Mikrotechnik für Raumfahrtanwendungen

This contribution describes the preparation, based upon a chemically amplified novolak resist (CAR), electron beam lithography, and ECR plasma etching, of structures with a high aspect ratio (10∶1) and lateral dimensions in the sub-micrometer range (150nm–300nm) which may serve as collector surfaces for sub-μm dust particles in a space experiment.     

The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis

Proinflammatory chemokine ligand 26 (CCL26, eotaxin-3) mediates transendothelial cell migration of eosinophils by binding and activating the G-protein-coupled (GPC) chemokine receptor 3 on the surface of eosinophilic cells. Here we have investigated the role of glycosaminoglycans (GAGs) as potential co-receptors in the process of CCL26-induced eosinophil chemotaxis. For this purpose, we have first identified the GAG-binding site of CCL26 by a site-directed mutagenesis approach in the form of an alanine screening. A panel of GAG-binding-deficient mutants has been designed, generated, and analyzed with respect to their binding affinities to heparan sulphate (HS) by isothermal fluorescence titration studies. This showed that basic amino acids in the α-helical part of CCL26 are strongly involved in GAG-binding. In chemotaxis experiments, we found that decreased GAG-binding affinity correlated with decreased chemotactic activity, which indicates an involvement of GAGs in eosinophil migration. This was further proven by the negative impact of heparinase III treatment and, independently, by the incubation of eosinophils with an anti heparan sulfate antibody. We finally investigated eosinophils’ proteoglycan (PG) expression patterns by real-time PCR, which revealed the highest expression level for serglycin. Including an anti-serglycin antibody in CCL26-induced eosinophil migration experiments reduced the chemotaxis of these immune cells, thereby proving the dependence of eosinophil mobilization on the proteoglycan serglycin.     

Predicting the Lifespan and Retweet Times of Tweets Based on Multiple Feature Analysis

In social network services, such as Facebook, Google+, Twitter, and certain postings attract more people than others. In this paper, we propose a novel method for predicting the lifespan and retweet times of tweets, the latter being a proxy for measuring the popularity of a tweet. We extract information from retweet graphs, such as posting times; and social, local, and content features, so as to construct prediction knowledge bases. Tweets with a similar topic, retweet pattern, and properties are sequentially extracted from the knowledge base and then used to make a prediction. To evaluate the performance of our model, we collected tweets on Twitter from June 2012 to October 2012. We compared our model with conventional models according to the prediction goal. For the lifespan prediction of a tweet, our model can reduce the time tolerance of a tweet lifespan by about four hours, compared with conventional models. In terms of prediction of the retweet times, our model achieved a significantly outstanding precision of about 50%, which is much higher than two of the conventional models showing a precision of around 30% and 20%, respectively.     

Crystallization of S-layer protein lattices on surfaces and interfaces

A review is given on the structure, chemistry, and assembly of crystalline bacterial cell surface layers (S-layers). S-layers composed of single protein or glycoprotein species represent the most common cell surface structures observed in prokaryotic organisms. Isolated S-layer proteins possess the intrinsic property for recrystallization into isoporous monomolecular arrays in suspension and at a broad spectrum of surfaces (e.g. silicon, metals, polymers) and interfaces (e.g. air–liquid interface or lipid films). The well-defined arrangement of functional groups on S-layer lattices allows the binding of functional molecules (e.g. enzymes, antibodies, ligands) and particles in defined regular arrays. S-layers also represent templates for the formation of inorganic nanocrystal superlattices (e.g. Au, CdS, Pt) as required for molecular electronics and non-linear optics. Finally, S-layers can be used as the structural basis for a biomolecular construction kit involving all major species of biological molecules for applications in molecular nanotechnology, nanobiotechnology and biomimetics.     

Self-assembled alpha-hemolysin pores in an S-layer-supported lipid bilayer

The mammalian distal colon, which is composed of different cell types, actively transports Na, K and Cl in absorptive and K and Cl in secretory directions. To further characterize the K absorption process and to identify the cells involved in K absorption, unidirectional Rb fluxes and luminal Rb uptake into different epithelial cell types were determined in isolated guinea-pig distal colon. Net Rb absorption (1.5-2.5 micromol.h-1.cm-2) was not influenced by inhibition of Na transport with amiloride or by incubating both sides of the epithelium with Na-free solutions, but was almost completely abolished by luminal ouabain, ethoxzolamide or by incubating both sides of the epithelium with Cl-free solutions. Luminal Rb uptake, blockable by luminal ouabain, preferentially occurred in columnar surface and neck cells, to a lesser extent in surface goblet cells and to an insignificant degree in lower crypt cells. Employing a luminal Rb-Ringer (5.4 mM Rb) the Rb concentration increased within 10 min in columnar surface and neck, surface goblet and lower crypt cells to 70, 32 and about 10 mmol. kg-1 wet weight, respectively. The presence of 5.4 mM K in the luminal incubation solution reduced Rb uptake almost completely indicating a much higher acceptance of the luminal H-K-ATPase for K than for Rb. The increase in Na and decrease in K concentrations in surface and neck cells induced by luminal ouabain might indicate inhibition of the basolateral Na-K-ATPase or drastic enhancement of cellular Na uptake by the Na-H exchanger. Bilateral Na-free incubation did not alter Rb uptake, but bilateral Cl-free incubation drastically reduced it. Inhibition of net Rb absorption by ethoxzolamide and inhibition of both Rb absorption and Rb uptake by bilateral Cl-free incubation support the notion that cellular CO2 hydration is a necessary prerequisite for K absorption and that HCO3 leaves the cell via a Cl-HCO3 exchanger. Since ouabain-inhibitable transepithelial Rb flux and luminal Rb uptake rate by surface and neck cells were about the same, Rb(K) absorption seems to be accomplished mainly by columnar surface cells.     

S-layers as patterning elements for application in nanobiotechnology

Two-dimensional bacterial cell surface layer protein crystals (S-layers) are the most commonly observed cell surface structure in bacteria and archaea. Isolated S-layer proteins have the intrinsic tendency to self-assemble into crystalline arrays in suspension and on various interfaces. Basic research on the structure, genetics, chemistry, morphogenesis and function of S-layers has led to a broad spectrum of applications in nanotechnology and biomimetics. The possibility to change the properties of S-layer proteins by genetic engineering opens new ways for tuning their functional and structural features. Functionalized S-layer proteins that maintain their ability to self-assemble have led to new affinity matrices, diagnostic tools, vaccines or biocompatible surfaces, as well as to biological templating or specific biomineralisation strategies at surfaces.