Are Angiotensin Converting Enzyme and von Willebrand factor circulating levels useful surrogate parameters to monitor disease activity in Kawasaki disease?

To verify if Angiotensin Converting Enzyme (ACE) and von Willebrand factor (vWF) may be used as a laboratory marker for the follow-up of endothelial derangement and therapeutic efficacy in Kawasaki disease (KD), circulating ACE, vWF routine hematological tests and cardiac involvement were assessed in 32 children with established diagnosis of KD before and up to six months after intravenous gamma-globulins (i.v.IG) treatment. I.v.IG treatment normalized progressively all the hematological parameters to levels comparable with healthy controls within 30 days. At baseline, ACE levels resulted significantly lower (1.8 +/- 1.3 pmol/ml/min), and vWF levels significantly increased (210.3 +/- 35.2%) when compared with controls (respectively 7.0 +/- 0.9 pm/ml/min and 99 +/- 17.9%). Seven days after the treatment vWF levels were decreased (188 +/- 18.4%) but still significantly higher than controls, and fully normalized after 15 days (104.8 +/- 14.3%). ACE levels were found progressively increased at 7, 15, and 30 days after the treatment (respectively 2.7 +/- 1.0, 3.7 +/- 0.4, 5.04 +/- 0.9 pm/ml/min) and reached the range of normality only after two months (7.74 +/- 2.46 pm/ml/min). The present study shows that ACE and vWF circulating levels are significantly modified during the acute phase of the disease.

Capsaicin stimulates the migration of human polymorphonuclear cells (PMN) in vitro.

Capsaicin, a homovanillic acid derivative in plants, has distinct pharmacological effects in vivo, e.g. it depletes primary afferent neurons of substance P and other tachykinins. The effect of capsaicin on the migration of human neutrophils was tested in concentrations ranging from 10(-8) M to 10(-3) M. In comparison to the control 10(-8) M capsaicin significantly enhanced the migration of PMN cells (CI 1.29; 2P < 0.009) and a peak migration activity was detected with 10(-6) M (CI 1.32; 2P < 0.01). With higher concentrations of capsaicin the CI was not significantly changed. These results show that capsaicin, a plant derived neurotoxin, exhibits a migration modifying activity on human neutrophils through a direct mechanism not mediated by neuropeptides. In addition capsaicin (10(-7) and 10(-5) M) did not affect the luminol-dependent chemiluminescence and therefore does not contribute to a superoxide anion generation in human PMN.

Synoviocytes from osteoarthritis and rheumatoid arthritis produce plasminogen activators and plasminogen activator inhibitor-1 and display u-PA receptors on their surface.

The production of plasminogen activators and their inhibitors was studied in vitro in osteoarthritic (OA) and rheumatoid arthritic (RA) synovial fibroblasts (SF), obtained from RA and OA patients undergoing joint surgery. Subcultured SF were cultivated for 2, 4, 6, 8, 10 and 13 days and the medium assayed for the presence of both plasminogen activators (PAs) and plasminogen activator inhibitor-1 (PAI-1). The presence of urokinase-Plasminogen Activator (u-PA) receptors (u-PAR) on the surface of synovial cells was investigated by radio-ligand binding assay and cross-linking and by transmission electron microscopy (TEM) of a gold-u-PA complex. Our results showed a low production of tissue-type-Plasminogen Activator (t-PA) in both OA and RA SF, but relatively high levels of u-PA, until confluence, both in OA and in RA. SF were also able to produce plasminogen activator inhibitor in large amounts, in particular in RA since the very beginning of the culture. Receptors for u-PA were evident on both RA and OA SF. Our data show that SF in vitro produce mainly u-PA, the most important plasminogen activator involved in tissue modifications. The demonstration of u-PA receptors on the surface of OA and RA SF represents a step forward in the understanding of the possible role of fibrinolytic and tissue destructive proteinase cascade in joint inflammation.

Low serum hyaluronan in psoriatic arthritis patients in comparison to rheumatoid arthritis patients.

