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Inhibition of gamma-irradiation induced adhesion molecules and NO production by alginate in human endothelial cells.

Inflammation is a frequent radiation-induced reaction following therapeutic irradiation. Treatment of human umbilical endothelial cells (HUVEC) with gamma-irradiation (gammaIR) induces the expression of adhesion proteins such as intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin. Since the upregulation of these proteins on endothelial cell surface has been known to be associated with inflammation, interfering with the expression of adhesion molecules is an important therapeutic target. In the present study, we demonstrate that high mannuronic acid-containing alginate (HMA) inhibits gammaIR induced expression of ICAM-1, VCAM-1, and E-selectin on HUVEC in a dose dependent manner. HMA also inhibited gammaIR induced production of Nitric oxide (NO). These data suggest that HMA has therapeutic potential for the treatment of various inflammatory disorder associated with an increase of endothelial leukocyte adhesion molecules.

Alginates↗

Modulation of TNF-alpha-induced ICAM-1 expression, NO and H2O2 production by alginate, allicin and ascorbic acid in human endothelial cells.

Plant nutrients are believed to provide protection against various diseases including inflammation. Since interactions of the cell adhesion molecules are known to play important roles in mediating inflammation, inhibiting adhesion protein upregulation is a possible therapeutic target. In this study, the interacellular adhesion molecule-1 (ICAM-1) was induced in human umbilical endothelial cells (HUVECs) after stimulation with TNF-alpha. In addition, alginate, ascorbic acid and allicin were demonstrated to inhibit the TNF-alpha induced expression of ICAM-1 on the HUVECs in a dose-dependent manner. These compounds also inhibited the production of NO and H2O2 induced by TNF-alpha, which suggests that the inhibition of ICAM-1 expression by the three compounds may be due to the modulated production of the reactive oxygen/nitrogen components. Overall, these results indicate that these dietary components have a therapeutic potential in the treatment of various inflammatory disorders associated with an increase in endothelial leukocyte adhesion molecules.

Alginates↗

Immunomodulatory function of murine NK cell activity by alginate.

The in vivo immunomodulatory function of the activity of murine natural killer (NK) cells induced by high mannuronic acid-containing alginate (HMA) was examined. HMA was injected i.p at doses of 25 and 100 mg/kg. The NK activity was 3 times higher with 100 mg/kg HMA than the baseline. In addition, in vitro studies of splenocytes cultured with HMA for 20 h showed a significant increase in NK activity at E:T ratio of 100:1; a 160% and 210% increase at 10 and 100 microg/mL, respectively. There was a six fold increase in interferon-gamma production in a postculture of splenocytes with 100 microg/mL HMA. HMA had no suppressive effects on the lymphocyte function in the presence or absence of mitogens. This suggests that HMA is useful in cancer immunotherapy.

Alginates↗

[Adult human chondrocytes in alginate culture. Preservation of the phenotype for further use in transplantation models].

Successful and cartilage-specific cultivation of chondrocytes requires a stable phenotype during the in vitro culture period. This is based on a differentiated extracellular matrix synthesis. The alginate system as a three-dimensional support is a useful system to culture chondrocytes and to analyze the biochemical processes in this system. Talar cartilage from the talocrural joints of 40 different donors were obtained through the Regional Organ Bank of Illinois within 24 hours of death. In 65% of the tissues the cartilage was classified as being undamaged. In these studies we were interested in the results of short-term culture over 14 days. Cell proliferation, total collagen content and total proteoglycan content were measured in the different matrix compartments and were visualized by histology and immunohistochemistry. Already after 7 days in culture the adult human chondroctes looked intact and formed a stable and cell-associated cartilage-specific extracellular matrix in the presence of 10% calf serum. This could be also demonstrated in the presence of IGF-I. With regards to the collagen content IGF-I at a concentration of 50 ng/ml seemed to induce an equal effect to 10% serum; with regards to the proteoglycan content IGF-I at a concentration of 20 ng/ml was equivalent. These encouraging preliminary results may lead to a new approach in tissue engineering for chondrocyte transplantation in combination with their extracellular matrix.

