Beta-lactoglobulin fibers under capillary flow

We describe the capillary flow behavior of gels of beta-lactoglobulin (beta-lg) containing droplets of fibrils and the shear flow alignment of beta-lg fibers in dilute aqueous solutions. Polarized optical microscopy and laser scanning confocal microscopy are used to show that capillary shear flow does not affect the fibril droplet sizes in the beta-lg gels, the system behaving in this respect as a solution of compact colloidal particles under shear flow. Small-angle X-ray scattering (SAXS) on dilute aqueous solutions indicates that the fibers can be initially aligned under capillary shear, but this alignment is lost after 18 min of shear. Transmission electron microscopy experiments on the samples studied by SAXS suggest that the loss of orientation is due to a shear-induced breakup of the swollen fibril network. Dynamic and static light scattering on dilute beta-lg fibril aqueous solutions are used to show that before shear beta-lg fibrils behave as strongly interacting semiflexible polymers, while they behave as weakly interacting rods after 18 min of capillary shear.

Spontaneous condensation in DNA-polystyrene-b-poly(l-lysine) polyelectrolyte block copolymer mixtures

We investigated the condensation of calf thymus DNA by amphiphilic polystyrene(m)-b-poly(l-lysine)(n) block copolymers (PSm-b- PLys(n), m, n = degree of polymerization), using small-angle X-ray scattering, polarized optical microscopy and laser scanning confocal microscopy. Microscopy studies showed that the DNA condenses in the form of fibrillar precipitates, with an irregular structure, due to electrostatic interactions between PLys and DNA. This is not modified by the presence of hydrophobic PS block. Scattering experiments show that the structure of the polyplexes corresponds to a local order of DNA rods which becomes more compact upon increasing n. It can be concluded that for DNA/ PSm-b- PLys(n) polyplexes, the balance between the PLys block length and the excess charge in the system plays an essential role in the formation of a liquid crystalline phase.

Polymer − surfactant vesicular complexes in aqueous medium

The introduction of ionic single-tailed surfactants to aqueous solutions of EO18BO10 [EO = poly(ethylene oxide), BO = poly(1,2-butylene oxide), subscripts denote the number of repeating units] leads to the formation of vesicles, as probed by laser scanning confocal microscopy. Dynamic light scattering showed that the dimensions of these aggregates at early stages of development do not depend on the sign of the surfactant head group charge. Small-angle X-ray scattering (SAXS) analysis indicated the coexistence of smaller micelles of different sizes and varying polymer content in solution. In strong contrast to the dramatic increase of size of dispersed particles induced by surfactants in dilute solution, the d-spacing of corresponding mesophases reduces monotonically upon increasing surfactant loading. This effect points to the suppression of vesicles as a consequence of increasing ionic strength in concentrated solutions. Maximum enhancements of storage modulus and thermal stability of hybrid gels take place at different compositions, indicating a delicate balance between the number and size of polymer-poor aggregates (population increases with surfactant loading) and the number and size of polymer−surfactant complexes (number and size decrease in high surfactant concentrations).

Uses of airborne scanning laser altimetry for river flood modelling and coastal geomorphology

Two ongoing projects at ESSC that involve the development of new techniques for extracting information from airborne LiDAR data and combining this information with environmental models will be discussed. The first project in conjunction with Bristol University is aiming to improve 2-D river flood flow models by using remote sensing to provide distributed data for model calibration and validation. Airborne LiDAR can provide such models with a dense and accurate floodplain topography together with vegetation heights for parameterisation of model friction. The vegetation height data can be used to specify a friction factor at each node of a model’s finite element mesh. A LiDAR range image segmenter has been developed which converts a LiDAR image into separate raster maps of surface topography and vegetation height for use in the model. Satellite and airborne SAR data have been used to measure flood extent remotely in order to validate the modelled flood extent. Methods have also been developed for improving the models by decomposing the model’s finite element mesh to reflect floodplain features such as hedges and trees having different frictional properties to their surroundings. Originally developed for rural floodplains, the segmenter is currently being extended to provide DEMs and friction parameter maps for urban floods, by fusing the LiDAR data with digital map data. The second project is concerned with the extraction of tidal channel networks from LiDAR. These networks are important features of the inter-tidal zone, and play a key role in tidal propagation and in the evolution of salt-marshes and tidal flats. The study of their morphology is currently an active area of research, and a number of theories related to networks have been developed which require validation using dense and extensive observations of network forms and cross-sections. The conventional method of measuring networks is cumbersome and subjective, involving manual digitisation of aerial photographs in conjunction with field measurement of channel depths and widths for selected parts of the network. A semi-automatic technique has been developed to extract networks from LiDAR data of the inter-tidal zone. A multi-level knowledge-based approach has been implemented, whereby low level algorithms first extract channel fragments based mainly on image properties then a high level processing stage improves the network using domain knowledge. The approach adopted at low level uses multi-scale edge detection to detect channel edges, then associates adjacent anti-parallel edges together to form channels. The higher level processing includes a channel repair mechanism.

