The effects of ß-glucan isolated from Pleurotus ostreatus on methotrexate treatment in rats with adjuvant arthritis.

The purpose of this study was to evaluate the effect of ß-(1,3/1,6)-D: -glucan isolated from Pleurotus ostreatus (ß-glucan-PO) on prophylactic treatment of adjuvant arthritis (AA) with methotrexate (MTX) in rats. Groups of rats with AA were treated with methotrexate (1 mg/kg/week), ß-glucan-PO (1 mg/kg every second day) or their combination for the period of 28 days from adjuvant application. Body mass, hind paw swelling, arthrogram scores and a level of serum albumin were measured as markers of inflammation and arthritis. Treatment with low dose of MTX significantly inhibited the markers of both inflammation and arthritis. MTX and its combination with ß-glucan-PO significantly increased body mass of arthritic rats. ß-glucan-PO administered alone significantly decreased both the hind paw swelling and arthritic score. In combination with MTX, ß-glucan-PO markedly potentiated the beneficial effects of MTX, which resulted in a more significant reduction of hind paw swelling and arthritic scores. The concentration of albumin in the serum of arthritic controls was significantly lower than in healthy controls. Both MTX alone and the combination treatment with MTX + ß-glucan-PO significantly inhibited the decrease in serum albumin. ß-Glucan-PO increased the treatment efficacy of basal treatment of AA with MTX.

Treatment of adjuvant-induced arthritis with the combination of methotrexate and probiotic bacteria Escherichia coli O83 (Colinfant).

A certain relationship was observed between the gastrointestinal system, arthritis and immune system. Patients with rheumatoid arthritis have an altered microflora composition and disturbed intestinal defensive barrier. Effect of probiotic bacteria (Colinfant; COL) with known favorable effect on intestinal microflora was determined on the methotrexate (MTX) treatment of adjuvant arthritis. Rats with adjuvant arthritis were administered methotrexate 0.5 mg/kg body mass 2-times weekly per os, COL 1 mL/kg body mass every second day per os, and a combination of MTX+COL for a period of 28 d from the immunization. Levels of serum albumin, body mass, changes in hind paw swelling, and arthrogram score were estimated in rats as variables of inflammation and destructive arthritis-associated changes. Treatment with MTX, as well as with the combination treatment with MTX+COL significantly inhibited both inflammation and destructive arthritis-associated changes. The combination treatment inhibited both the hind paw swelling and arthrogram score more remarkably than MTX alone; on the other hand, the difference between combination treatment and MTX alone was not significant. Treatment with COL alone had no effect on adjuvant arthritis in rats. Colinfant can increase the preventive effect of MTX treatment in rat adjuvant arthritis by improving its antiarthritic effects.

The effects of hyaluronan on bone resorption and bone mineral density in a rat model of estrogen deficiency-induced osteopenia.

Hyaluronan, or hyaluronic acid (HA), is an essential component of extracellular matrices. HA of appropriate molecular weight and concentration can induce osteoblast differentiation and bone formation in vitro. The aim of our study was to evaluate the effects of HA of different molecular weights on ovariectomy (OVX)-induced bone loss in rats. Adult female Sprague Dawley rats were subjected to bilateral OVX or sham surgical procedure (sham). OVX rats were treated with: HA of molecular weight of 0.75 MDa at a dose of 1 mg/kg/day and with HA of molecular weight of 1.62 MDa at a dose of both 0.5 mg/kg/day and 1 mg/kg/day. HA was applied orally once a day during the 8-week period after ovariectomy. Body weight, urinary pyridinoline (Pyr), deoxypyridinoline (DPyr) corrected for urinary creatinine, serum nitrite/nitrate concentrations and whole body and femoral bone mineral density (BMD) were measured. HA treatment had no effect on the body weight gain in OVX rats. Excretion of urinary Pyr and Dpyr significantly increased in OVX rats compared to sham controls. The higher molecular weight HA (1.62 MDa) significantly reduced urinary Pyr and DPyr concentrations measured on day 28 after ovariectomy (p < 0.001). Serum concentrations of nitric oxide metabolites, nitrite/nitrate significantly decreased in OVX rats in comparison with sham controls (p < 0.001). HA of both 0.75 MDa and 1.62 MDa molecular weights significantly enhanced serum nitrite/nitrate concentrations in OVX rats. There was a clear reduction of whole body and femoral BMD in untreated OVX rats. The higher molecular weight HA decreased both whole body and femoral BMD loss. Our results show that orally applied HA of high molecular weight (1.62 MDa) inhibits bone resorption and provides a protective effect on bone density in ovariectomized rats.