Adult↗

Immobilization of Candida krusei cells producing phytase in alginate gel beads: an application of the preparation of myo-inositol phosphates.

Cells of Candida krusei capable of producing phytase were immobilized in Ca-alginate gel beads and used for the preparation of myo-inositol phosphates. The immobilization yield was increased about 5-fold after the beads were treated for 96 h at pH 4.0, 4 degrees C. The increased yield was retained, even after 1 month, when the cells were kept at this temperature and pH. No shift in the pH optima of phytase of the immobilized cells was observed, compared with that of free cells. However, the optimum temperature for the enzyme of the immobilized cells was 55 degrees C, which was 15 degrees C higher than that of free cells. The degradation characteristics of the phytate in immobilized cells packed in a glass column (i.d. 1.2 cm, length 20 cm) were investigated. The variation in the composition of the products results from a change in the flow rate of phytate solution (5 mM). At a flow rate of 1.30 ml/min, a mixture of myo-inositol-2-monophosphate, myo-inositol-1,2,5-triphosphate and myo-inositol-1,2,5,6-tetrakisphosphate was produced, in which the latter two were physiologically active. Also, it was found by NMR analysis that the enzyme of this strain produced only one isomer of each of the inositol phosphates, with the exception of myo-inositol pentakisphosphate. Therefore, the pure isomers were easily isolated using ion-exchange chromatography.

6-Phytase↗

Silica-alginate composites for microencapsulation.

Optimisation of membrane properties of alginate microcapsules is a key factor for the application of microencapsulation techniques to bioartificial organ elaboration. Coacervation and layer-by-layer processes involving additional biopolymers have been extensively studied. Recently, the use of silica as a membrane-forming agent was investigated. This approach was rendered possible by the development of biocompatible routes to silica formation. The composites exhibit enhanced mechanical and thermal stability as well as suitable diffusion properties. Moreover, encapsulated enzymes and cells retain their biological activities. Similarly, silica can be associated to many other biopolymers, opening a promising route for new biocomposites design and biotechnology applications.

Alginates↗

Characterisation of the shrinkage of calcium alginate gel membrane with immobilised Lactobacillus rhamnosus.

To quantify the shrinkage of calcium alginate gel membrane as a support matrix for immobilising cells during the fermentation of Lactobacillus rhamnosus, factors including time, pH, membrane thickness, and the concentrations of immobilised cells, lactic acid, glucose, and calcium chloride were examined by statistical experimental design. A Plackett-Burman design was used for the first screening experiment to identify the important factors which caused the divergent effects. Uniform Design, a powerful modelling design technique, was thus chosen to design the modelling experiments. Through a non-linear step-wise regression analysis, the predictive mathematical model of the shrinkage in membrane thickness was established and the significant main effects and two-factor interactions were identified. However, no significant model equations could be obtained for the shrinkage in area and volume of gel membranes. The methodology developed can be extrapolated to the quantitative characterisation of shrinkage in other immobilised gel matrices, which will be very useful in mathematical modelling, design, operation and scale-up of gel immobilised cell systems.

Alginates↗

Differential rhizosphere establishment and cyanide production by alginate-formulated weed-deleterious rhizobacteria.

The effects of Pseudomonas putida ATH2-1RI/9 and Acidovorax delafieldii ATH2-2RS/1 on rhizosphere colonization, cyanide production, and growth of velvetleaf and corn was examined. When formulated in alginate beads and inoculated onto velvetleaf and corn plants (10(9) CFU/plant), only P. putida ATH2-1RI/9 consistently reduced velvetleaf growth. Neither isolate inhibited corn growth. Interestingly the levels of P. putida ATH2-1RI/9 in the velvetleaf rhizosphere were 1000-fold higher (7 x 10(7) CFU/g root) than the A. delafieldii ATH2-2RS/1 populations. Cyanide (53-68 mM/g root) was recovered from the P. putida ATH2-1RI/9-inoculated velvetleaf plants. In contrast both A. delafieldii ATH2-2RS/1 and P. putida ATH2-1RI/9 colonized the corn rhizosphere to the same extent (1-5 x 10(7) CFU/g root), producing 1 mM and 14 mM/g root respectively. These results suggest that bacterial formulation methods can influence the effectiveness of deleterious rhizobacteria in reducing weed growth.

