The Electrophoretic Patterns of Proteins in Synovial Fluid and Serum in Rheumatoid Arthritis12

Perlmann, Gertrude E.; Ropes, Marian W.; Kaufman, Donald B.; Bauer, Walter

doi:10.1172/jci102900

Cited by 41 publications

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“…During this study, joint fluids from six other patients with rheumatoid arthritis or neuroarthropathy were also fractionated. They were characterized by their low content of the a2-globulins and increased content of y-globulins, compared with the plasma of the same individuals (26). The results obtained from investigating the protein fractions of these joint fluids were essentially the same as those reported in the previous experiment.…”

Section: Resultssupporting

confidence: 69%

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Characterization of the Proteins of Human Synovial Fluid in Certain Disease States 1

Schmid¹,

MacNair²

1956

J. Clin. Invest.

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In search of the similarities and differences between blood plasma and synovial fluid, on one hand, and normal and pathological joint fluid on the other, it appeared important to investigate systematically the protein components of joint fluid. Investigations of the synovial fluid proteins by electrophoresis have been carried out by several authors (1-4). The identification of these proteins has been initiated (1,5). Fractionation with ammonium sulfate to obtain the albuminglobulin ratio (6) and demonstration of definite properties of certain proteins by immunochemical methods and enzymatic reactions have been reported (1,5).This publication presents a study on the characterization of the proteins in pathological synovial fluids. MATERIALS AND METHODSThe collection of synovial fluid and plasma as well as the procedure for the fractionation of their proteins have been described in a previous publication (7). All fluids were drawn from knee joints of patients with rheumatoid or traumatic arthritis or neuroarthropathy, and were obtained through the courtesy of the clinical staff of the Arthritis Unit. Most of the fluids deposited a precipitate within 20 hours although acid-citrate-dextrose (ACD) solution had been added. Prior to fractionation the joint fluids were treated with hyaluronidase (7).The electrophoretic and ultracentrifugal analyses were carried out in a Perkin-Elmer electrophoresis apparatus and a Spinco Model E ultracentrifuge, respectively. Paper electrophoresis was performed essentially according to the method of Grassmann and Hannig (8,9). After being heated at 110°for 10 minutes, the paper ing 2 per cent agar, for 17 hours at 180 V and 40 mA. The arrow "Electroph." indicates the direction of the electrophoretic movement of the proteins, and the arrow "Endosmosis" indicates the direction of the electroendosmosis. After the electrophoresis was completed, the troughs between the agar sections were filled with horse serum against human serum. Therefore, the precipitin zones formed symmetrical patterns. The positions of albumin and 7-globulin were found by applying these proteins as references on adjoining sections. The precipitin zones of synovial fluid (a) appeared less intense than those of the serum because of its lower protein concentration. The line corresponding to albumin, however, was very distinct due to the relatively high albumin concentration in this joint fluid.Whatman No. 1 paper (24 cm. wide) soaked in the borate buffer were used as electric connection between the ends of the agar plate and the adjoining borate buffer reservoirs. The buffer chambers, in turn, were again connected to holes filled with glass wool with a second set of buffer chambers in which the electrodes were placed. The electrodes were made by winding 120 cm. of stainless steel wire (0.05 cm.) around a glass tube with diameter of 1.5 cm. To secure an even level of the buffer in all 4 compartments, the two inner reservoirs were connected with each other by a thin rubber tubing. After half an hour of temperature equilibra...

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Section: Resultssupporting

confidence: 69%

“…The fluid clotted within 20 hours. The electrophoretic distribution of the proteins of the original synovial fluid, distinguished by the high content of albumin and y-globulin and low content of a2-globulin, is typical of traumatic joint fluid (26). The electrophoretic pattern of the serum was normal.…”

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confidence: 94%

Characterization of the Proteins of Human Synovial Fluid in Certain Disease States 1

Schmid¹,

MacNair²

1956

J. Clin. Invest.

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“…(1) ankylosing spondylitis; (2) osteoarthritis; and (3) rheumatoid arthritis, active, untreated. The serum of patients with rheumatoid arthritis gave positive latex fixation test.…”

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“…Liquid0 spinal e urina esseva obtenite ab patientes con arthritis. Le patientes esseva dividite in tres gruppos: (1) Spondylitis ankylosante; (2) It has been reported that in the serum and spinal fluid of patients with active rheumatoid arthritis, there was an increase in the relative concentration of the alpha-1, alpha-2, and gamma globulins and a decrease of the albumin fraction; the tendency being for the alpha globulins to rise early in the course of the disease, followed by au elevation of the gamma globulin. * Causes for alterations in an individual protein fraction, however, are not well known, although there is evidence that thc gamma globulins are increased whenever considerable inflammation or tissue destruction is present or when the gamma globulin antibody formation is increased.…”

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The Latex fixation test in the spinal fluid and urine of patients with rheumatoid arthritis