Quantitative assessment of markers of inflammation, arthritis and pain in rats with adjuvant arthritis following the treatment with ibuprofen cream and placebo.

When applied topically, nonsteroidal antiinflammatory drugs (NSAIDs) are absorbed locally and hence systemic adverse effects can be avoided due to very low plasma concentrations. In addition, direct local antiphlogistic treatment of the affected area is desirable. The present study was undertaken to assess markers of inflammation, arthritis and pain in rats with adjuvant arthritis undergoing treatment with ibuprofen cream (Dolgit cream) and placebo. Ibuprofen cream was applied (70 mg/hind paw) 3 times daily on hind paws from day 12 (after the initial signs of hind paw swelling appeared) for 3 weeks. Pain threshold, hind paw swelling, arthrogram score and serum albumin concentrations were measured in healthy controls, in rats with adjuvant arthritis and in rats treated with both placebo and active cream. Pain threshold increased in rats treated with ibuprofen cream (after 12 days of application), but this change was not significant. However, hind paw swelling significantly decreased as early as after 5 days of treatment with ibuprofen cream in comparison with both arthritic and placebo-treated rats and remained decreased throughout the study (an additional 16 days). Evaluation of the arthrogram score gave similar results. Serum albumin levels were unaffected by ibuprofen cream treatment. In conclusion, our results show that topical application of ibuprofen cream on inflamed hind paws produced significant local antiinflammatory and antiarthritic effects in rat adjuvant arthritis.

Eggshell calcium in the prevention and treatment of osteoporosis.

In this paper the most significant biological and clinical aspects of a biopreparation made of chicken eggshells are reviewed. Eggshell powder is a natural source of calcium and other elements (e.g. strontium and fluorine) which may have a positive effect on bone metabolism. Experimental and clinical studies performed to date have shown a number of positive properties of eggshell powder, such as antirachitic effects in rats and humans. A positive effect was observed on bone density in animal models of postmenopausal osteoporosis in ovariectomized female rats. In vitro eggshell powder stimulates chondrocyte differentiation and cartilage growth. Clinical studies in postmenopausal women and women with senile osteoporosis showed that eggshell powder reduces pain and osteoresorption and increases mobility and bone density or arrests its loss. The bioavailability of calcium from this source, as tested in piglets, was similar or better than that of food grade purified calcium carbonate. Clinical and experimental studies showed that eggshell powder has positive effects on bone and cartilage and that it is suitable in the prevention and treatment of osteoporosis.

Differences in hormonal and inflammatory parameters in male Lewis and Long Evans rats with adjuvant arthritis.

