Regulation of Protein Turnover during Hyper-osmotic Stress in Skeletal Muscle

The purpose of this study was to examine the effect of hyper-osmotic stress on protein turnover in skeletal muscle tissue using an established in-vitro model. Rat EDL muscles were incubated in either hyper-osmotic (400 ± 10 Osm) or isoosmotic (290 ± 10 Osm) custom-modified media (Gibco). L-[14C]-U-phenylalanine (n=8) and cycloheximide (n=8) were used to quantify protein synthesis and degradation, respectively. Western blotting analyses was performed to determine the activation of protein synthesis and degradation pathways. During hyperosmotic stress, protein degradation increased (p<0.05), while protein synthesis was decreased (p<0.05) as compared to the iso-osmotic condition. The decline in protein synthesis was accompanied by a decrease (p<0.05) in p70s6 kinase phosphorylation, while the increase in protein degradation was associated with an increase (p<0.05) in autolyzed calpain. Therefore, hyper-osmotic extracellular stress results in an intracellular catabolic environment in mammalian skeletal muscle tissue.

INFLUENCE OF INCREASED EXTRACELLULAR LEUCINE ON THE PROTEIN METABOLIC RESPONSES DURING OSMOTIC STRESS IN SKELETAL MUSCLE

The purpose of this study was to examine the effects of increased extracellular leucine concentration on protein metabolism in skeletal muscle cells when exposed to 3 different osmotic stresses. L6 skeletal muscle cells were incubated in either a normal or supplemental leucine (1.5mM) medium set to hypo-osmotic (230 ± 10 Osm), iso-osmotic (330 ± 10 Osm) or hyper-osmotic (440 ± 10 Osm) conditions. 3H-tyrosine was used to quantify protein synthesis. Western blotting analysis was performed to determine the activation of mTOR, p70S6k, ubiquitin, actin, and μ-calpain. Hypo-osmotic stress resulted in the greatest increase in protein synthesis rate under the normal-leucine condition while iso-osmotic stress has the greatest increase under the elevated-leucine condition. Elevated-leucine condition had a decreased rate in protein degradation over the normal condition within the ubiquitin proteasome pathway (p<0.05). Leucine and hypo-osmotic stress therefore creates a favourable environment for anabolic events to occur.

Effect of curing temperature , fibre loading and bonding agent on the equilibrium swelling of isora-natural rubbercomposites

Isora fibre-reinforced natural rubber (NR) composites were cured at 80, 100, 120 and 150°C using a low temperature curing accelerator system. Composites were also prepared using a conventional accelerator system and cured at 150°C. The swelling behavior of these composites at varying fibre loadings was studied in toluene and hexane. Results show that the uptake of solvent and volume fraction of rubber due to swelling was lower for the low temperature cured vulcanizates which is an indication of the better fibre/rubber adhesion. The uptake of aromatic solvent was higher than that of aliphatic solvent, for all the composites. As the fibre content increased, the solvent uptake decreased, due to the superior solvent resistance of the fibre and good fibre-rubber interactions. The bonding agent improved the swelling resistance of the composites due to the strong interfacial adhesion. Due to the improved adhesion between the fibre and rubber, the ratio of the change in volume fraction of rubber due to swelling to the volume fraction of rubber in the dry sample (V,) was found to decrease in the presence of bonding agent. At a fixed fibre loading, the alkali treated fibre composite showed a lower percentage swelling than untreated one for both systems showing superior rubber-fibre interactions.

Swelling behaviour of isora/natural rubber composites in oils used in automobiles

Natural rubber/isora fibre composites were cured at various temperatures. The solvent swelling characteristics of natural rubber composites containing both untreated and alkali treated fibres were investigated in aromatic and aliphatic solvents like toluene, and n-hexane. The diffusion experiments were conducted by the sorption gravimetric method. The restrictions on elastomer swelling exerted by isora fibre as well as the anisotropy of swelling of the composite have been confirmed by this study. Composite cured at 100°C shows the lowest percentage swelling. The uptake of aromatic solvent is higher than that of aliphatic solvent for the composites cured at all temperatures. The effect of fibre loading on the swelling behaviour of the composite was also investigated in oils like petrol, diesel, lubricating oil etc. The % swelling index and swelling coefficient of the composite were found to decrease with increase in fibre loading. This is due to the increased hindrance exerted by the fibres at higher fibre loadings and also due to the good fibre-rubber interactions. Maximum uptake of solvent was observed with petrol followed by diesel and then lubricating oil. The presence of bonding agent in the composites restrict the swelling considerably due to the strong interfacial adhesion. At a fixed fibre loading, the alkali treated fibre composite showed lower percentage swelling compared to the untreated one.