‘Symptomless’ infection by Botrytis cinerea

The study was carried out to clarify the nature of symptomless infection by Botrytis cinerea and to what extent it differs from aggressive necrotic infection in Lactuca sativa (lettuce) and Arabidopsis thaliana. Symptomless plants were produced by dry spore inoculation in plants growing in controlled environmental conditions or in glasshouses. Plating out of surface-disinfected and non-surface-disinfected samples of inoculated, apparently healthy, plants on selective medium revealed that the fungus was spreading from the initial inoculation site to newly developing plant organs both internally and externally. Similar findings were obtained in microscope experiments in which host plants were inoculated with GFP labelled B. cinerea and symptomless spreading was monitored under confocal laser scanning microscope. Spore germination on leaf surface was followed by development of sub-cuticular vesicles and plant cell damage in the infected epidermal cell and a few nearby cells. Sparsely branched long hyphae arose from the vesicles and spread on the leaf surface; spread was mostly on the outer surface of the epidermal layer but occasionally below the cuticle or epidermal cells. In the late symptomless phase, mycelium arising from single vesicles formed several mycelial networks on leaves. Experiments were carried out to compare the extent of gene expression in symptomless and necrotic infections, using RT-qPCR. Expression of selected genes was quantified in tissue samples based on the amount of mRNA of the respective genes found. In both host species, the mRNA concentration of signalling genes bcg1, bmp1 and calcineurin, and the pathogenicity genes bcsod1 and bcpg1 were similar to or slightly greater in symptomless samples than in necrotic samples. The mRNA of the signalling gene bac and pathogenicity genes bcbot1 and bcnep1, were not detected or detected in lower abundance than in necrosis. In lettuce, the leaves developing distant from the site of inoculation showed similar results to A. thaliana, but in healthy leaves close to the site of inoculation mRNA concentrations of bac and bcnep1 were similar to necrotic samples. Thus, in both host species, the fungus grew along with the plant and moved to newly growing plant parts without producing symptoms; during this growth some pathogenicity genes were less expressed than in necrotic infection.

Problems with determining the particle size distribution of chalk soil and some of their implications

Particle size distribution (psd) is one of the most important features of the soil because it affects many of its other properties, and it determines how soil should be managed. To understand the properties of chalk soil, psd analyses should be based on the original material (including carbonates), and not just the acid-resistant fraction. Laser-based methods rather than traditional sedimentation methods are being used increasingly to determine particle size to reduce the cost of analysis. We give an overview of both approaches and the problems associated with them for analyzing the psd of chalk soil. In particular, we show that it is not appropriate to use the widely adopted 8 pm boundary between the clay and silt size fractions for samples determined by laser to estimate proportions of these size fractions that are equivalent to those based on sedimentation. We present data from field and national-scale surveys of soil derived from chalk in England. Results from both types of survey showed that laser methods tend to over-estimate the clay-size fraction compared to sedimentation for the 8 mu m clay/silt boundary, and we suggest reasons for this. For soil derived from chalk, either the sedimentation methods need to be modified or it would be more appropriate to use a 4 pm threshold as an interim solution for laser methods. Correlations between the proportions of sand- and clay-sized fractions, and other properties such as organic matter and volumetric water content, were the opposite of what one would expect for soil dominated by silicate minerals. For water content, this appeared to be due to the predominance of porous, chalk fragments in the sand-sized fraction rather than quartz grains, and the abundance of fine (<2 mu m) calcite crystals rather than phyllosilicates in the clay-sized fraction. This was confirmed by scanning electron microscope (SEM) analyses. "Of all the rocks with which 1 am acquainted, there is none whose formation seems to tax the ingenuity of theorists so severely, as the chalk, in whatever respect we may think fit to consider it". Thomas Allan, FRS Edinburgh 1823, Transactions of the Royal Society of Edinburgh. (C) 2009 Natural Environment Research Council (NERC) Published by Elsevier B.V. All rights reserved.