The purpose of the present study was (i) to compare secretory responses of prolactin and corticosterone to the acute stress of immobilization in male rats of the Lewis and Long Evans strains and (ii) to compare secretion of the two hormones in rats with fully developed adjuvant arthritis (AA) and their relationship with the intensity of the inflammatory reaction. A short immobilization of 5 min induced equal elevations of both hormones in both strains, but 20-min immobilization produced significantly stronger responses in Long Evans rats than in Lewis rats. AA inhibited prolactin secretion equally in both strains (from 11.6 +/- 1.3 ng/ml to 4.2 +/- 0.6 ng/ml in Lewis rats, p < 0.01, and from 3.7 +/- 0.6 to 2.12 +/- 0.1 ng/ml in Long Evans rats, p < 0.05), but caused a conspiciously larger elevation of corticosterone in the Long Evans than in the Lewis animals (11.5 +/- 1.2 microg/dl in Long Evans rats versus 5.1 +/- 0.5 microg/dl in Lewis rats, p < 0.01) while basal levels were similar. The larger corticosterone response in the Long Evans rats was associated with a stronger inflammatory reaction assessed by hind paw swelling (2.3 +/- 0.1 ml for Long Evans rats versus 1.8 +/- 0.08 ml for Lewis rats, p < 0.01) and plasma levels of nitric oxide (47.5 +/- 5.7 microM for Long Evans rats versus 28.7 +/- 2.5 microM for Lewis rats, p < 0.01) than in the Lewis males with lower corticosterone levels. In conclusion, there are significant, obviously genetically based, differences in the corticosterone responses to both immobilization and AA between the two strains, with the Long Evans rats reacting more strongly than the Lewis rats. The lack of the expected inverse relationship between corticosterone levels and the intensity of the inflammation indicates that the activity of corticosterone is not its primary determinant and that other important factors are involved.

Treatment of experimental adjuvant arthritis with the combination of methotrexate and lyophilized Enterococcus faecium enriched with organic selenium.

The efficacy of combination therapy with methotrexate (MTX) and probiotic bacteria Enterococcus faecium enriched with organic selenium (EFSe) in rats with adjuvant arthritis was determined. Rats with adjuvant arthritis were given MTX (0.3 mg/kg 2-times weekly, orally); lyophilized E. faecium enriched with Se (15 mg/kg, 5 d per week, orally); and a combination of MTX plus EFSe for a period of 50 d from the immunization. Levels of serum albumin, serum nitrite/nitrate concentrations, changes in hind paw swelling, arthrogram score, bone erosions, whole body bone mineral density (BMD) and bone mineral content (BMC) were assayed in the rats as variables of inflammation and destructive arthritis-associated changes. Treatment with MTX and with the combination MTX + EFSe significantly inhibited markers of both inflammation and arthritis. Significant differences in favor of combination therapy with MTX + EFSe as compared to MTX alone were seen in serum albumin concentration, hind paw swelling and arthrogram score. Reductions in radiographic scores were also more pronounced in the combination therapy group. Combination therapy, but not MTX alone, inhibited the reduction of BMD and BMC; treatment with lyophilized EFSe alone had no significant effect on adjuvant arthritis in rats. The potent therapeutic effect of low dosage MTX therapy in combination with lyophilized EFSe on adjuvant arthritis in rats was shown.

Effects of chronic food restriction stress and chronic psychological stress on the development of adjuvant arthritis in male long evans rats.

The aim of the present experiments was to study the effect of stress of chronic food restriction (FR) and of repeated psychological challenge (PS) on the development of adjuvant arthritis in the Long Evans male rats. In the FR series, four groups of animals were compared: non-treated control (C) and arthritic (AA) rats, both with free access to food and water, and two analogous groups with a 40% food restriction-FR and AA-FR. All animals were killed 22 days after injection of cFA. In the PS series, stress was induced by random daily exposures of the rats to isolation, over-crowding, food/water deprivation, foot shock, tilting, fear for 14 days before cFA injection and 12 days thereafter (groups: C, AA, PS, and AA-PS). Arthritis causes swelling of the hindpaw, which was prevented in the AA-FR group. PS causes more severe disease symptoms: AA-PS rats had more severe hindpaw swelling than AA rats. Forty percent food restriction associated with elevated CORT levels mitigated inflammatory parameters activated during AA. PS worsened the disease. These results suggested that activated CORT is not the only cause of disease suppression, but some metabolic changes during FR play a role.

[Prediction of changes in bone density during alendronate treatment in postmenopausal women]. / Predikcia zmien kostnej denzity pri liecbe alendronátom u postmenopauzálnych zien.