Starnes

Ulloa

Holley

1961

Arthritis & Rheumatism

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The finding of an abnormal macroglobulin in the serum of patients with rheumatoid arthritis has been isolated and characterized by several groups of investigators. A study was undertaken to study other biological fluids as to the presence of this abnormal globulin. Spinal fluids and urines were obtained from patients with arthritis. The patients were divided into three groups:(1) ankylosing spondylitis; (2) osteoarthritis; and (3) rheumatoid arthritis, active, untreated. The serum of patients with rheumatoid arthritis gave positive latex fixation test. Latex fixation test on the spinal fluids from the three groups gave a negative latex agglutination. The urines and spinal fluids were concentrated to a known volume with polyvinyl pyrrolidone (P.V.P.). There was found an increase in the total protein bound hexoses in the three latex positive urine concentrates. It has been reported that in the serum and spinal fluid of patients with active rheumatoid arthritis, there was an increase in the relative concentration of the alpha-1, alpha-2, and gamma globulins and a decrease of the albumin fraction; the tendency being for the alpha globulins to rise early in the course of the disease, followed by au elevation of the gamma globulin.* Causes for alterations in an individual protein fraction, however, are not well known, although there is evidence that thc gamma globulins are increased whenever considerable inflammation or tissue destruction is present or when the gamma globulin antibody formation is increased.The rheumatoid factor, a hemoagglutinating substance, has been isolated or partially isolated and characterized from the serum and synovial fluid of patients with rheumatoid arthriti~.~ By means of electrophoresis of those fluids, the factor has been found to migrate within the gamma f r a~t i o n .~ However, by

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“…Synovial fluids from joints involved by rheumatoid arthritis contain a high concentration of protein, and electrophoretic studies reveal increased a2-globulins (1,2). Since this protein fraction contains the highest concentration of hexose among the, electrophoretically separated proteins of serum (3), determination of nondialyzable hexose concentration in synovial fluid was undertaken.…”

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Nondialyzable Hexose of Human Synovial Fluid*†

Sandson¹,

Hamerman²

1960

J. Clin. Invest.

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Studies that show differences in the components of synovial fluids in osteoarthritis and rheumatoid arthritis may be helpful diagnostically. Synovial fluids from joints involved by rheumatoid arthritis contain a high concentration of protein, and electrophoretic studies reveal increased a2-globulins (1, 2). Since this protein fraction contains the highest concentration of hexose among the, electrophoretically separated proteins of serum (3), determination of nondialyzable hexose concentration in synovial fluid was undertaken. In synovial fluid from osteoarthritic joints the nondialyzable hexose concentration, determined with the anthrone reagent, was similar to or lower than the normal level, while in synovial fluid from joints involved by rheumatoid arthritis it was elevated.The first step in this study was to identify the carbohydrate components of dialyzed synovial fluid that contributed to the anthrone reaction. Synovial fluid was dialyzed and then lyophilized. The lyophilized solids were hydrolyzed in sulfuric acid, and the carbohydrates in the hydrolysate were separated by a combination of zone electrophoresis in borate buffer and paper chromatography. Galactose, mannose, small amounts of fucose and glucose, and hyalobiuronic acid were found. MATERIALS AND METHODS I. Identification of carbohydrate componentsSynovial fluid from normal subjects. Synovial fluid was aspirated from apparently normal knee joints of deceased subjects. Fluids from 8 to 10 normal knees were pooled to provide about 10 ml. pools of normal synovial fluid were collected. The p-oled fluids were diluted with an equal volume of a buffer (0.03 M NaHCO3 and 0.15 M NaCl, pH 8.1), dialyzed against 500 ml of distilled water (changed twice daily) for 48 hours at 20 C, and lyophilized. Synovial fluid from patients with primary osteoarthritis or rheumatoid arthritis.' About 10 ml of synovial fluid was obtained from the involved knee joint of 1 patient with primary osteoarthritis and from 2 patients with rheumatoid arthritis. These fluids were treated in the same manner as the pooled normal synovial fluids.Normal serum. The total protein-bound hexose of pooled normal serum was precipitated with ethanol according to the method of Winzler (4), and dried in a vacuum over P205.Hydrolysis. Lyophilized solids (80 mg) of pooled normal synovial fluid were hydrolyzed in 1 ml of 2 N H2SO4 in a sealed tube for 2 hours at 1000 C. The hydrolysate was adjusted to pH 5.0 with Ba(OH)2, and the insoluble BaSO4 removed by centrifugation. Lyophilized solids of osteoarthritic synovial fluid (100 mg), rheumatoid synovial fluid (100 mg), and the dried precipitate of normal serum (100 mg) were similarly hydrolyzed and neutralized.Zone electrophoresis. The technique employed was modified from that of Miiller-Eberhard and Kunkel (5).Blocks (4 X 13 X 1/4 inches) of polyvinyl chloride (Geon Resin no. 435, obtained from B. F. Goodrich) were used. The hydrolysate of normal synovial fluid was applied to the block in a slit (1'2 X 1/4 X1/4 inches) 3 inches from the cathodal end...

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The Electrophoretic Patterns of Proteins in Synovial Fluid and Serum in Rheumatoid Arthritis12

Cited by 41 publications

References 9 publications

Characterization of the Proteins of Human Synovial Fluid in Certain Disease States 1

Characterization of the Proteins of Human Synovial Fluid in Certain Disease States 1

The Latex fixation test in the spinal fluid and urine of patients with rheumatoid arthritis

Nondialyzable Hexose of Human Synovial Fluid*†

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