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PubMed1986

[Fibrinolytic activity of the blood and synovial fluid in patients with rheumatoid arthritis].

A study of the blood fibrinolytic activity (BFA) in 95 patients with rheumatoid arthritis (RA) and in their 120 relatives and of the fibrinolytic activity of the synovial fluid (FASF) in 26 RA patients showed BFA and FASF suppression in most of them. A high activity of the contact factors and high concentrations of fibrinogen degradation products were revealed in the synovial fluid. BFA and RASF changes can contribute to the progression of rheumatoid synovitis.

Adolescent↗
PubMed1972

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

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 [(125)I]iodide and the following studies were done. (a) Ultracentrifugation in caesium chloride showed that the protein moiety ((125)I 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 ((125)I 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.

Centrifugation, Density Gradient↗
PubMed1994

The biased V gamma gene usage in the synovial fluid of patients with rheumatoid arthritis.

Taking advantage of the reverse transcriptase-polymerase chain reaction (RT-PCR), we have analyzed T cell receptor gamma-chain mRNA of synovial fluid gamma/delta T cells from patients with rheumatoid arthritis (RA) in comparison with those of peripheral blood mononuclear cells (PBMC) from RA patients and healthy individuals. The quantitative RT-PCR method in conjunction with nucleotide sequencing revealed the frequent usage of the V gamma 3 gene segment in RA synovial fluid mononuclear cells (SFMC) (p < 0.01) which in PBMC of healthy individuals occurred rarely. PBMC of most healthy individuals expressed the V gamma 9 gene predominantly (p < 0.01) as expected. However, only half of RA patients showed elevated levels of the V gamma 9 gene expression in their PBMC. The gamma-chain mRNA containing the V gamma 3 gene in RA SFMC showed no conserved junctional sequence (complementarity-determining region 3). To investigate the nature of ligands recognized by the V gamma 3-bearing T cells, we analyzed V gamma gene usage of RA SFMC, RA PBMC, and normal PBMC stimulated with Mycobacterium tuberculosis (MT) or MT plus interleukin-2 since there is mounting evidence of high reactivity of RA SFMC to MT and mycobacterial heat-shock protein 65. However, the V gamma usage appeared to be mostly V gamma 9 in RA SFMC, RA PBMC and normal PBMC. Taken together these results suggest that an as yet unknown antigen(s) (other than MT) might select gamma/delta T cells expressing the V gamma 3 gene in RA SFMC.

Adolescent↗
PubMedDate not supplied

Human platelets in synovial fluid. A focus on the effects of growth factors on the inflammatory responses in rheumatoid arthritis.

The alpha granules of platelets are a major storage site for peptide growth factors. The inflamed synovial fluid of rheumatoid arthritis contains a high number of platelets as well as platelet-derived growth factors. These platelets may apparently be acted upon to release their alpha granule-located substances. It can thus be suggested that platelets present in the synovial fluid express growth factors of significance to the local inflammatory responses of rheumatoid arthritis.

Arthritis, Rheumatoid↗
PubMed2003

Detection of antibodies against glucose 6-phosphate isomerase in synovial fluid of rheumatoid arthritis using surface plasmon resonance (BIAcore).

We have used a surface plasmon resonance biosensor (SPR, BIACORE 2000) to detect antibodies against glucose 6-phosphate isomerase (GPI) in synovial fluids of rheumatoid arthritis (RA) and osteoarthritis (OA). Recombinant human GPI proteins fused with or without NusA were expressed in E. coli, purified to homogeneity and immobilized in flow cells of CM5 sensor chips. The flow cells immobilized with NusA protein or bovine serum albumin were used to monitor non-specific binding. Synovial fluid samples from RA patients showed a significantly higher level of binding to recombinant GPI proteins than samples from OA patients. Proteins which bound to the recombinant GPI proteins were confirmed to be immunoglobulin through the administration of anti-human immunoglobulin. NusA fusion protein was excellent for this assay because of a low background binding activity in the SPR analysis and its advantage of increased solubility in recombinant protein production. These results suggested a useful utilization of recombinant NusA-GPI fusion protein for the detection of autoantibodies against GPI in RA patients.

Aged↗
PubMedDate not supplied

Metastatic carcinomatous arthritis and carcinoma of the lung. A report of two cases diagnosed by synovial fluid cytology.