Alendronate is an aminobisphosphonate, a selective inhibitor of osteoclast-mediated bone resorption. Due to its influence a decline of markers of bone turnover occurs. The latter react much sooner than it is possible to detect significant changes of bone density. In the submitted trial the authors investigated changes of selected markers: total alkaline phosphatase (ALP), osteocalcin (OC), N-terminal telopeptide fragment of collagen type I (NTx) after 3 months' treatment with alendronate, the influence on bone density after one year's treatment in 50 postmenopausal women with densitometrically verified osteoporosis. After one-year treatment in the whole group a significant increase of bone density occurred in the area L2-L4 by 4.52% (SD = 3.9), neck of the femur by 2.24% (SD = 3.6), trochanter by 2.81% (SD = 3.0) and total by 1.89% (SD = 2.7). Total ALP and OC in serum, similarly as NTx in urine declined significantly already after 3 months treatment and the decline persisted also after one year of treatment. With the change of bone density after one year correlated significantly only NTx. A decline of NTx after 3 months by more than 30% as compared with the baseline value was recorded in 81% patients and this change predicted a significant rise of the bone density in the area of the neck of the femur, on average by 30. Urinary NTx is a promising predictor of the effect of alendronate treatment. Its drop by more than 30% after 3 months justifies the assumption that bone density increased after one year's treatment.

Evidence for increased pyridinoline concentration in fibrotic tissues in diffuse systemic sclerosis.

Systemic sclerosis is a generalized disease characterized mainly by the accumulation of collagen in the skin and internal organs. The aim of our study was to determine the amount of collagen cross-link pyridinoline (Pyd) in a variety of fibrotic tissues (skin, fascia, endocardium, bladder) from an autopsy patient with diffuse systemic sclerosis, and to compare these with normal tissues from the same patient. Mean concentrations of Pyd in the fibrotic skin samples (66 mmol/mol collagen) were more than two-times greater than those in the uninvolved normal samples (27 mmol/mol collagen). The increase of Pyd in the endocardium, fascia, and bladder was also markedly higher (1.41 x, 1.26 x and 2.64 x higher than normal samples). The increased deposition of collagen in systemic sclerosis is accompanied by a significantly increased amount of Pyd in the collagen of fibrotic tissues.

Stress of chronic food restriction attenuates the development of adjuvant arthritis in male Long Evans rats.

OBJECTIVE: To study the effect of the stress of chronic food restriction on the development of adjuvant arthritis in Long Evans male rats. METHODS: Four groups of animals were compared: non-treated control (C) and arthritic (AA) rats, both with free access to food and water and two analogous groups with a 40% food restriction, i.e. non-treated (FR) and arthritic (AA-FR) animals. All rats were killed 22 days following the injection of complete Freund's adjuvant. The parameters measured were: serum levels of albumin (ALB), nitrate, glucose, insulin, corticosterone (CORT), prolactin (PRL) and PRL mRNA in the adenopituitaries. In addition the activity of gamma-glutamyl transpeptidase (GGTP) was measured in the spleen. The pain threshold was determined by the tailflick method. The body weight of the animals was recorded on day 0, 3, 7, 11, 15 and 18 of the disease. RESULTS: Arthritis caused swelling of the hind paw (2.37 +/- 0.15 ml vs 1.1 +/- 0.05ml in controls, p < 0.01) which was prevented in the AA-FR group (1.44 +/- 0.13 ml, not significant against controls). Arthritis increased serum NO and reduced ALB levels; both changes were significantly restored in the FR-AA group. Food restriction did not alter the activation of GGTP, or the decrease of PRL mRNA observed in the AA group. Serum CORT was elevated in rats with food restriction (15.49 +/- 2.1 vs. 3.7 +/- 0.87 microg/dL) and remained enhanced to the same extent in AA and FR-AA groups. The tailflick latency prolonged in the AA group was reduced by food restriction. CONCLUSION: These results show that 40% food restriction associated with elevated CORT levels mitigated the inflammatory parameters activated during AA.

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.

Inhibitory effect of enzyme therapy and combination therapy with cyclosporin A on collagen-induced arthritis.

