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.

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.

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.

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.

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.

Possible role of sweating in the pathophysiology of panic attacks.

In recent years the role of hyperventilation in the generation of panic attacks has attracted a considerable amount of interest. According to these studies hyperventilation can elicit the somatic symptoms of panic due to systemic alkalosis. We suggest that since in the case of panic, sweating might cause alkalosis, it could also contribute to the generation of panic attacks. In light of this hypothesis we made a statistical analysis of the panic symptoms of 111 panic patients diagnosed according to DSM-III criteria. The analysis revealed that: (1) there was a well identified group of panic patients who had minor breathing difficulties with heavy sweating; and (2) that all the patients sampled had either severe breathing, or sweating symptoms, or both. We conclude that in the absence of the intensive physical activity of the 'flight or fight' reaction, sweating as well as hyperventilation can cause alkalosis, which in turn might generate panic attacks.