Arthritis is rarely caused by metastatic carcinomatous deposits in and about joints. Two patients with bronchogenic carcinoma and metastatic carcinomatous arthritis (MCA) are described. The diagnosis was established by the demonstration of malignant cells in the synovial fluid. Histology of the synovium, in 1 case, revealed several metastatic carcinomatous deposits. This report emphasizes the value of synovial fluid exfoliative cytology in the early recognition of MCA so that appropriate palliative therapy can be implemented.

Adenocarcinoma↗
PubMed1993

Increased RAHA titer and interleukin-6 levels in the synovial fluid in a patient with polymyalgia rheumatica.

We report a patient with polymyalgia rheumatica (PMR) accompanied by an increased Rheumatoid Arthritis Hemagglutinin Test (RAHA) titer and interleukin-6 level in the synovial fluid. A 60-year-old female was admitted because of polymyalgia, a body temperature of 39.2 degrees C, and an erythrocyte sedimentation rate increased to 94 mm/h. Since a muscle biopsy failed to show a specific finding, she was diagnosed as PMR. The titer of RAHA and the interleukin-6 level were increased only in the synovial fluid; prednisolone treatment decreased both. The present case raised the possibility that a similar mechanism in rheumatoid arthritis may involve the development of synovitis in PMR.

Arthritis, Rheumatoid↗
PubMedDate not supplied

Boundary lubricating ability of synovial fluid in degenerative joint disease.

The boundary lubricating ability of eleven synovial fluids was measured in a miniaturized latex--glass test system. The specimens were obtained at necropsy from knees in which the degree of degenerative joint disease varied from none to very severe. The lubricating ability of the fluid was independent of the viscosity over a wide range of shear rates. It was not diminished even in advanced lesions. In two additional fluids, the mucin clot was poor; the lubricating ability of one of these was compromised. Thus, although degenerative joint disease, during its quiescent stages, is not associated with defective synovial lubrication, the possibility that transient defects might lead to cartilage wear during life has not been excluded. The measurements are believed to be valid indicators of boundary lubricating ability under physiological conditions despite the fact that the test surfaces were not cartilaginous and the loading was relatively low (up to 47 pounds per square inch).

Adult↗
PubMed1992

Vancomycin kinetics in plasma and synovial fluid following intravenous administration in horses.

Vancomycin hydrochloride was infused intravenously (i.v.) over a 30-min period in five horses at doses of 6.6, 11.0 and 15.4 mg/kg. Vancomycin concentration in plasma and synovial fluid samples was measured using a polarization immunoassay. A pharmacokinetic model was developed to accommodate the special features of the present study. The data were described by a two compartment open model with synovial fluid as an additional compartment in exchange with plasma. Minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) were measured for Staphylococcus aureus and Enterococcus sp. using isolates from hospital patients. Based on the pharmacokinetic model and MIC/MBC data, a practical therapeutic protocol for vancomycin administration was established at doses of 4.3-7.5 mg/kg given as a 1-h infusion every 8 h.

Animals↗
PubMed1988

Changes in lipoxygenase products from synovial fluid in carrageenan induced arthritis in dogs.

A non-suppurative chronic arthritis was induced in the juvenile dog knee by intra-articular instillations with Carrageenan. Lipoxygenase products of arachidonic acid were separated from synovial fluid by reversed-phase high-performance liquid chromatography (RP-HPLC). After ten weeks we observed an accumulation of leukotriene B4 (LTB4) in synovial fluid in five out of six experimental knees (0.94 to 5.5 ng/ml), as judged by integrated optical density. Biological activity of LTB4 was confirmed by chemokinesis. LTB4 was not detected in control knees. The 15-lipoxygenase products, 15-hydroxyeicosatetraenoic acid (15-HETE) and 13-hydroxy-9,11-octadecadienoic acid (13-HODD), being inhibitors of 5-lipoxygenase, were found in relatively high levels in the control knees compared to the arthritic knees. The results denote LTB4 as a pro-inflammatory local mediator during carrageenan-induced arthritis; possibly, the decreased levels of 15-HETE and 13-HODD in the arthritic knees may have a regulatory function, thus facilitating LTB4 generation.

Animals↗
PubMedDate not supplied

Levels of soluble CD27 in sera and synovial fluid and its expression on memory T cells in patients with juvenile idiopathic arthritides.

