Discovery of Affinity-Based Probes for Btk Occupancy Assays

Bruton's tyrosine kinase (Btk) is an attractive target for the treatment of a wide array of B-cell malignancies and autoimmune diseases. Small-molecule covalent irreversible Btk inhibitors targeting Cys481 have been developed for the treatment of such diseases. In clinical trials, probe molecules are required in occupancy studies to measure the level of engagement of the protein by these covalent irreversible inhibitors. The result of this pharmacodynamic (PD) activity provides guidance for appropriate dosage selection to optimize inhibition of the drug target and correlation of target inhibition with disease treatment efficacy. This information is crucial for successful evaluation of drug candidates in clinical trials. Based on the pyridine carboxamide scaffold of a novel solvent-accessible pocket (SAP) series of covalent irreversible Btk inhibitors, we successfully developed a potent and selective affinity-based biotinylated probe 12 (2-[(4-{4-[5-(1-{5-[(3aS,4S,6aR)-2-oxo-hexahydro-1H-thieno[3,4-d]imidazol-4-yl]pentanamido}-3,6,9,12-tetraoxapentadecan-15-amido)pentanoyl]piperazine-1-carbonyl}phenyl)amino]-6-[1-(prop-2-enoyl)piperidin-4-yl]pyridine-3-carboxamide). Compound 12 has been used in Btk occupancy assays for preclinical studies to determine the therapeutic efficacy of Btk inhibition in two mouse lupus models driven by TLR7 activation and type I interferon.     

The third order nonlinear optical properties of gold nanoparticles in glasses, part II

The optical properties of gold nanoparticles have been known for a number of years and recent advances in laser power have now allowed the non-linear optical properties to be studied. In this short review paper, the various theories that have been used to describe the optical properties of gold nanoparticles are presented. Methods of preparing gold nanoparticles in glasses are explained briefly, as well as characterization techniques. The optical properties of gold nanoparticles are reviewed, as well as the effects of particle size, shape, concentration and c.     

Barium aluminate hydrates. IV. α-BaO.Al2O3.4H2O

α-BaO.Al₂O₃.4H₂O has been synthesised and studied by infrared, X-ray and thermal analytical techniques. The compound of approximate formula BaO.Al₂O₃.0.5H₂O, described in Part III, forms as a dehydration product, and appears to be identical with the compound β-BAH₂ described by other workers. A possible explanation for the discrepancy in water content is discussed.     

Quantitative31P nuclear magnetic resonance analysis of the phospholipids of erythrocyte membranes using detergent

³¹P Nuclear magnetic resonance (NMR) spectra of human erythrocyte lysates dissolved in sodium cholate were acquired. The narrow resonances of phospholipids were mostly well resolved, allowing identification and accurate quantitative analysis of phospholipid classes of the erythrocyte membranes. The ether-linked phosphatidylethanolamine components of the erythrocyte membranes were identified, based on the removal of plasmalogens by acidolysis and of diacyl phospholipid species by degradation using phospholipase A₁. It was also shown that the introduction of double bonds on the acyl chains of phosphatidylcholine shifted the³¹P NMR resonances to lower frequencies. Quantitative analyses of phospholipids from the spectra were based on their apparent molar concentrations. The recoveries of phospholipids from erythrocytes were significantly higher than those using conventional extraction procedures.     

Effect of Visual Cues and a Mixture of Isothiocyanates on Trap Capture of Cabbage Seed Weevil, Ceutorhynchus assimilis

The effect of trap design, trap color, and a mixture of isothiocyanates on the capture of Ceutorhynchus assimilis (Paykull) was studied in a series of field experiments. Unbaited yellow water traps, a yellow sticky box trap, and a yellow sticky card trap, mounted vertically, or at 45° to the vertical with the yellow card facing upwards were effective for capturing seed weevils, but a horizontal yellow sticky card trap caught few. White, green, and black traps were unattractive. During migratory periods, trap catch could be enhanced two to four times with a lure consisting of a mixture of allyl, 3-butenyl, 4-pentenyl, and 2-phenylethyl isothiocyanates, but this bait did not attract in a trap with ineffective design. The sticky card trap, mounted at 45° to the vertical and baited with the isothiocyanate mixture, may be useful for monitoring movement of C. assimilis during migratory periods. However, during the colonization phase, lures of either a mixture of isothiocyanates or of allyl isothiocyanate alone were not attractive.     

