Synthetic 6-O-methylglucose-containing polysaccharides (sMGPs): design and synthesis

With the hope of mimicking the chemical and biological properties of natural 6-O-methyl-D-glucose-containing polysaccharides (MGPs), synthetic 6-O-methyl-D-glucose-containing polysaccharides (sMGPs) were designed and synthesized from alpha-, beta-, and gamma-cyclodextrins (CDs). The synthetic route proved to be flexible and general, to furnish a series of sMGPs ranging from 6-mer to 20-mer. A practical and scalable method was discovered selectively to cleave the CD derivatives and furnish the linear precursors to the glycosyl donors and acceptors. The Mukaiyama glycosidation was adopted to couple the glycosyl donors with the glycosyl acceptors. Unlike in the 3-O-methyl-D-mannose-containing polysaccharide (sMMP) series, the amount of the Mukaiyama acid required in the sMGP series increased with an increase of substrate size; that is, for large oligomers, more than one equivalent of the acid was required.     

Few-cycle-driven XUV laser harmonics: generation and focusing

We have investigated the use of sub-10-fs near-infrared laser pulses to generate high-order harmonic radiation efficiently in the wavelength region between 30 to 10 nm. The ultrashort rise time of the driver pulses allows harmonics to be produced at low ionization levels and hence to grow coherently over propagation lengths becoming comparable to the XUV absorption lengths in the noble gas medium. As a result, absorption-limited harmonic generation has been extended to the 10-nm range for the first time. Harmonic conversion efficiencies of (3-4)᎒^ъ in the range of 10-13 nm in neon and some two orders of magnitude higher at around 30 nm in argon have been obtained in simple gas tube targets under these conditions. Preliminary focusing tests with 13-nm harmonic radiation have been carried out with a specially designed zoneplate and a spherical Mo/Si multilayer mirror and have resulted in spot sizes of about 2 microns. Our experiments aim at paving the way to nonlinear optics in the soft-X-ray regime.     

Evaluation of light scattering data from bimodal turbid suspensions: a simple fit procedure

Whereas correlation spectroscopy gives reliable information on the size of immersed particles in those cases where the size distribution is narrow, large problems arise for more complex particle distributions. For instance, samples containing distinctly different particles of rather similar size lead to correlation functions which are very close to those of monodisperse samples. We present a measurement technique which is based on angle dependent measurements of 3D cross correlation functions and an evaluation scheme which uses the results of the Mie theory. The experimental technique warrants applications to strongly scattering samples. Having tested this procedure with mixtures containing standard latex particles we applied it to a sample of skimmed, homogenized milk.     

Reliability optimization of friction-damped systems using nonlinear modes

A novel probabilistic approach for the design of mechanical structures with friction interfaces is proposed. The objective function is defined as the probability that a specified performance measure of the forced vibration response is achieved subject to parameter uncertainties. The practicability of the approach regarding the extensive amount of required design evaluations is strictly related to the computational efficiency of the nonlinear dynamic analysis. Therefore, it is proposed to employ a recently developed parametric reduced order model (ROM) based on nonlinear modes of vibration, which can facilitate a decrease of the computational burden by several orders of magnitude.     

A Flow Cytometry-Based Ultrahigh-Throughput Screening Method for Directed Evolution of Oxidases

Oxidases are of interest to chemical and pharmaceutical industries because they catalyze highly selective oxidations. However, oxidases found in nature often need to be re-engineered for synthetic applications. Herein, we developed a versatile and robust flow cytometry-based screening platform “FlOxi” for directed oxidase evolution. FlOxi utilizes hydrogen peroxide produced by oxidases expressed in E. coli to oxidize Fe²⁺ to Fe³⁺ (Fenton reaction). Fe³⁺ mediates the immobilization of a His₆-tagged eGFP (eGFP^His) on the E. coli cell surface, ensuring the identification of beneficial oxidase variants by flow cytometry. FlOxi was validated with two oxidases—a galactose oxidase (GalOx) and a D-amino acid oxidase (D-AAO)—yielding a GalOx variant (T521A) with a 4.4-fold lower K_m value and a D-AAO variant (L86M/G14/A48/T205) with a 4.2-fold higher k_cat than their wildtypes. Thus, FlOxi can be used for the evolution of hydrogen peroxide-producing oxidases and applied for non-fluorescent substrates.     

Optimization of Fishing Strategies in Space and Time as a Non-convex Optimal Control Problem

The behavior of a fishing fleet and its impact onto the biomass of fish can be described by a nonlinear parabolic diffusion–reaction equation. Looking for an optimal fishing strategy leads to a non-convex optimal control problem with a bilinear control action. In this work, we present such an optimal control formulation, prove its well-posedness and derive first- and second-order optimality conditions. These results provide a basis for tailored finite element discretization as well as for Newton type optimization algorithms. First numerical test problems show typical features as so-called No-Take-Zones and maximal fishing quota (total allowable catches) as parts of an optimal fishing strategy.     

Structural characterization of ß-amyloid oligomer-aggregates by ion mobility mass spectrometry and electron spin resonance spectroscopy

Formation and accumulation of fibrillar plaques and aggregates of ß-amyloid peptide (Aß) in brain have been recognized as characteristics of Alzheimer’s disease (AD). Oligomeric aggregates of Aß are considered critical intermediates leading to progressive neurodegeneration; however, molecular details of the oligomerization and aggregation pathway and the structures of Aß-oligomers are hitherto unclear. Using an in vitro fibril formation procedure of Aß(1–40), ß-amyloid aggregates were prepared and insoluble aggregates separated from soluble products by centrifugation. In this study, ion mobility mass spectrometry (IM-MS) was applied in combination with electron paramagnetic resonance spectroscopy (EPR) to the identification of the components of Aß-oligomers, and to their structural and topographical characterization. The formation of Aß-oligomers and aggregates was monitored by gel electrophoresis, and Aß-oligomer bands were identified by in-gel tryptic digestion and matrix-assisted laser desorption ionization–mass spectrometry (MALDI-MS) to consist predominantly of Aß(1–40) peptide. First, ion mobility-MS studies of soluble Aß-aggregates prepared by incubation for 5 days were performed on a quadrupole time-of-flight mass spectrometer and revealed (1) the presence of at least two different conformational states, and (2), the formation of Met-35 oxidized products. For estimation of the size of Aß-aggregates using EPR spectroscopy, a modified Aß(1–40) peptide containing an additional N-terminal cysteine residue was prepared, and a 3-(2-iodoacetamido)-2,2,5,5-tetramethyl-1-pyrrolidinyloxy radical spin label derivative (IPSL) was coupled by S-alkylation. The EPR spectra of the spin-labeled Cys-Aß(1–40) oligomers were matched with spectra simulations using a multi-component simulation strategy, resulting in complete agreement with the gel electrophoresis results.