Koji Nishiya

Serum paraoxonase activity decreases in rheumatoid arthritis

OBJECTIVE To estimate the alterations of paraoxonase 1 (PON1) and high-density lipoprotein (HDL) in rheumatoid arthritis (RA). DESIGN AND METHODS We investigated the serum enzyme activity and concentration of PON1 and their relationship with serum lipids, high-density lipoprotein (HDL) parameters, and acute phase reactants of serum amyloid A (SAA) and C-reactive protein (CRP) in patients with RA. RESULTS Serum paraoxonase (PON) activity was significantly decreased in RA patients (n = 64, 131 +/- 53 micro mol/min/L) compared with healthy subjects (n = 155, 164 +/- 59) despite the absence of any difference in serum lipid levels between the two groups. This decrease of serum PON activity in RA patients was found in every genotype (Q/Q, Q/R, R/R) of PON1 at 192 Q/R. There was a different distribution in PON1 Q/R genotypes between RA patients and healthy subjects, and RA patients exhibited less (44%) positive PON1-Q than did the healthy subjects (66%). In a further investigation of age- and gender-matched subgroups of RA (n = 25) and healthy subjects (n = 25), not only serum PON activity, but also lecithin-cholesterol acyltransferase (LCAT) was found to be significantly decreased in RA patients (125 +/- 61 micro mol/min/L, 63.2 +/- 17.2 nmol/ml/hr/37 degrees C) than in healthy subjects (169 +/- 67, 74.7 +/- 19.5), respectively. PON1 and LCAT as well as HDL constituent apolipoprotein (apo) AI and apo AII, were altered significantly in RA patients. CONCLUSIONS Acute-phase HDL, which is remodeled structurally and functionally in RA, might be less anti-atherogenic due to the impairment of original HDL function. These alterations of HDL in RA patients may explain in part the reported increase in cardiovascular mortality in patients with RA.

Immunoglobulin Isotypes of Anti-Myeloperoxidase and Anti-Lactoferrin Antibodies in Patients with Collagen Diseases

To investigate the prevalence and clinical relevance of immunoglobulin (Ig) isotypes of antimyeloperoxidase (MPO) and antilactoferrin (LF) antibodies in collagen diseases, enzyme-linked immunosorbent assay was employed to detect the Ig isotypes of both antibodies. The purified proteins of MPO and LF were used as two major representative antigens for anti-neutrophil cytoplasmic antibodies (ANCA) with a perinuclear staining pattern by an indirect immunofluorescent technique. We examined 131 serum samples from 79 patients with rheumatoid arthritis (RA), 32 with systemic lupus erythematosus (SLE), 14 with progressive systemic sclerosis (PSS), 6 with polymyositis/dermatomyositis (PM/DM), and 5 with idiopathic crescentic glomerulonephritis who served as positive controls and 36 healthy subjects who served as controls. A limited number of patients with RA (4–10%), SLE (6–9%), and PSS (7–14%) but not PM/DM showed positive IgG or IgA anti-MPO antibody (MPO-ANCA) but not IgM MPO-ANCA. However, 10–20% of RA, 40–60% of SLE, 20–36% of PSS but none of the PM/DM patients showed positive IgG, IgA, or IgM anti-LF antibody (LF-ANCA). MPO- and LF-ANCA positivity in RA patients was correlated with markers of disease activity such as the erythrocyte sedimentation rate, C-reactive protein, and serum Ig levels. IgG LF-ANCA but not MPO-ANCA positivity in SLE patients also was correlated with the disease activity index but not with clinical features. Neither MPO- nor LF-ANCA positivity in PSS patients was correlated with any clinical features. Overall, both MPO- and LF-ANCA were found mainly in RA, SLE, and PSS patients but not in PM/DM patients. The Ig isotypes of MPO- and LF-ANCA frequently belonged to both IgG and IgA and rarely to the IgM class. Both MPO- and LF-ANCA positivity reflected disease activity in RA and SLE rather than specific organ involvement.

Ferritin correlates with C-reactive protein and acute phase serum amyloid A in synovial fluid, but not in serum.

