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.

The Binding of Lithium Carmine to a2-Macroglobulin

1969 ◽  
Vol 24 (11) ◽  
pp. 1442-1447 ◽  
Author(s):  
J. J. Picard ◽  
J. F. Heremans
Keyword(s):  
Human Serum ◽  
Density Gradient ◽  
Protein Complex ◽  
Gel Filtration ◽  
Heavy Fraction ◽  
Normal Human Serum ◽  
Density Gradient Ultracentrifugation ◽  
Γ Globulins ◽  
Normal Human

The colloidal dye lithium carmine was added in vitro to normal human serum. Electrophoretic experiments showed that the dye was associated mainly with α2-globulins, small amounts with the albumin and only traces with the γ-globulins. The main complex was eluted with the macroglobulin peak obtained by gel filtration on Sephadex G-200 and sedimented in the heavy fraction on density gradient ultracentrifugation. The dye-protein complex could be precipitated with an antiserum specific for a2-macroglobulin. Gel filtration of a solution of pure a2-macroglobulin, to which lithium carmine was added, demonstrated that the dye was bound to this protein.

Capillary electrophoresis of normal human synovial fluid: comparison with fluid from the disease states of osteoarthritis and rheumatoid arthritis

1994 ◽  
Vol 31 (9) ◽  
pp. 257 ◽  
Author(s):  
John M. Duffy ◽  
James Grimshaw ◽  
Aine Kane ◽  
Raymond A. B. Mollan ◽  
Peter L. Spedding ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Capillary Electrophoresis ◽  
Synovial Fluid ◽  
Disease States ◽  
Normal Human ◽  
Human Synovial Fluid

Normal human synovial fluid

1985 ◽  
Vol 67 (9) ◽  
pp. 1336-1339 ◽  
Author(s):  
M Baumgarten ◽  
R D Bloebaum ◽  
S D Ross ◽  
P Campbell ◽  
A Sarmiento
Keyword(s):  
Synovial Fluid ◽  
Normal Human ◽  
Human Synovial Fluid

scholarly journals The presence of polysaccharide in normal human gastric mucus

1983 ◽  
Vol 215 (2) ◽  
pp. 421-423 ◽  
Author(s):  
J R Clamp ◽  
B Cooper ◽  
J M Creeth ◽  
D Ene ◽  
J Barrett ◽  
...  
Keyword(s):  
Amino Acid ◽  
Molecular Mass ◽  
Density Gradient ◽  
Elemental Analysis ◽  
Density Gradient Ultracentrifugation ◽  
Gastric Mucus ◽  
Average Molecular Mass ◽  
Normal Human ◽  
Beta Elimination

Polysaccharide material was found in the proteolysis glycopolypeptide fraction from normal human gastric mucus. The polysaccharide was identified by carbohydrate and amino acid analyses, by elemental analysis and from its behaviour on density-gradient ultracentrifugation. The polysaccharide is polydisperse with a weight-average molecular mass of 300 000 Da. Over 85% of the polysaccharide consists of galactose, and this represents 26% of all the galactose present in the fractions after beta-elimination with reduction of the glycopolypeptide material.

scholarly journals THE ORIGIN AND NATURE OF NORMAL HUMAN SYNOVIAL FLUID 12

1940 ◽  
Vol 19 (6) ◽  
pp. 795-799 ◽  
Author(s):  
Marian W. Ropes ◽  
Elsie C. Rossmeisl ◽  
Walter Bauer
Keyword(s):  
Synovial Fluid ◽  
Normal Human ◽  
Human Synovial Fluid

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 Isolation of different high-Mr mucin species from human whole saliva

1992 ◽  
Vol 283 (3) ◽  
pp. 807-811 ◽  
Author(s):  
E C I Veerman ◽  
P A M van den Keybus ◽  
M Valentijn-Benz ◽  
A V Nieuw Amerongen
Keyword(s):  
Monoclonal Antibodies ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Buoyant Density ◽  
Density Gradient Ultracentrifugation ◽  
Immunochemical Analysis ◽  
Guanidinium Chloride ◽  
Whole Saliva ◽  
Cscl Density Gradient

By using CsCl-density-gradient ultracentrifugation, two high-Mr mucin species were isolated from human whole saliva, having buoyant densities in 0.2 M-guanidinium chloride of approx. 1.56 g/ml (pool IA) and 1.48 g/ml (pool IIA). Analytical density-gradient centrifugation of submandibular, sublingual, labial and palatal saliva, followed by immunochemical analysis with anti-mucin monoclonal antibodies, indicated immunochemical and physicochemical similarities between the high-density mucins of pool IA and mucins from palatal salivary glands. Chemical analysis indicated that the putative palatal mucin was rich in sulphate, but poor in sialic acid. The lower-density mucins of pool IIA equated with the high-Mr mucins of submandibular-sublingual saliva, both immunochemically and physicochemically (buoyant density).

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