OBJECTIVE: CD27 is a member of tumour necrosis factor receptor family. Its expression is predominantly confined to mature lymphocytes and is strongly enhanced after cell activation. Shedding of the CD27 from the surface of activated cells is related to their effector phase. The aim of the present study was to evaluate the levels of soluble CD27 in sera and synovial fluids, together with its expression on circulating and synovial fluid (SF) memory T cells, in children with JIA. METHODS: Sera from 40 patients with active JIA were studied for soluble CD27. Paired SF samples were available in 20 patients. Sera from 12 age-matched patients affected with various acute infectious diseases and 12 age-matched healthy subjects were used as controls. In 8 JIA patients freshly isolated circulating and SF lymphocytes were stained for CD27 in CD4+CD45 RO+ T cell subpopulation and analyzed by cytometry. RESULTS: Soluble CD27 serum levels were significantly higher in patients with polyarticular JIA and acute systemic infectious diseases than in patients with active oligoarticular or healthy controls. Both polyarticular and oligoarticular JIA patients showed increased levels of soluble CD27 in SF when compared with paired serum samples (p = 0.01). In all the patients tested a significant enrichment of CD27- T cells was seen in the SF (median 39.5%, range 18-56%) when compared to paired CD4+CD45RO+ peripheral lymphocytes (median 19.5%, range 5-43%; p = 0.01). CONCLUSIONS: A clear enrichment of CD4+ memory SF T cells with a CD27-phenotype is observed when compared to correspondent circulating T lymphocytes. This issue is conceivably related to re-activation and recruitment of memory T cells to the site of inflammation, and to the subsequent expansion of a subpopulation of "effector" memory T cells.

Adolescent↗
PubMed1988

Interleukin-2 secretion by synovial fluid lymphocytes in rheumatoid arthritis.

Interleukin-2 (IL-2) receptor bearing cells and soluble IL-2, measured in a bioassay with IL-2 dependent human T-cell blasts, were recognized in synovial fluid, but not in the peripheral blood of patients with rheumatoid arthritis (RA). After stimulation in vitro with appropriate concentrations of the mitogen concanavalin A (Con-A), comparable proportions of IL-2 receptor (IL-2R) bearing cells were seen in cultures of synovial fluid lymphocytes (SFL) and in cultures of peripheral blood lymphocytes (PBL). On the other hand, higher levels of secreted IL-2 were found in SFL cultures compared to corresponding PBL cultures of RA patients and normal donors. Specificity of the IL-2 bioassay was confirmed by blocking the T-cell blast proliferation (induced by SFL culture supernatants), by 83 +/- 4%, after addition of a monoclonal anti-IL-2R antibody. Despite the high levels of soluble IL-2, only a weak proliferative response was observed in the corresponding SFL cultures.

Adult↗
PubMed1997

Symmetrical synovial fluid cell cytokine messenger RNA expression in rheumatoid arthritis: analysis by reverse transcription/polymerase chain reaction.

To investigate the complex intra-articular immune activity in rheumatoid arthritis (RA), we analysed the expression of a wide range of cytokine mRNAs in synovial fluid cells from patients with rheumatoid arthritis. To minimize in vitro artefact, mRNA was rapidly extracted from synovial fluid leucocytes taken from single joints of seven patients and simultaneously from both knee joints of four patients. Expression of interleukin (IL) 1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, granulocyte-macrophage colony-stimulating factor, tumour necrosis factor-alpha and interferon-gamma (IFN-gamma) was detected using the reverse transcription/polymerase chain reaction. The expression of cytokines varied between patients. IFN-gamma mRNA was detected in 60% of the patients and IL-4 mRNA in 10%. Cytokine expression in both knees was very similar. These results suggest that T-cell activity in RA is detectable using sensitive techniques and that the intra-articular immunopathology of RA is systemically very similar.

Adult↗
PubMedDate not supplied

Protein-corticosteroid interactions in peripheral plasma and synovial fluid of patients with rheumatoid arthritis and osteoarthrosis.

The individual protein fractions, together with the unbound (biologically active) and specifically globulin-bound 11-hydroxysteroids, were determined in the plasma and synovial fluid of patients with rheumatoid arthritis (RA). The values were compared with those measured in control subjects and in patients with severe osteoarthrosis (OA). Hypoalabuminaemia and hyperglobulinaemia, as well as an increase in the unbound and a decrease in the specifically bound corticosteroids were found in the plasma of the RA patients. The protein and corticosteroid levels were lower, and the level of the protein-bound corticosteroid fraction higher, in the synovial fluid than in the plasma. Significant differences between the RA and OA patients in respect of these parameters were also observed. The findings indicate that the altered hormone pattern is not a nonspecific result of the permanent stress situation to which RA patients are subjected. They point out the existence of a fairly specific but still unknown mechanism that compensates the high level of unbound corticosteroids, thus accounting for the absence of any clinical signs of hypercortisonism.