OBJECTIVE To evaluate ferritin concentration in serum and synovial fluid (SF) as a marker of activity of arthritis in comparison with C-reactive protein (CRP) and acute-phase serum amyloid A protein (A-SAA). METHODS We determined the concentrations of ferritin, CRP and A-SAA in paired serum and SF in 34 rheumatoid arthritis (RA) and 21 osteoarthritis (OA) patients. The erythrocyte sedimentation rate (ESR) was also measured. RESULTS The serum concentrations of ferritin, CRP and A-SAA were 93 +/- 76 (mean +/- SD) ng/ml, 4 +/- 5 mg/ml, 8 +/- 4 mg/ml in OA and 140 +/- 227, 59 +/- 34, 289 +/- 223 in RA, respectively. There was no significant difference in serum ferritin levels between OA and RA, and serum ferritin did not correlate with ESR, CRP or A-SAA. Both serum CRP and A-SAA levels were significantly higher in RA than in OA (p < 0.0001, p < 0.0001), and correlated with ESR in all arthritis (r = 0.658, p < 0.0001, r = 0.404, p < 0.01), respectively. Serum CRP levels correlated with A-SAA levels in serum (r = 0.727, p < 0.0001). In SF, the concentrations of ferritin, CRP and A-SAA in RA (421 +/- 307, 25 +/- 20 and 39 +/- 41) were significantly higher (p < 0.01, p < 0.0001, p < 0.001) than those in OA (202 +/- 220, 2 +/- 2 and 2 +/- 2), respectively. There were significant correlations among SF ferritin, CRP and A-SAA. CONCLUSION Ferritin levels in SF but not in serum are significantly elevated in RA more than in OA, and ferritin correlated with CRP or A-SAA in SF, but not in serum. Higher levels of SF ferritin, as well as SF CRP and SF A-SAA, seem to reflect greater degrees of joints inflammation in RA and OA.

Down regulation by iron of prostaglandin E2 production by human synovial fibroblasts.

OBJECTIVE To examine the effect of iron on the prostaglandin (PG) E2 production by human synovial fibroblasts in vitro. METHODS Human synovial fibroblasts were isolated from synovial tissue of rheumatoid arthritis (RA) and osteoarthritis (OA) patients and cultured in medium. Synovial fibroblasts were stimulated by human recombinant interleukin (IL) 1β (0.1–10 ng/ml) with or without ferric citrate (Fe-citrate, 0.01–1 mM). The amount of PGE2 in the culture medium was measured by an enzyme linked immunosorbent assay. RESULTS The production of PGE2 by the synovial fibroblasts was increased by stimulation with IL1β at all concentrations tested. Fe-citrate but not sodium citrate (Na-citrate) down regulated the production of PGE2 by the synovial fibroblasts, both with and without stimulation by IL1β. Fe-citrate inhibited the spontaneous PGE2 production by the cells in a dose dependent manner, and a maximum inhibition by Fe-citrate was observed at the concentration of 0.1 mM with IL1β stimulation. The down regulation by iron was reversed by the co-addition of desferrioxamine (100 μg/ml), an iron chelator. CONCLUSION Iron down regulates the PGE2 production by synovial fibroblasts in vitro.

Collagenofibrotic Glomerulonephropathy Associated with Immune Complex Deposits

A 66-year-old Japanese male, who suffered from persistent proteinuria and leg edema, underwent renal biopsy. Light microscopy revealed marked narrowing of the glomerular capillary lumen with a diffuse accumulation of weakly PAS-positive material. By electron microscopy, abundant abnormal collagen fibers were observed predominantly in the subendothelial space and occasionally in the mesangial matrix. The fibers had a periodicity of about 60 nm and were immunoreactive for anti-type III collagen. Subendothelial electron-dense deposits were also found in some of the capillary walls. The serum level of procollagen III peptides was elevated and changed in parallel with the amount of proteinuria during the patient’s clinical course. On the basis of these findings, a diagnosis of the collagenofibrotic glomerulonephropathy was made. A review of the literature, including 29 similar or identical cases, failed to reveal the etiology and pathogenesis of this disease. We suggest that this disease may be divided into two different clinical subtypes, an adult-onset type and a pediatric type.

Serum and synovial fluid levels of interleukin-5 in a patient with eosinophilic fasciitis.

We did not suggest that any of the authors had substituted genetic screening for the classification criteria. As shown in our paper, patients who have one or more copies of the shared epitope do have a worse prognosis. However, those who are shared epitope negative do not universally have a good prognosis. Between 19% and 30% develop radiological erosions. One could not, therefore, advocate withholding potential disease modifying agents from this group. We are delighted that Professor Emery and his group are examining the value of stratification by HLA-DRB1 genotype in a controlled clinical trial. We look forward to the publication of the results of these studies. In the meantime we are all agreed that testing for the possession of the shared epitope should remain a research tool and should not become part of routine clinical practice.

