Purification and structural characterisation of lipid transfer protein from red wine and grapes

Lipid transfer proteins (LTP) play a major role in plant defence and are of particular interest due to their known ability to cause allergic reactions. These proteins are expressed in grapes and also remain detectable after vinification, especially in red wine. However, it remains unknown whether the protein undergoes any changes during the vinification process. Here, we present a purification method for LTPs from Dornfelder grapes and wine. By liquid-chromatography–mass spectroscopy (LC–MS/MS) we identified LTPs from two different species (Vitis vinifera and Vitis aestivalis). Additionally, the purified LTPs were characterised using spectrometric methods, confirming their high purity and structural stability during vinification. We conclude that LTPs are resistant to the alcohol content (13.5 vol%), acidic milieu of wine and other ingredients present during the vinification process, indicating that the allergenic potential of grape LTP is not diminished by the vinification process.     

Characterization of glow‐discharge–treated cellulose acetate membrane surfaces for single‐layer enzyme electrode studies

Cellulose acetate membranes (CA) were modified by means of plasma polymerization of ethylene diamine (EDA) and n‐butylamine (n‐BA). The motivation for this work was the application of a modified membrane for the single‐layer enzyme electrode. A tubular reactor with the external radiofrequency (13.56 MHz) excitation was used. Surface modification was performed at 5, 10, and 15 W power (at 27 Pa working pressure) for 5, 10, 15 min. Modified surfaces were characterized in detail by FTIR–ATR, XPS (ESCA), contact angle, and enzyme immobilization activity. The best treatment results were obtained for EDA with 5 W and 30 min and 15 W and 10 min. These results are discussed using surface analysis data. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 81: 1341–1352, 2001     

Use of the Cone Calorimeter and ConeTools software for development of innovative intumescing graphite systems

The Cone Calorimeter test method has been one of the most used small‐scale fire test method for years now and is at present widely spread over the world. In contrast to many other fire test methods, the Cone Calorimeter provides a range of data with sound scientific basis, which allows a wide range of applications. It can be used for modelling and also for enhanced product development. This paper describes the use of the Cone Calorimeter for the development of new innovative materials in combination with a mathematical model. As example, the cost effective development of an innovative intumescing graphite system for protecting particle board is explained. The performance goal of the project is to obtain the threshold values for a B class in the Single Burning Item (SBI) test method used for the newly developed Euroclass system. The focus of this paper is on the development tools and not on the chemical development of the protective system. During the research it was necessary to develop a new sample holder for the Cone Calorimeter. The results from the project show that the industry can save development time and resources by using the Cone Calorimeter in combination with a simple mathematical model. Copyright © 2010 John Wiley & Sons, Ltd.     

Factorial interlaboratory tests for microbiological measurement methods

This paper presents a validation approach for microbiological methods based on a combination of interlaboratory tests and factorial experiments. It requires not more than 4 participants but is achieving comparable statistical confidence as in method validation studies with 8–12 participants, if properly designed. The approach is illustrated by a comprehensive validation of the Arxula adeninivorans yeast estrogen screen (A-YES) assay for the detection of estrogenic activity in mineral water.     

Agent-based simulation for science,technology, and innovation policy

Scientometrics - Policymaking implies planning, and planning requires prediction—or at least some knowledge about the future. This contribution starts from the challenges of complexity,...     

Germanium-68 as a tracer for silicon fluxes in freshwater sediment

The sorption of Si may decisively influence the P dynamics in surface sediment through competitive ligand exchange mechanisms. Many aspects of the process, such as the impact of Si dissolving from diatoms, are both insufficiently known and difficult to monitor by quantitative analyses due to the ubiquitousness of Si in sediment. Since the radioactive isotopes of Si have impractical half-times, the applicability of 68Ge as a tracer for Si additions in both inorganic and biogenic form to freshwater sediment (Lake Vesij?rvi, Southern Finland) was studied in a 24-h laboratory experiment. The 68Ge-label was added to the sediment with either inorganic Si (940mg l(-1)) or diatoms (3.2 x 10(6) cellsl(-1)), and the distributions of the 68Ge-label and the different forms of added Si between the interstitial water and sequentially extracted, solid-phase sediment pools were critically examined. The inorganic Si addition significantly increased the amount of Si in the interstitial water and in the reversibly bound fraction, while the diatom addition had no discernible effects. The relative distribution of Si and 68Ge between the various sediment pools indicated that the first sorption phase of the added inorganic Si was similar to that reported for P. The high concentration of diatom-derived 68Ge-labelled Si in the interstitial water and in the easily soluble, reversibly bound pool indicated rapid dissolution of the added diatoms. The comparable distributions of the diatom-derived and inorganic Si-derived 68Ge-label within the studied pools indicated that the sorption of Si dissolving from diatoms in surface sediment closely resembles that of an inorganic addition of Si. Caution about the chemical speciation of the 68Ge-label is, however, advisable in sediment environments. Fractionation procedures designed for e.g. P may also be of limited use when applied to a different element, such as Si.     