Arthritis, Rheumatoid↗
PubMed1986

Serum and synovial fluid antibodies to collagen in rheumatic diseases: a review.

The literature on the occurrence and significance of antibodies to native or denatured collagen in rheumatic diseases is reviewed. Mainly type I and type II collagen have been investigated, both in serum and synovial fluid, with special reference to their possible role in rheumatoid arthritis. Brief results of the analysis of 40 synovial fluid samples, using an ELISA technique are included.

Arthritis, Rheumatoid↗
PubMed1979

Effect of leukocyte hydrolases on bacteria. XIV. Bacteriolytic effects of human sera, synovial fluids, and purulent exudates on Staphylococcus aureus and Streptococcus faecalis: modulation by Cohn's fraction II and by polyelectrolytes.

Normal sera and plasma, derived from humans, calves, rats, rabbits, horses, human synovial fluids, inflammatory exudates, and leukocyte extracts, when sufficiently diluted are highly bacteriolytic for Staph, aureus, Strep. faecalis, B. sutilis and to a variety of gram-negative rods. On the other hand, concentrated serum or the other body fluids are usually not bacteriolytic for these bacterial species. While the lysis of Staph, aureus and B. subtilis by diluted serum is not lysozyme dependent, lysis of Strep. faecalis is absolutely dependent on the concentration of lysozyme. The lytic factor in human serum is present in Cohn's fractions III, IV, and V. It is nondialyzable, resistant to heating for 75 degrees C and 20 min, and acts optimally at pH 5.0. Like leukocyte extracts, synovial fluids, and inflammatory exudates, it lyses only young staphylococci. The inability of concentrated serum to lyse Staph. aureus and Strep. faecalis is due to the presence in the gamma globulin fraction of a potent inhibitor, which can be partly removed by dilution of by adsorption upon the homologous bacteria. Lysis of the bacteria is also strongly inhibited by Cohn's fraction II (gamma globulin) by high-molecular-weight DNA, heparin, liquoid, and histone. The possible role played by serum globulin in the protection of bacteria against degradation by leukocyte is discussed.

Animals↗
PubMed1993

Heterogeneity of the TCR repertoire in synovial fluid T lymphocytes responding to BCG in a patient with early rheumatoid arthritis.

T lymphocytes have been implicated in the pathogenesis of rheumatoid arthritis. Interestingly, many of the activated T cells isolated from the synovial fluid of individuals with rheumatoid arthritis react with antigens from Mycobacterium tuberculosis or BCG. This response is seen to a much lesser extent in the peripheral blood of these patients. To investigate the nature of the T-cell response to BCG in RA, we isolated T cells from the synovial fluid of a patient with early-stage rheumatoid arthritis, stimulated them with BCG and cloned by limiting dilution. Staining with monoclonal antibodies specific for different V beta gene families revealed a statistically significant greater proportion of synovial-derived T-cell clones expressing the V beta 8 gene family product compared with peripheral blood clones. While the antigen specificity of some of the clones could not be determined, several of the clones displayed distinct antigen reactivities. Sequencing the TCR beta chain genes of these T cells suggested that although the V beta 8 gene products appeared to be over-represented in these BCG-specific clones, each clone utilized distinct J beta gene segments and used N segment addition to different extents. In addition, no common motifs were identified in the beta chain CDR3s of the clones sequenced. Analysis of bulk cultured BCG-specific SF T cells and unstimulated peripheral blood T cells for V beta 8 gene expression also revealed a large amount of diversity within the CDR3 region. Thus, the T-lymphocyte response to BCG in this patient with early rheumatoid arthritis appears to be quite heterogeneous.

Adult↗
PubMed1983

Ferroxidase and ascorbate oxidase activities of caeruloplasmin in synovial fluid from rheumatoid patients.

1. The protein caeruloplasmin inhibits certain free radical reactions, in part probably due to its ability to oxidize Fe2+ into Fe3+ (ferroxidase activity). Since caeruloplasmin is present in synovial fluid from rheumatoid patients, we investigated its properties in relation to protection of the joint from damage by oxygen radicals produced by activated phagocytes. 2. The ferroxidase and ascorbate oxidase activities of serum from both normal and rheumatoid patients could be accounted for by the caeruloplasmin present, as determined immunologically. More caeruloplasmin is present in the serum of rheumatoid patients than in normal serum. 3. Synovial fluid from rheumatoid patients contains caeruloplasmin protein but its ferroxidase and ascorbate oxidase activities are abnormally low. It is suggested that ferroxidase deficiency contributes to radical damage in the rheumatoid joint.

Arthritis, Rheumatoid↗