Contributions of matric and osmotic potentials to the unfrozen water content of frozen soils

Recent reports show that biogeochemical processes continue when the soil is frozen, but are limited by water availability. However, there is little knowledge about the interactive effects of soil and environmental variables on amounts of unfrozen water in frozen soils. The aims of this study were to determine the contributions of matric and osmotic potentials to the unfrozen water content of frozen soil. We determined the effects of matric and osmotic potential on unfrozen water contents of frozen mineral soil fractions (ranging from coarse sand to fine silt) at -7 degrees C, and estimated the contributions of these potentials to liquid water contents in samples from organic surface layers of boreal soils frozen at -4 degrees C. In the mineral soil fractions the unfrozen water contents appeared to be governed solely by the osmotic potential, but in the humus layers of the sampled boreal soils both the osmotic and matric potentials control unfrozen water content, with osmotic potential contributing 20 to 69% of the total water potential. We also determined pore size equivalents, where unfrozen water resides at -4 degrees C, and found a strong correlation between these equivalents and microbial CO2 production. The larger the pores in which the unfrozen water is found the larger the microbial activity that can be sustained. The osmotic potential may therefore be a key determinant of unfrozen water and carbon dynamics in frozen soil. (C) 2008 Elsevier B.V. All rights reserved.

Dual-specificity phosphatases in the hypo-osmotic stress response of keratin-defective epithelial cell lines

Although mutations in intermediate filament proteins cause many human disorders, the detailed pathogenic mechanisms and the way these mutations affect cell metabolism are unclear. In this study, selected keratin mutations were analysed for their effect on the epidermal stress response. Expression profiles of two keratin-mutant cell lines from epidermolysis bullosa simplex patients (one severe and one mild) were compared to a control keratinocyte line before and after challenge with hypo-osmotic shock, a common physiological stress that transiently distorts cell shape. Fewer changes in gene expression were found in cells with the severely disruptive mutation (55 genes altered) than with the mild mutation (174 genes) or the wild type cells (261 genes) possibly due to stress response pre-activation in these cells. We identified 16 immediate-early genes contributing to a general cell response to hypo-osmotic shock, and 20 genes with an altered expression pattern in the mutant keratin lines only. A number of dual-specificity phosphatases (MKP-1, MKP-2, MKP-3, MKP-5 and hVH3) are differentially regulated in these cells, and their downstream targets p-ERK and p-p38 are significantly up-regulated in the mutant keratin lines. Our findings strengthen the case for the expression of mutant keratin proteins inducing physiological stress, and this intrinsic stress may affect the cell responses to secondary stresses in patients' skin.

Osmotic upshift transiently inhibits uptake via ABC transporters in gram-negative bacteria

ATP-binding cassette transporters from several rhizobia and Salmonella enterica serovar Typhimurium, but not secondarily coupled systems, were inhibited by high concentrations (100 to 500 mM) of various osmolytes, an effect reversed by the removal of the osmolyte. ABC systems were also inactivated in isolated pea bacteroids, probably due to the obligatory use of high-osmolarity isolation media. Measurement of nutrient cycling in isolated pea bacteroids is impeded by this effect.

Osmotic pressure and aggregate shape in BSA/poly (ethylene glycol)-lipid/Dextran solutions

In this work we study the colloidal osmotic pressure (COP) and aggregate shape in phosphate saline buffer solutions (PH 7.4) containing bovine serum albumin (BSA), poly(ethylene glycol) lipid (PEG(2000)-PE) and Dextran (Dx). Dx was added to the BSA/PEG(2000)-PE system in order to increase the COP of the solution to levels comparable to the COP of healthy adults, with the aim of using the solution as a blood COP regulator. Dynamic light scattering and small angle X-ray scattering results shown the formation of BSA/PEG(2000)-PE/Dx aggregates in the solution. Osmometry results shown that the addition of Dx to the BSA/PE2000-PE system could successfully increase the COP, through the formation of BSA/PEG(2000)-PE/Dx aggregates. The BSA/PEG(2000)-PE/Dx solutions attained COP= 15 mm Hg, representing 60% of COP measured for healthy adults. (c) 2008 Elsevier B.V. All rights reserved.