OBJECTIVE: There is increasing interest in the use of combination therapy for rheumatoid arthritis and in the possibility of combining the conventional drug approach with newer antirheumatic therapy. The present study investigates the efficacy of long-term prophylactic enzyme therapy and combination therapy with cyclosporin A in rats with collagen-induced arthritis. METHODS: Rats with collagen-induced arthritis were administered the following drugs: cyclosporin A (5 mg/kg/day and 10 mg/kg/day orally); a mixture of enzymes containing pure substances (bromelain, trypsin, rutin) in the same ratio as in Phlogenzym (PHL, 150 mg/kg, twice daily intrarectally); and a combination of 5 mg/kg/day cyclosporin A plus 300 mg/kg/day PHL for a period of 50 days from the immunization. Levels of serum albumin, serum nitrite/nitrate concentrations, changes in hind paw swelling and bone erosions were measured in the rats as variables of inflammation and destructive arthritis-associated changes. RESULTS: Treatment with 10 mg/kg cyclosporin A, as well as combination therapy with half dosages of cyclosporin A (5 mg/kg) plus PHL significantly inhibited both inflammation and destructive arthritis-associated changes. Significant differences in favor of combination therapy with 5 mg/kg CsA + 300 mg/kg PHL as compared to 5 mg/kg CsA alone were seen in hind paw swelling. Also, reduction of the radiographic scores was more significant in the combination therapy group. Five mg cyclosporin A or PHL alone reduced the disease markers studied to a lesser extent, and in the case of enzyme therapy this occurred at a later stage of arthritis development. CONCLUSION: Our results show the inhibitory effect of enzyme therapy on collagen-induced arthritis in rats, as well as the efficacy of cyclosporin A given in low doses in combination with enzyme therapy, which may be useful in the treatment of rheumatoid arthritis.

Suppression of adjuvant arthritis in rats by boar seminal immunosuppressive fraction.

OBJECTIVE: The effect of the immunosuppressive fraction of boar seminal vesicle fluid (ISF) was tested on the manifestation of adjuvant arthritis (AA) in rats. METHODS: The inhibitory effect of ISF on mitogen-stimulated proliferation of rat lymphocytes was evaluated by immunoassay using bromodeoxyuridine incorporation. Adjuvant arthritis was induced in male Long Evans rats with Mycobacterium butyricum in adjuvant. ISF was administered at the time of the induction of arthritis. At the time of maximal manifestation of the disease, the hind paw swelling and thymus weight were estimated. IgM and IgG in the rat blood sera were quantified by sandwich ELISA. Serum corticosterone was analyzed by radioimmunoassay. Serum NO2-/NO3-were estimated by diazotation. Serum albumin was measured spectrophotometrically. The expression of IL-6 mRNA in peritoneal macrophages was estimated by dot-blot hybridization. RESULTS: Treatment of arthritic rats with ISF attenuated hind paw edema. The production of IgG subclasses dropped in ISF-treated AA rats. The thymus mass and serum albumin concentration were partially restored due to the ISF treatment. Serum corticosterone as well as NO2-/NO3- concentrations were reduced by the ISF effect. The expression of IL-6 in peritoneal macrophages was inhibited in AA rats after ISF treatment. CONCLUSION: ISF attenuated the manifestation of AA in rats and mitigated the inflammation. Immunoglobulin production was most probably inhibited by the decreased proliferation of B lymphocytes.

Increased urinary pyridinoline cross-link compounds of collagen in patients with systemic sclerosis and Raynaud's phenomenon.

