287 research outputs found
2D DIGE analysis of protein extracts from muscle tissue
No full text2D DIGE, two-dimensional difference gel electrophoresis, is a technology used to study the protein expression on two-dimensional gels. Protein samples are labeled with different color fluorescent dyes designed not to affect the relative migration of proteins during electrophoresis. Here, we describe the practical procedures necessary to perform a 2D DIGE experiment for a muscle tissue protein extract followed by CyDye DIGE fluors minimal labeling and the analysis of 2D DIGE gels for the assessment of quantitative differences
Gel gradient electrophoresis, isoelectric focusing and two-dimensional techniques in horizontal, ultrathin polycrylamide layers
No full textAn ultrathin layer, horizontal polyacrylamide gel system for electrophoresis, isoelectric focusing and two-dimensional techniques is described. Gel slabs 240 micron thin for unidimensional, or 360 micron thin for two-dimensional runs are cast on cellophane foils as support. The sample is loaded in pockets pre-cast in the gel (2--3 microliter size) or in trenches for two-dimensional experiments. The second dimension is routinely performed in concave exponential gel gradients, spanning an acrylamide concentration from 4% to 22.5%. The sensitivity with the common Coomassie Blue stain is very high, well below 0.1 microgram protein/band. Zymogram detections can be developed within a few minutes, thus retaining the band sharpness of the focused zones or of the bands separated in pore gradient electrophoresis. Sample handling, staining and destaining and gel drying and storage are greatly simplified and performed in a fraction of the time needed for conventional, thick gels in the 1-2 mm thickness range
Isoelectric focusing in immobilized pH gradients: principle, methodology and some applications
No full textA new technique for generating pH gradients in isoelectric focusing is described, based on the principle that the buffering groups are covalently linked to the matrix used as anticonvective medium. For the generation of this type of pH gradient in polyacrylamide gels, a set of buffering monomers, called Immobiline (in analogy with Ampholine), is used. The pH gradient gels are cast in the same way as pore gradient gels, but instead of varying the acrylamide content, the light and heavy solutions are adjusted to different pH values with the aid of the Immobiline buffers. Available Immobiline species make it possible to generate any narrow linear pH gradient between pH 3 and 10. The behaviour of these types of gradients in isoelectric focusing is described. Immobilized pH gradients show a number of advantages compared with carrier ampholyte generated pH gradients. The most important are: (1) the cathodic drift is completely abolished; (2) they give higher resolution and higher loading capacity; (3) they have uniform conductivity and buffering capacity; (4) they represent a milieu of known and controlled ionic strength
Sectoral job effects of trade: An input-output analysis for Germany
No full textThe current globalisation process is characterized by the emergence of global value chains. That is, production processes are becoming increasingly geographically fragmented. Not only are final goods traded internationally, but in particular, trade in intermediate goods and services has increased significantly over time. In the industrialised countries, the manufacturing sectors were the first that were compelled to face the challenges of globalisation. When services were still considered non-tradable, manufacturing firms had already decided to relocate their production sites to developing or emerging economies, due to lower wage levels abroad and increasing price competition domestically. In this paper, we use input-output analysis to explore the relationship between trade and both job creation and job destruction in the German manufacturing industry in 2005. The results show that being integrated into the world economy is advantageous for the German economy. In 2005, the net exports of the manufacturing industries led to trade-induced job gains of around 2,400,000. This figure is equivalent to 6.2 per cent of total German employment. Furthermore, the job effects of trade were positive for a large majority of countries. The greatest job gains resulted from trade with the United States, the United Kingdom and France. Interestingly, even trade with the new EU Member States is beneficial in terms of job creation. --input-output analysis,international trade,employment
Evaluating the efficacy of subcellular fractionation of blast cells using live cell labeling and 2D DIGE
No full textLabeling of exposed cell surface proteins of live cells using CyDye DIGE fl uor minimal dyes is an efficient strategy for cell surface proteome profi ling and quantifying differentially expressed proteins in diseases. Here we describe a strategy to evaluate a two-step detergent-based protein fractionation method using live cell labeling followed by visualization of the fluorescently labeled cell surface proteins and fractionated proteins within a single 2D gel.Yin Ying Ho, Megan Penno, Michelle Perugini, Ian Lewis and Peter Hoffman
Comparative 2D DIGE analysis of the depleted serum proteome for biomarker discovery
