Thermodynamics of binding of regulatory ligands to tissue transglutaminase

The transamidating activity of tissue transglutaminase is regulated by the ligands calcium and GTP, via conformational changes which facilitate or interfere with interaction with the peptidyl-glutamine substrate. We have analysed binding of these ligands by calorimetric and computational approaches. In the case of GTP we have detected a single high affinity site (K (D) approximately 1 muM), with moderate thermal effects suggestive that binding GTP involves replacement of GDP, normally bound to the protein. On line with this possibility no significant binding was observed during titration with GDP and computational studies support this view. Titration with calcium at a high cation molar excess yielded a complex binding isotherm with a number of "apparent binding sites" in large excess over those detectable by equilibrium dialysis (6 sites). This binding pattern is ascribed to occurrence of additional thermal contributions, beyond those of binding, due to the occurrence of conformational changes and to catalysis itself (with protein self-crosslinking). In contrast only one site for binding calcium with high affinity (K (D) approximately 0.15 muM) is observed with samples of enzyme inactivated by alkylation at the active site (to prevent enzyme crosslinkage and thermal effects of catalysis). These results indicate an intrinsic ability of tissue transglutaminase to bind calcium with high affinity and the necessity of careful reassessment of the enzyme regulatory pattern in relation to the concentrations of ligands in living cells, taking also in account effects of ligands on protein subcellular compartimentation.

The behaviour of polymer quenchants

The internationally accepted Wolfson Heat Treatment Centre Engineering Group test was used to evaluate the cooling characteristics of the most popular commercial polymer quenchants: polyalkylene glycols, polyvinylpyrrolidones and polyacrylates. Prototype solutions containing poly(ethyloxazoline) were also examined. Each class of polymer was capable of providing a wide range of cooling rates depending on the product formulation, concentration, temperature, agitation, ageing and contamination. Cooling rates for synthetic quenchants were generally intermediate between those of water and oil. Control techniques, drag-out losses and response to quenching in terms of hardness and residual stress for a plain carbon steel, were also considered. A laboratory scale method for providing a controllable level of forced convection was developed. Test reproducibility was improved by positioning the preheated Wolfson probe 25mm above the geometric centre of a 25mm diameter orifice through which the quenchant was pumped at a velocity of 0.5m/s. On examination, all polymer quenchants were found to operate by the same fundamental mechanism associated with their viscosity and ability to form an insulating polymer-rich-film. The nature of this film, which formed at the vapour/liquid interface during boiling, was dependent on the polymer's solubility characteristics. High molecular weight polymers and high concentration solutions produced thicker, more stable insulating films. Agitation produced thinner more uniform films. Higher molecular weight polymers were more susceptible to degradation, and increased cooling rates, with usage. Polyvinylpyrrolidones can be cross-linked resulting in erratic performance, whilst the anionic character of polyacrylates can lead to control problems. Volatile contaminants tend to decrease the rate of cooling and salts to increase it. Drag-out increases upon raising the molecular weight of the polymer and its solution viscosity. Kinematic viscosity measurements are more effective than refractometer readings for concentration control, although a quench test is the most satisfactory process control method.

Properties and performance of polymer modified cements and mortars

Polymer modified cements and mortars have become popular for use as patch repair materials. General evidence suggests that these materials offer considerable improvements compared to traditional mortars although the mechanisms for this are not fully understood. This work elucidates the factors which govern some properties and performance of different polymer systems. In view of the wide range of commercial systems available, investigations concentrated on the use of three of the most commonly available groups of polymers. These were: (1) Styrene Butadiene Rubber (SBR), (2) Acrylics and, (3) Ethylene Vinyl Acetates (EVA). The later two were in the form of both emulsions and redispersible powders. Experiments concentrated on: (1) Rheological behaviour of polymer modified cement pastes; (2) Workability of polymer modified mortars; (3) Influence of curing conditions on the pore size distribution and diffusion of chloride ions; (4) Bond strength of polymer modified cement and mortar patches; and (5) Microscopic examination and semi-quantitative analyses of the bulk and interfacial microstructures. The following main conclusions were reached: (1) The addition of polymer emulsions have a considerable influence on the workability of fresh cement pastes, the extent of this depending on the type of system used. (2) The rheological parameters of fresh polymer modified mortars can be established using a two-point workability test which may be used when comparing the properties of different systems at constant workability. (3) Curing conditions affect the properties of polymer modified systems and a wet/dry curing regime was essential for good adhesion of these materials to mortar substrates. (4) In contrast, the wet/dry curing regime resulted in a curing affected zone at the surface of patch materials. This can result in a much coarser pore structure and enhanced diffusion of e.g. chloride ions. (5) The microstructure of polymer modified systems was very different compared with the unmodified cement/mortar and varied depending on curing conditions.

