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Large-scale microbial industrial fermentations have significantly higher absolute pressure and dissolved CO2 concentrations than otherwise comparable laboratory-scale processes. Yet the effect of increased dissolved CO2 (dCO2) levels is rarely addressed in the literature. In the current work, we have investigated the impact of industrial levels of dCO2 (measured as the partial pressure of CO2, pCO2) in an Escherichia coli-based fed-batch process producing the human milk oligosaccharide 2′-fucosyllactose (2′-FL). The study evaluated the effect of high pCO2 levels in both carbon-limited (C-limited) and carbon/nitrogen-limited (C/N-limited) fed-batch processes. High-cell density cultures were sparged with 10%, 15%, 20%, or 30% CO2 in the inlet air to cover and exceed the levels observed in the industrial scale process. While the 10% enrichment was estimated to achieve similar or higher pCO2 levels as the large-scale fermentation it did not impact the performance of the process. The product and biomass yields started being affected above 15% CO2 enrichment, while 30% impaired the cultures completely. Quantitative proteomics analysis of the C-limited process showed that 15% CO2 enrichment affected the culture on the protein level, but to a much smaller degree than expected. A more significant impact was seen in the dual C/N limited process, which likely stemmed from the effect pCO2 had on nitrogen availability. The results demonstrated that microbial cultures can be seriously affected by elevated CO2 levels, albeit at higher levels than expected.

RNA-protein interactions are often mediated by dedicated canonical RNA binding domains. However, interactions through non-canonical domains with unknown specificity are increasingly observed, raising the question how RNA targets are recognized. Knowledge of the intrinsic RNA binding specificity contributes to the understanding of target selectivity and function of an individual protein.The presented in vitro RNA immunoprecipitation assay (vitRIP) uncovers intrinsic RNA binding specificities of isolated proteins using the total cellular RNA pool as a library. Total RNA extracted from cells or tissues is incubated with purified recombinant proteins, RNA-protein complexes are immunoprecipitated and bound transcripts are identified by deep sequencing or quantitative RT-PCR. Enriched RNA classes and the nucleotide frequency in these RNAs inform on the intrinsic specificity of the recombinant protein. The simple and versatile protocol can be adapted to other RNA binding proteins and total RNA libraries from any cell type or tissue.Graphic abstract:Figure 1. Schematic of the in vitro RNA immunoprecipitation (vitRIP) protocol

Two new modifications of the pentafluoridoaluminate K2AlF5 were obtained from ammonothermal synthesis at 753 K, 224 MPa and 773 K, 220 MPa, respectively. Both crystallize in the orthorhombic space group type Pbcn, with close metric relations and feature kinked chains of cis-vertex-connected AlF6 octahedra resulting in the Niggli formula ∞1{[AlF2/2eF4/1t]2−}. The differences lie in the number of octahedra necessary for repetition within the chains, which for K2AlF5-2 is realized after four and for K2AlF5-3 after eight octahedra. As a result, the orthorhombic unit cell for K2AlF5-3 is doubled in chain prolongation direction [001] as compared to K2AlF5-2 (1971.18(4) pm versus 988.45(3) pm, respectively), while the unit cell parameters within the other two directions are virtually identical. Moreover, the new elpasolite Rb2KAlF6 is reported, crystallizing in the cubic space group Fm3¯m with a = 868.9(1) pm and obtained under ammonothermal conditions at 723 K and 152 MPa.

The ammonothermal synthesis of three ammoniates of indium, namely InAlF6(NH3)2, [In(NH3)6][AlF6], and [In2F(NH3)10]2[SiF6]5 ∙ 2 NH3 was successful from near-ammononeutral conditions in the presence of fluoride ions. Initially, all these compounds were obtained upon corrosion of the applied liner and crucible material Si3N4, which also contains small amounts of aluminum. The syntheses were performed in supercritical ammonia (T = 753 K, p up to 307 MPa). The crystal structures were solved and refined from single crystal X-ray diffraction intensity data. InAlF6(NH3)2 crystallizes as a typical layer-type structure with corner-sharing [InF4(NH3)2]– and [AlF6]3− octahedra. [In(NH3)6][AlF6] features isolated [In(NH3)6]3+ and [AlF6]3− octahedra. The crystal structure of [In2F(NH3)10]2[SiF6]5 ∙ 2 NH3 contains [(NH3)5In–F–In(NH3)5]5+ octahedra doubles next to [SiF6]2− octahedra and ammonia molecules. All intermediates have strong hydrogen bonding systems. The results from vibrational spectroscopy are reported.

Earth observation imagery have traditionally been expensive, difficult to find and access, and required specialized skills and software to transform imagery into actionable information. This has limited adoption by the broader science community. Changes in cost of imagery and changes in computing technology over the last decade have enabled a new approach for how to organize, analyze, and share Earth observation imagery, broadly referred to as a data cube. The vision and promise of image data cubes is to lower these hurdles and expand the user community by making analysis ready data readily accessible and providing modern approaches to more easily analyze and visualize the data, empowering a larger community of users to improve their knowledge of place and make better informed decisions. Image data cubes are large collections of temporal, multivariate datasets typically consisting of analysis ready multispectral Earth observation data. Several flavors and variations of data cubes have emerged. To simplify access for end users we developed a flexible approach supporting multiple data cube styles, referencing images in their existing structure and storage location, enabling fast access, visualization, and analysis from a wide variety of web and desktop applications. We provide here an overview of that approach and three case studies.

Scaling limits of continuous-time random walks are used in physics to model anomalous diffusion in which particles spread at a different rate than the classical Brownian motion. In this paper, we characterize the scaling limit of the average of multiple particles, independently moving as a continuous-time random walk. The limit is taken by increasing the number of particles and scaling from microscopic to macroscopic view. We show that the limit is independent of the order of these limiting procedures and can also be taken simultaneously in both procedures. Whereas the scaling limit of a single-particle movement has quite an obscure behavior, the multiple-particle analogue has much nicer properties.

Rezensiertes Werk:Luca Mannori (Hg.), Kataster und moderner Staat in Italien, Spanien und Frankreich (18. Jh.) / Cadastre et Etat moderne en Italie, Espagne et France (18e s.) (Jahrbuch für europäische Verwaltungsgeschichte 13), Baden-Baden: Nomos 2001, VII, 368 S., ISBN 3-7890-7557-4