No abstract

Co-stimulation via receptors of the tumor necrosis factor superfamily (TNFSF) emerges as promising strategy to support antitumor immune responses. Targeted strategies with antibody-fusion proteins composed of a tumor-directed antibody part and the extracellular domain of a co-stimulatory ligand of the TNFSF constitute an attractive option to focus the co-stimulatory activity to the tumor site. Since TNFSF members intrinsically form functional units of non-covalently linked homotrimers, the protein engineering of suitable antibody-fusion proteins is challenging. Aiming for molecules of simple and stable configuration, we used TNFSF ligands in a single-chain format (scTNFSF), i.e., three units of the ectodomain connected by polypeptide linkers, folding into an intramolecular trimer. By fusing tumordirected scFv antibody fragments directed against EpCAM or FAP to co-stimulatory scTNFSF molecules (sc4-1BBL, scOX40L, scGITRL or scLIGHT), a set of monomeric scFv-scTNFSF fusion proteins was generated. In comparison to the scFv-TNFSF format, defined by intermolecular homotrimerization via the TNFSF part, scFv-scTNFSF showed equal or enhanced co-stimulatory activity despite reduced avidity in antibody binding. In addition, enhanced serum stability and improved bioavailability in mice were observed. We show that the scFv-scTNFSF format can be applied to various members of the TNFSF, presenting targeting-dependent co-stimulatory activity. Hence, this format exhibits favorable properties that make it a promising choice for further therapeutic fusion protein development.Abbreviations: EpCAM, epithelial cell adhesion molecule; ED-A, fibronectin extradomain A; FAP, fibroblast activation protein; GITRL, glucocorticoid-induced tumor necrosis factor receptor (GITR) ligand; LIGHT, homologous to lymphotoxins, shows inducible expression and competes with herpes simplex virus glycoprotein D for herpesvirus entry mediator (HVEM), a receptor expressed by T lymphocytes; mAb, monoclonal antibody; scFv, single-chain fragment variable; TNF, tumor necrosis factor; TNFSF, tumor necrosis factor superfamily; TRAIL, TNF-related apoptosis inducing ligand

During the last years GaN‐technology has proven to fulfill the requirements of solid state lighting. Lighting requirements are mainly driven by brightness, operation voltage and lifetime. Brigthness is determined by internal efficiency as well as extraction efficiency whereas the ohmic losses determining the operating voltage are dominated by series resistance and contact resistance. Both, brightness and voltage, strongly depend on the device structure as well as the chip design. SiC based [1, 2] as well as Sapphire based LEDs [3] have proven their capability for high brightness devices, still suffering from various compromises such as cost, ESD‐stability, high series resistance etc. Recently OSRAM‐OS has demonstrated its newly developed product line based on the so called ThinGaN™ technology, a true thinfilm approach that overcomes most of the compromises mentioned. The technology allows highest brightness levels at lowest operating voltage, is scalable and supports all wavelengths. The devices act as true surface emitters with a lambertian emission pattern. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)

Because only 0.01% of prokaryotic genospecies can be cultured and in situ observations are often impracticable, culture-independent methods are required to understand microbial life and harness potential applications of microbes. Here, we report a methodology for the production of proteins with desired functions based on single amplified genomes (SAGs) from unculturable species. We use this method to resurrect an alcohol dehydrogenase (ADH/D1) from an uncharacterized halo-thermophilic archaeon collected from a brine pool at the bottom of the Red Sea. Our crystal structure of 5,6-dihydroxy NADPH-bound ADH/D1 combined with biochemical analyses reveal the molecular features of its halo-thermophily, its unique habitat adaptations, and its possible reaction mechanism for atypical oxygen activation. Our strategy offers a general guide for using SAGs as a source for scientific and industrial investigations of "microbial dark matter."

A feasibility study using an ion mobility spectrometer coupled with a multi-capillary column (MCC) was started to identify characteristic peaks of volatile compounds in exhaled human breath samples of 10 ml volume. The breath of 20 patients with sarcoidosis and suspicion of sarcoidosis because of mediastinal lymph node enlargement was investigated. Using a set of procedures for data processing and scoring a sector of interest was determined within the IMS-chromatogram. It could be shown that a procedure related to a single peak in the IMS-chromatogram delivers differentiation into the two groups of patients with confirmed sarcoidosis and those suffering no sarcoidosis. The potential biomarker is characterized by the following parameters: inverse mobility (1/K(0)) 0.53 ± 0.01 V s cm(-2)-retention time 22 ± 5 s. These results are a first step in breath analysis by MCC/IMS in patients with sarcoidosis by an automated procedure applied to IMS-chromatograms directly.