The use of proton transfer reaction mass spectrometry for high throughput screening of terpene synthases

In this work, we introduce the application of proton transfer reaction mass spectrometry (PTR-MS) for the selection of improved terpene synthase mutants. In comparison with gas chromatography mass spectrometry (GC-MS)-based methods, PTR-MS could offer advantages by reduction of sample preparation steps and analysis time. The method we propose here allows for minimal sample preparation and analysis time and provides a promising platform for the high throughput screening (HTS) of large enzyme mutant libraries. To investigate the feasibility of a PTR-MS-based screening method, we employed a small library of Callitropsis nootkatensis valencene synthase (CnVS) mutants. Bacterial cultures expressing enzyme mutants were subjected to different growth formats, and headspace terpenes concentrations measured by PTR-Qi-ToF-MS were compared with GC-MS, to rank the activity of the enzyme mutants. For all cultivation formats, including 96 deep well plates, PTR-Qi-ToF-MS resulted in the same ranking of the enzyme variants, compared with the canonical format using 100 mL flasks and GC-MS analysis. This study provides a first basis for the application of rapid PTR-Qi-ToF-MS detection, in combination with multi-well formats, in HTS screening methods for the selection of highly productive terpene synthases.     

Numerical Simulation of a Turbulent Flow in a Channel with Surface Mounted Cubes

In this paper we report on a fourth-order, spectro-consistent simulation of a complex turbulent flow. A spatial discretization of a convection-diffusion equation is termed spectro-consistent if the spectral properties of the convective and diffusive operators are preserved, i.e. convection skew-symmetric; diffusion symmetric positive definite. We consider a fully developed flow in a channel, where a matrix of cubes is placed at a wall of the channel. The Reynolds number (based on the channel width and the mean bulk velocity) is equal to Re = 13,000. The three-dimensional flow around the surface mounted cubes has served at a test case at the 6th ERCOFTAC/IAHR/COST workshop on refined flow modeling (Delft, June 1997). Here, mean velocity profiles as well as Reynolds stresses at various locations in the channel have been computed without using any turbulence models. The results agree well with the available experimental data.     

A symmetry-preserving discretisation and regularisation model for compressible flow with application to turbulent channel flow

Most simulation methods for compressible flow attain numerical stability at the cost of swamping the fine turbulent flow structures by artificial dissipation. This article demonstrates that numerical stability can also be attained by preserving conservation laws at the discrete level. A new mathematical explanation of conservation in compressible flow reveals that many conservation properties of convection are due to the skew-symmetry of the convection operator. By preserving this skew-symmetry at the discrete level, a fourth-order accurate collocated symmetry-preserving discretisation with excellent conservation properties is obtained. Also a new symmetry-preserving regularisation subgrid-scale model is proposed. The proposed techniques are assessed in simulations of compressible turbulent channel flow. The symmetry-preserving discretisation for compressible flow has good stability without artificial dissipation and yields acceptable results already on coarse grids. Regularisation does not consistently improve upon no-model results, but often compares favourably with eddy-viscosity models.     

Multiplying after the turn

In education multiplying is usually viewed as repeated joining together and dividing as repeated taking away or, which comes to the same thing, as an equal distribution. This presentation springs from Antiquity, when thought was mostly concrete. In modern mathematics we have relation-numbers instead of, image-numbers and likewise multiplying is a facet of relational thinking. The view that children merely can learn through the concrete is often biassedly understood in the sense that the concrete has to be abstracted, which characterizes substantial thinking. However, in the case of relational thinking, learning through the concrete means that to achieve insight the mathematical activities have to be applied to reality, a crucial point, for most people have difficulties with applying multiplication, much more than with, the inherent algorithms. It appears that they do not really know what multiplication is, particularly not its space structure. The more general the structure the more and wider the applications. This thesis infers that multiplying as multiple of classes is much less useful than multiplying as space form. Questing for the essence of multiplication is the major topic of this paper. Which changes has its structure undergone and how can education deal with them? At the end it is illustratively explained why probability, based on the established multiplication, is usually such a tough domain.     

Sulfur substituted Small-ring Heterocycles

The synthesis of epoxy sulfines and sulfonyl azirines is described