scholarly journals Fractionation of a hyaluronic acid preparation in a density gradient. Some properties of the hyaluronic acid

1968 ◽  
Vol 109 (1) ◽  
pp. 43-50 ◽  
Author(s):  
Panee Silpananta ◽  
J. R. Dunstone ◽  
A. G. Ogston
Keyword(s):  
Hyaluronic Acid ◽  
Density Gradient ◽  
Chondroitin Sulphate ◽  
Sedimentation Equilibrium ◽  
Partial Specific Volume ◽  
Low Velocity ◽  
Molecular Configuration ◽  
Caesium Chloride ◽  
A Value ◽  
Couette Viscometer

1. Hyaluronic acid was isolated from ox synovial fluid by sedimentation equilibrium in a caesium chloride density gradient (Silpananta, Dunstone & Ogston, 1967). The product was almost free from chondroitin sulphate and from protein. 2. Its composition did not differ significantly from that of the carbohydrate part of the protein-containing material isolated by filtration. Its physicochemical properties and molecular configuration were similar, except for its viscosity, which showed markedly reduced concentration-dependence and shear-dependence. This suggests that the associated protein tends to form links between molecules of hyaluronic acid. 3. The accurate measurement of viscosity at very low velocity gradient, by use of the damping of oscillations in a Couette viscometer, is described. 4. A method is described for measuring, approximately, the thermodynamic non-ideality of a solute from the shape of its schlieren curve at sedimentation equilibrium in a density gradient. 5. A value for the partial specific volume of hyaluronic acid in dilute salt solution was calculated from its isopycnic density in a caesium chloride gradient.

scholarly journals Isolation and properties of hyaluronic acid from bovine heart valves

1969 ◽  
Vol 113 (3) ◽  
pp. 559-563 ◽  
Author(s):  
F. A. Meyer ◽  
B. N. Preston ◽  
D. A. Lowther
Keyword(s):  
Hyaluronic Acid ◽  
Synovial Fluid ◽  
Heart Valves ◽  
Glucuronic Acid ◽  
Molecular Size ◽  
Sedimentation Equilibrium ◽  
Molar Ratio ◽  
Soluble Extract ◽  
Caesium Chloride ◽  
Bovine Heart

1. A soluble extract of bovine heart valves was obtained after the tissue had been pulverized at liquid-nitrogen temperatures in a mill. 2. Hyaluronic acid was isolated from the crude extract by sedimentation equilibrium in a caesium chloride density gradient (Franek & Dunstone, 1966). 3. Analysis of the product indicated that it contained 15% of protein and the molar ratio of glucuronic acid to glucosamine was 1·27. 4. Its physicochemical properties, as determined by lightscattering, viscosity and sedimentation studies, suggested that its molecular size and configuration were similar to those of hyaluronic acid isolated from ox synovial fluid (Preston, Davies & Ogston, 1965).

scholarly journals The use of equilibrium-density-gradient methods for the preparation and characterization of blood-group-specific glycoproteins

1970 ◽  
Vol 117 (5) ◽  
pp. 879-891 ◽  
Author(s):  
J. M. Creeth ◽  
M. A. Denborough
Keyword(s):  
Molecular Weight ◽  
Density Gradient ◽  
Blood Group ◽  
Buoyant Density ◽  
Gradient Methods ◽  
Apparent Molecular Weight ◽  
Sedimentation Equilibrium ◽  
Equilibrium Density ◽  
Caesium Chloride ◽  
Selective Solvation

1. The method of sedimentation equilibrium in a gradient of caesium chloride has been applied to the preparation of blood-group-specific glycoproteins from human ovarian-cyst fluids: it is shown that virtually complete separation from contaminating protein is easily accomplished in a single step. 2. The glycoproteins isolated in this way have been characterized by analytical density-gradient experiments in both caesium chloride and caesium sulphate and values of the buoyant density, selective solvation and apparent molecular weight have been obtained. 3. In some cases, materials prepared from the same cysts by solvent extraction methods have also been characterized in these terms. 4. The selective solvation values are about 0.1 and 0.5g of water/g of glycoprotein in caesium chloride and caesium sulphate respectively. 5. The apparent molecular-weight values are much lower than the weight-average molecular weights, and it is shown that the origin of the discrepancy is heterogeneity in density of the glycoproteins. 6. Some sources of error in the interpretation of density-gradient schlieren patterns are examined.

scholarly journals Studies on the composition of egg-white ovomucin

1971 ◽  
Vol 121 (3) ◽  
pp. 537-547 ◽  
Author(s):  
D. S. Robinson ◽  
J. B. Monsey
Keyword(s):  
Density Gradient ◽  
Guanidine Hydrochloride ◽  
Egg White ◽  
Sedimentation Equilibrium ◽  
Mild Acid Hydrolysis ◽  
Acetylneuraminic Acid ◽  
Glycoprotein Complex ◽  
Caesium Chloride ◽  
Hydrolysis Of ◽  
Mild Acid