No full textSerum is unarguably the most used diagnostic fluid. As it circulates throughout the body, leakage peptides/proteins from damaged and dying cells, host-response proteins including inflammatory mediators, and aberrant secretions from tumors and diseased tissues are released into serum, potentially providing a rich source of disease biomarkers. Here, a method for extending access to the serum proteome by removing highly abundant proteins prior to comparative two-dimensional difference gel electrophoresis (2D DIGE) and subsequent protein digestion for identification by mass spectrometry is described.Megan Penno, Matthias Ernst, and Peter Hoffman
Jobs gained and lost through trade: The case of Germany
No full textThe nature of international trade has changed significantly. For centuries, trade concentrated on the exchange of finished goods. It now increasingly involves bits of value that are added at different locations to combine into one final product. Therefore, trade in functions or tasks are of growing importance and exports of final goods are no longer an appropriate indicator of the international competitiveness of countries. The process of globalisation has an impact on domestic labour markets. Due to the increasing integration of the world economy, some jobs are gained and others lost in any open economy. Concerns about German workers losing jobs to foreign competition dominate many political debates. Many people fear that being integrated in the world economy is disadvantageous for Germany. In this paper, we use input-output analysis to explore the relationship between trade and both job creation and job destruction in Germany over the period 1995-2006. We present two main findings. First, in an autarkic situation, around 7.0 per cent of total German jobs would not have existed at all in 2006. The job effect of trade was positive in every reporting year. Second, the manufacturing sector contributed most to this positive job effect, but also in the service sector, many jobs were retained through trade. --input-output analysis,international trade,employment
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Application of DIGE and mass spectrometry in the study of type 2 Diabetes Mellitus (T2DM) mouse models
No full textKnowledge of the differences between the amounts and types of protein that are expressed in diseased compared to healthy subjects may give an understanding of the biological pathways that cause disease. This is the reasoning behind the presented protocol, which uses difference gel electrophoresis to discover up‐ or down‐regulated proteins between mice of different genotypes, or
of those fed on different diets, that may thus be prone to develop diabetes‐like phenotypes. Subsequent analysis of these proteins by tandem mass spectrometry typically facilitates their identification with a high degree of confidence
Der Netz-Cent: Ein Instrument zur Finanzierung des Breitbandausbaus
No full textDer Breitbandausbau soll auch in wirtschaftlich unattraktiven Gebieten vorangetrieben werden. Vor allem ländliche Gebiete sind mangels Wirtschaftlichkeit noch unterversorgt. Der Autor schlägt die Einführung eines Netz-Cents vor, um die notwendigen Finanzierungsmittel sicherzustellen. Der Netz-Cent ist ein Konzept, mit dem positive externe Effekte bei den Endkunden internalisiert werden können.To benefit from the digital revolution, broader investments in the digital infrastructure are needed. But private investments are not sufficient to build up the infrastructure needed for the digital revolution. Especially in rural areas, the return on investments is rather low or negative. The author suggests imposing a fee of e.g. one cent per gigabyte used. The money collected from the consumer would then be used to fund private investments. The fee can be justified with positive external effects. Every internet user benefits from a broad expansion of fast internet connections but does not pay for it. The fee internalises those positive external effects
Secondary zoospores in the algal endoparasite Maullinia ectocarpii (Plasmodiophorea).
No full textThe present paper deals with the ultrastructure of zoospores produced by the plasmodiophorid Maullinia ectocarpii , living in the marine algal host Ectocarpus siliculosus. The zoospores described here are very similar to secondary zoospores of Polymyxa graminis and Phagomyxa sp. (the latter an algal endopara- site, also). Our results indicate that M. ectocarpii produces two types of plasmodia, and suggest that is a species with a complete life cycle, as it is known for all the Plasmodiophormycota that have been studied. Sporogenic and sporangial plasmodia produce, respectively, primary zoospores with parallel flagella within thick walled resting sporangia, and secondary zoospores with opposite flagella within thin walled sporangia.Fil: Parodi, Elisa Rosalia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional del Sur; ArgentinaFil: Caceres, Eduardo Jorge. Universidad Nacional del Sur. Departamento de Biología, Bioquímica y Farmacia. Laboratorio de Ficología y Micología; ArgentinaFil: Westermeier, Renato. Universidad Austral de Chile; ChileFil: Muller, Dieter G.. Universität Konstanz; Alemani
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