OBJECTIVE: Serum hyaluronan (HA) was determined in 37 patients suffering from psoriatic arthritis (PSA), 39 patients with rheumatoid arthritis (RA), 31 with osteoarthritic joint disease (OA) and 26 healthy controls (C) in order to examine earlier reports that HA levels are increased in the serum of RA and to assess whether this finding is also relevant for PSA, another inflammatory joint disease, since HA in serum is considered a sign of inflammation in general. METHOD: HA in the serum samples was measured with an enzyme linked microplate assay. RESULTS: Sera from PSA, RA and OA patients showed a significantly higher HA concentration than those of healthy controls (56.0 +/- 16.0 micrograms/l). The serum HA concentration in PSA patients amounted to 107.8 +/- 57.2 micrograms/l, which was not significantly different from OA patients (104.9 +/- 16 micrograms/l). A significant difference, however, could be observed between the HA concentrations of the PSA subgroups: the mean HA level of patients suffering from symmetrical polyarthritis was 134 +/- 79.6 micrograms/l, which turned out to be significantly higher than in patients suffering from symmetrical oligoarthritis (89.9 +/- 42.8 micrograms/l; P < 0.04), but was insignificantly increased in comparison to patients with ankylosing spondylitis as the predominant feature (109 +/- 27.8 micrograms/l; P = 0.49). The mean HA concentration for RA sera was 197.1 +/- 122.9 micrograms/l, which was statistically significantly increased compared to PSA (P < 0.001) and OA (P < 0.001) sera. The sera of seropositive RA patients showed significantly higher HA levels than PSA patients with symmetrical polyarthritis (P < 0.04). CONCLUSION: The data obtained support recent studies which have shown HA levels to be higher in RA patients than in OA patients. Seronegative and seropositive RA patients showed the same HA concentrations, while patients suffering from "seronegative" PSA were found to have lower HA concentrations. Therefore, HA serum levels may reflect cartilage degradation in general or the degree of articular inflammatory processes, indicating different pathogenetic pathways.

Evidence for hyaluronan production in the air pouch model in rats.

OBJECTIVE: The aim of our work was to investigate the presence of hyaluronan (HA) in the rat air pouch and its behaviour in response to inflammatory stimuli. METHODS: HA levels (by a microplate assay) and the leucocyte count were determined in the fluid obtained from air pouches in which acute or subacute inflammation had been induced by the injection of monosodium urate crystals (MSU) or high density polyethylene (HDPE) debris respectively and in relative controls. RESULTS: In control pouches of both groups, remarkable levels of HA were found; these levels were higher in the very first hours (2475 and 1850 micrograms/l at 6 hrs) and then gradually decreased. In pouches injected with MSU, HA moderately increased (p < 0.001) after 6 hrs, reached a peak after 12 hrs (p < 0.001) and began to taper at 24 hrs (p < 0.001). The leucocyte count was also increased at 6 hrs (p < 0.001), became higher at 12 hrs (p < 0.001) and tapered at 24 hrs (p < 0.001). In the HDPE pouches, HA levels were significantly reduced with respect to controls after 6 hours (p < 0.001), increasing later (p < 0.001) to reach a peak at 24 hrs (p < 0.001), and returning to the original levels, or even below, in the following 72 hours. CONCLUSIONS: These data confirm that the pouch lining produces fair amounts of HA and provide evidence that, in this system, HA levels seem to be influenced by the degree of inflammation even if with variable behaviour in relation to the different characteristics and phases of phlogosis. The present data suggest that the air pouch is a useful experimental model for studies on HA metabolism in either acute or chronic inflammation.

Auranofin and its combination with LTB4 influences ATP level and migration of human polymorphonuclear cells in vitro.

Auranofin (AF), an orally administered antirheumatic drug, reduces the ATP level of PMN cells in vitro in a dose-dependent manner and provokes various effects on PMN migration. Under the experimental conditions, AF in concentrations between 10(-8) M and 10(-3) M produced a statistically significant (P < 0.05) dose-related reduction in ATP level, which ranged from 89% of the control value with 10(-8) M AF to 46.8% of the control with 10(-3) M AF. On the other hand, the combination of AF and the chemoattractant LTB4 (1 ng/ml) shows agonistic effects on the intracellular ATP level. AF at 10(-5) M significantly increases the ATP (33%; P < 0.03). In general migration of PMN cells is stimulated by 10(-7) M AF [chemotactic index (CI) = 1.26], but inhibited by 10(-5) M (CI = 0.65), 10(-4) M (CI = 0.09) and 10(-3) M AF (CI = 0.01). These effects were statistically significant at P < 0.05. In the presence of LTB4 (1 ng/ml), which resulted in an average CI of 2.9, AF also inhibits the chemotactic effect of the chemoattractant, with the CI being significantly reduced at 10(-6) M AF (CI = 2.3) and 10(-4) M AF (CI = 0.05). In the latter case the effect was also confirmed by the leading-front technique. AF at 10(-6) M is a level that could be reached in the blood after continuous therapy regimens, and these results are therefore of practical interest. They expand our knowledge of the effects of AF on PMN cells, whereby reducing effects on intracellular ATP were observed with AF alone and stimulating effects in combination with LTB4. With low AF concentrations, the reduction of the ATP level is only a part of its action that seems to be independent of its effect on cell migration and chemotaxis.

