The Yariv reagent: behaviour in different solvents and interaction with a gum arabic arabinogalactan-protein.

The ß-D-Glc Yariv reagent is frequently used to isolate and to study the structure of arabinogalactan-proteins with the arabinogalactan type II structure. The present paper describes the aggregation features of the Yariv reagent in water, salt solutions and in organic solvents as determined by NMR, absorption spectroscopy and light scattering experiments. The results indicate that in water the Yariv reagent forms aggregates of up to 300 units and in 1% aqueous NaCl the degree of aggregation is approx. 150. The aggregates are formed both by H-bonds and hydrophobic interactions, the former appearing to be of most importance in water. The interaction between the Yariv reagent and an AGP fraction from gum arabic, showed a degree of aggregation of the Yariv reagent when using 1% NaCl to be of approx. 150 units, whereas disruption of the aggregate took place in 10% NaCl with an aggregation number of approx. 100. Partial acid hydrolysis of an AGP from gum Arabic (Acacia Senegal) and analyses of the linkage types remaining indicated that a certain length of (1→3)-ß-linked galactose units was necessary for binding between the Yariv reagent and the AGP. This is in accordance to what also was recently observed by Kitazawa et al. (2013).

Elasticity of polymer solutions in Couette flow measured by fluorescence resonance energy transfer (FRET).

Polymer solutions are complex fluids that show elasticity and deformation in response to shear flows. A fluorescence resonance energy transfer (FRET) technique has been applied to measure the end-to-end distances of individual polymer molecules in Couette flow, using end-tagged reversible-addition fragmentation chain transfer (RAFT) polymerised poly(methyl methacrylate) (PMMA). Real-time rheofluorescence measurements on these polymers in solution above the critical overlap concentration are reported at several shear rates. The PMMA in Couette flow shows a systematic decrease in fluorescence, corresponding to a reduction in end-to-end distance of the polymer molecules with shear exposure. Full reversibility of the fluorescence signal is observed after the cessation of shear. These results show that polymer solution elasticity arises from compressive deformation of the polymer molecules in Couette flow. At polymer concentrations above the critical overlap, the polymer molecules are restricted by their neighbours and the net hydrodynamic forces are compressive rather than extensive.

Aging and yielding in a sheared AOT/iso-octane/water lyotropic lamellar phase.

We define a creep-flow-based measurement procedure to allow reliable and reproducible results on aging and yielding materials to be obtained. Investigation of the effects of different parameter such as the pre-shear time, the recovery time and the applied stress magnitude on the viscoelastic properties of a lyotropic liquid crystal phase is reported. Cryo-TEM observations indicate the formation of multiconnected bilayers at rest. Shearing the investigated material shows a propensity to acquire all the macroscopic properties of "soft jammed systems". These properties are then interpreted in terms of shear-induced structural rearrangement on the basis of cryofracture observation obtained at different times after the preshear imposed.

Comparison of casein micelles in raw and reconstituted skim milk.

During the manufacture of skim milk powder, many important alterations to the casein micelles occur. This study investigates the nature and cause of these alterations and their reversibility upon reconstitution of the powders in water. Samples of skim milk and powder were taken at different stages of commercial production of low-, medium-, and high-heat powders. The nature and composition of the casein micelles were analyzed using a variety of analytical techniques including photon correlation spectroscopy, transmission electron microscopy, turbidity, and protein electrophoresis. It was found that during heat treatment, whey proteins are denatured and become attached to the casein micelles, resulting in larger micelles and more turbid milk. The extent of whey protein attachment to the micelles is directly related to the severity of the heat treatment. It also appeared that whey proteins denatured during heat treatment may continue to attach to casein micelles during water removal (evaporation and spray-drying). The process of water removal causes casein and Ca in the serum to become increasingly associated with the micelles. This results in much larger, denser micelles, increasing the turbidity while decreasing the viscosity of the milk. During reconstitution, the native equilibrium between colloidal Ca and serum Ca is slowly reestablished. The reequilibration of the caseins and detachment of the whey proteins occur even more slowly. The rate of reequilibration does not appear to be influenced by shear or temperature in the range of 4 to 40 degrees C.

Avoiding "mud" cracks during drying of thin films from aqueous colloidal suspensions.