OBJECTIVE: To study concentration changes in collagen degradation markers in patients with diffuse and limited cutaneous systemic sclerosis and patients with scleroderma-related diseases. METHODS: Pyridinoline cross-link compounds were analysed in urine samples using high-performance liquid chromatography. Samples were analysed for pyridinoline (Pyr), deoxypyridinoline (Dpyr) and soft-tissue pyridinoline (stPyr) in patients with diffuse cutaneous systemic sclerosis (dcSSc, n=23) and limited cutaneous systemic sclerosis (lcSSc, n=48) and in patients with scleroderma-related diseases such as primary Raynaud's phenomenon (pRP, n=16) and secondary Raynaud's phenomenon (sRP, n=14). Healthy controls (n=18) and patients with post-menopausal osteoporosis (OP, n=35) were also investigated. RESULTS: Urinary Pyr, Dpyr and stPyr concentrations were significantly higher in patients with Raynaud's phenomenon and systemic sclerosis than in healthy controls. The highest concentrations (two to three times greater than in healthy controls) were found in patients with dcSSc. The stPyr concentration was significantly higher in patients with dcSSc than in those with lcSSc, sRP and pRP. No significant difference in stPyr concentration was found between the healthy controls and the OP group, suggesting that stPyr is derived from soft tissues rather than bone. The extent and severity of skin involvement, measured as a skin score, significantly correlated with the concentrations of stPyr and Pyr, whereas no such correlation was found for Dpyr. CONCLUSIONS: Increased urinary concentrations of piridinoline cross-links reflect alterations in collagen turnover in both Raynaud's phenomenon and systemic sclerosis. The close correlation between stPyr concentration and the extent of skin involvement in systemic sclerosis suggests that this parameter may be useful in monitoring ongoing fibrosis in this disease.

Suppression of adjuvant arthritis in rats with chronic bromocriptine treatment does not prevent associated oxidative stress.

We studied the effect of prolactin (PRL) inhibition by bromocriptine (BRC) in the first phase of adjuvant induced arthritis (AA), up to day 11(BRCl-AA), and in the whole time course of AA, up to day 23 (BRC-AA), on the development of the disease in male Lewis rats. On day 24, arthritic rats showed inhibition of PRL secretion, but not PRL mRNA expression in adenopituitaries. BRC treatment suppressed PRL serum levels and PRL mRNA expression in adenopituitaries. In BRC/-AA group PRL levels and PRLmRNA were at the level of rats with AA. Serum corticosterone (CORT) was stimulated by AA from 16.9+/-5.8 to 59.1+/-8.7 ngml(-1), p<0.05, to the same level in BRC-control (BRC-C) and BRC-AA group and further potentiated in BRCI-AA group (148.2+/-33.1 ngml(-1), p<0.05 vs. group with AA). Hind paw swelling was reduced but not completely inhibited in BRC1-AA group and totally prevented in BRC-AA rats as was the core temperature (36.5+/-0.1 degrees C vs. 37.4+/-.0.1 degrees C in AA rats on day 23, p<0.01). Serum concentration of NO-ZNO-3 rose in rats with AA to 28.7+/-2.5 &mgr;mo1L-1 against. 13.9+/-1.9 &mgr;molL(-1) in controls (p<0.01), remained elevated in BRC-AA group and was potentiated in BRC1-AA group (48.2+/-3.5 &mgr;mol(-1), p<0.01 vs. AA or BRC1-AA group) Thiobarbituric acid reactive substances (TBARS) and antioxidant capacity in the spleen were enhanced in rats with AA and to the same extent in BRC-AA or BRC1-AA groups. These results show a discrepancy between the suppression of clinical symptoms and persisting oxidative stress in AA rats after the BRC induced PRL inhibition. The potentiation of nintric oxide (NO-) production after the sudden cessation of PRL inhibition during the disease may promote further joint damage.

[Collagen in the treatment of rheumatic diseases--oral tolerance]. / Kolagén v liecbe reumatických chorôb--orálna tolerancia.

