One Question,Multiple Answers: Biochemical and Biophysical Screening Methods Retrieve Deviating Fragment Hit Lists

Fragment‐based lead discovery is gaining momentum in drug development. Typically, a hierarchical cascade of several screening techniques is consulted to identify fragment hits which are then analyzed by crystallography. Because crystal structures with bound fragments are essential for the subsequent hit‐to‐lead‐to‐drug optimization, the screening process should distinguish reliably between binders and non‐binders. We therefore investigated whether different screening methods would reveal similar collections of putative binders. First we used a biochemical assay to identify fragments that bind to endothiapepsin, a surrogate for disease‐relevant aspartic proteases. In a comprehensive screening approach, we then evaluated our 361‐entry library by using a reporter‐displacement assay, saturation‐transfer difference NMR, native mass spectrometry, thermophoresis, and a thermal shift assay. While the combined results of these screening methods retrieve 10 of the 11 crystal structures originally predicted by the biochemical assay, the mutual overlap of individual hit lists is surprisingly low, highlighting that each technique operates on different biophysical principles and conditions.     

Characterization of piezoelectric ceramics using the burst/transient method with resonance and antiresonance analysis

In this paper, a comprehensive methodology for characterizing the high power resonance behavior of bulk piezoelectric ceramics using the burst method is described. In the burst method, the sample is electrically driven at its resonance frequency, and then either a short circuit or an open circuit condition is imposed, after which the vibration decays at the resonance or antiresonance frequency, respectively. This decay can be used to measure the quality factor in either of these conditions. The resulting current in the short circuit vibration condition is related to the vibration velocity through the “force factor.” The generated voltage in the open circuit vibration condition corresponds to the displacement by the “voltage factor.” The force factor and the voltage factor are related to material properties and physical dimensions of the sample. Using this method, the high power behavior of the permittivity, compliance, effective piezoelectric charge constant, electromechanical coupling factor, and material losses can be determined directly by measuring the resonance (short circuit) and antiresonance (open circuit) frequencies, their corresponding quality factors, the force factor A, and the voltage factor B. The experimental procedure to apply this method is described and demonstrated on commercially available hard and semi‐hard PZT materials of geometry.     

Production of Polyunsaturated Fatty Acids by Mortierella alpina Using Submerse and Solid State Fermentation

A new isolate of Mortierella alpina, > 98 % identical with M. alpina ATCC 16266, was cultivated in a defined glucose‐based medium with three organic nitrogen sources (glycine, urea and Na‐L‐glutamate) at three different concentrations in shaking flasks at 20 °C. The results were compared to the cultivation in complex medium with yeast extract as nitrogen source. In the defined media, high yields of polyunsaturated fatty acids (PUFAs) and arachidonic acid (ARA), respectively, were obtained with Na‐L‐glutamate. However, the absolute highest yields of PUFA and ARA were measured with the yeast extract medium. An optimized yeast extract complex medium was used for a submerse bioreactor cultivation in a 45‐L scale. Furthermore, M. alpina was cultivated in a solid state fermenter, using an oat bran water mixture as substrate.     

Potential and Limitations of Natural Chabazite for Selective Catalytic Reduction of NOx with NH3

The potential of the natural chabazite for the selective catalytic reduction (SCR) of NOx with NH₃ is evaluated in the present work. Activity tests were performed under technically relevant reaction and temperature conditions for the fresh and hydrothermally aged catalysts. The natural chabazite before and after alkaline removal as well as after iron and copper addition were compared. The structural as well as surface and bulk properties were elucidated by a variety of complementary characterization techniques, i.e. XRD, XPS, EPR, BET, NH₃‐TPD, ex situ and in situ XAS. The results indicate that an important facet for using the natural chabazite for the standard and fast SCR reactions is the removal of alkaline metals, which at the same time also leads to a partial change of the structure and the size of the iron‐containing particles. The performance and especially the hydrothermal stability can be further improved by copper addition.     

