From microvasculature to fibroblasts: Contribution of anti-endothelial cell antibodies in systemic sclerosis.

Systemic sclerosis (SSc) is an autoimmune disease characterized by skin and internal organ fibrosis, caused by microvascular dysfunction. In recent years, the hypothesis that anti-endothelial cell antibodies (AECA) play a key role in microvascular damage seems to be increasingly convincing. In fact, AECA can induce antibody-dependent cellular apoptosis and stimulate the microvasculature to release pro-inflammatory and pro-fibrotic cytokines. Human-microvascular-endothelial-cells (MVECs) were stimulated with SSc sera (with and without AECA) and with sera from healthy donors. The conditioned MVEC culture media were then added to fibroblast cultures obtained from control skin (CTR), non-affected skin of SSc patients (NA), and affected skin of the same sclerodermic (SSc) patients, respectively. AECA contributed to the MVEC increased release of endothelin-1 (ET-1) in the culture medium and to MVEC apoptosis. Fibroblast (CTR, NA, and SSc) proliferation was increased after treatment with AECA-positive conditioned media, compared to AECA-negative and control conditioned media. Furthermore, both AECA-positive (in major contribution) and AECA-negative conditioned media were responsible for alpha-smooth-muscle-actin (αSMA) over-expression in all fibroblast cultures, compared to control conditioned media. Fibroblast type I collagen synthesis was upregulated by both SSc conditioned media (with and without AECA). Finally, the synthesis of fibroblast transforming-growth-factor-beta (TGF-ß) was statistically higher in AECA-positive conditioned media, compared to AECA-negative and control conditioned media. These findings support the concept that AECA may mediate the crosstalk between endothelial damage and dermal-fibroblast activation in SSc.

Effects of intravenous zoledronate and ibandronate on carotid intima-media thickness, lipids and FGF-23 in postmenopausal osteoporotic women.

OBJECTIVE: Osteoporosis and atherosclerosis are interconnected entities and share also some pathophysiological mechanisms. Moreover, recent literature data have supported the hypothesis that bisphosphonates (BPs) may have some antiatherogenic actions. This study aimed to evaluate the effects of one year with zoledronate or ibandronate given intravenously on lipid profile and on carotid artery intima-media thickness (CA-IMT). METHODS: Sixty postmenopausal osteoporotic women (mean age: 66.6±7.8years) were randomly assigned to 1-year treatment with zoledronate 5mg i.v. annually or ibandronate 3mg i.v. every 3 months. In all patients at baseline and after 12months we measured CA-IMT, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), 25-hydroxyvitamin D (25OHD), bone alkaline phosphatase (B-ALP), type I collagen ß carboxy telopeptide (ßCTX), osteocalcin (OC), fibroblast growth factor 23 (FGF-23) and sclerostin. RESULTS: The osteoporotic women treated with zoledronate showed a greater reduction in CA-IMT than those treated with ibandronate. HDL-C and HDL-C/LDL-C ratio showed a significant (p<0.01) increase in the 2 groups, whereas, LDL-C showed a reduction in the two groups which, however, reached statistical significance (p<0.05) only in the zoledronate group. FGF-23 serum levels showed a similar and significant decrease in both the women treated with zoledronate and in those treated with ibandronate. At the end of the study period sclerostin serum levels showed a higher increase in the patients treated with zoledronate than in those treated with ibandronate. CONCLUSION: In osteoporotic women both zoledronate and ibandronate given intravenously resulted in an increase in HDL-C/LDL-C ratio and a reduction of CA-IMT which was significant only for zoledronate. Further prospective studies are needed to clarify whether the change in FGF-23 and sclerostin levels is a marker or a potential mechanism of the action of BPs at a vascular level.

The association of body composition and sex hormones with quantitative ultrasound parameters at the calcaneus and phalanxes in elderly women.

We investigated the associations of body composition and sex hormones with quantitative ultrasound (QUS) parameters carried out at different skeletal sites. In 897 postmenopausal women (64.1 ± 6.6 years) we measured QUS at the calcaneus (stiffness) by Achilles-GE and at phalanxes (amplitude-dependent speed of sound [AD-SOS], bone transmission time [BTT], and ultrasound bone profile index [UBPI]) by Bone Profiler-IGEA. In all subjects we measured fat mass (FM), lean mass (LM), android fat, and gynoid fat by DXA. In all subjects we also assessed serum testosterone (T), estradiol (E(2)), sex-hormone binding globulin, free estrogen index (FEI), free androgen index, 25-hydroxyvitamin D (25OHD), bone alkaline phosphatase (B-ALP), and type I collagen ß carboxy telopeptide. Both E(2) and FEI showed weak but significant correlations with stiffness and QUS parameters at phalanxes. No significant relationships were found between T and QUS. BMI and LM were positively correlated with stiffness (r = 0.14 and r = 0.17, respectively), whereas BMI and FM showed negative correlations with AD-SOS, BTT, and UBPI. 25OHD showed positive relationships with stiffness and QUS at phalanxes. In multivariate models LM and age were associated with stiffness whereas E(2) and age were significant predictors of BTT. AD-SOS was negatively associated with FM, B-ALP, and age but positively with E(2) and 25OHD. In postmenopausal women QUS parameters at the calcaneus and at phalanxes are significantly, but diversely, associated with body composition, sex hormones, 25OHD, and bone turnover markers.