Microblock ionomers: a new concept in high temperature, swelling-resistant membranes for PEM fuel cells

A novel series of polyaromatic ionomers with similar equivalent weights but very different sulphonic acid distributions along the ionomer backbone has been designed and prepared. By synthetically organising the sequence-distribution so that it consists of fully defined ionic segments (containing singlets, doublets or quadruplets of sulphonic acid groups) alternating strictly with equally well-defined nonionic spacer segments, a new class of polymers which may be described as microblock ionomers has been developed. These materials exhibit very different properties and morphologies from analogous randomly substituted systems. Progressively extending the nonionic spacer length in the repeat unit (maintaining a constant equivalent weight by increasing the degree of sulphonation. of the ionic segment) leads to an increasing degree of nanophase separation between hydrophilic and hydrophobic domains in these materials. Membranes cast from ionomers with the more highly phase-separated morphologies show significantly higher onset temperatures for uncontrolled swelling in water. This new type of ionomer design has enabled the fabrication of swelling-resistant hydrocarbon membranes, suitable for fuel cell operation, with very much higher ion exchange capacities (>2 meq g(-1)) than those previously reported in the literature. When tested in a fuel cell at high temperature (120 degrees C) and low relative humidity (35% RH), the best microblock membrane matched the performance of Nafion 112. Moreover, comparative low load cycle testing of membrane -electrode assemblies suggests that the durability of the new membranes under conditions of high temperature and low relative humidity is superior to that of conventional perfluorinated materials.

Stress generation in the tension wood of poplar is based on the lateral swelling power of the G-layer

The mechanism of active stress generation in tension wood is still not fully understood. To characterize the functional interdependency between the G-layer and the secondary cell wall, nanostructural characterization and mechanical tests were performed on native tension wood tissues of poplar (Populus nigra x Populus deltoids) and on tissues in which the G-layer was removed by an enzymatic treatment. In addition to the well-known axial orientation of the cellulose fibrils in the G-layer, it was shown that the microfibril angle of the S2-layer was very large (about 36 degrees). The removal of the G-layer resulted in an axial extension and a tangential contraction of the tissues. The tensile stress-strain curves of native tension wood slices showed a jagged appearance after yield that could not be seen in the enzyme-treated samples. The behaviour of the native tissue was modelled by assuming that cells deform elastically up to a critical strain at which the G-layer slips, causing a drop in stress. The results suggest that tensile stresses in poplar are generated in the living plant by a lateral swelling of the G-layer which forces the surrounding secondary cell wall to contract in the axial direction.

Assessment of wheat cultivars for drought tolerance via osmotic stress imposed at early seedling growth stages

A study was conducted in the Department of Plant Breeding and Genetics,Sindh Agriculture University, Tandojam, Pakistan during the year 2009. Sixteen spring wheat cultivars (Triticum aestivum L.) were screened under osmotic stress with three treatments i.e. control-no PEG (polyethylene glycol), 15 percent and 25 percent PEG-6000 solution. The analysis of variance indicated significant differences among treatments for all seedling traits except seed germination percentage. Varieties also differed significantly in germination percentage, coleoptile length, shoot root length, shoot weight, root/shoot ratio and seed vigour index. However, shoot and root weights were non-significant. Significant interactions revealed that cultivars responded variably to osmotic stress treatments; hence provided better opportunity to select drought tolerant cultivars at seedling growth stages. The relative decrease over averages due to osmotic stress was 0.8 percent in seed germination, 53 percent in coleoptile length 62.9 percent in shoot length, 74.4 percent in root length, 50.6 percent in shoot weight, 45.1 percent in root weight, 30.2 percent in root/shoot ratio and 68.5 percent in seed vigour index. However, relative decrease of individual variety for various seedling traits could be more meaningful which indicated that cultivar TD-1 showed no reduction in coleoptile length, while minimum decline was noted in Anmol. For shoot length, cultivar Sarsabz expressed minimum reduction followed by Anmol. However, cultivars Anmol, Moomal, Inqalab-91, and Pavan gave almost equally lower reductions for root length suggesting their higher stress tolerance. In other words, cultivars Anmol, Moomal, Inqalab-91, Sarsabz, TD-1, ZA-77 and Pavan had relatively longer coleoptiles, shoots and roots, and were regarded as drought tolerant. Correlation coefficients among seedlings traits were significant and positive for all traits except germination percentage which had no significant correlation with any of other trait. The results indicated that increase in one trait may cause simultaneous increase in other traits; hence selection for any of these seedling attributes will lead to develop drought tolerant wheat cultivars.

