The combi-CLEA approach: enzymatic cascade synthesis of enantiomerically pure (S)-mandelic acid

Enantiomerically pure (S)-mandelic acid was synthesised from benzaldehyde by sequential hydrocyanation and hydrolysis in a bienzymatic cascade at starting concentrations up to 0.25 M. A cross-linked enzyme aggregate (CLEA) composed of the (S)-selective oxynitrilase from Manihot esculenta and the non-selective nitrilase from Pseudomonas fluorescens EBC 191 was employed as the biocatalyst. The nitrilase produces approx. equal amounts of (S)-mandelic acid and (S)-mandelic amide from (S)-mandelonitrile under standard conditions, but we surprisingly found that high (up to 0.5 M) concentrations of HCN induced a marked drift towards amide production. By including the amidase from Rhodococcus erythopolis in the CLEA we obtained (S)-mandelic acid as the sole product in 90% yield and >99% enantiomeric purity.     

Unsupervised cell identification on multidimensional X‐ray fluorescence datasets

A novel approach to locate, identify and refine positions and whole areas of cell structures based on elemental contents measured by X‐ray fluorescence microscopy is introduced. It is shown that, by initializing with only a handful of prototypical cell regions, this approach can obtain consistent identification of whole cells, even when cells are overlapping, without training by explicit annotation. It is robust both to different measurements on the same sample and to different initializations. This effort provides a versatile framework to identify targeted cellular structures from datasets too complex for manual analysis, like most X‐ray fluorescence microscopy data. Possible future extensions are also discussed.     

A new profile likelihood confidence interval for the mean probability of detection in collaborative studies of binary test methods

A confidence interval for the probability of detection across laboratories (LPOD) for qualitative methods, described in the AOAC INTERNATIONAL guidelines for validation of microbiological methods for food and environmental surfaces, is considered. It is demonstrated that under certain conditions, the observed confidence of this confidence interval can be rather low, so that statistical minimum requirements are not fulfilled. A new profile likelihood confidence interval based on a latent random laboratory effect approach is proposed. Observed confidence levels for this confidence interval demonstrate its applicability already for a small number of laboratories.     

A fixed‐grid b‐spline finite element technique for fluid–structure interaction

We present a fixed‐grid finite element technique for fluid–structure interaction problems involving incompressible viscous flows and thin structures. The flow equations are discretised with isoparametric b‐spline basis functions defined on a logically Cartesian grid. In addition, the previously proposed subdivision‐stabilisation technique is used to ensure inf–sup stability. The beam equations are discretised with b‐splines and the shell equations with subdivision basis functions, both leading to a rotation‐free formulation. The interface conditions between the fluid and the structure are enforced with the Nitsche technique. The resulting coupled system of equations is solved with a Dirichlet–Robin partitioning scheme, and the fluid equations are solved with a pressure–correction method. Auxiliary techniques employed for improving numerical robustness include the level‐set based implicit representation of the structure interface on the fluid grid, a cut‐cell integration algorithm based on marching tetrahedra and the conservative data transfer between the fluid and structure discretisations. A number of verification and validation examples, primarily motivated by animal locomotion in air or water, demonstrate the robustness and efficiency of our approach. Copyright © 2013 John Wiley & Sons, Ltd.