A Click-Chemistry-Based Enrichable Crosslinker for Structural and Protein Interaction Analysis by Mass Spectrometry

Mass spectrometry is the method of choice for the characterisation of proteomes. Most proteins operate in protein complexes, in which their close association modulates their function. However, with standard MS analysis, information on protein–protein interactions is lost and no structural information is retained. To gain structural and interactome data, new crosslinking reagents are needed that freeze inter- and intramolecular interactions. Herein, the development of a new reagent, which has several features that enable highly sensitive crosslinking MS, is reported. The reagent enables enrichment of crosslinked peptides from the majority of background peptides to facilitate efficient detection of low-abundant crosslinked peptides. Due to the special cleavable properties, the reagent can be used for MS² and potentially for MS³ experiments. Thus, the new crosslinking reagent, in combination with high-end MS, should enable sensitive analysis of interactomes, which will help researchers to obtain important insights into cellular states in health and diseases.     

Nitroreductase-Mediated Release of Inhibitors of Lysine-Specific Demethylase 1 (LSD1) from Prodrugs in Transfected Acute Myeloid Leukaemia Cells

Lysine-specific demethylase 1 (LSD1) has evolved as a promising therapeutic target for cancer treatment, especially in acute myeloid leukaemia (AML). To approach the challenge of site-specific LSD1 inhibition, we developed an enzyme-prodrug system with the bacterial nitroreductase NfsB (NTR) that was expressed in the virally transfected AML cell line THP1-NTR⁺. The cellular activity of the NTR was proven with a new luminescent NTR probe. We synthesised a diverse set of nitroaromatic prodrugs that by design do not affect LSD1 and are reduced by the NTR to release an active LSD1 inhibitor. The emerging side products were differentially analysed using negative controls, thereby revealing cytotoxic effects. The 2-nitroimidazolyl prodrug of a potent LSD1 inhibitor emerged as one of the best prodrug candidates with a pronounced selectivity window between wild-type and transfected THP1 cells. Our prodrugs are selectively activated and release the LSD1 inhibitor locally, proving their suitability for future targeting approaches.     

Extruded meat alternatives made from Maillard-reacted beef bone hydrolysate and plant proteins: part I – Effect of moisture content

This study investigated the effects of moisture content (MC) on the physicochemical properties of extruded meat alternatives made from Maillard-reacted beef bone hydrolysate and plant proteins. Samples were extruded at 170 °C (maximum barrel temperature), at 3.6 kg h⁻¹ (liquid feed rate) and at 1.8, 2.2, 2.6 and 3.0 kg h⁻¹ (dry feed rates) to obtain MC of 60%MC, 56%MC, 52%MC and 49%MC, respectively. Meat alternatives at 52%MC showed the greatest degree of texturisation. However, meat alternatives at 49%MC were the closest in terms of both textural and microstructural properties to reference sample, boiled chicken breast. Results from protein solubility suggested that a large amount of aggregated proteins were associated with hydrogen bonds, while disulphide bonds were the main contributor in the formation of fibrous structure in meat alternatives. Results showed that the change in MC as process parameter played an important role in the formation of fibrous structure in extruded meat alternatives.     

A trout (Oncorhynchus mykiss) perfusion model approach to elucidate the role of blood removal for lipid oxidation and colour changes in ice-stored fish muscle

Whole body saline-perfused rainbow trout (Oncorhynchus mykiss) was ice-stored for 4 weeks and compared with unwashed/washed minces from unbled and bled trout in terms of rancid odour, peroxide value (PV), thiobarbituric acid reactive substances (TBARS) and redness loss. Muscle from saline-perfused fish, which had 72% less total haem, was deficient in rancid odour during the whole storage, while bled (54% less haem) and unbled samples developed rancid odour already after ~4 and 2 days; higher intensity without bleeding. PV/TBARS also developed in the order unbled > bled > perfused samples; however, PV/TBARS were not as completely prevented as rancid odour after perfusion. Saline washing (3 × 3 volumes) of unbled mince removed 84% haem and yielded the second most stable sample while saline washing (1 × 1 volumes) destabilised unbled mince, despite 64% haem removal. Concurrent antioxidant removal during washing of minces obviously counteracted the effect of blood removal and washing fish mince with small volumes of solution should be used with great care.     

Performance evaluation of a C-RAN supporting a mixture of random and quasi-random traffic

Wireless Networks - In this paper we consider a cloud radio access network (C-RAN) where the remote radio heads (RRHs) are separated from the baseband signal processing servers, named baseband...     

Stretchable Composite Acoustic Transducer for Wearable Monitoring of Vital Signs

A highly flexible, stretchable, and mechanically robust low‐cost soft composite consisting of silicone polymers and water (or hydrogels) is reported. When combined with conventional acoustic transducers, the materials reported enable high performance real‐time monitoring of heart and respiratory patterns over layers of clothing (or furry skin of animals) without the need for direct contact with the skin. The approach enables an entirely new method of fabrication that involves encapsulation of water and hydrogels with silicones and exploits the ability of sound waves to travel through the body. The system proposed outperforms commercial, metal‐based stethoscopes for the auscultation of the heart when worn over clothing and is less susceptible to motion artefacts. The system both with human and furry animal subjects (i.e., dogs), primarily focusing on monitoring the heart, is tested; however, initial results on monitoring breathing are also presented. This work is especially important because it is the first demonstration of a stretchable sensor that is suitable for use with furry animals and does not require shaving of the animal for data acquisition.     

Notes for a Substantive Theory of Rural Planning: Evidence from the US Experience

Rural areas are the locus of many pressing planning issues, pointing to the need for and possibility of a substantive theory of rural planning. In this article we develop a framework for such a theory. We begin by outlining the emergence of planning and the marginalization of ‘the rural,’ by investigating aspects of existing planning theory. Then we explain the framework and apply it to a small set of critical cases in deep rural areas of the U.S. To conclude, we summarize our findings and propose future directions for theory in rural planning.     

Stratification Dynamics and Geothermal Potential of a Deep Shaft in the Flooded Wolf Mine,Siegerland/Germany

Mine Water and the Environment - Mine water hydraulics and geothermal potential of a deep shaft of the flooded Wolf mine in the Siegerland ore district of the Rhenish Massif in Germany were...