Tagging Glycoproteins with Fluorescently Labeled GDP‐Fucoses by Using α1,3‐Fucosyltransferases

Fucose‐containing glycans mediate a variety of biological processes, but there is little information on reaction processes and mechanisms mediated by fucosyltransferases. We recently reported on fluorescently labeled GDP‐β‐L ‐fucose‐ATTO 550, which enabled monitoring of α1,3‐fucosyltransferase activity. Here we present an extension to the previously described results, based on the synthesis of a fluorescein‐isothiocyanate (FITC)‐labeled and two carboxyfluorescein‐labeled (FAM‐labeled) NDP‐β‐L ‐fucose derivatives, and applied all four compounds in labeling of different glycoproteins with the aid of four different fucosyltransferases. The labeling processes were analyzed by in‐gel fluorescence and fluorescence polarization measurements. Comparison with the ATTO‐labeled sugar revealed that the FITC‐labeled fucose was the best of these substrates, and that the bacterial enzyme HP‐FucT tolerated the fluorescent substrates better than human fucosyltransferases.     

Begrenzung der Rissbreite nach Eurocode 2

Crack width control according to Eurocode 2 – Explanatory notes on factor k _c Reinforced concrete members are designed now according to EC2 [1]. A significant aspect is the determining of the minimum reinforcement area specified in section 7.3.2. Due to the shortened illustrations of the equations (7.1) and (7.2), the mechanical background is difficult to be highlighted. This often results in inaccurate use of the model parameters, which might lead to an unsafe design, particularly for members predominantly reinforced only by the minimum reinforcement. Therefore the mechanical background of the equations (7.1) and (7.2) is focused on in this contribution. Proper discussions and explanations might help the structural engineer in the practical design.     

A comparison of PCB bioaccumulation factors between an arctic and a temperate marine food web

To test how environmental conditions in the Arctic and the resulting ecological adaptations affect accumulation of persistent organic pollutants (POPs) in the marine food web, bioaccumulation of four polychlorinated biphenyls (PCBs) in an arctic (Barents Sea 77 °N-82 °N) and a temperate marine (Baltic Sea 54 °N-62 °N) food web were compared. Three different trophic levels were studied (zooplankton, fish, and seal), representing the span from first-level consumer to top predator. Previously published high-quality data on PCB water concentrations in the two areas were used for calculation of bioaccumulation factors (BAF). BAF was calculated as the ratio of the PCB concentration in the organism ([PCB]_org; pg/kg lipid) to the dissolved water concentration (C_w; pg/L). The BAF_Arctic:BAF_Temperate ratios were above 1 for all four PCB congeners in zooplankton (6.4-13.8) and planktivorous fish (2.9-5.0)), whereas the ratios were below 1 in seal. The mean ratio between arctic and temperate BAFs for all trophic levels and congeners (BAF_Arcti:BAF_Temperate) was 4.8. When the data were corrected for the seawater temperature difference between the two ecosystems, the ratio was 2.0. We conclude that bioaccumulation differences caused by ecological or physiological adaptations of organisms between the two ecosystems were well within a water concentration variability of 50%. Further, our data support the hypothesis that lower seawater temperature lead to a thermodynamically favoured passive partitioning to organic matrices and thus elevated ambient BAFs in the Arctic compared to the Baltic Sea. This would imply that bioaccumulation in the Arctic may be described in the same way as bioaccumulation in temperate regions, e.g. by the use of mechanistic models parameterised for the Arctic.     

Efficiency improvement of dye-sensitized solar cells based on electrodeposited TiO2 films by low temperature post-treatment

Porous crystalline TiO₂ films can be prepared at low temperatures (80 °C) by surfactant-assisted electrodeposition from TiCl₃ solution. Nevertheless, up to now calcination at high temperatures (typically 450 °C) was still necessary to establish a good performance of these films in dye-sensitized solar cells (DSSC). With this study we report that water vapour treatment at much lower temperatures (150 °C) for 1 week improves the performance of the films in DSSC to the same degree as calcination although the overall crystallinity remains lower. Reason for the good efficiency is that the porous structure stays intact and thus the dye molecules can be better adsorbed. Avoiding high temperatures during the preparation process of TiO₂ films for the application in DSSC enables the use of polymer substrates for the fabrication of flexible solar cells.     

