Antibody engineering reveals the important role of J segments in the production efficiency of llama single-domain antibodies in Saccharomyces cerevisiae

Variable domains of llama heavy-chain antibodies (VHH) are becoming a potent tool for a wide range of biotechnological and medical applications. Because of structural features typical of their single-domain nature, they are relatively easy to produce in lower eukaryotes, but it is not uncommon that some molecules have poor secretion efficiency. We therefore set out to study the production yield of VHH. We computationally identified five key residues that are crucial for folding and secretion, and we validated their importance with systematic site-directed mutations. The observation that all key residues were localised in the V segment, in proximity of the J segment of VHH, led us to study the importance of J segment in secretion efficiency. Intriguingly, we found that the use of specific J segments in VHH could strongly influence the production yield. Sequence analysis and expression experiments strongly suggested that interactions with chaperones, especially with the J segment, are a crucial aspect of the production yield of VHH.     

A Method for the Stress Analysis of Lap Joints

A theory is presented for the adhesive stresses in single and double lap joints under tensile loading, while subjected to thermal stress. The formulation includes the effects of bending, shearing, stretching and hygrothermal deformation in both the adherend and adhesive. All boundary conditions, including shear stress free surfaces, are satisfied. The method is general and therefore applicable to a range of material properties and joint configurations including metal-to-metal, metal-to-CFRP or CFRP-to-CFRP. The solution is numerical and is based on an equilibrium finite element approach. Through the use of an iterative procedure, the solution has been extended to cater for non-linear adhesive materials.     

Nucleation and crystallization of polypropylene by mineral fillers: relationship to impact strength

The mechanical properties of mineral-filled polypropylene (PP) are determined not only by the size, shape and modulus of the filler particles, but also by microstructure. For example, poor impact strength is correlated with a high capacity for nucleation of crystallization. In the present study, optical microscopy has been used to measure the growth rate of spherulites in PP. In tandem with this, isothermal d.s.c. measurements have been made of the crystallization of the PP filled with talc, calcium carbonate and stearate-coated carbonate at different loading levels. Computer simulation of spherulite growth has been used to derive the number of nucleating sites per unit volume of polymer and, using surface area measurements, the number of sites per unit area of mineral surface was obtained. Values for talc were, as expected, considerably higher than those for carbonate (and especially coated carbonate) fillers. The presence of filler affects not only the nucleation and kinetics of the crystallization process but also the crystallinity and orientation indices and the proportion of β-phase crystallites present. The connection between these factors and impact strength is discussed. It is concluded that impact properties are determined by inter alia crack pinning and blocking by filler particles, stress concentrations at the edges of the filler particles, and the nucleating ability of the filler.     

Polyurethane‐infiltrated carbon foams: A coupling of thermal and mechanical properties

The thermal and mechanical properties of polyurethane‐infiltrated carbon foam of various densities were investigated. By combining the high thermal conductivity of the carbon foam with the mechanical toughness of the pure polyurethane, a mechanically tough composite (relative to the unfilled foam) that could be used at higher temperatures than the polyurethane's degradation was formed. Both the tensile strength and the modulus increased by an order of magnitude for the composites compared to unfilled foam, while the compressive and shear strengths and moduli of the composites approached values exhibited by pure polyurethane. At both 300 and 400°C, the rectangular blocks of pure polyurethane lost their mechanical integrity due to decomposition in air. Thermogravimetric analysis confirms substantial initial weight loss above 290°C. Filled carbon foam blocks, however, maintain their mechanical integrity at both 300 and 400°C indefinitely, although the bulk of the rectangular block mass is polyurethane. Three different carbon foam densities are examined. As expected, the higher density foams show greater heat transfer. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 87: 2348–2355, 2003     

Non-equilibrium particle motion in the vicinity of a single blade

The motion of sand particles close to a single moving blade was investigated using Positron Emission Particle Tracking (PEPT) during the period in which the free bed surface profile was evolving to an equilibrium shape. The area affected by the blade was divided into active and inactive regions and these were analysed separately. The characteristic heart-shape of the active region in the plan view was determined. An approximately 10-particle-diameter wide velocity transition zone is found between the two regions. While the tracer particle is in the inactive region moving away from the blade, the time dependence of the axial displacement is well described by a logarithmic relationship. The probability of particle movement towards the centre of the blade was quantified using a “central tendency” index. The calculated central tendency shows maxima at each side of the blade. The separation of the two maxima, which indicates the width of the active region, increases with fill level but is independent of rotational speed.     