In situ studies of phase separation and crystallization directed by Marangoni instabilities during spin-coating

Results of a pioneering study are presented in which for the first time, crystallization, phase separation and Marangoni instabilities occurring during the spin-coating of polymer blends are directly visualized, in real-space and real-time. The results provide exciting new insights into the process of self-assembly, taking place during spin-coating, paving the way for the rational design of processing conditions, to allow desired morphologies to be obtained. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

An investigation into the wavelength stability of polymer optical fibre Bragg gratings

The inscription of Bragg gratings has been demonstrated in PMMA-based polymer optical fibre. The water affinity of PMMA can introduce significant wavelength change in a polymer optical fibre Bragg grating (POFBG). In polymer optical fibre losses are much higher than with silica fibre. Very strong absorption bands related to higher harmonics of vibrations of the C-H bond dominate throughout the visible and near infrared. Molecular vibration in substances generates heat, which is referred to as the thermal effect of molecular vibration. This means that a large part of the absorption of optical energy in those spectral bands will convert into thermal energy, which eventually drives water content out of the polymer fibre and reduces the wavelength of POFBG. In this work we have investigated the wavelength stability of POFBGs in different circumstances. The experiment has shown that the characteristic wavelength of a POFBG starts decreasing after a light source is applied to it. This decrease continues until equilibrium inside the fibre is established, depending on the initial water content inside the fibre, the surrounding humidity, the optical power applied, and the fibre size. Our investigation has shown that POFBGs operating at around 850 nm show much smaller wavelength reduction than those operating at around 1550 nm in the same fibre; POFBGs with different diameters show different changes; POFBGs powered by a low level light source, or operating in a very dry environment are least affected by this thermal effect.

Investigation into time response of polymer fiber Bragg grating based humidity sensors

In this work we experimentally investigate the response time of humidity sensors based on polymer optical fiber Bragg gratings. By the use of etching with acetone we can control the poly (methyl methacrylate) based fiber in order to reduce the diffusion time of water into the polymer and hence speed up the relative wavelength change caused by humidity variations. A much improved response time of 12 minutes for humidity decrease and 7 minutes for humidity increase, has been achieved by using a polymer optical fiber Bragg grating with a reduced diameter of 135 microns.

Influence of mounting on the hysteresis of polymer fiber Bragg grating strain sensors

Fiber Bragg grating sensors recorded in poly(methyl methacrylate) fiber often exhibit hysteresis in the response of Bragg wavelength to strain, particularly when exposed to high levels of strain. We show that, when such a fiber grating sensor is bonded directly to a substrate, the hysteresis is reduced by more than 12 times, compared to the case where the sensor is suspended freely between two supports. © 2013 Optical Society of America.

Enhancing the humidity response time of polymer optical fiber Bragg grating by using laser micromachining

The humidity sensors constructed from polymer optical fiber Bragg gratings (POFBG) respond to the water content change in the fiber induced by varying environmental condition. The water content change is a diffusion process. Therefore the response time of the POFBG sensor strongly depends on the geometry and size of the fiber. In this work we investigate the use of laser micromachining of D-shaped and slotted structures to improve the response time of polymer fiber grating based humidity sensors. A significant improvement in the response time has been achieved in laser micromachined D-shaped POFBG humidity sensors. The slotted geometry allows water rapid access to the core region but this does not of itself improve response time due to the slow expansion of the bulk of the cladding. We show that by straining the slotted sensor, the expansion component can be removed resulting in the response time being determined only by the more rapid, water induced change in core refractive index. In this way the response time is reduced by a factor of 2.5.

Combustion of fuel blends containing digestate pyrolysis oil in a multi-cylinder compression ignition engine

