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31.
32.
Werner E. G. Müller 《Die Naturwissenschaften》1998,85(1):11-25
Geodia cydonium , which code for proteins. The analyses of their deduced amino acid sequences allowed a molecular biological approach to solve
the problem of monophyly of Metazoa. Molecules of the extracellular matrix/basal lamina, with the integrin receptor, fibronectin,
and galectin as prominent examples, cell-surface receptors (tyrosine kinase receptor), elements of sensory systems (crystallin,
metabotropic glutamate receptor), and homologs/modules of an immune system (immunoglobulin like molecules, scavenger receptor
cysteine-rich, and short consensus repeats, rhesus system) classify the Porifera as true Metazoa. As living fossils, provided
with simple, primordial molecules allowing cell-cell and cell-matrix adhesion as well as processes of signal transduction
as known in a more complex manner from higher Metazoa, they also show peculiarities not known in other metazoan phyla. Tissues
of sponges are rich in telomerase activity, suggesting a high plasticity in the determination of cell lineages. It is concluded
that molecular biological studies with sponges as model will not only help to understand the evolution of Protoctista to Metazoa
but also the complex, hierarchial regulatory network of cells in higher Metazoa. 相似文献
33.
34.
Sperm competition is thought to be an important selective pressure shaping sperm form and function. However, few studies have
moved beyond gross examinations of sperm morphology. Sperm length is subject to sexual selection via sperm competition in
the scarab beetle Onthophagus taurus. Here, the structure and ultrastructure of spermatozoa in this species were investigated using light and electron microscopy.
Spermatozoa were found to be filiform, measuring about 1,200 mm in length. The sperm head consists of a three-layered acrosome
and a nuclear region bearing the anterior extension of the centriole adjunct. Acrosome and nuclear regions are bilaterally
symmetric, with their axes of symmetry being orthogonal to each other. Head and flagellar structures are connected by a well-developed
centriole adjunct. The sperm heads are asymmetrically surrounded by accessory material and embedded into the cytoplasm of
the spermatocyst cell. The accessory material is produced inside the spermatids and then transferred to the outside due to
a new membrane formed around the sperm’s organelles. The old spermatid membrane separates the accessory material from the
cyst cell. The flagellum contains a 9+9+2 axoneme, two accessory bodies, and two mitochondrial derivatives of unequal size.
The major mitochondrial derivative is significantly larger than the minor one. The axoneme is arranged in a sinusoidal manner
parallel along the major mitochondrial derivative. The spermatozoa show no progressive motility when released in buffer solution
which is likely to be the result of the flagellar arrangement and the structure of the major mitochondrial derivative. The
cross-sectional area of the minor and the major mitochondrial derivatives show different patterns of genetic variation. The
data provide the first estimates of genetic variation in sperm ultrastructure for any species, and give evidence for the persistence
of genetic variation in ultrastructure required for the rapid and divergent evolution that characterizes spermatozoa generally. 相似文献
35.
36.
Thomas Schwartz Christina Jungfer Stefan Heißler Frank Friedrich Werner Faubel Ursula Obst 《Chemosphere》2009,77(2):249-257
DNA-based population analysis was applied in combination with Raman spectrometry and Environmental Scanning Electron Microscopy for the characterisation of natural biofilms from sand and activated carbon filters operated for a long term at a municipal waterworks. Whereas the molecular biology polymerase chain reaction combined with denaturing gradient gel electrophoresis approach provides a deeper insight into the bacterial biofilm diversities, Raman spectrometry analyses the chemical composition of the extracellular polymer substances (EPS), microorganisms embedded in EPS as well as other substances inside biofilm (inorganic compounds and humic substances). Microscopy images the spatial distribution of biofilms on the two different filter materials. In addition, bacterial bulk water populations were compared with biofilm consortia using the molecular fingerprint technique mentioned.Population analysis demonstrated the presence of more diverse bacterial species embedded in a matrix of EPS (polysaccharides, peptides, and nucleic acids) on the sand filter materials. In contrast to this, activated carbon granules were colonised by reduced numbers of bacterial species in biofilms. Besides α-, β-, and γ-Proteobacteria, a noticeable specific colonisation with Actinobacteria was found on activated carbon particles. Here, the reduced biofilm formation came along with a decreased EPS synthesis. The taxonomy profiles of the different biofilms revealed up to 60% similarity on the same filter materials and 32% similarity of different materials. Similarity of adherent communities from filter materials and bulk water populations from the filter effluent varied between 36% and 58% in sand filters and 6–40% in granular activated carbon filters.The biofilm investigation protocols are most crucial to subsequent acquisition of knowledge on biofilm dynamics and bacterial contributions to transformation or adsorption processes in waterworks facilities. 相似文献
37.