Effects of variation in the palm stearin: palm olein ratio on the crystallisation of a low-trans shortening

Changes in the theological properties during crystallisation and in the crystal size and morphology of blends containing rapeseed oil with varying percentages of palm stearin (POs) and palm olein (POf) have been studied. The crystals formed from all three blends were studied by confocal laser scanning microscopy, light microscopy and environmental scanning electron microscopy, which revealed the development of clusters of 3-5 individual elementary "spherulites" in the early stages of crystallisation. The saturated triacylglycerol content of the solid crystals separated at the onset of crystallisation was much greater than that in the total fat. Fat blends with a higher content of palm stearin had a more rapid nucleation rate when observed by light microscopy, and this caused an earlier change in the rheological properties of the fat during crystallisation. Using a low torque amplitude (0.005 Pa, which was within the linear viscoelastic region of all samples studied) and a frequency of 1 Hz, the viscoelastic properties of melted fat during cooling were studied. All samples, prior to crystallisation, showed weak viscoelastic liquid behaviour (G '', loss modulus >G', storage modulus). After crystallisation a more "solid like" behaviour was observed (G' similar to or greater than G ''). The blend having the highest concentration of POs was found to have the earliest onset of crystallisation (27% w/w POs; 12 mins, 22% w/w POs; 13.5 mins, 17% w/w POs, 15 mins, respectively). However, there were no significant differences in the time to the point when G' became greater than G' among the three blends. (c) 2006 Elsevier Ltd. All rights reserved.

Production and evaluation of dry alginate-chitosan microcapsules as an enteric delivery vehicle for probiotic bacteria

This study investigates the production of alginate microcapsules, which have been coated with the polysaccharide chitosan, and evaluates some of their properties with the intention of improving the gastrointestinal viability of a probiotic (Bifidobacterium breve) by encapsulation in this system. The microcapsules were dried by a variety of methods, and the most suitable was chosen. The work described in this Article is the first report detailing the effects of drying on the properties of these microcapsules and the viability of the bacteria within relative to wet microcapsules. The pH range over which chitosan and alginate form polyelectrolyte complexes was explored by spectrophotometry, and this extended into swelling studies on the microcapsules over a range of pHs associated with the gastrointestinal tract. It was shown that chitosan stabilizes the alginate microcapsules at pHs above 3, extending the stability of the capsules under these conditions. The effect of chitosan exposure time on the coating thickness was investigated for the first time by confocal laser scanning microscopy, and its penetration into the alginate matrix was shown to be particularly slow. Coating with chitosan was found to increase the survival of B. breve in simulated gastric fluid as well as prolong its release upon exposure to intestinal pH.

Mozzarella-type curd made from buffalo, cows’ and ultrafiltered cows’ milk. 1. Rheology and microstructure

Rennet-induced curd was made from both natural buffalo and cows’ milk, and ultrafiltered cows’ milk (cows’ milk was concentrated such that it had a chemical composition approximately equivalent to that of the buffalo milk). These milk samples were compared on the basis of their rheology, physicochemical characteristics and curd microstructure. The ionic and soluble calcium contents were found to be similar in all milk samples studied. The total and casein bound calcium were higher in concentrated cows’ milk than in standard cows’ milk. Both cows’ milk types were found to have lower total and casein bound calcium than the buffalo milk. This is probably due to concentration of the colloidal part of milk (casein), during the ultrafiltration (UF) process. The rennet coagulation time was similar in UF cows’ and buffalo milk while both were shorter when compared with that of the cows’ milk. The dynamic moduli (G′, G″) values were higher in both the buffalo and UF cows’ milk than in the cows’ milk after 90 min coagulation. The loss tangent, however, was found to be similar in both the UF cows’ and buffalo milk curds and was lower than that observed for the cows’ milk (0.42, 0.42 and 0.48, respectively). The frequency profile of each type of curd was recorded 90 min after the enzyme addition (0.1–10 Hz); all samples were found to be “weak” viscoelastic, frequency dependent gels. The yield stress was also measured 95 min after the enzyme addition, and a higher value was observed in buffalo milk curd when compared with other curd samples made from both the natural cows’ milk and the UF cows’ milk. The cryo-scanning electron and confocal laser scanning micrographs showed that curd structure appeared to be more “dense” and less porous in buffalo milk than cows’ milk even after concentration to equivalent levels of protein/total solids to those found in the buffalo milk.