The term "oral tolerance" means antigen specific suppression of immune response after oral application of antigen. Primary mechanisms by which oral tolerance is mediated include: deletion, anergy and active cellular suppression. The determining factor in this process is the dose of applied antigen. High doses of antigen develop deletion and anergy of cells while low doses of antigen result in bystander suppression. Recently bystander suppression has attracted attention in the treatment of autoimmune diseases. This process is connected with induction of regulatory T cells of Th2/Th3 phenotypes in gut with characteristic profile of anti-inflammatory cytokines as IL-4, IL-10 and TGF-beta. By means of circulation the lymphocytes enter the affected place and when meeting again with the antigen, they produce the same profile of cytokines which they originally made in the gut. These cytokines then suppress local autoimmune and inflammatory reaction independently of the antigen type. After successful trials of treatment with low doses of orally applied collagen type II in animal models of experimental arthritis, this treatment was also studied in clinical trials in humans with rheumatoid arthritis. Although the results obtained to this date are very promising they can not be considered final. Several questions still need to be solved: identification of responders, determination of character and amount of collagen applied as well as the route of application. Another promising therapeutic approach could be the simultaneous application of collagen and the compounds enhancing the cell response of Th2 or Th3 lymphocytes such as TGF-beta, IL-2, antibodies to IL-12 which can augment the oral tolerance. In clinical praxis the treatment of osteoarthrosis with collagen type I has also been successfully applied. Induction of oral tolerance is new approach in the treatment of rheumatoid arthritis and as each new therapy, it requires refinement. In the future it is expected that an improved study design and a better understanding of the underlying mechanisms of oral tolerance will lead to an increased efficacy of the therapy in humans similar to the effectiveness previously demonstrated in animal models.

Enzyme and combination therapy with cyclosporin A in the rat developing adjuvant arthritis.

Recent knowledge of the pathophysiology of rheumatoid arthritis and the mechanism of drug effects have enabled the use of new drugs and drug combinations in rheumatoid arthritis therapy. This study investigates the efficacy of both enzyme therapy and combined therapy with cyclosporin in rats with adjuvant arthritis. Rats with adjuvant-induced arthritis were administered either cyclosporin A (2.5 or 5.0 mg/kg/day per os), a mixture of enzymes (Phlogenzym (PHL); 45 mg/kg twice daily intrarectally), or a combination of 2.5 mg cyclosporin A and 90 mg PHL for a period of 40 days from the adjuvant application. Levels of serum albumin, changes in hind paw swelling and bone erosions were measured in rats as variables of inflammation and arthritis-associated destructive changes. Treatment with 5 mg of cyclosporin A, as well as with the combination therapy with cyclosporin A plus PHL, significantly inhibited both the inflammation and destructive arthritis-associated changes. However, 2.5 mg of cyclosporin A and PHL alone inhibited these disease markers, although to a lesser extent and at a later stage of arthritis development. The results show the inhibitory effect of enzyme therapy on rat adjuvant arthritis, as well as the efficacy of a low dose of cyclosporin A given in combination with enzyme therapy, which may be useful in the treatment of rheumatoid arthritis.

Serum adenosine deaminase activity and its isoenzyme pattern in patients with systemic lupus erythematosus.

OBJECTIVE: To study the correlation between serum adenosine deaminase (ADA) activity and clinical activity in patients with systemic lupus erythematosus (SLE). The patterns of two ADA isoenzymes, ADA1 and ADA2, were also analysed in healthy controls and patients to determine the source of increased ADA activity in patients. METHODS: Total serum ADA activity (tADA) was measured spectrophotometrically. The isoenzyme pattern of ADA was analysed by the HPLC method using a specific inhibitor of ADA1, erytro-9-(2-hydroxy-3-nonyl)adenine (EHNA). Disease activity was assessed using the European Consensus Lupus Activity Measurement index (ECLAM). RESULTS: Serum tADA activity was significantly increased in patients compared to healthy controls (mean +/- SD; 476.9 +/- 145.3 vs 254.0 +/- 98.9 ncat/L, p < 0.001). The isoenzyme analyses showed that the increased total ADA activity in the patients was mainly due to increased ADA2 activity (371.3 +/- 154.8 vs 214.2 +/- 47.9 ncat/L in healthy controls, p < 0.001). The mean values for ADA1 activity in the patients (64.6 +/- 37.9 ncat/L) and healthy controls (69.2 +/- 26.9 ncat/L) were similar. A strong correlation was found between serum ADA activity and disease activity as measured by ECLAM (Spearman's rank correlation coefficient 0.74, p < 0.0001, linear regression coefficient 0.68, p < 0.01). CONCLUSION: Serum ADA activity is closely associated with SLE activity and appears to be useful as a new disease activity parameter of SLE.