Replication of sub‐micron features using amorphous thermoplastics

A comprehensive experimental study was carried out to replicate sub‐micron features using the injection molding technique. For the experiments, five different plastic materials were selected according to their flow properties. The materials were polycarbonate (PC), styrene‐butadiene block copolymer (SBS), impact modified poly(methyl methacrylate), methyl methacrylate‐acrylonitrile‐butadiene‐styrene polymer (MABS), and cyclic olefin copolymer (COC). Nanofeatures down to 200‐nm line width and with aspect ratios (aspect ratio = depth/width) of 1:1 could be replicated. In all selected materials, the greatest differences between the materials emerged when the aspect ratio increased to 2:1. The most favorable results were obtained with the use of high flow polycarbonate as the molding material. The best replication results were achieved when melt and mold temperatures were higher than normal values.     

Aerobic Oxidation of Benzylic Alcohols in Water by 2,2,6,6‐Tetramethylpiperidine‐1‐oxyl (TEMPO)/Copper(II) 2‐N‐Arylpyrrolecarbaldimino Complexes

Novel copper(II) 2‐N‐arylpyrrolecarbaldimine‐based catalysts for the aerobic oxidation of benzylic alcohols mediated by the 2,2,6,6‐tetramethylpiperidine‐1‐oxyl (TEMPO) radical are reported. The catalytic activity for both synthesized and in situ made complexes in alkaline water solutions was studied revealing high efficiency and selectivity (according to GC selectivity always >99%) for both of these catalytic systems. For example, quantitative conversion of benzyl alcohol to benzaldehyde can be achieved with the in situ prepared bis[2‐N‐(4‐fluorophenyl)‐pyrrolylcarbaldimide]copper(II) catalysts in 2 h with atmospheric pressure of O₂ at 80 °C. Interestingly, these catalysts can utilize dioxygen as well as air or hydrogen peroxide as the end oxidants, producing water as the only by‐product.     

Electrochemical reduction of oxygen on thin-film Pt electrodes in acid solutions

The electrochemical reduction of oxygen on thin-film platinum electrodes in 0.1 M HClO₄ and 0.05 M H₂SO₄ solutions has been investigated using the rotating disk electrode (RDE) method. Thin films of Pt (0.25-20 nm thick) were prepared by vacuum evaporation onto glassy carbon substrate. The surface morphology of Pt films was examined by transmission electron microscopy (TEM). The specific activity of O₂ reduction was higher in HClO₄ and decreased with decreasing film thickness. In H₂SO₄, the specific activity was lower and appeared to be independent of the Pt loading. The values of Tafel slopes close to −120 mV dec⁻¹ in high current density range and −60 mV dec⁻¹ in low current density range were obtained for all electrodes in both solutions, indicating that the mechanism of O₂ reduction is the same for thin-film electrodes as for bulk Pt. The number of electrons transferred per O₂ molecule was close to four for all thin Pt films studied.     

Reconfiguring flexible machine modules of a PCB assembly line

Assembly of electronic components to Printed Circuit Boards (PCB) is a complicated manufacturing process and therefore its control is usually divided into several subtasks which are handled separately. We consider the combined task of determining a machine configuration and line balancing for a single assembly line of interconnected, reconfigurable machine modules and one PCB type in production. The modules can be tailored to the needs of each PCB type by suitable assignments for placement heads, nozzles and feeders. Out of these, the component-to-machine assignment appears to be most difficult and we propose five different solution methods for it; brute force, random, greedy, local search and genetic algorithm. The genetic algorithm outperformed the other methods in practical test.     

Chemoenzymatic Asymmetric Synthesis of Pyridine-Based α-Fluorinated Secondary Alcohols

Fluoro-substituted and heteroaromatic compounds are valuable intermediates for a variety of applications in pharma- and agrochemistry and synthetic chemistry. This study investigates the chemoenzymatic preparation of chiral alcohols bearing a heteroaromatic ring with an increasing degree of fluorination in α-position. Starting from readily available picoline derivatives prochiral α-halogenated acyl moieties were introduced with excellent selectivity and 64–95 % yield. The formed carbonyl group was subsequently reduced to the corresponding alcohols using the alcohol dehydrogenase from Lactobacillus kefir, yielding an enantiomeric excess of 95–>99 % and up to 98 % yield.