Digestate from the anaerobic digestion conversion process is widely used as a farm land fertiliser. This study proposes an alternative use as a source of energy. Dried digestate was pyrolysed and the resulting oil was blended with waste cooking oil and butanol (10, 20 and 30 vol.%). The physical and chemical properties of the pyrolysis oil blends were measured and compared with pure fossil diesel and waste cooking oil. The blends were tested in a multi-cylinder indirect injection compression ignition engine.Engine combustion, exhaust gas emissions and performance parameters were measured and compared with pure fossil diesel operation. The ASTM copper corrosion values for 20% and 30% pyrolysis blends were 2c, compared to 1b for fossil diesel. The kinematic viscosities of the blends at 40 C were 5–7 times higher than that of fossil diesel. Digested pyrolysis oil blends produced lower in-cylinder peak pressures than fossil diesel and waste cooking oil operation. The maximum heat release rates of the blends were approximately 8% higher than with fossil diesel. The ignition delay periods of the blends were higher; pyrolysis oil blends started to combust late and once combustion started burnt quicker than fossil diesel. The total burning duration of the 20% and 30% blends were decreased by 12% and 3% compared to fossil diesel. At full engine load, the brake thermal efficiencies of the blends were decreased by about 3–7% when compared to fossil diesel. The pyrolysis blends gave lower smoke levels; at full engine load, smoke level of the 20% blend was 44% lower than fossil diesel. In comparison to fossil diesel and at full load, the brake specific fuel consumption (wt.) of the 30% and 20% blends were approximately 32% and 15% higher. At full engine load, the CO emission of the 20% and 30% blends were decreased by 39% and 66% with respect to the fossil diesel. Blends CO2 emissions were similar to that of fossil diesel; at full engine load, 30% blend produced approximately 5% higher CO2 emission than fossil diesel. The study concludes that on the basis of short term engine experiment up to 30% blend of pyrolysis oil from digestate of arable crops can be used in a compression ignition engine.

Effects of annealing, pre-tension and mounting on the hysteresis of polymer strain sensors

When exposed to high levels of strain, polymer optical fibre grating sensors recorded in poly(methyl methacrylate) based fibre often exhibit hysteresis in the response of their Bragg wavelength to strain. We demonstrate that the application of pre-tension and annealing of the polymer fibre can reduce this hysteresis when the fibre is suspended freely between two supports, but much better performance is obtained when the sensor is attached directly to a substrate. In this case, the hysteresis can be lessened by more than a factor of 12. © 2014 IOP Publishing Ltd.

An investigation into the wavelength stability of polymer optical fibre Bragg gratings

The inscription of Bragg gratings has been demonstrated in PMMA-based polymer optical fibre. The water affinity of PMMA can introduce significant wavelength change in a polymer optical fibre Bragg grating (POFBG). In polymer optical fibre losses are much higher than with silica fibre. Very strong absorption bands related to higher harmonics of vibrations of the C-H bond dominate throughout the visible and near infrared. Molecular vibration in substances generates heat, which is referred to as the thermal effect of molecular vibration. This means that a large part of the absorption of optical energy in those spectral bands will convert into thermal energy, which eventually drives water content out of the polymer fibre and reduces the wavelength of POFBG. In this work we have investigated the wavelength stability of POFBGs in different circumstances. The experiment has shown that the characteristic wavelength of a POFBG starts decreasing after a light source is applied to it. This decrease continues until equilibrium inside the fibre is established, depending on the initial water content inside the fibre, the surrounding humidity, the optical power applied, and the fibre size. Our investigation has shown that POFBGs operating at around 850 nm show much smaller wavelength reduction than those operating at around 1550 nm in the same fibre; POFBGs with different diameters show different changes; POFBGs powered by a low level light source, or operating in a very dry environment are least affected by this thermal effect.

Optimisation of polymer optical fibre based interferometric sensors

A numerical model for studying the performance of polymer optical fibre-based interferometric sensors is presented. The strain sensitivity of Fabry-Perot and two-beam interferometric sensors is investigated by varying the physical and optical properties corresponding to frequently used wavelengths. The developed model was used to identify the regimes in which these devices offer enhanced performance over their silica counterparts when used for stress sensing. © (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.

Facile synthesis of polymer-peptide conjugates via direct amino acid coupling chemistry

Polymer-peptide conjugates (also known as biohy-brids) are attracting considerable attention as injectable materials owing to the self-assembling behavior of the peptide and the ability to control the material properties using the polymer component. To this end, a simple method for preparing poly(ethylene oxide)-oligophenylalanine polymer-peptide conjugates (mPEOm-F n-OEt) using isobutylchloroformate as the activating reagent has been identified and developed. The synthetic approach reported employs an industrially viable route to produce conjugates with high yield and purity. Moreover, the approach allows judicious selection of the precursor building blocks to produce libraries of polymer-peptide conjugates with complete control over the molecular composition. Control over the molecular make-up of the conjugates allows fine control of the physicochemical properties, which will be exploited in future studies into the prominent self-assembling behavior of such materials. © 2013 Wiley Periodicals, Inc.