Post-frying oil drainage from potato chips and french fries: a comparative study of atmospheric and vacuum drainage

This paper represents a study of the transient changes occurring in temperature, and moisture and oil contents during the so called “post-frying drainage”—which is the duration for which a product is held in the head space of the fryer after it is removed from the oil. Since most of the oil adhering to the product penetrates into the structure during this period, this paper examines the effects of applying vacuum during drainage (1.33 kPa) to maintain the product temperature consistently above the water saturation temperature corresponding to the prevailing pressure (11 °C), which potentially eliminates water condensation and prevents the occluded surface oil from penetrating into the product structure. Draining under vacuum significantly lowers the oil content of potato chips by 38% compared to atmospheric drainage. This phenomenon can be further confirmed by confocal laser scanning microscopy (CLSM) images, which show that the boundary between the core and the crust regions is clearly visible in the case of vacuum drainage, whereas in the case of atmospheric drainage, the oil is distributed throughout the structure. Unfortunately, the same approach did not reduce the oil content of French fries—the oil content of vacuum-drained product was found similar to the product obtained by draining under atmospheric pressure. This is because the reduction in oil content only occurs when there is net moisture evaporation from the product and the evaporation rate is sufficient to force out the oil from the product; this was clearly not the case with French fries. The CLSM images show that the oil distribution in the products drained under atmospheric pressure and vacuum was similar.

Layer-by-layer coating of alginate matrices with chitosan–alginate for the improved survival and targeted delivery of probiotic bacteria after oral administration

The oral administration of probiotic bacteria has shown potential in clinical trials for the alleviation of specific disorders of the gastrointestinal tract. However, cells must be alive in order to exert these benefits. The low pH of the stomach can greatly reduce the number of viable microorganisms that reach the intestine, thereby reducing the efficacy of the administration. Herein, a model probiotic, Bifidobacterium breve, has been encapsulated into an alginate matrix before coating in multilayers of alternating alginate and chitosan. The intention of this formulation was to improve the survival of B. breve during exposure to low pH and to target the delivery of the cells to the intestine. The material properties were first characterized before in vitro testing. Biacore™ experiments allowed for the polymer interactions to be confirmed; additionally, the stability of these multilayers to buffers simulating the pH of the gastrointestinal tract was demonstrated. Texture analysis was used to monitor changes in the gel strength during preparation, showing a weakening of the matrices during coating as a result of calcium ion sequestration. The build-up of multilayers was confirmed by confocal laser-scanning microscopy, which also showed the increase in the thickness of coat over time. During exposure to in vitro gastric conditions, an increase in viability from <3 log(CFU) per mL, seen in free cells, up to a maximum of 8.84 ± 0.17 log(CFU) per mL was noted in a 3-layer coated matrix. Multilayer-coated alginate matrices also showed a targeting of delivery to the intestine, with a gradual release of their loads over 240 min.

CLSM method for the dynamic observation of pH change within polymer matrices for oral delivery

If acid-sensitive drugs or cells are administered orally, there is often a reduction in efficacy associated with gastric passage. Formulation into a polymer matrix is a potential method to improve their stability. The visualization of pH within these materials may help better understand the action of these polymer systems and allow comparison of different formulations. We herein describe the development of a novel confocal laser-scanning microscopy (CLSM) method for visualizing pH changes within polymer matrices and demonstrate its applicability to an enteric formulation based on chitosan-coated alginate gels. The system in question is first shown to protect an acid-sensitive bacterial strain to low pH, before being studied by our technique. Prior to this study, it has been claimed that protection by these materials is a result of buffering, but this has not been demonstrated. The visualization of pH within these matrices during exposure to a pH 2.0 simulated gastric solution showed an encroachment of acid from the periphery of the capsule, and a persistence of pHs above 2.0 within the matrix. This implies that the protective effect of the alginate-chitosan matrices is most likely due to a combination of buffering of acid as it enters the polymer matrix and the slowing of acid penetration.

