Isolation and properties of chondroitin sulphates from bovine heart valves

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.

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Isolation and properties of hyaluronic acid from bovine heart valves

Biochemical Journal ◽

10.1042/bj1130559 ◽

1969 ◽

Vol 113 (3) ◽

pp. 559-563 ◽

Cited By ~ 14

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).

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Some Physicochemical Properties of Erysimum Latent Virus

Australian Journal of Biological Sciences ◽

10.1071/bi9820005 ◽

1982 ◽

Vol 35 (1) ◽

pp. 5

Author(s):

Keith H Gough ◽

Glenn G Lilley ◽

Dharma D Shukla ◽

Frank Woods

Keyword(s):

Molecular Weight ◽

Buoyant Density ◽

Latent Virus ◽

Sedimentation Equilibrium ◽

Protein Subunit ◽

Molecular Weights ◽

Caesium Chloride ◽

Rna Content ◽

Velocity Diffusion ◽

Phosphorus Determination

Sedimentation velocity, diffusion coefficient and sedimentation equilibrium measurements gave a molecular weight of 5 �90 x 106 for the intact Erysimum latent virus. The molecular weight of the empty shell was estimated to be 3�92 X 106 and the protein subunit to be 21 600. The RNA content calculated from the molecular weights of the full and empty particles is 33 %, in agreement with that estimated from the buoyant density in caesium chloride. However, a direct phosphorus determination gave an RNA content of only 28 %.

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

Biochemical Journal ◽

10.1042/bj1090043 ◽

1968 ◽

Vol 109 (1) ◽

pp. 43-50 ◽

Cited By ~ 39

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.

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

Veterinary and Comparative Orthopaedics and Traumatology ◽

10.1055/s-0038-1632504 ◽

1996 ◽

Vol 09 (02) ◽

pp. 60-5 ◽

Cited By ~ 9

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.

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Subunit structures of sonicated .ALPHA. and .BETA.-ovomucin and their molecular weights estimated by sedimentation equilibrium.

Agricultural and Biological Chemistry ◽

10.1271/bbb1961.42.957 ◽

1978 ◽

Vol 42 (5) ◽

pp. 957-961 ◽

Cited By ~ 3

Author(s):

Shigeru HAYAKAWA ◽

Yasushi SATO

Keyword(s):

Sedimentation Equilibrium ◽

Molecular Weights

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Molecular weights from approach-to-sedimentation equilibrium data using nonlinear regression analysis

Biophysical Chemistry ◽

10.1016/0301-4622(79)85038-3 ◽

1979 ◽

Vol 10 (2) ◽

pp. 183-185 ◽

Cited By ~ 3

Author(s):

Leslie A. Holladay

Keyword(s):

Regression Analysis ◽

Nonlinear Regression ◽

Sedimentation Equilibrium ◽

Nonlinear Regression Analysis ◽

Molecular Weights ◽

Equilibrium Data

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Methods for the evaluation of ideal molecular weights from single sedimentation equilibrium experiments

FEBS Letters ◽

10.1016/0014-5793(70)80331-3 ◽

1970 ◽

Vol 9 (2) ◽

pp. 124-126 ◽

Cited By ~ 3

Author(s):

H.J. Rowe ◽

A.J. Rowe

Keyword(s):

Sedimentation Equilibrium ◽

Molecular Weights

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Identification of unusual phospholipids from bovine heart mitochondria by HPLC-MS/MS

The Journal of Lipid Research ◽

10.1194/jlr.ra120001044 ◽

2020 ◽

Vol 61 (12) ◽

pp. 1707-1719

Author(s):

Junhwan Kim ◽

Charles L. Hoppel

Keyword(s):

Acyl Chain ◽

Heart Mitochondria ◽

Ether Linkage ◽

Molecular Weights ◽

Bovine Heart ◽

Class Separation ◽

Normal Phase Hplc ◽

Acyl Chains ◽

Clear Identification ◽

Fragmentation Patterns

Phospholipids, including ether phospholipids, are composed of numerous isomeric and isobaric species that have the same backbone and acyl chains. This structural resemblance results in similar fragmentation patterns by collision-induced dissociation of phospholipids regardless of class, yielding complicated MS/MS spectra when isobaric species are analyzed together. Furthermore, the presence of isobaric species can lead to misassignment of species when made solely based on their molecular weights. In this study, we used normal-phase HPLC for ESI-MS/MS analysis of phospholipids from bovine heart mitochondria. Class separation by HPLC eliminates chances for misidentification of isobaric species from different classes of phospholipids. Chromatography yields simple MS/MS spectra without interference from isobaric species, allowing clear identification of peaks corresponding to fragmented ions containing monoacylglycerol backbone derived from losing one acyl chain. Using these fragmented ions, we characterized individual and isomeric species in each class of mitochondrial phospholipids, including unusual species, such as PS, containing an ether linkage and species containing odd-numbered acyl chains in cardiolipin, PS, PI, and PG. We also characterized monolysocardiolipin and dilysocardiolipin, the least abundant but nevertheless important mitochondrial phospholipids. The results clearly show the power of HPLC-MS/MS for identification and characterization of phospholipids, including minor species.

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