Structural parameters of polyacrylamide hydrogels obtained by the Equilibrium Swelling Theory

In this paper, the synthesis and structural characterization of a series of polyacrylamide hydrogels with different degrees of reticulation are reported. Although the Equilibrium Swelling Theory was recognized as a simple and reliable tool for the determination of structural hydrogels network parameters like equilibrium degree of swelling, cross-link ratio and mesh size, this is the first application of this methodology for polyacrylamide hydrogels. By changing the total monomer content in the synthesis solution (%T) from 5 to 30%, at a fixed value of cross-linker content in the total monomer amount (%C) of 5%, the final parameter obtained, the mesh size, can be tuned from 2 to 0.3 nm. It was also possible to change the mesh size (0.19-0.35) by varying %C from 5 to 12% (at %T = 20%). Scanning Electron Microscopy images for the most different formulations are shown and corroborate data obtained from the theory. (c) 2008 Elsevier Ltd. All rights reserved.

Cellulose swelling by aprotic and protic solvents: What are the similarities and differences?

The swelling of microcrystalline, native and mercerized cotton and eucalyptus celluloses by 16 aprotic solvents was investigated. The number of moles of solvent/anhydroglucose unit, nSw, correlates well with solvent molar volume, basicity and dipolarity/polarizability. Swelling is sensitive to cellulose crystallite size, surface area and the presence of its chains in parallel or anti-parallel arrangements. Use of solvatochromic parameters is a superior alternative to the use of other descriptors, such as Hildebrand`s solubility parameters and Gutmann`s donor numbers. The calculated nSw for 28 protic and aprotic solvents correlated well with their experimental counterparts, although hydrogen bond donation by the solvent was not included.

Cellulose swelling by protic solvents: which properties of the biopolymer and the solvent matter?

The question posed in the title has been addressed by studying the swelling of celluloses at 20 C by twenty protic solvents, including water; linear- and branched-chain aliphatic alcohols; unsaturated aliphatic alcohols, and alkoxyalcohols. The biopolymers investigated included microcrystalline cellulose, MC, native and never-dried mercerized cotton cellulose, cotton and M-cotton, and native and never-dried mercerized eucalyptus cellulose, eucalyptus and M-eucalyptus, respectively. In most cases, better correlations with the physico-chemical properties of the solvents were obtained when the swelling was expressed as number of moles of solvent/anhydroglucose unit, nSw, rather than as % increase in sample weight. The descriptors employed in these correlations included, where available, Hildebrand`s solubility parameters, Gutmann`s acceptor and donor numbers, solvent molar volume, V(S), as well as solvatochromic parameters. The latter, employed for the first time for correlating the swelling of biopolymers, included empirical solvent polarity, E(T)(30), solvent ""acidity"", alpha(S), ""basicity"", beta(S), and dipolarity/polarizability, pi(S)*, respectively. Small regression coefficients and large sums of the squares of the residues were obtained when values of nSw were correlated with two solvent parameters. Much better correlations were obtained with three solvent parameters. The most statistically significant descriptor in the correlation equation depends on the cellulose, being pi(S)* for MC, cotton, and eucalyptus, and V(S) for M-cotton and M-eucalyptus. The best correlations were obtained with the same set of four parameters for all celluloses, namely, solvent pKa (or alpha(S)) beta(S), pi(S)*, and V(S), respectively. These results indicate that the supra-molecular structure of the biopolymer, in particular the average sizes of crystallites and micro-pores, and the presence of its chains in parallel (cellulose I) or anti-parallel (cellulose II) arrangements control its swelling. At least for the present biopolymer/solvent systems, use of solvatochromic parameters is a superior alternative to Hildebrand`s solubility parameters and/or Gutmann`s acceptor and donor numbers. The relevance of these results to the accessibility of the hydroxyl groups of cellulose, hence to its reactivity, is briefly discussed.

Optimization of osmotic dehydration of papaya followed by air-drying

FERNANDES, Fabiano A. N. et al. Optimization of Osmotic Dehydration of Papaya of followed by air-drying. Food Research Internation, v. 39, p. 492-498, 2006.