Simple method for sub-diffraction resolution imaging of cellular structures on standard confocal microscopes by three-photon absorption of quantum dots

This study describes a simple technique that improves a recently developed 3D sub-diffraction imaging method based on three-photon absorption of commercially available quantum dots. The method combines imaging of biological samples via tri-exciton generation in quantum dots with deconvolution and spectral multiplexing, resulting in a novel approach for multi-color imaging of even thick biological samples at a 1.4 to 1.9-fold better spatial resolution. This approach is realized on a conventional confocal microscope equipped with standard continuous-wave lasers. We demonstrate the potential of multi-color tri-exciton imaging of quantum dots combined with deconvolution on viral vesicles in lentivirally transduced cells as well as intermediate filaments in three-dimensional clusters of mouse-derived neural stem cells (neurospheres) and dense microtubuli arrays in myotubes formed by stacks of differentiated C2C12 myoblasts.

Optimization of a sponge cake formulation with inulin as fat replacer: structure, physicochemical, and sensory properties

The effects of several fat replacement levels (0%, 35%, 50%, 70%, and 100%) by inulin in sponge cake microstructure and physicochemical properties were studied. Oil substitution for inulin decreased significantly (P < 0.05) batter viscosity, giving heterogeneous bubbles size distributions as it was observed by light microscopy. Using confocal laser scanning microscopy the fat was observed to be located at the bubbles’ interface, enabling an optimum crumb cake structure development during baking. Cryo-SEM micrographs of cake crumbs showed a continuous matrix with embedded starch granules and coated with oil; when fat replacement levels increased, starch granules appeared as detached structures. Cakes with fat replacement up to 70% had a high crumb air cell values; they were softer and rated as acceptable by an untrained sensory panel (n = 51). So, the reformulation of a standard sponge cake recipe to obtain a new product with additional health benefits and accepted by consumers is achieved.

Velocities of compressional and shear waves in limestones

Carbonate rocks are important hydrocarbon reservoir rocks with complex textures and petrophysical properties (porosity and permeability) mainly resulting from various diagenetic processes (compaction, dissolution, precipitation, cementation, etc.). These complexities make prediction of reservoir characteristics (e.g. porosity and permeability) from their seismic properties very difficult. To explore the relationship between the seismic, petrophysical and geological properties, ultrasonic compressional- and shear-wave velocity measurements were made under a simulated in situ condition of pressure (50 MPa hydrostatic effective pressure) at frequencies of approximately 0.85 MHz and 0.7 MHz, respectively, using a pulse-echo method. The measurements were made both in vacuum-dry and fully saturated conditions in oolitic limestones of the Great Oolite Formation of southern England. Some of the rocks were fully saturated with oil. The acoustic measurements were supplemented by porosity and permeability measurements, petrological and pore geometry studies of resin-impregnated polished thin sections, X-ray diffraction analyses and scanning electron microscope studies to investigate submicroscopic textures and micropores. It is shown that the compressional- and shear-wave velocities (V-p and V-s, respectively) decrease with increasing porosity and that V-p decreases approximately twice as fast as V-s. The systematic differences in pore structures (e.g. the aspect ratio) of the limestones produce large residuals in the velocity versus porosity relationship. It is demonstrated that the velocity versus porosity relationship can be improved by removing the pore-structure-dependent variations from the residuals. The introduction of water into the pore space decreases the shear moduli of the rocks by about 2 GPa, suggesting that there exists a fluid/matrix interaction at grain contacts, which reduces the rigidity. The predicted Biot-Gassmann velocity values are greater than the measured velocity values due to the rock-fluid interaction. This is not accounted for in the Biot-Gassmann velocity models and velocity dispersion due to a local flow mechanism. The velocities predicted by the Raymer and time-average relationships overestimated the measured velocities even more than the Biot model.