Binding of external ligands onto an engineered virus capsid

The development of novel delivery systems for therapeutic substancesincludes targeting of the carriers to a specific site or tissuewithin the body of the recipient. This can be accomplished byappropriate receptor-binding domains and requires linking ofthese domains to the carrier. We have used recombinantly expressedpolyomavirus-like particles as a model system and inserted thesequence of a WW domain into different surface loops of theviral capsid protein VP1. In one variant, the WW domain maintainedits highly selective binding properties of proline-rich ligandsand showed an increased affinity but also an accelerated association/dissociationequilibrium compared to the isolated WW domain, thus allowinga short-term coupling of external ligands onto the surface ofthe virus-like particles.     

Polyionic fusion peptides function as specific dimerization motifs

The de novo design of a molecular adapter for directed associationand covalent linkage of two polypeptides is presented. Usingpeptides containing charged amino acid residues and an additionalcysteine residue (AlaCysLys₈ and AlaCysGlu₈) we demonstratethat the electrostatic interaction promotes the associationof two synthetic peptides and, subsequently, disulfide bondformation. The reaction depends on both the redox potentialand on the ionic strength of the buffer. Varying the redox potential,the interaction of the peptides was quantified by a G^0' of 6.6± 0.2 kcal/mol. Heterodimerization of the peptides ishighly specific, a competition of association by other cysteinecontaining compounds could not be observed. Two proteins comprisingcysteine-containing polyionic fusion peptides, a modified Fabfragment and an -glucosidase fusion, could be specifically conjugatedby directed association and subsequent disulfide bond formation.Both proteins retain their functional characteristics withinthe bifunctional conjugate: enzymatic activity of the glucosidaseand antigen-binding capacity of the Fab fragment are equivalentto the non-conjugated components.     

Synthesis of Hard Materials by Field Activation: The Synthesis of Solid Solutions and Composites in the TiB2–WB2–CrB2 System

The synthesis of solid solutions of (Ti,W,Cr)B₂ from elemental reactants using the field-activated, pressure-assisted synthesis method and employing the SPS apparatus was investigated. The nature of the products depended on temperature; they were nearly pure solid solutions at 1900°C with minor amounts of β-WB. The product density and microhardness depended on the temperature of synthesis for the same value of applied pressure (64 MPa). Samples with the highest density (94%) corresponded to a hardness of 22.7 GPa. When annealed at 1500°C, the solid solutions decomposed, precipitating a (W,Ti,Cr)B₂ phase in a spinodal form. In addition, β-WB precipitates in the form of thin (0.4–5.3 nm) layers were observed. They existed in a 60°/120° orientation to the (Ti,W,Cr)B₂ matrix, in agreement with previous observations. Highly faceted, small (nanosized) pores associated with the β-WB precipitates were also observed.     

Frequency and phase estimation in time series with quasi periodic components

In this article, we consider frequency and phase estimation in a noisy oscillation with potentially non‐constant phase increments resulting from an underlying non‐constant frequency. A maximum periodogram method on segments is used to estimate the time‐varying frequency and a subsequent least squares approach to estimate the phase. A key problem addressed in this article is the question how to set up a meaningful concept of asymptotic statistics for this model. This problem is solved by a special infill asymptotics concept. We use this concept to prove consistency and asymptotic normality of the estimates. Furthermore, the phase estimate is compared to the Hilbert transform in a simulation.     

Artificial Light Regulation of an Allosteric Bienzyme Complex by a Photosensitive Ligand

The artificial regulation of proteins by light is an emerging subdiscipline of synthetic biology. Here, we used this concept to photocontrol both catalysis and allostery within the heterodimeric enzyme complex imidazole glycerol phosphate synthase (ImGP‐S). ImGP‐S consists of the cyclase subunit HisF and the glutaminase subunit HisH, which is allosterically stimulated by substrate binding to HisF. We show that a light‐sensitive diarylethene (1,2‐dithienylethene, DTE)‐based competitive inhibitor in its ring‐open state binds with low micromolar affinity to the cyclase subunit and displaces its substrate from the active site. As a consequence, catalysis by HisF and allosteric stimulation of HisH are impaired. Following UV‐light irradiation, the DTE ligand adopts its ring‐closed state and loses affinity for HisF, restoring activity and allostery. Our approach allows for the switching of ImGP‐S activity and allostery during catalysis and appears to be generally applicable for the light regulation of other multienzyme complexes.     

NONLINEAR PI CONTROLLER FOR pH PROCESS

The control of pH is one of the most difficult challenges in the process industry because of the severe nonlinearities and high precision required in manipulating the flow rate. The Wiener model, which consists of a linear dynamic element followed by a nonlinear static element, is used for representing such nonlinear processes. Piecewise continuous polynomials are used for mapping the nonlinear static gain accurately. A nonlinear PI controller was designed based on the Wiener model. Simulation results on the nonlinear mathematical model are presented to highlight the superior performance of the Wiener model based nonlinear PI controller in comparison to that of the local linear PI controller. The performance of the nonlinear PI controller was further improved upon by using the method of inequalities to obtain a single set of PI controller settings that takes into account the parametric variations in the linear dynamic element at different operating points. Simulation and experimental results are presented to support the work.     

Controlled reversible debundling of single-walled carbon nanotubes by photo-switchable dendritic surfactants

Stimulus responsive surfactants based on dendritic glycerol azobenzene conjugates were used to solubilize and debundle single-walled carbon nanotubes in aqueous media. Their debundling property as well as their reaggregation behavior upon irradiation with light was examined and light triggered reversible bundling and precipitation are shown.     

Evaluation of polycaprolactone − poly‐D,L‐lactide copolymer as biomaterial for breast tissue engineering

The potential of the copolymer polycaprolactone‐co‐ poly‐d ,l ‐lactic acid (PCLLA ) as a biomaterial for scaffold‐based therapy for breast tissue engineering applications was assessed. First, the synthesized PCLLA was evaluated for its processability by means of additive manufacturing (AM ). We found that the synthesized PCLLA could be fabricated into scaffolds with an overall gross morphology and porosity similar to that of polycaprolactone. The PCLLA scaffolds possessed a compressive Young's modulus (ca 46 kPa ) similar to that of native breast (0.5 ? 25 kPa ), but lacked thermal stability and underwent thermal degradation during the fabrication process. The PCLLA scaffolds underwent rapid degradation in vitro which was characterized by loss of the scaffolds' mechanical integrity and a drastic decrease in mass‐average molar mass (M _w) and number‐average molar mass (M _n) after 4 weeks of immersion in phosphate buffer solution maintained at 37 °C. The tin‐catalysed PCLLA scaffold was also found to have cytotoxic effects on cells. Although the initial mechanical properties of the PCLLA scaffolds generally showed potential for applications in breast tissue regeneration, the thermal stability of the copolymer for AM processes, biocompatibility towards cells and degradation rate is not satisfactory at this stage. Therefore, we conclude that research efforts should be geared towards fine‐tuning the copolymer synthesizing methods. © 2016 Society of Chemical Industry