Effect of substance P and somatostatin on migration of polymorphonuclear (PMN) cells in vitro.

The effects of the two neuropeptides, substance P (SP) and somatostatin (SOM), on the migration of polymorphonuclear cells derived from 13 volunteers were investigated. The neuropeptides were applied in concentrations between 10(-12) and 10(-6) M. Only at a concentration of 10(-6) M SP did the chemotaxis of PMN cells increase slightly but statistically significantly. In contrast to SP, SOM showed a significant dose-dependent stimulation of chemotaxis, which was first traceable at 10(-10) M and increased up to 10(-6) M. Although it is uncertain whether in vivo SP and SOM contribute directly to the invasion of PMN cells into the joint cavity, the influence of these neuropeptides on PMN migration in vitro is a further indication of the neuropeptide involvement in the genesis of inflammation.

Gold salts and somatostatin: a new combined analgesic treatment for psoriatic arthritis.

In a group of patients affected with psoriatic arthritis the effects of the association between gold salts (GS) and somatostatin (SOM), in comparison with two groups treated with SOM and GS respectively, were investigated. Sixty patients with psoriatic arthritis were selected and randomly allocated in three groups of twenty patients each. Group 1 received SOM infusion (250 micrograms/h for 96 h) and was assessed at baseline and 1, 15, 30, 60, 90 and 120 days after; Group 2 received intramuscular GS and was assessed at baseline, four months later, and then every month for four months; Group 3 received GS for 8 months; at the fourth month SOM was infused (as in Group 1) and the patients assessed at baseline four months later and then as Group 1. Assessment was made with the Ritchie index, pain scale and morning stiffness evaluation. Growth hormone was assayed in Group 1 every 4 h for 24 h the day before and the day after SOM infusion. The association between GS and SOM demonstrated a particular analgesic activity, effective on joint pain and tenderness, that lasted for all four months of follow-up. SOM showed a good response only after 15 and 30 days, and GS proved to be effective at about the sixth month of treatment. Side effects were reported in Group 1 (abdominal cramps, mild erythrodermia and supraventricular arrhythmia). A growth hormone circadian rhythm was found in psoriatic patients both before and after SOM treatment. The beneficial effect of the SOM/GS combination is demonstrated in psoriatic arthritis.(ABSTRACT TRUNCATED AT 250 WORDS)

Pharmacokinetic studies on diponium bromide.

The body elimination kinetics and the metabolic pathway of intravenously administered diponium bromide ((2-[alpha,alpha-dicyclopentylacetoxy)- ethyl] triethylammonium bromide) were studied. The rapid alpha- distribution phase had a t 1/2 of 4 to 11 minutes. The elimination rate varied between 2.3 and 7.7 hours. The distribution volume was 29.64 +/- 15.03 l, the total body clearance was 72.99 +/- 28.52 ml/min, and the renal clearance showed a mean value of 45.03 +/- 12.51 ml/minute. Thin layer chromatography of extracts derived from urine and faeces showed unchanged DB and a metabolite in urine; in the faeces of some volunteers three metabolites were detected, which were characterized by their Rf-values. DB does not bind to erythrocytes but to plasma proteins.

[Plasma amino acids in ankylosing spondylitis (author's transl)]. / Die Plasma-Aminosäuren bei der Spondylitis ankylosans.

The plasma amino acids of male patients with ankylosing spondylitis (ASp) and male controls were analysed and the concentrations statistically evaluated. The total concentration of all 28 amino acids in ASp-patients did not differ from the controls, but there were some distinct differences in the levels of individual amino acids, e. g. arginine and isoleucine which showed raised concentration (29% and 27%). A higher concentration was also detectable with rare amino acids, e. g. alpha-amino-adipinic acid (23%) and 1-methyl-histidine (32%). Correlation between the amino acid concentration and age was detected only in the case of citrulline. Some amino acids showed a significant correlation to one another which was sometimes evident in both groups and other times was noted in the control- or ASp-group only. If such correlations were found to be disease-dependent this finding could be helpful in the diagnosis of certain diseases.