Supporting the consumption and co-authoring of locative media experiences for a rural village community: design and field trial evaluation of the SHARC2.0 framework

Locative Media Experiences (LMEs) have significant potential in enabling visitors to engage with the places that they visit through an appreciation of local history. For example, a visitor to Berlin that is exploring remnants of the Berlin Wall may be encouraged to appreciate (or in part experience) the falling of the Berlin wall by consuming multimedia directly related to her current location such as listening to audio recordings of the assembled crowds on 10th November 1989. However, despite the growing popularity of enabling technologies (such as GPS-equipped smart phones and tablets), the availability of tools that support the authoring of LMEs is limited. In addition, mobile apps that support the consumption of LMEs typically adopt an approach that precludes users from being able to respond with their own multimedia contributions. In this article we describe the design and evaluation of the SHARC2.0 framework that has been developed as part of our long-term and participatory engagement with the rural village of Wray in the north of England. Wray has very limited cellular data coverage which has placed a requirement on the framework and associated tools to operate without reliance on network connectivity. A field study is presented which featured a LME relating to Wray’s local history and which contained multimedia content contributed by members of the community including historic photos (taken from an existing ‘Digital Noticeboard’ system), audio-clips (from a local historian and village residents) and video (contributed during a design workshop). The novelty of our approach relates to the ability of multiple authors to contribute to a LME in-situ, and the utilisation of personal cloud storage for storing the contents associated with a multi-authored LME.

     

Recombinant antibody fragments that detect enoyl acyl carrier protein reductase in Brassica napus

Purified Brassica napus enoyl acyl carrier protein reductase (ENR) was used to select specific antibodies from a library of antibody fragments, single-chain F_v (scF_v), displayed on filamentous phage. Analysis of the selected clones by BstNl fingerprinting and nucleotide sequencing showed that the scF_v were derived from three different human V_H germline genes. The binding specificities were confirmed by Western blots and ELISA. The scF_v preparations reacted with B. napus ENR, but not with β-keto reductase, nor enoyl reductase from Escherichia coli. Analysis of fragments generated by CNBr treatment indicates that the scF_v 3.13 recognizes an epitope located within the n-terminal 80 amino acids of the enzyme molecule. The scF_v were used to detect ENR directly in extracts of B. napus seeds.     

The roles of embedded monitoring requests and questions in improving mental models of computer-based scientific text

Prior research has shown that people are likely to skim information presented digitally with the resultant deleterious effect on accurate mental models of the text. Teaching monitoring strategies and presenting text with adjunct questions are effective strategies for improving the mental models of readers of scientific text, but the two strategies have not been directly compared for studying text read from a computer screen. In the present study, participants were individually tested in two experiments in which they studied computer-based text on the heart and circulatory system while either reading the text with no interruption, answering adjunct inference questions, answering adjunct factual questions, or answering prompts for global or specific monitoring. Reading time was untimed but looking back was not available. Prompting for global monitoring was most effective for improvement of mental models as measured by scores in diagram drawing, concept map construction, factual memory, and generating inferences. Further, global monitoring did not require more time-on-task than the other methods.     

Electrostatic Control of Structure in Self‐Assembled Membranes

Self‐assembling peptide amphiphiles (PAs) can form hierarchically ordered membranes when brought in contact with aqueous polyelectrolytes of the opposite charge by rapidly creating a diffusion barrier composed of filamentous nanostructures parallel to the plane of the incipient membrane. Following this event, osmotic forces and charge complexation template nanofiber growth perpendicular to the plane of the membrane in a dynamic self‐assembly process. In this work, we show that this hierarchical structure requires massive interfacial aggregation of PA molecules, suggesting the importance of rapid diffusion barrier formation. Strong PA aggregation is induced here through the use of heparin‐binding PAs with heparin and also with polyelectrolytes of varying charge density. Small angle X‐ray scattering shows that in the case of weak PA‐polyelectrolyte interaction, membranes formed display a cubic phase ordering on the nanoscale that likely results from clusters of PA nanostructures surrounded by polyelectrolyte chains.