Modelling collisions of soft agglomerates at the continuum length scale

Growth and breakdown mechanisms in granulation processes may involve collisions between soft plastically deforming agglomerates. It has been established previously that the flow stress in such collisions increases with the strain rate, which is dependent on the impact velocity and the size of the agglomerates. In the current paper, a scaling model is described that is based on a continuum constitutive relationship and formulated in terms of accessible experimental parameters. It is an extension of an existing contact mechanics model for elastoplastic nonadhesive collisions and therefore limited to deformations in which the contact radius is less than about 40% of the agglomerate radius. In addition, it is assumed that the elastic strains are small compared to the maximum value. Finite element simulations were carried out for a range of impact velocities and material parameters associated with an elastoviscoplastic constitutive relationship of the type used in the scaling model. The results were employed to validate the scaling model. Within the specified limits of applicability, it was found that the coefficient of restitution, contact area, loading and unloading curves and also the time evolution of the compressive displacement could be calculated with relatively high accuracy. Moreover, it was found that for a viscoplastic material the rate of decrease of the coefficient of restitution with increasing impact velocity is greater than for plastic deformation. The model should prove useful in understanding collision processes in granulation systems, particularly those occurring at relatively high impact velocities when the main energy dissipation process arises from viscoplastic deformation. Under this circumstance, the influence of adhesion is negligible and coalescence may be taken to occur when the coefficient of restitution is small.     

Evolution of dithiane-based strategies for the construction of architecturally complex natural products

Umpolung-based strategies play a significant role in organic synthesis. Particularly important are 1,3-dithiane linchpins, which serve as convenient acyl anion equivalents. The general synthetic accessibility and impressive reactivity of 1,3-dithianes have thus led to widespread application. Since the late 1970s, dithianes have featured prominently in our program directed toward the synthesis of complex natural and unnatural products, both for effective union of advanced fragments and for multicomponent linchpin couplings. In this Account, we present the evolution of dithiane chemistry in our laboratory.     

DEM‐CFD modeling of particle systems with long‐range electrostatic interactions

To investigate dynamic behaviors of monocharged particle systems, a direct truncation (DT) method and a hybrid particle‐cell (HPC) method are implemented into the discrete element method coupled with computational fluid dynamics (DEM‐CFD) with defined cutoff distances. The DT method only considers electrostatic interactions between particles within the cutoff distance while the HPC method computes electrostatic interactions in the entire computational domain. The deposition process of monocharged particles in a container in air was simulated using the developed DEM‐CFD. It was found that using the DT method, the macrostructure, evolution of granular temperature, and radial distribution function of the particle system were sensitive to the specified cutoff distance. In contrast, using the HPC method, these results were independent of the specified cutoff distance, as expected. This implies that, although electrostatic interactions between particles with large separation distances are weak, they should be considered in DEM‐CFD for accurate modeling of charged particle systems. © 2015 The Authors AIChE Journal published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers AIChE J, 61: 1792–1803, 2015     

Evaluation of intermodel agreement using ISO 13655 M0, M1, and M2 measurement modes in commercial spectrophotometers

Spectrophotometers are routinely used for color measurement and color management in many commercial printing and proofing workflows. In the case of media containing optical brightening agents, ultraviolet (UV)‐induced fluorescence has led to poor levels of agreement between models from different manufacturers, and different models from the same manufacturer. A relevant standard, ISO 13655, has been revised and now clearly defines measurement modes and conditions for the UV component in spectrophotometers. ISO 13655:2009—Graphic Technology—Spectral Measurement and Colorimetric Computation for Graphic Arts Images now defines four measurement modes: M0, M1, M2, and M3. The intermodel difference between 10 commercially available spectrophotometers is evaluated for different substrate types in measurement modes (M0, M1, and M2) as allowed by each instrument. In particular, the authors compare devices using M0 legacy mode versus newer instruments that are compliant with the new M1 and M2 (UV‐included and UV‐excluded) measurement modes. A finding with significant practical applications is that there is greatly improved intermodel agreement between the new generation of ISO 13655‐compliant instruments in M1 (D50) mode when compared with the previous generation of hand‐held spectrophotometers. © 2016 Wiley Periodicals, Inc. Col Res Appl, 42, 27–37, 2017     

Semicontinuous separation of dimethyl ether (DME) produced from biomass

A ternary semicontinuous system for the separation of bio‐dimethyl ether from methanol and water is presented. The performance of eight potential control configurations, including the application of temperature inferential control, is evaluated. Dynamic simulations of the semicontinuous system and associated control scheme demonstrate that the temperature inferential control configuration is effective in achieving the separation objectives while remaining within operational limits. The semicontinuous system using the inferential temperature control scheme is simulated and shown to be economically preferable to the traditional continuous process for a range of production rates. © 2013 Canadian Society for Chemical Engineering