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
38.
Urbatzka R van Cauwenberge A Maggioni S Viganò L Mandich A Benfenati E Lutz I Kloas W 《Chemosphere》2007,67(6):1080-1087
The river Lambro is the most polluted tributary of the river Po in North Italy and was chosen as a representative water course discharging industrialized areas. Water and sediment samples of the river Lambro were investigated regarding the presence of endocrine disrupting compounds. A combined procedure was used consisting of solid-phase extraction and HPLC based fractionation of samples, of screening for (anti)androgenic activity using the yeast androgen screen (YAS) and of chemical analysis using HPLC-MS/MS and GC-MS. Androgenic and antiandrogenic activities were found in specific fractions of the water and sediment while the total extracts showed antiandrogenic activity only. The chemical analysis of the fractions and total extracts with antiandrogenic activities revealed the presence of compounds with suspected antiandrogenic potency such as bisphenol A, iprodione, nonylphenol, p,p'-DDE and tert-octylphenol but other unknown compounds contributed mainly to the observed antiandrogenic activities. The antiandrogenic load of the river Lambro ranged between 1.34 and 17.1 microM flutamide-equivalents and may pose a risk to aquatic environments. Future screenings for EDC in the environment that have the potential to interfere with reproduction of aquatic organisms should be extended to different modes of actions including (anti)androgenic ones. 相似文献
39.
de Souza Pereira M Heitmann D Reifenhäuser W Meire RO Santos LS Torres JP Malm O Körner W 《Chemosphere》2007,67(9):1736-1745
Monitoring of immission of persistent organic pollutants in the industrialized area of Volta Redonda (V.R.) and in the National Park of Itatiaia (PNI) in southeast Brazil was performed using an endemic bromeliad species as biomonitor and measuring bulk deposition rates of polychlorinated biphenyls (PCB) and polycyclic aromatic hydrocarbons (PAH). For the sum of PCB, overall deposition rates were between 17 and 314 ng/(m2 day) in winter and between 43 and 81 ng/(m2 day) in summer, respectively. Deposition rates of dioxin-like PCBs ranged from 0.14 to 2.8 pg WHO-TEQ/(m2 day) in winter and from 0.90 to 4.3 pg WHO-TEQ/(m2 day) in summer. PCB deposition rates (total PCB and WHO-TEQ) were in the same range in winter in V.R. and PNI. In summer, contamination levels in V.R. were 6-10-folds higher than in PNI. PCB concentrations in biomonitor samples from V.R. and PNI were in the same range in summer and in winter. Concentrations of total PCB ranged from 14 to 95 microg/kg dry matter (d.m.) in winter and from 18 to 27 microg/kg d.m. in summer, respectively. The TEQ values were between 1.7 and 4.1 ng WHO-TEQ/kg d.m. in winter and between 1.9 and 2.9 ng WHO-TEQ/kg d.m. in summer. PCB concentrations of di-ortho PCB but not of non-ortho PCB were a factor of 2-4 lower in summer in both areas. PCB congener profiles resembled those from technical formulations. The profiles shifted to the higher chlorinated congeners in summer, probable due to revolatilisation of the lighter components at higher temperatures. PCB profiles in biomonitor resembled those from deposition samples and the shift to the heavier congeners in summer was even more pronounced. PAH deposition rates were in a similar range in both areas (131-2415 ng/(m2 day)). PAH levels in biomonitor samples from V.R. were about one order of magnitude higher than in samples from PNI indicating the impact of local sources. PAH profiles revealed stationary thermal processes as main source of contamination in V.R. whereas in PNI, biomass burning seems to be the main contamination source. 相似文献
40.