencies of gaseous mercury for several types of APCDs of municipal solid waste incineration (MSWI) were offered using a statistical method. 534 data of mercury removal efficiencies for APCDs used in MSWI were collected. APCDs were categorized as fixed-bed absorber (FA), wet scrubber (WS), electrostatic precipitator (ESP), and fabric filter (FF), and their hybrid systems. Data series of all APCD types had Gaussian log-normality. The average removal efficiency with a 95% confidence interval for each APCD was estimated. The FA, WS, and FF with carbon and/or dry sorbent injection systems had 75% to 82% average removal efficiencies. On the other hand, the ESP with/without dry sorbent injection had lower removal efficiencies of up to 22%. The type of dry sorbent injection in the FF system, dry or semi-dry, did not make more than 1% difference to the removal efficiency. The injection of activated carbon and carbon-containing fly ash in the FF system made less than 3% difference. Estimation errors of removal efficiency were especially high for the ESP. The national average of removal efficiency of APCDs in Japanese MSWI plants was estimated on the basis of incineration capacity. Owing to the replacement of old APCDs for dioxin control, the national average removal efficiency increased from 34.5% in 1991 to 92.5% in 2003. This resulted in an additional reduction of about 0.86Mg emission in 2003. Further study using the methodology in this study to other important emission sources like coal-fired power plants will contribute to better emission inventories.

Existence state of bromine as an indicator of the source of brominated flame retardants in indoor dust.

Indoor dust is an important medium for human exposure to brominated flame retardants (BFRs) such as polybrominated diphenyl ethers (PBDEs). In this study, we used micro X-ray fluorescence spectrometry (XRFS), digital optical microscopy, and gas chromatography-high resolution mass spectrometry to investigate the existence state of bromine as an indicator of the source of BFRs in indoor dusts and in dusts from the interior of televisions collected in Japan. By means of micro XRFS bromine mapping conducted at a 0.5-s dwell time, we were able to detect bromine levels as low as about 0.1% at each point of about a beam diameter of 50 microm across. The presence of fragments containing 1.0% or more bromine was confirmed in 27 of the 48 dust samples tested. Using magnified images of the fragments, we classified them roughly into particulates and fibrous substances. We analyzed PBDEs in the fragments containing high concentrations of bromine (> or = 0.1%) and confirmed that the fragments contained PBDEs, mainly BDE 209. Furthermore, to detect bromine concentrations < or = 0.1% in the dust samples, we analyzed the samples at a dwell time of 100 s to enhance the detection sensitivity of mapping; atthis dwell time, we confirmed the presence of bromine in the dust coating. Our results suggest that bromine is transferred from products to dust matrixes not only through miniaturization and subsequent direct migration into dust as plastic and textile fragments but also through other pathways such as vaporization and airborne transfer of microparticulates.

Effect of exposure test conditions on leaching behavior of inorganic contaminants from recycled materials for roadbeds.

Throughout the utilization of recycled materials, weathering factors such as humidity, gas composition and temperature have the potential to change the material properties and enhance the release of inorganic contaminants. In this research, the effects of weathering factors on recycled gravel materials for roadbeds were evaluated by applying three kinds of accelerating exposure tests: freezing-melting cycle test, carbonation test, and dry-humid cycle test. The effects of exposure tests were determined by X-ray diffraction (XRD) analysis and serial batch leaching test, making it possible to identify the change in release mechanisms. Sixteen elements, mainly metals, were investigated. Tested samples were molten slag from municipal solid waste, molten slag from automobile shredded residue, and crushed natural stone. After the exposure tests, the increase of cumulative release in the leaching test was generally less than 2.0 times that of the samples without the exposure test. Among the three test conditions, freezing-melting showed a slightly higher effect of enhancing the release of constituents. XRD analysis showed no change in chemical species. From these results, it was determined that the stony samples were stable enough so that their properties were not significantly changed by the exposure tests.

[Relationship between fetal lead exposure and birth weight--evaluation using deciduous incisor enamel].

OBJECTIVE: The purpose of this study was to examine the relationship between fetal lead exposure and birth weight, using deciduous incisor enamel as a biomarker and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) for analysis of lead. METHODS: We asked dental clinics (n=11) and kindergartens (n=11), mainly in the Tokyo metropolitan area and Miyagi Prefecture, and parents to enroll donors of exfoliate deciduous incisors. Donors who consented to this study voluntarily sent deciduous incisors and questionnaires to the authors between March and October, 2003. The Pb/Ca ratios of inner enamel samples were determined by LA-ICP-MS. The relationship between the lead-to-calcium ratio in the enamel and the birth weight was evaluated by stepwise multiple linear regression analysis. RESULTS: One hundred forty-one subjects (male, 72; female, 69) were enrolled in this study. The mean Pb/Ca ratio in the enamel was as low as 0.24 mg-Pb/kg-Ca (SD=0.17). Stepwise multiple linear regression analysis extracted the following maternal variables with a positive coefficient with birth weight: gestational age (p < 0.01), weight gain during pregnancy (p < 0.01), prepregnancy BMI (body mass index, p < 0.01), and parity (p < 0.05). Lead concentration in the enamel was significantly negatively associated with birth weight (p < 0.05). CONCLUSIONS: Our study indicated that lead concentration in the enamel was inversely related to birth weight. This association was found in a population of which the mean blood lead level was estimated to be as low as about 1 microg/dl. We concluded that in utero low lead exposure decreased birth weight, within the normal range of birth weight.