Prozessorientierte Wirtschaftlichkeitsuntersuchung für E-Government

E-Government economy needs to be dealt with in a multidimensional and multiperspective way. Existing evaluation concepts, though, hardly meet these pre-requisites. In this paper we give an overview of approaches and difficulties in evaluating IT-projects in public administration. Current E-Government evaluation approaches and surveys are compared as to their fitness to judge on an actual E-Government project. Based on the E-Government evaluation criteria used for the assessment of the evaluation approaches we develop a process-oriented E-Government evaluation concept which is illustrated in a mini-case.     

The absence of the Isw2p-Itc1p chromatin-remodelling complex induces mating type-specific and Flo11p-independent invasive growth of Saccharomyces cerevisiae

The Isw2p-Itc1p chromatin remodelling complex of Saccharomyces cerevisiae is a member of the ISWI class of ATPases with a nucleosome spacing activity, involved in regulation of expression of a broad spectrum of genes. Its absence causes derepression of a-specific genes and aberrant morphology in alpha-mating type cells. We report here that the deletion of the ISW2 gene in the originally non-invasive BY strain induces mating type-specific invasive growth strongly affected by nitrogen starvation. Although the Flo11 protein was postulated to be critical for haploid invasive growth, we showed that the invasive growth caused by the isw2 and itc1 deletions in alpha-mating type cells was Flo11p-independent. This type of invasive growth was proved to be a consequence of the activation of the pheromone response pathway. Our results suggest that Isw2 and Itc1 proteins do not have the same impact on the described phenomenon.     

Development of a class-specific ELISA for sulfonylurea herbicides (sulfuron screen)

The development of a direct competitive ELISA for the detection of a broad range of sulfonylurea herbicides (SUs) is described. Polyclonal antibodies were generated in rabbits using three different immunizing haptens. Antiserum with the broadest specificity was obtained with a mesosulfuronbenzylamine derivative which was coupled via a succinic acid spacer to keyhole limpet hemocyanine. A heterologous enzyme tracer which did not contain the succinic acid bridge was prepared using activated horseradish peroxidase. The direct competitive ELISA was optimized and applied for spiked tap and surface water samples. From 30 SUs, 8 compounds showed a molar cross-reactivity (CR) higher than 100% (this value was set for the hapten) and 11 compounds CRs between 10% and 100%. The ELISA can detect 16 SUs at a concentration of 0.1 microg/L or lower. Different surface and tap water samples were spiked with chlorimuron ethyl, metsulfuron methyl, or primisulfuron methyl at concentrations of 100, 200, or 500 ng/L and subsequently analyzed by both ELISA and HPLC-UV. Correlation analysis revealed good agreement between both methods (r2 = 0.983/0.948/0.982; n = 21 for each analyte). Using ELISA, no sample pretreatment other than filtration was necessary.     

Ultrasensitive detection of unstained proteins in acrylamide gels by native UV fluorescence

Visualization of proteins inside acrylamide and other gels usually relies on different staining methods. To omit the protein-staining procedure, we visualized unstained proteins inside acrylamide gels by laser excitation with ultraviolet (UV) light (280 nm, 35 mJ/cm2) and directly detected native UV fluorescence. In one-dimensional gels, a detection limit as low as 1 ng for bovine serum albumin and 5 ng for other proteins with a linear dynamic range (2.7 orders of magnitude) comparable to state of the art fluorescent dyes could be achieved. In addition, the application of this method to 20 microg of a whole cell lysate separated in a two-dimensional gel showed more than 600 spots. Since protein labeling always represents a serious obstacle in protein identification technologies, the working efficiency with our procedure can be considered as a significant improvement for protein visualization and reproducibility in proteomics.