[In vitro investigations on the influence of various antirheumatic drugs on the enzymes of the purine salvage pathway (author's transl)]. / In-vitro-Untersuchungen über den Einfluss verschiedener Antirheumatika auf die Enzyme des Purin-Wiederverwertungsstoffwechsels

On testing the effect of 5 drugs (azapropazone, flufenamic acid, indomethacin, oxyphenbutazone and prednisolone) commonly used in the therapy of rheumatic diseases on 3 enzymes of the purine salvage pathway (adenine-phosphoribosyltransferase, guanine-phosphoribosyltransferase and hypoxanthine-phosphoribosyltransferase) a distinct inhibitory effect of oxyphenbutazone and flufenamic acid was noted. A particularly marked effect was observed on adenine-phosphoribosyltransferase at the applied concentration (50 mug%) by both drugs but also guanine-phosphoribosyltransferase and hypoxanthine-phosphoribosyltransferase indicated reduced activities.

[The effect of oxyphenbutazone and flufenamic acid in vitro on the purine salvage pathway of human lymphocytes (author's transl)]. / Einfluss von Oxyphenbutazon und Flufenaminsäure in vitro auf den Purin-Wiederverwertungsstoffwechsel menschlicher Lymphozyten

The effect of oxyphenbutazone and flufenamic acid was investigated on the enzymes of the purine salvage pathway of human lymphocytes in vitro. A distinct decrease in adenine-phosphoribosyltransferase activity and in guanine-hypoxanthine-phosphoribosyltransferase activity was observed following the administration of either pharmacological preparation. The loss of enzyme activity was 23 to 39%. The pharmaceuticals were used at a concentration of 50 mug%.

[Antinuclear antibodies in ankylosing spondylitis (author's transl)]. / Antinukleäre Antikörper bei Spondylitis ankylosans (Morbus Bechterew).

Antinuclear antibodies (ANA) were found in 19.1% of patients suffering from ankylosing spondylitis (n = 320). This figure is significantly higher than the results obtained in a control group. ANA were present in all stages of the disease, including the abortive form of isolated sacroiliitis. ANA were also found in different stages of activity. In addition, ANA-positive cases showed a higher inflammatory activity. In females ANA were more frequently found than in males. There was a positive correlation between ANA and HLA B27 in 87.2%.

[Immunological features in ankylosing spondylitis (author's transl)]. / Immunphänomene bei Spondylitis ankylosans (Morbus Bechterew).

Sera from 50 patients with various stages of ankylosing spondylitis were tested for the presence of antinuclear antibodies (ANA), anti-DNA antibodies, rheumatoid factor and antistreptolysin. Antinuclear antibodies (immunofluorescent technique) were detected in the sera of 10 patients (20%), associated in one case with anti-DNA antibodies (immunofluorescent technique). The disease activity in ANA-positive cases was low to moderate.

[Cellular and humoral immune phenomena in psoriatic arthritis]. / Untersuchungen über zelluläre und humorale Immunphänomene bei der Arthritis psoriatica.

The function of cellular immunity factors (lymphocyte transformation and phagocytosis by polymorphonuclear leucocytes [PMN] and monocytes in connection with the concentration of intracellular ATP) and humoral immunity factors (serum concentration of immunoglobulin and complement factors C'3 and C'4) was investigated in 16 controls, 21 patients with psoriatic arthritis and 19 with psoriasis vulgaris. The results were compared with the clinical and anamnestic data of the patients. PMN phagocytosis of zymosan opsonized with rabbit standard serum was decreased in psoriasis vulgaris in comparison with the controls. Also, monocyte phagocytosis of non-opsonized zymosan was decreased in psoriatic arthritis, as compared with psoriasis vulgaris. Furthermore, in PMNs intracellular ATP was elevated in psoriatic arthritis as compared with the controls, but decreased in comparison with patients with psoriasis vulgaris. The intracellular ATP in monocytes was decreased in psoriasis vulgaris as compared with the controls. Humoral immunological findings: serum IgG concentration was higher in psoriatic arthritis than in controls and in psoriasis vulgaris. Elevated C'3 and decreased C'4 serum concentrations in psoriatic arthritis indicate an activation of the complement system.

The effect of a cartilage bone marrow extract on experimentally induced osteoarthrosis in the knee joints of rabbits. A scanning electron microscopic study.

The development of osteoarthrosis following a partial meniscectomy on the knee cartilage of rabbits (Chinchilla hybrids) was monitored with a scanning electron microscope. Simultaneously, a study was made of the effect of the cartilage bone marrow extract Rumalon trademark on the development of the osteoarthrotic changes. Twelve days after the operation, osteoarthrotic changes were evident in the untreated operated joints. After 36 days the damage caused to the cartilage was already radical. The immobility of the operated joint also gave rise to obvious changes in the cartilage of the knee joints which had not undergone an operation. The irregular weight distribution due to the fixation of one joint was apparently enough to provoke degenerative processes on the other side. When the cartilage bone marrow extract Rumalon trademark was administered three times weekly (0.5 mg/kg body weight i.m.) a distinct retardation of the osteoarthrotic development in the early stages was observed. Where the changes had penetrated to the inner cartilage layers, no difference could be established compared to the untreated animals.