Enhancement by iron of interleukin 1 induced granulocyte macrophage colony stimulating factor (GM-CSF) production by human synovial fibroblasts

Iron infusion activates synovium and induced joint inflammation in experimental animals and causes the flaring up of arthritis in patients with rheumatoid arthritis (RA). Marked iron deposition in RA synovia has been reported over the past 30 years and has also recently been demonstrated by quantitative photometric assessment and is correlated with exudative and proliferative histological features.1 It has been reported that the amount of iron deposition in RA synovial tissue is correlated with disease activity and severity. Iron has an important role in RA synovitis through the formation of radical oxygen species, and the enhancement of collagen synthesis and synovial fibroblast proliferation2 possibly owing to down regulation of prostaglandin E2 (PGE2) production.3 Synovial fibroblasts produce a number of inflammatory mediators including cytokines such as interleukin (IL)1, IL6, IL8, fibroblast growth factor, vascular endothelial growth factor, tumour necrosis factor, and granulocyte macrophage colony stimulating factor (GM-CSF). GM-CSF produces the progenitor cells of macrophage lineage stem cells and stimulates mature granulocytes and macrophages. GM-CSF is produced by T cells, macrophages, and fibroblasts and has been found in synovial fluid and tissue from patients with RA. GM-CSF has an important role in type II collagen induced arthritis in rats and in the acute methylated bovine …

Enlarged spleen detected by abdominal ultrasonography in patients with RA

Rheumatoid arthritis (RA) is a systemic, chronic inflammatory disease of unknown cause. Approximately 5–10% of patients with RA have an enlarged spleen on manual palpation1 or on isotope scanning.2 We measured the size of the spleen in 50 patients with RA (nine men, 41 women; average age 55.8 years (range 25–78)) and 14 healthy control subjects (no men, 14 women; average age 47.8 years (range 25–80)) by abdominal ultrasonography (Aloka, Japan). This examination was done by one skilful examiner (TD) in all cases, and comparisons were made with clinical profile and disease activity. The patients with a diagnosis of Feltys syndrome, categorised as the triad, RA, leucopenia, and splenomegaly, and patients with complications of any viral or bacterial infections were excluded from this study. Mean (SD) disease duration was 6.2 (6.6) years. The disease stage was I (13 patients), II (11), III (nine), IV (17) and the functional class was I (seven patients), II (40), III (three), IV (0), assigned according to Steinbrocker criteria.3 The following treatment …

Suppressive Effect of Iron on Concanavalin A‐Induced Multinucleated Giant Cell Formation by Human Monocytes

Immune dysfunction in patients with iron overload has been reported. Iron disturbed CD2 expression on T‐cells, cell‐mediated immunity by Th1 cells and monocyte functions including phagocytosis and natural killer activity. In the present study, we examined the effects of iron and desferrioxamine (DFX, an iron chelator) on generation of multinucleated giant cells (MGC) by human monocytes in vitro. Human monocytes were isolated from venous blood and cultured with concanavalin A (Con A) stimulation with additives, ferric citrate (Fe‐citrate) or sodium citrate (Na‐citrate) or DFX for 4 days. The cells were fixed and subjected to Wright staining. MGC formation was observed under light microscopy. Con A induced MGC formation in a dose‐dependent manner, and reached a plateau after 3 days of incubation. MGC formation was suppressed when Con A‐stimulated monocytes were cultured with the co‐addition of Fe‐citrate but not Na‐citrate only in the early phase of culture (less than 24 hours). DFX also suppressed MGC formation in a dose‐dependent manner. Using flow cytometry analysis, the co‐addition of Fe‐citrate significantly suppressed CD18 (ß2 integrin) and CD54 (ICAM‐1) but not CD11a (α integrin) expression on Con A‐stimulated monocytes. Iron supressed the generation of MGC by human monocytes in vitro. These observations suggested that iron might affect MGC generation by down‐regulation of adhesion molecule expression on monocytes.

Gastrointestinal symptoms associated with enteric-coated sulfasalazine (Azulfidine EN tablets)

Abstract To investigate both the incidence and the dosage used to treat gastrointestinal (GI) symptoms associated with enteric-coated sulfasalazine (Azulfidine EN, AZL) in patients with rheumatoid arthritis (RA), we studied the clinical history of 153 RA patients, and any available data on GI symptoms that might have been associated with AZL. GI symptoms appeared in 64 (42.5%) of the 153 cases. There were 19 events of nausea, vomiting, or dyspepsia, 14 events each of epigastric discomfort and reduction or loss of appetite, 10 events of epigastric, stomach, or abdominal pain, 9 events of heartburn, 8 events of mouth ulcer, 3 events each of loss of taste and abdominal bloating or borborygmus, 2 events each of diarrhea or loose stools, hematemesis or melanemia, and gastric or esophageal ulcer, and 1 event of stomatitis. These results indicate that GI symptoms associated with AZL are usually mild and treatment can continue, with almost all cases responding to a reduction in dose or drug cessation. In some cases, a histamine receptor-2 blocker or proton pump inhibitor is also required.