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Generally accepted quality criteria for the comparison of immunoassays are still missing and assay conditions vary greatly between different laboratories. To address this problem, the influence of different parameters on the overall assay performance was assessed, specifically for different enzyme immunoassays (EIAs) for the anthropogenic markers caffeine (CAF) and carbamazepine (CBZ). Special emphasis was dedicated to the parameters temperature, assay format and enzyme-substrate combination. The temperature parameter was systematically studied for all incubation steps of the direct EIA formats employing the photometric horseradish peroxidase (HRP) substrate 3,3’,5,5’- tetramethylbenzidine (TMB) and the fluorometric HRP substrate 3-(4-hydroxyphenyl)propionic acid for both analytes. A temperature decrease only during the competition step led to an increase in assay sensitivity by a factor of 10 to 15 for CBZ and CAF, respectively, independent of the enzyme substrate used. Room temperature experiments yielded the smallest coefficients of variations, minimizing the edge effect. The influence of the assay format on different performance parameters was studied with the determination of CAF in consumer products. In addition to the HRP substrates, the enzyme alkaline phosphate (AP) and its chromogenic substrate para-nitrophenyl phosphate and a fluorescent substrate, 4-methylumbelliferyl phosphate, were employed. Seven quality criteria were defined and validated to compare these immunoassays. The evaluation of the four criteria (sensitivity, measurement range, relative dynamic range and goodness of fit) for the standard curves revealed that the direct format is superior to the indirect format, with the HRP TMB format showing the best performance. Three additional criteria for an applicationdriven analysis of real samples, in this case CAF-containing beverages and cosmetics, confirmed this result in terms of accuracy as well as intra- and inter-plate precision. The enzyme-substrate combination was investigated when several direct CBZ assays were applied to the analysis of water samples; here, three HRP assays and four AP assays were studied, along with luminescence detection. The HRP assays reached better sensitivities and lower quantifiable concentrations compared to the AP assays. Only the HRP assays and the chemiluminescent AP juice assay fulfilled the requirements for the four criteria applied to standard curves; all other AP assays were not considered for application to real samples based on these criteria. The AP juice assay can only be employed for influent samples whereas all HRP assays are applicable to influent and effluent wastewater samples according to intra- and inter-plate precision. Furthermore, the HRP assays alone are suitable for surface water analysis; here, the chromogenic HRP TMB assay yielded the best results, as any type of water sample can be quantified with high precision. Whether these quality criteria, derived here for standard curves as well as their application to real samples, can be transferred to other immunoassay formats for quality assurance remains to be shown.
The CuII complex of H4TETP (H4TETP = 1,4,8,11-tetraazatetradecane-1,4,8,11-tetrapropionic acid) is five-coordinate with a distorted square-pyramidal structure (τ = 0.45; i.e. the geometry is nearly half-way between square-pyramidal and trigonal-bipyramidal) and a relatively long Cu–N and a short Cu–O bond; the comparison between powder and solution electronic spectroscopy, the frozen solution EPR spectrum and ligand-field-based calculations (angular overlap model, AOM) indicate that the solution and solid state structures are very similar, i.e. the complex has a relatively low 'in-plane' and a significant axial ligand field with a dx²-y² ground state. The ligand-enforced structure is therefore shown to lead to a partially quenched Jahn–Teller distortion and to a relatively low complex stability, lower than with the corresponding acetate-derived ligand H4TETA. This is confirmed by potentiometric titration and by the biodistribution with 64Cu-labeled ligands which show that the uptake in the liver is significantly increased with the H4TETP-based system.
Polyoxometalates (POMs) are inorganic cluster compounds that have been shown to possess a number of pharmacological properties, including antidiabetic, antibacterial, antiprotozoal, antiviral and anticancer activities. Their molecular mechanism of action is largely unknown. However, several studies indicate that many of their activities may be due to the inhibition of enzymes, in particular, of those enzymes that are accessible from the extracellular space and do not require the penetration of cell membranes. In this review, we describe the recent progress in the preparation and optimization of POMs, and an evaluation of their use as inhibitors of different families of enzymes. The next important steps in this area of research will be to gain a better understanding of the interactions of POMs with enzymes on a structural level through an X-ray crystallographic study of enzyme–POM complexes and the analysis of structure–activity relationships. Furthermore, POMs with increased stability and in vivo half-lives have to be prepared. Surface modification may allow the targeting of POM drugs at their sites of action.