Electrospinning of solvent‐resistant nanofibers based on poly(acrylonitrile‐co‐glycidyl methacrylate)

This article reports on the preparation of novel solvent‐resistant nanofibers by electrospinning of poly(acrylonitrile‐co‐glycidyl methacrylate) (PANGMA) and subsequent chemical crosslinking. PANGMA nanofibers with diameters ranging from 200 to 600 nm were generated by electrospinning different solutions of PANGMA dissolved in N,N‐dimethylformamide. Different additives were added to reduce the fiber diameter and improve the morphology of the nanofibers. The as‐spun PANGMA nanofibers were crosslinked with 27 wt % aqueous ammonia solution at 50°C for 3 h to gain the solvent resistance. Swelling tests indicated that the crosslinked nanofibers swelled in several solvents but were not dissolved. The weight loss of all the crosslinked nanofibrous mats immersed in solvents for more than 72 h was very low. The characterization by electron microscopy revealed that the nanofibrous mats maintained their structure. This was also confirmed by the results of the pore size measurements. These novel nanofibers are considered to have a great potential as supports for the immobilization of homogeneous catalysts and enzymes. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

The advantage of using in-situ methods for studying hydrogen mass transport: Neutron radiography vs. carrier gas hot extraction

Neutron radiography (NR) is compared with the commonly used carrier gas hot extraction (CGHE) technique. We performed isothermal hydrogen effusion experiments at 623 K to study the mass transport kinetics. The investigated material was technical iron. The quantification of the hydrogen mass flow is done for NR by using concentration standards. The temporal hydrogen concentration evolution in the sample coincides well for both methods, i.e. NR and CGHE, and is in good agreement with literature. The advantages of the NR method are the non-destructive nature of measuring and the in-situ determination of hydrogen concentrations with high spatial and temporal resolution. Remaining hydrogen inside the sample can be identified directly by the NR method.     

A computer-based system to support forensic studies on handwritten documents

Computer-based forensic handwriting analysis requires sophisticated methods for the pre-processing of digitized paper documents, in order to provide high-quality digitized handwriting, which represents the original handwritten product as accurately as possible. Due to the requirement of processing a huge amount of different document types, neither a standardized queue of processing stages, fixed parameter sets nor fixed image operations are qualified for such pre-processing methods. Thus, we present an open layered framework that covers adaptation abilities at the parameter, operator, and algorithm levels. Moreover, an embedded module, which uses genetic programming, might generate specific filters for background removal on-the-fly. The framework is understood as an assistance system for forensic handwriting experts and has been in use by the Bundeskriminalamt, the federal police bureau in Germany, for two years. In the following, the layered framework will be presented, fundamental document-independent filters for textured, homogeneous background removal and for foreground removal will be described, as well as aspects of the implementation. Results of the framework-application will also be given. Received July 12, 2000 / Revised October 13, 2000     

Interaction of hIAPP with Model Raft Membranes and Pancreatic β‐Cells: Cytotoxicity of hIAPP Oligomers

Type II diabetes mellitus (T2DM) is associated with β‐cell failure, which correlates with the formation of pancreatic islet amyloid deposits. The human islet amyloid polypeptide (hIAPP) is the major component of islet amyloid and undergoes structural changes followed by self‐association and pathological tissue deposition during aggregation in T2DM. There is clear evidence that the aggregation process is accelerated in the presence of particular lipid membranes. Whereas hIAPP aggregation has been extensively studied in homogeneous model membrane systems, especially negatively charged lipid bilayers, information on the interaction of hIAPP with heterogeneous model raft membranes has been missing until now. In the present study, we focus on the principles of aggregation and amyloid formation of hIAPP in the presence of model raft membranes. Time‐lapse tapping mode AFM and confocal fluorescence microscopy experiments followed membrane permeabilization and localization of hIAPP in the raft membrane. Together with the ThT and WST‐1 assay, the data revealed elevated cytotoxicity of hIAPP oligomers on INS‐1E cells.     

Application of cross-flow microfiltration to semi-hard cheese production from milk retentates

In the cheese industry, the concentration of milk using ultrafiltration for continuous soft and fresh cheese production is standard technology. The object of the work presented here was to produce a semi-hard cheese of quality and composition comparable to that of traditionally made cheese from highly concentrated microfiltered milk retentate. Two different membrane systems were tested for the production of high viscous milk retentate with high dry matter content. For milk containing 3.2% fat and skim milk, a concentration factor of 6.6 and 9.1 respectively was obtained using the MF/UF/UF pilot plant fitted with cassette modules. Milk containing 3.2% fat was concentrated in batches by a factor of 5.7 in the pilot plant using a ceramic membrane. Using minimal curd separation, a semi-hard one day old cheese with a dry matter of 533 g/kg, moisture on a fat-free basis (MFFB) of 626 g/kg and fat on a dry basis (FDB) value of 478 g/kg was made from the milk retentate produced with the ceramic module. The ripened cheese fulfilled the legal requirements of a traditionally produced semi-hard cheese with superb sensory qualities. Using the MF/UF/UF plant, a dry matter of 495 g/kg (MFFB 669 g/kg, FDB 493 g/kg) was achieved in a semi-hard cheese made from skim milk retentate. Our results suggest that by using a larger spacer distance in the last loop of the MF/UF/UF plant, combined with new hybrid technologies, semi-hard cheese production from full concentrate milk will soon become possible.