[show abstract][hide abstract] ABSTRACT: Plant cell walls are structurally diverse macromolecular composites. One of our best methodologies to determine the temporal and spatial regulation of cell wall polysaccharides in relation to development are monoclonal antibody (MAB) and carbohydrate-binding module (CBM) probes and their detection by immunofluorescence microscopy. Here we describe resin embedding, sectioning, and in situ chemical and enzymatic cell wall disassembly and their use with immunocytochemical analyses as a means to unravel the complexity of cell wall molecular architecture in hard tissues and seeds.
Methods in molecular biology (Clifton, N.J.) 01/2014; 1080:41-52.
[show abstract][hide abstract] ABSTRACT: Surface plasmon resonance (SPR) spectroscopy is a rapidly developing technique for the study of ligand binding interactions with membrane proteins, which are the major molecular targets for validated drugs and for current and foreseeable drug discovery. SPR is label-free and capable of measuring real-time quantitative binding affinities and kinetics for membrane proteins interacting with ligand molecules using relatively small quantities of materials and has potential to be medium-throughput. The conventional SPR technique requires one binding component to be immobilised on a sensor chip whilst the other binding component in solution is flowed over the sensor surface; a binding interaction is detected using an optical method that measures small changes in refractive index at the sensor surface. This review first describes the basic SPR experiment and the challenges that have to be considered for performing SPR experiments that measure membrane protein-ligand binding interactions, most importantly having the membrane protein in a lipid or detergent environment that retains its native structure and activity. It then describes a wide-range of membrane protein systems for which ligand binding interactions have been characterised using SPR, including the major drug targets G protein-coupled receptors, and how challenges have been overcome for achieving this. Finally it describes some recent advances in SPR-based technology and future potential of the technique to screen ligand binding in the discovery of drugs.
Biochimica et Biophysica Acta (BBA) - Biomembranes 01/2014; 1838(1):43-55.
[show abstract][hide abstract] ABSTRACT: The mean average life is increasing; therefore, there is a need to increase the lifetime of the prostheses. To fulfil this requirement, new prosthetic designs and materials are being introduced. Two of the design parameters that may affect wear of total knee replacements, and hence the expected lifetime, are the insert conformity and material. Computational models have been used extensively for wear prediction and optimisation of artificial knee designs. The objective of the present study was to use a previously validated non-dimensional wear coefficient-based computational wear model to investigate the effect of insert conformity and material on the predicted wear in total knee replacements. Four different inserts (curved, lipped, partial flat and custom flat), with different conformity levels, were tested against the same femoral and under two different kinematic inputs (intermediate and high), with different levels of cross-shear. The insert bearing materials were either conventional or moderately cross-linked ultra-high molecular weight polyethylene (UHMWPE). Wear predictions were validated against the experimental data from Leeds knee simulation tests. The predicted wear rates for the curved insert (most conformed) were more than three times those for the flat insert (least conformed). In addition, the computationally predicted average volumetric wear rates for moderately cross-linked UHMWPE bearings were less than half of their corresponding conventional UHMWPE bearings. Moreover, the wear of the moderately cross-linked UHMWPE was shown to be less dependent on the degree of cross-shear, compared to conventional UHMWPE. These results along with supporting experimental studies provide insight into the design variables, which may reduce wear in knee replacements.
Proceedings of the Institution of Mechanical Engineers. Part H, Journal of engineering in medicine. 12/2013;
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