A novel Gram-positive, aerobic, actinobacterial strain, CF6/1T, was isolated in 2007 during environmental screening of arid desert soil in the Sahara near to Ourba, Chad. The isolate was found to grow best in a temperature range of 20–37 °C and at pH 6.0–8.5 and showed no NaCl tolerance, forming black-coloured and nearly circular colonies on GYM agar. Chemotaxonomic and molecular characteristics determined for the isolate match those previously described for members of the genus Geodermatophilus. The DNA G + C content of the novel strain was determined to be 74.9 mol %. The peptidoglycan was found to contain meso-diaminopimelic acid as the diagnostic diamino acid. The main phospholipids were determined to be phosphatidylethanolamine, phosphatidylinositol, phosphatidylcholine, diphosphatidylglycerol and traces of phosphatidylglycerol; MK-9(H4) was identified as the dominant menaquinone and galactose as the diagnostic sugar. The major cellular fatty acids were found to be the branched-chain saturated acids iso-C16:0 and iso-C15:0, as well as C17:1ω8c. The 16S rRNA gene sequence shows 97.5–97.9 % sequence identity with the four validly named or at least effectively published members of the genus: Geodermatophilus obscurus (97.5 %), Geodermatophilus arenarius (97.7 %), Geodermatophilus ruber (97.9 %) and Geodermatophilus nigrescens (97.9 %). Based on the results from this polyphasic taxonomic analysis and DNA–DNA hybridizations with all type strains of the genus, we propose that strain CF6/1T represents a novel species, Geodermatophilus siccatus, with the type strain CF6/1T = DSM 45419T = CCUG 62765T = MTCC 11414T.
Analyse lokaler Verformungsfelder mittels DIC-Technik während der Heißrissbildung beim Schweißen
(2013)
Für die Gewährleistung der Bauteilsicherheit ist während des Fertigungsprozesses das Vermeiden von Imperfektionen maßgeblich. Zu diesen Imperfektionen zählen auch beim Schweißen auftretende Heißrisse, die insbesondere die Bauteileigenschaft beeinflussen und demzufolge ausgeschlossen werden müssen. Aufgrund der hohen Komplexität, hervorgerufen durch das Zusammenwirken zahlreicher Einflussgrößen, existieren überwiegend vereinfachte Werkstoffmodelle, die das Heißrissphänomen nur unzureichend beschreiben. Da bislang eine Charakterisierung dieser Heißrissentstehung überwiegend auf spannungs- und dehnungsbasierten Modellen erfolgt, wurde in diesem Fall speziell geprüft, in wie weit vereinfachend eine Quantifizierung des Heißrissverhaltens anhand von Verschiebungen bzw. Verschiebungsraten möglich ist. Die Tatsache, dass Heißrisse in Werkstoffen auftreten, die aus metallurgischer Sicht als heißrissunkritisch eingestuft werden können, zeigt, dass weitere Faktoren, wie der konstruktive Einfluss, betrachtet werden müssen. In der vorliegenden Arbeit wurde dieser Aspekt unter Nutzung des Controlled-Tensile- Weldability(CTW)-Tests untersucht. Hierfür wurde die Digital-Image-Correlation( DIC)-Technik erfolgreich zur In-situ-Analyse der lokalen Querverschiebungen dy und Querverschiebungsraten dy/dt in direkter Schmelzbadnähe während des Schweißens eingesetzt.-----------------------------------------------------------------------------------------------------------------------------------------------------------------
In order to guarantee the component safety, it is essential to avoid imperfections during the production process. These imperfections also include hot cracks which arise during welding, exert an influence on the component property in particular and must consequently be excluded. Due to the high complexity caused by the interaction of a large number of influencing variables, there are mostly simplified material models which describe the hot cracking phenomenon only insufficiently. Since this hot crack initiation has, until now, been characterised mostly on models based on stresses and strains, it was especially checked in this case the extent to which the hot cracking behaviour can be quantified using a simplifying method taking account of displacements and displacement rates. The fact that hot cracks occur in materials that, from the metallurgical point of view, cannot be classified as hot-crack-critical shows that further factors such as the influence of design need to be considered. This aspect was investigated in the present study by using the Controlled Tensile Weldability (CTW) test. For this, the Digital Image Correlation (DIC) technique was successfully applied for the in situ analysis of the local transverse displacements dy and transverse displacement rates dy/dt in the direct weld pool vicinity during welding.