Alginates↗

Immobilization of urease by using chitosan-alginate and poly(acrylamide-co-acrylic acid)/kappa-carrageenan supports.

Jack bean urease (urea aminohydrolase, E.C. 3.5.1.5) was entrapped into chitosan-alginate polyelectrolyte complexes (C-A PEC) and poly(acrylamide-co-acrylic acid)/kappa-carrageenan (P(AAm-co-AA)/carrageenan) hydrogels for the potential use in immobilization of urease, not previously reported. The effects of pH, temperature, storage stability, reuse number, and thermal stability on the free and immobilized urease were examined. For the free and immobilized urease into C-A PEC and P(AAm-co-AA)/carrageenan, the optimum pH was found to be 7.5 and 8, respectively. The optimum temperature of the free and immobilized enzymes was also observed to be 55 and 60 degrees C, respectively. Michaelis-Menten constant (K(m)) values for both immobilized urease were also observed smaller than free enzyme. The storage stability values of immobilized enzyme systems were observed as 48 and 70%, respectively, after 70 days. In addition to this, it was observed that, after 20th use in 5 days, the retained activities for immobilized enzyme into C-A PEC and P(AAm-co-AA)/carrageenan matrixes were found as 55 and 89%, respectively. Thermal stability of the free urease was also increased by a result of immobilization.

Acrylamides↗

Morphology and metabolism of Ba-alginate encapsulated hepatocytes with galactosylated poly(allyl amine) and poly(vinyl alcohol) as extracellular matrices.

Lactobionic acid, bearing a beta -galactose group, was coupled with poly(allyl amine) to provide synthetic extracellular matrices together with poly(vinyl alcohol) (PVA). The hepatocytes were encapsulated in Ba-alginate capsules with galactosylated poly(allyl amine) (GA) and PVA as extracellular matrices. From microscopic observation, it was revealed that the microcapsule prepared has a highly porous structure with interconnected pores and pore sizes ranging between 50-150 nm on both the surface and the cross-section. It was found, from the permeability experiment of microcapsules using FITC-dextrans with different molecular weights, that the capsule has a molecular weight cut off (MWCO) of 120 kDa, showing the potential that it can function as an immunoprotecting wall. The hepatocytes, cultured with GA and PVA in the core of the microcapsule, rapidly aggregated within a day, thus resulting in good metabolic functions such as albumin synthesis and ammonia removal.

Albumins↗

Dimensional changes of alginate dental impression materials.

The weight loss and corresponding dimensional changes of two dental alginate impression materials have been studied. The weight loss kinetics indicate this to be a diffusion controlled process, but with a boundary condition at the surface of the concentration decreasing exponentially with time. This is in marked contrast to most desorption processes, where the surface concentration becomes instantaneously zero. The appropriate theory has been developed for an exponential boundary condition, and its predictions compared with experimental data; the agreement was satisfactory. The diffusion coefficients for two thicknesses of the same material were not identical as predicted by theory; the possible reasons for this are discussed.

Alginates↗

Control of alginate synthesis in Pseudomonas aeruginosa: regulation of the algR1 gene.

The triggering of mucoidy (formation of the exopolysaccharide alginate) in Pseudomonas aeruginosa is accomplished primarily in Cystic Fibrosis lung environment through activation of the promoter of a gene algD, which encodes GDP-mannose dehydrogenase, by the product of a regulatory gene algR1. Both algD and algR1 promoter regions have significant homology, including the presence of sequences recognized by the RNA polymerase sigma 54. We demonstrate that the algR1 promoter is partly constitutive and its activation, similar to that of algD, is dependent on high osmolarity as well as the presence of its own gene product and is repressed by high concentrations of AlgR1. The RpoN sigma factor also plays a critical role in the transcription of both algD and algR1 genes.

Alginates↗

The molecular size and shape of xanthan, xylinan, bronchial mucin, alginate, and amylose as revealed by electron microscopy.