1. Purified ovomucin was isolated as an insoluble glycoprotein complex from thick egg white. 2. A homogeneous glycoprotein, designated α-ovomucin, of molecular weight 210000 and containing N-acetylglucosamine (6.7%, w/w), N-acetylgalactosamine (0.6%, w/w), galactose (1.8%, w/w), mannose (4.6%, w/w), N-acetylneuraminic acid (1.0%, w/w) and sulphate (0.7%, w/w), was isolated from preparations of reduced ovomucin by sedimentation equilibrium in a density gradient of caesium chloride formed in the presence of 4m-guanidine hydrochloride. 3. A carbohydrate-rich fraction, designated β-ovomucin (which is homogeneous by sedimentation-velocity analysis in 5m-guanidine hydrochloride but which is heterogeneous by analytical sedimentation equilibrium in a density gradient of caesium chloride in the presence of 4m-guanidine hydrochloride), containing N-acetylglucosamine (11.0%, w/w), N-acetylgalactosamine (8.7%, w/w), galactose (19.2%, w/w), mannose (4.1%, w/w), N-acetylneuraminic acid (13.8%, w/w) and sulphate (2.7%, w/w), was also obtained from preparations of reduced ovomucin by the density-gradient method. 4. Mild acid hydrolysis of the unfractionated ovomucin complex showed that N-acetylneuraminic acid occupied a terminal position of the oligosaccharide chains. 5. Alkaline β-elimination reactions with the unfractionated ovomucin complex indicated that N-acetylgalactosamine was linked by alkali-labile bonds to hydroxy amino acids.

scholarly journals Protein Associated with Hyaluronic Acid in Ox Synovial Fluid

1969 ◽  
Vol 22 (4) ◽  
pp. 1031 ◽  
Author(s):  
Panee Silpananta ◽  
JR Dunstone ◽  
AG Ogston
Keyword(s):  
Hyaluronic Acid ◽  
Synovial Fluid ◽  
Density Gradient ◽  
Caesium Chloride ◽  
Equilibrium Sedimentation ◽  
Molecular Sizes

Protein which is associated with hyaluronic acid, prepared in a physico-chemically unchanged state from ox synovial fluid by ultrafiltration, has been separated by preparative equilibrium sedimentation in a caesium chloride density gradient and by chromatography on DEAE-Sephadex. Earlier work has been confirmed by showing that the protein consists of at least two chemically distinct lipoprotein, components. It has also been confirmed that its removal from the hyaluronic acid causes marked changes of viscosity, without the occurrence of degradation of the hyaluronic acid. The separated proteins appeared in a number of differently aggregated forms, and this fact has prevented our drawing conclusions about their molecular sizes in synovial fluid.

scholarly journals The state of aggregation of red deer (Cervus elaphus L.) β-lactoglobulin preparations near neutral pH

1977 ◽  
Vol 167 (1) ◽  
pp. 45-51 ◽  
Author(s):  
E I McDougall ◽  
J C Stewart
Keyword(s):  
Cervus Elaphus ◽  
Red Deer ◽  
Guanidine Hydrochloride ◽  
Second Virial Coefficient ◽  
The State ◽  
Sedimentation Equilibrium ◽  
Partial Specific Volume ◽  
A Value ◽  
Recovered Material ◽  
Beta Lactoglobulin

1. The state of aggregation of four red-deer (Cervus elaphus L.) beta-lactoglobulin preparations and a control ox beta-lactoglobulin A preparation was studied by sedimentation-equilibrium experiments at pH 6.5 and 20 degrees C. 2. Three of the deer preparations and the ox control each behaved as a monomer-dimer system, with a value of log K (where K is the association constant in litres/mol) in the range 5.4-5.5. 3. When one of these deer preparations was examined in the presence of dithiothreitol, log K appeared to decrease to 4.5.4. One deer preparation, comprising recovered material, appeared to have undergone irreversible changes and to behave like a non-equilibrating system containing monomer, dimer and trimer. 5. The sedimentation-equilibrium properties of the deer monomer was studied in 6M-guanidine hydrochloride at pH 7.0; the mol.wt. was 17600, the second virial coefficient was 3.4 × 10(-3) ml - mol - g-2, and the apparent partial specific volume 0.724 ml/g, a value indicating an appreciable decrease in volume on dissociation and denaturation.

scholarly journals Isolation and properties of chondroitin sulphates from bovine heart valves