The critical cracking thickness of films obtained by drying aqueous alumina suspensions has been investigated. The effects of solution chemistry, binder and binder crosslinking were studied. Films formed from flocculated and dispersed suspensions are compared. The influence of the addition of the polymeric binder, poly(vinyl alcohol) (PVA) was also investigated. In addition, in some of the dispersed suspensions the PVA was covalently crosslinked. The critical cracking thickness is found to be 3 times greater for the films obtained from dispersed suspensions than for the films obtained from flocculated suspensions. The superior mechanical properties are primarily due to the higher final solids concentration in the films obtained from dispersed suspensions. Addition of PVA leads to an increase of the critical cracking thickness by a factor of two for both dispersed and flocculated systems. When the PVA is crosslinked, the mechanical properties of the gel during drying are improved and the critical cracking thickness is increased 10 fold with respect to the suspensions with uncrosslinked PVA.

Atomic force microscopy imaging of DNA-cationic liposome complexes optimised for gene transfection into neuronal cells.

BACKGROUND: Cationic liposomes represent an important gene delivery system due to their low immunogenicity, but are relatively inefficient, with optimisation of DNA-liposome complexes (lipoplexes) for transfection necessary for each cell type of interest. There have been few studies examining optimisation in neuronal cell types or determining how the structure of lipoplexes affects transfection efficiency. METHODS: Four commercially available cationic liposome formulations were used to optimise transfection efficiency in neuronal cells. The DNA to liposome ratio and the amount of DNA used in transfections were varied. Transfection efficiency was determined by the percentage of cells positive for the micro-galactosidase reporter gene product. The structure of lipoplexes was studied using atomic force microscopy. Lipoplexes were characterised further using dynamic light scattering to determine size and fluorescence techniques to show DNA compaction. RESULTS: Optimal transfection conditions were found to differ between immortalised cell lines and primary cells. High transfection efficiencies in immortalised cell lines were achieved predominantly with multivalent cationic liposomes while primary neuronal cells showed optimal transfection efficiency with monovalent cationic liposomes. The structure of lipoplexes was observed with atomic force microscopy and showed globular complexes for multivalent cationic liposomes, while monovalent liposomes gave less compact structures. In support of this finding, high levels of DNA compaction with multivalent liposomes were observed using fluorescence quenching measurements for all DNA to liposome ratios tested. One monovalent liposome showed increasing levels of compaction with increasing liposome amount. Dynamic light scattering showed little change in complex size when the different lipoplexes were studied. CONCLUSIONS: Optimisation of transfection efficiency was different for cell lines and primary neurons. Immortalised cells showed optimal transfection with multivalent liposomes while primary neurons showed optimal transfection with monovalent liposomes. The charge ratio of the monovalent liposome was below one, suggesting a different mechanism of lipoplex binding and uptake in primary neurons. The structure of lipoplexes, as

Diffusing probe measurements in Newtonian and elastic solutions.

The diffusion coefficients of polystyrene latex spheres and hematite particles in both Newtonian and elastic liquids have been measured using dynamic light scattering. The diffusion coefficients of the latex particles measured in glycerol/water (Newtonian) solutions obey Stokes-Einstein behaviour over a range of solvent viscosities and temperatures. Two apparent diffusion coefficients for the particles are measured in visco-elastic polyacrylamide and polyacrylate solutions and are designated Dfast and Dslow. The apparent fast diffusion coefficients measured in the elastic solutions show an increase to a maximum, above that measured in the solvent water, with increasing polyelectrolyte concentration. At higher polyelectrolyte concentrations the observed Dfast values decrease below the value obtained in the solvent water. Dfast increases with the scattering vector squared (q2) while Dslow, is independent of q2.

Cytoskeletal regulation of the platelet glycoprotein Ib/V/IX-von willebrand factor interaction.

Shear-induced binding of von Willebrand factor (vWf) to the platelet glycoprotein (GP) Ib/V/IX complex plays a key role in initiating platelet adhesion and aggregation at sites of vascular injury. This study demonstrated that pretreating human platelets with inhibitors of actin polymerization, cytochalasin D or latrunculin B, dramatically enhances platelet aggregation induced by vWf. The effects of these inhibitors were specific to the vWf-GPIbalpha interaction because they enhanced vWf-induced aggregation of Glanzmann thrombasthenic platelets and Chinese hamster ovary (CHO) cells transfected with GPIb/V/IX. Moreover, cytochalasin D enhanced the extent of platelet aggregation induced by high shear stress (5000 s(-1)) and also lowered the shear threshold required to induce aggregation from 3000 s(-1) to as low as 500 s(-1). Studies of CHO cells expressing GPIbalpha cytoplasmic tail truncation mutants that failed to bind actin-binding protein-280 (deletion of residues 569-610 or 535-568) demonstrated that the linkage between GPIb and actin-binding protein-280 was not required for vWf-induced actin polymerization, but was critical for the enhancing effects of cytochalasin D on vWf-induced cell aggregation. Taken together, these studies suggest a fundamentally important role for the cytoskeleton in regulating the adhesive function of GPIb/V/IX.