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- 17α-ethinylestradiol (EE2) (1)
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Organisationseinheit der BAM
Metaproteomics has matured into a powerful tool to assess functional interactions in microbial communities. While many metaproteomic workflows are available, the impact of method choice on results remains unclear. Here, we carry out a community-driven, multi-laboratory comparison in metaproteomics: the critical assessment of metaproteome investigation study (CAMPI). Based on well-established workflows, we evaluate the effect of sample preparation, mass spectrometry, and bioinformatic analysis using two samples: a simplified, laboratory-assembled human intestinal model and a human fecal sample. We observe that variability at the peptide level is predominantly due to sample processing workflows, with a smaller contribution of bioinformatic pipelines. These peptide-level differences largely disappear at the protein group level. While differences are observed for predicted community composition, similar functional profiles are obtained across workflows. CAMPI demonstrates the robustness of present-day metaproteomics research, serves as a template for multi-laboratory studies in metaproteomics, and provides publicly available data sets for benchmarking future developments.
Für die Entfernung von VOC (volatile organic compounds) aus der Luft von Fertigungsstätten werden vielfach mikroporöse Adsorbentien eingesetzt. Die gereinigte Luft wird rezyklisiert. Für die mit VOC beladenen Adsorbentien wird ein Regenerierungsweg für den mehrfachen Gebrauch gesucht. Bei der Regenerierung sollen die VOC möglichst vollständig desorbiert werden und mit einer geringen Gasmenge in einer katalytischen Verbrennung zu CO2 und H2O umgewandelt werden. Die Wirtschaftlichkeit der Luftreinigung wird wesentlich bestimmt durch die Sorptionskapazität des Adsorbens, die erforderliche Energie für die thermische Regenerierung und die Menge des Spülgases bei der Regenerierung. In dieser Arbeit wird der Einsatz von Typ X-Zeolithpulver in der Luftreinigung einer Lackiererei beschrieben und dessen Regenerierung im Labor mittels DTA/TG und Isothermenmessung der Schlüsselkomponenten untersucht. Ferner wurde die Regenerierung des beladenen Zeoliths in einem Labor-Drehrohrofen unter Variation der Betriebsparameter untersucht und anschließend mit ausgewählten Parametern in einem kleintechnischen Drehrohrofen durchgeführt. Die Verfahrensvarianten werden diskutiert.
Inflammatory bowel diseases (IBD) comprise mainly ulcerative colitis (UC) and Crohn´s disease (CD). Both forms present with a chronic inflammation of the (gastro) intestinal tract, which induces excessive changes in the composition of the associated extracellular matrix (ECM). In UC, the inflammation is limited to the colon, whereas it can occur throughout the entire gastrointestinal tract in CD. Tools for early diagnosis of IBD are still very limited and highly invasive and measures for standardized evaluation of structural changes are scarce. To investigate an efficient non-invasive way of diagnosing intestinal inflammation and early changes of the ECM, very small superparamagnetic iron oxide nanoparticles (VSOPs) in magnetic resonance imaging (MRI) were applied in two mouse models of experimental colitis: the dextran sulfate sodium (DSS)-induced colitis and the transfer model of colitis. For further validation of ECM changes and inflammation, tissue sections were analyzed by immunohistochemistry. For in depth ex-vivo investigation of VSOPs localization within the tissue, Europium-doped VSOPs served to visualize the contrast agent by imaging mass cytometry (IMC). VSOPs accumulation in the inflamed colon wall of DSS-induced colitis mice was visualized in T2* weighted MRI scans. Components of the ECM, especially the hyaluronic acid content, were found to influence VSOPs binding. Using IMC, colocalization of VSOPs with macrophages and endothelial cells in colon tissue was shown. In contrast to the DSS model, colonic inflammation could not be visualized with VSOP-enhanced MRI in transfer colitis. VSOPs present a potential contrast agent for contrast-enhanced MRI to detect intestinal inflammation in mice at an early stage and in a less invasive manner depending on hyaluronic acid content.
Multiple anthropogenic stressors cause worldwide amphibian declines. Among several poorly investigated causes is global pollution of aquatic ecosystems with endocrine disrupting compounds (EDCs). These substances interfere with the endocrine system and can affect the sexual development of vertebrates including amphibians. We test the susceptibility to an environmentally relevant contraceptive, the artificial estrogen 17α-ethinylestradiol (EE2), simultaneously in three deeply divergent systematic anuran families, a model-species, Xenopus laevis (Pipidae), and two non-models, Hyla arborea (Hylidae) and Bufo viridis (Bufonidae). Our new approach combines synchronized tadpole exposure to three EE2-concentrations (50, 500, 5,000 ng/L) in a flow-through-system and pioneers genetic and histological sexing of metamorphs in non-model anurans for EDC-studies. This novel methodology reveals striking quantitative differences in genetic-male-to-phenotypic-female sex reversal in non-model vs. model species. Our findings qualify molecular sexing in EDC-analyses as requirement to identify sex reversals and state-of-the-art approaches as mandatory to detect speciesspecific vulnerabilities to EDCs in amphibians.