1970 ◽  
Vol 118 (4) ◽  
pp. 595-601 ◽  
Author(s):  
D. A. Lowther ◽  
B. N. Preston ◽  
F. A. Meyer
Keyword(s):  
Heart Valves ◽  
Chondroitin Sulphate ◽  
Sedimentation Equilibrium ◽  
Structural Studies ◽  
Variable Size ◽  
Keratan Sulphate ◽  
Molecular Weights ◽  
Caesium Chloride ◽  
Bovine Heart ◽  
Basic Model

1. Several protein–polysaccharides were isolated from the soluble extracts of bovine heart valves by sedimentation equilibrium in a caesium chloride density gradient (Meyer, Preston & Lowther, 1969). 2. Compositional and structural studies indicated that the polysaccharide moiety was chondroitin sulphate. Differences in the protein content of the products were observed. There was no evidence suggesting the presence of keratan sulphate. 3. Sedimentation studies indicated that the molecular weights of the samples were between 4.2×104 and 6.5×104. The results are discussed in terms of a basic model for the protein–polysaccharides of two polysaccharide chains linked by a protein of variable size.

Healing of Circular Defects in the Rabbit Medial Meniscus Can Occur Spontaneously and Is not Improved by Intra-Articular Hyaluronic Acid

1996 ◽  
Vol 09 (02) ◽  
pp. 60-5 ◽  
Author(s):  
N. Hope ◽  
P. Ghosh ◽  
S. Collier
Keyword(s):  
Hyaluronic Acid ◽  
Medial Meniscus ◽  
Chondroitin Sulphate ◽  
Rabbit Model ◽  
Type Ii Collagen ◽  
Type Ii ◽  
Keratan Sulphate ◽  
Meniscal Healing ◽  
Meniscus Healing ◽  
Made In

SummaryThe aim of this study was to determine the effects of intra-articular hyaluronic acid on meniscal healing. Circular defects, 1.0 mm in diameter, were made in the anterior third of the medial meniscus in rabbits. In one joint, 0.4 ml hyaluronic acid (Healon®) was instilled, and in the contralateral (control) joint, 0.4 ml Ringer’s saline. Four rabbits were killed after four, eight and 12 weeks and the menisci examined histologically. By eight weeks most of the lesions had healed by filling with hyaline-like cartilage. Healing was not improved by hyaluronic acid treatment. The repair tissue stained strongly with alcian blue, and the presence of type II collagen, keratan sulphate, and chondroitin sulphate was demonstrated by immunohistochemical localisation. In contrast to the circular defects, longitudinal incisions made in the medial menisci of a further six rabbits did not show any healing after 12 weeks, indicating that the shape of the lesion largely determined the potential for healing.The effect of hyaluronic acid on meniscal healing was tested in a rabbit model. With one millimeter circular lesions in the medial meniscus, healing by filling with hyalinelike cartilage was not significantly affected by the application of hyaluronic acid intra-articularly at the time of surgery, compared to saline controls, as assessed histologically four, eight and 12 weeks after the operation.

scholarly journals Isolation of human synovial-fluid hyaluronate by density-gradient ultracentrifugation and evaluation of its protein content

1972 ◽  
Vol 126 (5) ◽  
pp. 1073-1080 ◽  
Author(s):  
Irwin Scher ◽  
David Hamerman
Keyword(s):  
Synovial Fluid ◽  
Density Gradient ◽  
Density Gradient Ultracentrifugation ◽  
Proteolytic Digestion ◽  
Peptide Fragments ◽  
Guanidinium Chloride ◽  
Caesium Chloride ◽  
Normal Human ◽  
Protein Moiety ◽  
Human Synovial Fluid

1. A compound of hyaluronate and protein, called hyaluronate–protein was isolated from pooled human synovial fluids by caesium chloride density-gradient ultracentrifugation. 2. The isolated hyaluronate–protein was labelled with [125I]iodide and the following studies were done. (a) Ultracentrifugation in caesium chloride showed that the protein moiety (125I counts) and hyaluronate (hexuronate) sedimented together in the middle of the gradient. (b) The labelled hyaluronate–protein was treated with trypsin, and ultracentrifugation showed that peptide fragments (125I counts) were dispersed throughout the gradient, indicating proteolytic digestion. Hyaluronate sedimented in the middle of the gradient. (c) The labelled hyaluronate–protein was digested with streptococcal hyaluronidase, and ultracentrifugation showed that hyaluronate fragments were dispersed throughout the gradient, indicating digestion of the polysaccharide. The protein moiety, without attached hyaluronate, now sedimented at the top of the gradient. (d) Ultracentrifugation of labelled hyaluronate–protein in 4m-guanidinium chloride showed that protein and hyaluronate sedimented together. 3. These studies confirm that hyaluronate is combined with a small quantity of protein in normal human synovial fluid. A mild method for the rapid isolation of hyaluronate–protein in good yield is described.

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