Electron microscopy of some selected, vacuum-dried and rotary-shadowed, polyelectrolytic polysaccharides and glycoproteins adsorbed to mica indicates that this technique can yield reliable information about polymer conformation for chains with persistence lengths q exceeding about 10 nm. Statistical analyses of the local polymer tangent-direction yield q = 150 nm for double-stranded xanthan, q = 60 nm for single-stranded xanthan, q = 45 nm for xylinan, q = 16 nm for alginate (90% beta-D-mannuronic acid), and q = 15 nm for human-bronchial mucin. These values are all in adequate agreement with values of q obtained by using other techniques. Amylose, on the other hand, appears as non-randomly aligned chains. The observed contour lengths of amylose indicate a mass per unit length of 1440 dalton/nm, consistent with a pseudo-helical conformation.

Alginates↗

Structural determination of alginic acid and the effects of calcium binding as determined by high-field n.m.r.

The nature of the solution conformations of the alginic acid components D-mannuronan (poly-ManA) and L-guluronan (poly-GulA) from Azotobacter vinelandii were investigated by both one- and two-dimensional n.m.r. methods. Unequivocal proton assignments for both polymers as well as their constituent monomer units were made based on chemical-shift theory, coupling constant analysis, and nuclear Overhauser enhancement measurements. These data were used to investigate the interactions of poly-GulA and poly-ManA with Ca2+ ion in aqueous medium. Based on relative crosspeak integrals measured in two-dimensional phase-sensitive NOESY spectra of free and calcium-bound polymer, a model for calcium binding is proposed.

Alginates↗

Nitric oxide and proteoglycan biosynthesis by human articular chondrocytes in alginate culture.

Interleukin-1 alpha and beta induced the production of large amounts of nitric oxide by normal, human articular chondrocytes in alginate culture; at the same time the biosynthesis of proteoglycan was strongly suppressed. In a dose-dependent manner, NG-monomethyl-L-arginine both inhibited nitric oxide formation and relieved the suppression of proteoglycan synthesis. However concentrations of NG-monomethyl-L-arginine which completely prevented nitric oxide production only partially restored proteoglycan biosynthesis, even at low doses of interleukin-1 where suppression of proteoglycan synthesis was modest. The organic donor of nitric oxide, S-nitrosyl-acetyl-D,L- penicillamine also inhibited proteoglycan biosynthesis, but not as extensively as interleukin-1. These data suggest that interleukin-1 suppresses synthesis of the cartilaginous matrix through more than one mechanism, at least one of which is dependent upon the production of nitric oxide.

Adult↗

Diffusion of proteases in calcium alginate beads.

Diffusion of proteases from Bacillus subtilis and Serratia marcescens within calcium alginate beads has been assayed, and the experimental data fitted into a mathematical model for diffusion into a finite volume liquid medium. Values of effective diffusivity were calculated for the studied proteases and compared with the available data in the literature for molecules of lower molecular weight.

Alginates↗

Utilization of DNA as functional materials: preparation of filters containing DNA insolubilized with alginic acid gel.

Thin films composed of DNA and alginic acid were prepared by casting their mixed solution on glass plate followed by coagulation with aqueous solution of calcium chloride. DNA could be conveniently insolubilized by this method. DNA in the films adsorbed intercalating materials, such as ethidium bromide. This phenomenon was successfully applied to the preparation of filters for the selective removal or accumulation of harmful intercalating pollutants.

Adsorption↗

Sorbsan calcium alginate fibre dressings in footcare.

This report outlines the successful application of Sorbsan-calcium alginate fibre dressings in footcare. This biomaterial has been found in clinical trials to have quite outstanding merit and can be used for sinus drainage and in the treatment of fissures, hypergranulation tissue, interdigital maceration, heloma molle and other lesions. Sorbsan bonds with and aids natural healing scabs and effects drainage of moist sites thus assisting in creating an environment conducive to the healing process. Similar promise has been demonstrated for this material in ongoing trials in hospitals in Sunderland where Sorbsan has been used effectively in the treatment of diabetic and trophic ulcers.

Adolescent↗