Trypsiligase-Catalyzed Labeling of Proteins on Living Cells

Fluorescent fusion proteins are powerful tools for studying biological processes in living cells, but universal application is limited due to the voluminous size of those tags, which might have an impact on the folding, localization or even the biological function of the target protein. The designed biocatalyst trypsiligase enables site-directed linkage of small-sized fluorescence dyes on the N terminus of integral target proteins located in the outer membrane of living cells through a stable native peptide bond. The function of the approach was tested by using the examples of covalent derivatization of the transmembrane proteins CD147 as well as the EGF receptor, both presented on human HeLa cells. Specific trypsiligase recognition of the site of linkage was mediated by the dipeptide sequence Arg-His added to the proteins’ native N termini, pointing outside the cell membrane. The labeling procedure takes only about 5 minutes, as demonstrated for couplings of the fluorescence dye tetramethyl rhodamine and the affinity label biotin as well.     

Deliberately Losing Control of C−H Activation Processes in the Design of Small-Molecule-Fragment Arrays Targeting Peroxisomal Metabolism

Combined photochemical arylation, “nuisance effect” (S_NAr) reaction sequences have been employed in the design of small arrays for immediate deployment in medium-throughput X-ray protein–ligand structure determination. Reactions were deliberately allowed to run “out of control” in terms of selectivity; for example the ortho-arylation of 2-phenylpyridine gave five products resulting from mono- and bisarylations combined with S_NAr processes. As a result, a number of crystallographic hits against NUDT7, a key peroxisomal CoA ester hydrolase, have been identified.     

Atomistic simulations of grain boundary migration in copper

While the motion of twist boundaries can be readily studied by atomistic simulations with molecular dynamics (MD) under the action of an elastic driving force, the approach fails for tilt boundaries. This is due to the interaction of the elastic stress with the grain boundary (GB) structure, which causes plastic strain by GB sliding. A novel concept, the orientation correlated driving force, is introduced to circumvent this problem. It is shown that this concept can be successfully applied to the study of the migration of tilt boundaries. The migration behavior of several twist and tilt GBs was investigated. The transition from low-to high-angle boundaries can be captured, and a structural transition of tilt boundaries was found at high temperatures, which also affected the migration behavior. The results compare well with experimental results of the motion high-angle boundaries, but for low-angle boundaries, the agreement is poor. This article is based on a presentation made in the “Hillert Symposium on Thermodynamics & Kinetics of Migrating Interfaces in Steels and Other Complex Alloys,” December 2–3, 2004, organized by The Royal Institute of Technology in Stockholm, Sweden.     

Sensor guided handling and assembly of active micro-systems

Positioning accuracies in the range of a few micrometers and below are necessary for the assembly of active micro-systems. In order to reach these accuracies, an assembly system for sensor guided micro-assembly is developed in the Collaborative Research Centre 516 “Design and manufacturing of active micro-systems”. The combination of a parallel robot with an integrated 3D vision sensor and a micro-gripper enables to reach relative positioning accuracies below 1 μm.     

Surfactant replacement therapy in acute respiratory distress syndrome from viral pneumonia

A modified natural surfactant was administered to a patient with life-threatening adult respiratory distress syndrome caused by viral pneumonia. Subsequently, there was a marked improvement in gas exchange. In order to assess the mechanism for improved oxygenation, computed tomography of the lungs was done. Quantitative analysis of the scans taken before and after surfactant administration indicates that improvement in gas exchange was largely due to the expansion of underinflated and collapsed lung areas. Although this is a single case report, it provides insight into the possible beneficial effect of instilled surfactant in severe respiratory distress from viral pneumonia.     

Growth Stresses in the Oxide Scales on TiAl Alloys at 800 and 900°C

In the oxidation of TiAl alloys, the role of scale-growth stresses formed during oxidation has, thus far, been unknown. In the present paper the oxide-growth stresses were investigated by the deflection-test method in monofacial oxidation (DTMO) accompanied by acoustic-emission measurements. On unmodified surfaces the growth stresses are compressive and reach levels of around –100 MPa. At the same time, significant acoustic emission occurs indicating that even under isothermal conditions, stresses are relieved by a scale-cracking mechanism. For oxide scales on TiAl surfaces, which had been ion implanted with chlorine before oxidation, a very thin protective alumina layer is formed which, however, develops growth stresses in the range of several GPa, accompanied by intensive acoustic emission. In all stress–time curves, a dynamic situation is observed. This consists of phases of stress relief by scale microcracking and phases of stresses increase due to crack healing and further oxide growth. As a result, the level of stress as a function of oxidation time, is characterized by an oscillating course.     

Thomas Fincke und die <Emphasis Type="Italic">Geometria rotundi</Emphasis>

Thomas Fincke (January 6^th, 1561–April 24^th, 1656), born in Flensburg (Germany), was one of the very most important and significant scientists in Denmark during the seventeenth century, a mathematician and astrologer and physician in the beginning of modern science, a representative of humanism and an influentual academic organizer. He studied in Strasbourg (since 1577) and Padua (since 1583) and received his M.D. in Basel (1587), he practised as a physician throughout his life (since 1587 or 1590) and became a professor at Copenhagen (1591). But he was best known because of his Geometriae rotundi libri XIIII (1583), a famous book on plane and spherical trigonometry, based not on Euclid but on Petrus Ramus. In this influentual work, in which Fincke introduced the terms tangent and secant and probable first noticed the Law of Tangents and the so-called Newton-Oppel-Mauduit-Simpson-Mollweide-Gauss-formula, he showed himself to be „abreast of the mathematics of his time“.