Structural Insight into IAPP-Derived Amyloid Inhibitors and Their Mechanism of Action

Designed peptides derived from the islet amyloid polypeptide (IAPP) cross-amyloid interaction surface with Aβ (termed interaction surface mimics or ISMs) have been shown to be highly potent inhibitors of Aβ amyloid self-assembly. However, the molecular mechanism of their function is not well understood. Using solution-state and solid-state NMR spectroscopy in combination with ensemble-averaged dynamics simulations and other biophysical methods including TEM, fluorescence spectroscopy and microscopy, and DLS, we characterize ISM structural preferences and interactions. We find that the ISM peptide R3-GI is highly dynamic, can adopt a β-like structure, and oligomerizes into colloid-like assemblies in a process that is reminiscent of liquid–liquid phase separation (LLPS). Our results suggest that such assemblies yield multivalent surfaces for interactions with Aβ40. Sequestration of substrates into these colloid-like structures provides a mechanistic basis for ISM function and the design of novel potent anti-amyloid molecules.     

Model simulation and experimental verification of a cation-exchange IgG capture step in batch and continuous chromatography

The cation-exchange capture step of a monoclonal antibody (mAb) purification process using single column batch and multicolumn continuous chromatography (MCSGP) was modeled with a lumped kinetic model. Model parameters were experimentally determined under analytical and preparative conditions: porosities, retention factors and mass transfer parameters of purified mAb were obtained through a systematic procedure based on retention time measurements. The saturation capacity was determined through peak fitting assuming a Langmuir-type adsorption isotherm. The model was validated using linear batch gradient elutions. In addition, the model was used to simulate the start-up, cyclic steady state and shut down behavior of the continuous capture process (MCSGP) and to predict performance parameters. The obtained results were validated by comparison with suitable experiments using an industrial cell culture supernatant. Although the model was not capable of delivering quantitative information of the product purity, it proved high accuracy in the prediction of product concentrations and yield with an error of less than 6%, making it a very useful tool in process development.     

Dynamic Nuclear Polarization in the solid state: a transition between the cross effect and the solid effect

Proton Dynamic Nuclear Polarization (DNP) experiments were conducted on a 3.4 T homebuilt hybrid pulsed-EPR-NMR spectrometer, on static samples containing 10 mM or 40 mM TEMPOL in frozen glassy solutions of DMSO/water. During DNP experiments proton-NMR signals are enhanced with the help of microwave (MW) irradiation on or close to the Electron Paramagnetic Resonance (EPR) spectrum of the free radicals in the sample, transferring polarization from the free electrons to the nuclei. In the solid state a distinction is made between three DNP enhancement mechanisms: the Solid Effect (SE), the Cross Effect (CE) and Thermal Mixing (TM). In an effort to determine the dominant DNP mechanisms responsible for the enhancement of the nuclear signals, electron and nuclear spin-lattice relaxation rates, enhancement buildup times and microwave (MW) swept DNP spectra were measured as a function of temperature and MW irradiation strength. We observed lineshape variations of the DNP spectra that indicated changes in the relative contributions of SE-DNP and CE-DNP with temperature and MW power. Using a theoretical model describing the SE-DNP and CE-DNP the DNP spectra could be analyzed without involving the TM-DNP mechanism and the relative SE-DNP and CE-DNP contributions to the nuclear enhancement could be determined. From this analysis it follows that lowering the temperature beyond 20 K increases the SE-DNP and decreases the CE-DNP contributions. Possible explanations for this behavior are suggested.     

Minimal hyperspace actions of homeomorphism groups of h-homogeneous spaces

Suppose that X is an h-homogeneous zero-dimensional compact Hausdorff space, i.e., X is a Stone dual of a homogeneous Boolean algebra. Using the dual Ramsey theorem and a detailed combinatorial analysis of what we call stable collections of subsets of a finite set, we obtain a complete list of the minimal sub-systems of the compact dynamical system (Exp(Exp(X)), Homeo(X)), where Exp(X) denotes the hyperspace comprising the closed subsets of X equipped with the Vietoris topology. The importance of this dynamical system stems from Uspenskij’s characterization of the universal ambit of G = Homeo(X). The results apply to the Cantor set, the generalized Cantor sets X = {0,1} ^κ for noncountable cardinals κ, and to several other spaces. A particular interesting case is X = ω* = βω \ ω, where βω denotes the Stone- ?ech compactification of the natural numbers. This space, called the corona or the remainder of ω, has been extensively studied in the fields of set theory and topology.     

Synthesen mit Nucleins?urebausteinen. 4. Mitt. Die sequenzspezifische Cooligokondensation von Nucleins?ure-Bausteinen mit Affinit?tsschutzgruppen. Ein neuer Weg zur Synthese von Poly nucleotidfragmenten

Ohne Zusammenfassung     

Invarianten Kegelpunktfreier (k+1)-Gratregelflächen

We extend striction and, parameter of distribution to (k+1)-dimensional generalized ruled surfaces with edge ruled surfaces.