Amphibians are undergoing a global decline. One poorly investigated reason could be the pollution of aquatic habitats by endocrine disrupting compounds (EDCs). We tested the susceptibility to the synthetically stabilized estrogen 17α-ethinylestradiol (EE2) in three deeply diverged anuran species, differing in sex determination systems, types of gonadogenesis and larval ecologies. To understand whether data from the amphibian model Xenopus laevis (Pipidae) are analogous and applicable to only distantly related non-model amphibians, tadpoles of X. laevis, Hyla arborea (Hylidae) and Bufo viridis (Bufonidae) were simultaneously exposed to 50, 500 and 5000 ng/L EE2 from hatching until completion of metamorphosis, using a flow-through-system under identical experimental conditions. Comparing molecularly established genetic with histologically assessed phenotypic sex in all species, we have recently shown that EE2 provoked numerous genetic-male-to-phenotypic-female sex reversals and mixed sex individuals, confirming overall its expected feminizing effect. In the present study, we focus on the influence of EE2 on gonadal and somatic development. Anatomy and histology revealed several species-specific effects. In both non-model species, H. arborea and B. viridis, high numbers of anatomically impaired gonads were observed. In H. arborea, exposed to 5000 ng/L EE2, numerous underdeveloped gonads were detected. Whereas EE2 did not alter snout-to-vent length and body weight of X. laevis metamorphs, H. arborea showed a treatment-dependent decrease, while B. viridis exhibited an increase in body weight and snout-to-vent length. Apart from a concentration-dependent occurrence of yellowish skin color in several H. arborea, no organ-specific effects were detected. Since EE2 ubiquitously occurs in many aquatic ecosystems and affects sexual and somatic development, among EDCs, it may indeed contribute to amphibian decline. The inter-species variation in developmental EE2-effects corroborates species-specific vulnerability differences towards EDCs between deeply diverged amphibian groups
A method for the analysis of Pt–protein complexes in biological samples, previously subjected to cisplatin treatment, has been developed. Proteins were separated by gel electrophoresis, and those bound to Pt were detected with high sensitivity by LA-ICP-(SF)-MS. Pt-containing spots were in-gel digested with trypsin, and the peptides produced identified using nHPLC-ESI-LTQ-FT-MS/MS. The influence of protein separation conditions, staining and gel processing prior to laser ablation on Ptprotein bonds preservation have been evaluated using standard proteins incubated with cisplatin. 2-DE separation under non-reducing conditions followed by either Coomassie blue brilliant or silver staining is appropriate for Pt–protein complexes, achieving a good separating resolution of the proteins in biological samples. Direct LA-ICP-MS analysis of glycerol-treated dried gels for Pt–protein monitoring resulted in better sensitivity, more reliable relative Pt signals and a simpler and less time-consuming approach compared to the analysis of blotted membranes. Ablation of gels allowed tackling protein identification of Pt-spots in the remaining non-ablated material in the gel, making it unnecessary to run several gels in parallel for separate Pt detection and protein identification. By using this approach, Pt coordinated to proteins, such as α-2-macroglobulin, transferrin, albumin or hemoglobin, was detected in the serum from a rat treated in vivo with cisplatin after nrSDS-PAGE separation. Furthermore, the first complete LA-ICP-MS metalloprotein contour map in a 2-DE gel has been produced, in this case for the detection of Pt–protein complexes in renal proximal tubule epithelial cells (RPTECs) incubated with cisplatin. Several proteins were identified in those spots containing Pt, which may have a connection with the drug-induced nephrotoxicity mainly affecting this cell type in the kidney.
Synchrotron radiation-induced X-ray fluorescence (SR-XRF) spectrometry was applied to determine iodine concentrations in alluvial Pt–Pd aggregates with delicate morphological features from Córrego Bom Sucesso, Minas Gerais, Brazil. The millimetre-sized botryoidal and rod-shaped grains of alluvial Pt–Pd–Hg intermetallic compounds have surprisingly high concentrations of iodine, in the range from 10 to ~ 120 μg/g. Because iodine is a strongly biophile element, known to be enriched in peatlands and plant remains in soils by microbial activity, its concentration in the Pt–Pd nuggets suggests biogenic precious-metal fixation in the aqueous alluvial milieu. Biogenic and inorganic processes, such as bioreduction and electrochemical metal accretion, could mutually have contributed to the growth of nanoparticles, formed on organic templates, to millimetric Pt–Pd nuggets.
The second part of this paper looks continues the comparative investigation of lean duplex stainless steels, manganese alloyed austenitic and duplex stainless steels and ferritic stainless steels. These new materials were investigated in many different test procedures and in different conditions focused on the application in civil engineering and common use. In addition to electrochemical investigations, all materials were exposed to atmospheric conditions in different surface states. In Part 2, Prof. Gümpel and his team examine the effects of coastal and urban atmospheres on the samples, and presents their final conclusions.
Rising prices of nickel and molybdenum in the recent past years have led to unprecedented interest in substituting leaner-content alloys for standard 300-series austenitic stainless steels in many applications. Due to the price volatility of various alloying elements, many new materials have entered the market, yet no direct comparison of corrosion resistance of these new materials exists. In this project, comparative investigations were carried out and in each case one or several representatives of a material group were taken into account. The material groups covered were: lean duplex stainless steels, manganese alloyed austenitic and duplex stainless steels and ferritic stainless steels. In addition to electrochemical investigations, all materials were exposed to various atmospheric conditions in different surface states.