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851.
Robert Kase Peter D. Hansen Birgit Fischer Werner Manz Peter Heininger Georg Reifferscheid 《Environmental science and pollution research international》2009,16(1):54-64
Background, aim, and scope The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor
binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents
(HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness
against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to
the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA
was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The
applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor
for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different
levels to characterise its influence on elution and binding processes of receptor-binding substances.
Materials and methods The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup
known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically
relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA
conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing
step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic
receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity
at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of
sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the
elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option
to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method.
Results This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference
substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l
to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay
against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations
of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For
sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation
of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could
be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard
deviation in the salinity adaptation procedure of the elutriates was below 5%.
Discussion Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different
salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with
regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate
both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given
the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance
to discriminate receptor-binding potentials in environmental samples.
Conclusions The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput
screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust
against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods.
The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish
samples can be measured up to salinity levels of 20.5‰.
Recommendations and perspectives In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test
method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen
and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part
of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information
about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening
tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments. 相似文献
852.
853.
A simple modeling concept was previously applied to study air-foliage exchange of both polybrominated diphenyl ethers (PBDEs) and polycyclic aromatic hydrocarbons (PAHs) using field measurements. In this paper, the predictive capabilities of this framework were tested using an independent set of data collected at the same sampling location for a different time period. Firstly, atmospheric (particle-bound and gaseous phase) PBDE and PAH concentrations were used to predict their accumulation in spruce needles. Conversely, we used PBDE and PAH concentrations in vegetation to predict their atmospheric concentrations. In both cases, calculated and measured values showed good agreement, providing evidence that the developed framework and our derived deposition velocities can be used as a predictive tool for these two different classes of semi-volatile organic compounds. 相似文献
854.
The Sambhar Salt Lake hydrological system, including river waters, groundwaters, evaporating pans and sub-surface brines, has been analyzed for the salt content (TDS) and naturally occurring radionuclides (210Po, 210Pb and 226,228Ra). The abundance of these radionuclides and their activity ratios show a wide variation in different hydrological regimes, which helps to geochemically characterize the lake system. A significantly lower Ra to total dissolved solids (TDS) ratio in the brines (by two to three orders of magnitude), when compared to the groundwaters and river waters, suggests removal of dissolved Ra by co-precipitation with Ca–Mg minerals at an early stage of the brine evolution. The concentration of Ra in evaporating lake/pan waters saturates at a value of about 35 Bq L−1 over the salinity range of 100–370 g L−1; attributable to its equilibration with the clay minerals. The two distinct regimes, saline lake system (lake water, evaporating pans and sub-surface brines) and groundwaters have been identified based on their differences in the distribution of 226,228Ra isotopes. This observation points to the conclusion that the groundwaters and the lake brines are not intimately coupled in terms of their origin and evolution. The abundances of 210Po and 210Pb along with their activity ratios (210Po/210Pb) are markedly different among the surface lake waters/evaporating pans, sub-surface lake brines and groundwaters. These differences are explained in terms of different geochemical behaviour of these nuclides in presence of algae and organic matter present in these water regimes. 相似文献
855.
S.J. Sartandel S.K. Jha S.V. Bara R.M. Tripathi V.D. Puranik 《Journal of environmental radioactivity》2009,100(10):831-834
The understanding and evaluation of the possible interactions of various naturally occurring radionuclides in the world's third largest man-made dam, Nagarjuna Sagar located in Andhra Pradesh, India and built on river Krishna assumed significance with the finding of uranium deposits in locations near the dam. For the present work, surface soil samples from the mineralized area of Lambapur, Mallapuram, Peddagattu and sediment core samples from the Nagarjuna Sagar dam were analyzed for naturally occurring radionuclides namely uranium and thorium using gamma spectrometric technique. Also toxic elements lead and chromium were analysed by the Energy Dispersive X-ray Fluorescence Spectrometer (EDXRF) technique. Surface soil samples show a variation from 25 to 291 Bq/kg (2.02–23.5 mg/kg) for 238U and 32–311 Bq/kg (7.9–76.9 mg/kg) for 232Th. U/Th concentration ratio in surface soil samples ranged from 0.19 to 0.31 and was found comparable with the nation wise average of 0.26. The study of sediment core samples reflected higher U/Th concentration ratio of 0.30–0.33 in the bottom section of the core as compared to 0.22–0.25 in the upper section. The concentration ratio in the upper section of the core was similar to the ratio 0.23 found in the western Deccan Basalt region through which the river originates. A higher concentration of lead and chromium was observed in the upper section of the core compared to bottom section indicating the impact of river input on the geochemical character of dam sediment. 相似文献
856.
857.
Typologically identical (dwarf birch-herb-dwarf shrub-moss) open and closed larch forests growing on the same altitudinal transect have proved to differ in the structural-functional organization of lower vegetation layers. Coverage, general species composition, and species richness of the herb-dwarf shrub layer are higher in the open forest than in the closed forest. Correlations between individual species of vascular plants weaken upon transition from the open to the closed forest. Conversely, the coverage of the lichen-moss layer increases in the closed forest, which contributes to its role as a factor of selection of vascular plant species against the background of the prevailing influence of the tree layer. 相似文献
858.
E. L. Zheleznaya 《Russian Journal of Ecology》2009,40(1):39-43
The overgrowing of meadow-bog communities by shrubs and trees (age 5–40 years, crown closure 0.4–0.9) leads to a decrease in incident illumination. As a consequence, juvenile and generative plants disappear from D. incarnata population loci, their average ecological density decreases, and the loci enter the state of regression. The digging activity of wild boars disrupts phytocenotically closed groups of longirhizomatous herbs, thereby creating favorable conditions for seed reproduction of D. incarnata. The species begins to form population loci with a complete ontogenetic spectrum and high ecological density, eventually restoring the normal (definitive) population structure. 相似文献
859.
860.
Specific ecogenetic features of solods in the forest-steppe zone has been studied, and the role of biotic and abiotic factors of soil formation has been estimated. Differentiation of soil horizons with respect to the physical clay fraction accounts for specific features of their moistening, which is reflected in the ratio of strongly and weakly crystallized forms of iron. Periodic changes in redox conditions results in iron segregation, and a high content of fulvic acids accounts for its eluvial migration. These processes are controlled by soil geochemical barriers. 相似文献