Induction of arthritic processes by synovial fluids of rheumatoid arthritis patients in long-term experiments with mice.

Diluted synovial fluids derived from three patients with rheumatoid arthritis (RA) and applied subcutaneously once as a single dose in Swiss mice caused arthritic processes after a long period of latency (10 months). The arthritis-inducing effect was enhanced by storing the original synovial fluids at 4 degrees C for three months. The effects were followed histologically for at least 18 months. In contrast to the theory that RA is based on immunological processes, these preliminary observations support the finding of other authors that a transmissible agent which is rather slow-acting exists in the synovial fluid of RA patients.

The influence of synovial fluids from patients with rheumatic diseases on chick embryo fibroblasts.

Primary chick-embryo fibroblasts (PCEF) were used as target cells to measure the influence of synovial fluids of patients suffering from osteoarthritis (OA; n = 5), rheumatoid arthritis (RA; n = 12) and psoriatic arthritis (PA; n = 2). The following metabolic cell products were measured: DNA, RNA, glycosaminoglycans (GAG), sulfated glycosaminoglycans, protein and collagen, with the same joint effusions being used in each test. Since it is not a single substance that provokes a stimulating or inhibiting effect in the joint, the crude synovial fluids were applied in these preliminary experiments. It was found that each type of synovial fluid showed an influence on the biological processes in the PCEF. The DNA, RNA and GAG syntheses were strongly influenced by the joint effusions, in contrast to the protein, collagen and sulfated glycosaminoglycan syntheses which were less affected. Generally, the nucleic acid synthesis differed significantly between the OA, RA and PA synovial fluids. The addition of heparin to the synovial fluids caused an additive inhibiting effect on the DNA synthesis but did not influence the other biochemical parameters. The synovial fluids of RA patients, and to a much greater extent those of PA patients, inhibited the thymidine incorporation whereas OA synovial fluids had a less pronounced effect. This result indicates a disease-dependent composition of the synovial fluids. RNA synthesis was diminished in all three groups, but again this effect was strongest in the case of the PA synovial fluids. GAG synthesis was markedly stimulated by the PA synovial fluids and somewhat, though to a lesser extent, by the OA and RA synovial fluids. The sulfated glycosaminoglycan synthesis in the PCEF, as revealed by 35S incorporation into the GAG, was less influenced and on the whole stimulated by the OA and RA synovial fluids. The same trend could be observed with regard to the collagen synthesis. The intracellular protein synthesis was less influenced by the OA (91.9%) and more strongly suppressed by the RA (78.7%) and the PA (76.7%) synovial fluids. PCEF therefore appear to be a convenient and sensitive target cell system to study alterations of biochemical processes caused by crude synovial fluids and also of different origin by individual factors isolated from synovial fluids.

[The enzymatic mechanisms involved in the pathogenesis of rheumatoid arthritis and arthrosis. The role of metalloproteases and serine proteases in the breakdown of articular cartilage]. / I meccanismi enzimatici coinvolti nella patogenesi dell'artrite reumatoide e dell'artrosi. Ruolo delle metallo-proteasi e delle serino-proteasi nel deterioramento della cartilagine articolare.

Rheumatoid arthritis and osteoarthritis, expression, respectively, of inflammatory and degenerative articular involvement, are the most important diseases affecting joint cartilage. Proteolytic enzymes are the effectors of the articular damage: their increased production by chondrocytes and synoviocytes leads to cartilage breakdown. These enzymes, whose structure and specific activities have been defined in the recent years, carry out their action in the extracellular matrix. They are characterized by the presence of a particular element at the active site, that allows to distinguish four different families. The most studied and the best known among them are metalloproteinases, represented by several enzymes with common features, and serinoproteinases, some members of which, particularly urokinase and tissue-type plasminogen activator, are recently indicated as potential responsible for articular destruction. Activators, and inhibitors of proteinases play a fundamental role: a fine balance, that controls the mechanisms of activation and inhibition, seems to take place at transcriptional level. Any factor able to modify the cellular shape and the cytoskeleton, gives rise to lytic enzyme expression at the genomic level. With the progress of knowledges, serinoproteinases are assuming an increasing relevance, particularly the components of the fibrinolytic pathway.