The relationship of ghrelin and adiponectin with bone mineral density and bone turnover markers in elderly men.

Body weight is commonly considered a significant predictor of bone mineral density (BMD). Adiponectin, an adipocyte-derived hormone, could modulate BMD. Moreover, recent studies have reported that ghrelin is able to stimulate bone formation. In this study, we investigated any associations of adiponectin and ghrelin serum levels with bone turnover markers and BMD in elderly men. In 137 men aged 55 years and older (mean age 67.4 +/- 5.4 years, mean body mass index [BMI] 26.6 +/- 3.4 kg/m2), we evaluated serum adiponectin, serum ghrelin, body composition (fat mass and lean mass), BMD, bone alkaline phosphatase (ALP), and the carboxy-terminal telopeptide of type I collagen (betaCTX). Ghrelin showed significant correlations with BMD at the femoral neck (r = 0.25, P < 0.01), total femur (r = 0.22, P < 0.05), and whole body (r = 0.18, P < 0.05). However, after adjusting for age, BMI, and calcium intake, the correlation remained significant only for femoral neck BMD. Ghrelin showed a significant correlation with lean mass but not with fat mass and bone turnover markers. Adiponectin showed a positive association with both bone ALP and betaCTX; the correlation between adiponectin and bone ALP (r = 0.25, P < 0.01) remained significant after adjusting for confounding variables. No significant correlations between adiponectin and BMD at all skeletal sites were observed. In conclusion, our study suggests that in elderly Italian men serum ghrelin was significantly associated with femoral neck BMD and that adiponectin was positively associated with bone ALP. Further studies are needed to elucidate the role of adipocytokines in bone metabolism.

Bone ultrasonography at phalanxes in patients with Rett syndrome: a 3-year longitudinal study.

Osteopenia is a frequent and early complication of Rett syndrome. This study aimed to evaluate the usefulness of Quantitative Ultrasonography (QUS) at phalanxes in the assessment and monitoring of bone status in Rett patients. We studied 109 girls (10.1+/-6.1 years; range 3-25 years) and 101 age-matched controls. Serum calcium (Ca), bone alkaline phosphatase (B-ALP), parathyroid hormone (PTH), 25-hydroxyvitamin D (25OHD) and QUS parameters at phalanxes by Bone Profiler-IGEA (amplitude dependent speed of sound: AD-SoS and bone transmission time: BTT) were measured. At baseline both QUS parameters and 25OHD levels were significantly lower in Rett patients than in controls. Serum 25OHD was inversely correlated with serum PTH and BTT Z-score and BTT Z-score was significantly lower (p<0.05) in the girls with a 25OHD serum levels

Prevalence of Helicobacter pylori infection in male patients with osteoporosis and controls.

Cytokines that regulate bone turnover (tumor necrosis factor-alpha, interleukin-6, etc.) may influence the pathogenesis of skeleton disorders, such as osteoporosis. Since Helicobacter pylori infection increases the systemic levels of inflammatory cytokines, we investigated the possibility that this infection increases the risk of developing osteoporosis and affects the bone metabolism in a group of male patients with osteoporosis. We examined 80 osteoporotic male patients and 160 controls for serum antibodies to H. pylori and the CagA protein and determined, in patients alone, the most important biochemical and instrumental parameters of the disease. Fifty-one patients (63.7%) and 107 controls (66.8%) were seropositive for H. pylori infection (nonsignificant); 30 infected patients (58.8%) and 43 infected controls (40.1%) were positive for anti-CagA antibodies (P = 0.028; OR = 2.13). Levels of estradiol in infected CagA-positive patients were significantly lower than in infected CagA-negative patients (28.5 [SD = 10.18] vs. 39.5 [SD = 14.50] pg/ml; P = 0.002) and uninfected patients (35.2 [SD = 12.7] pg/ml; P = 0.028). Levels of urinary cross-laps(a marker of bone resorption) were increased in patients infected by CagA-positive strains compared to patients infected by CagA-negative strains (282.9 [SD = 103.8] vs. 210.5 [SD = 150.1]microg/mmol; P = 0.048) and uninfected patients (204.3 [SD = 130.1] microg/mmol; P = 0.016). Differences among uninfected and infected patients, independent of CagA status, were observed for other markers of bone turnover, but they did not reach statistical significance. Infection by CagA-positive H. pylori strains is more prevalent in men with osteoporosis, who show reduced systemic levels of estrogens and increased bone turnover. H. pylori infection by strains expressing CagA may therefore be considered a risk factor for osteoporisis in men.

Leptin and body composition in healthy postmenopausal women.

BACKGROUND: Leptin has been proposed to be involved in central control of adiposity and fat distribution but the role of this peptide is controversial. The aim of our study was to test the relationship between serum leptin and body composition, fat distribution, and some biochemical markers such as fasting insulinemia and lipoproteins in a population of healthy Italian postmenopausal women. METHODS: One hundred and twenty-three postmenopausal women (62.1+/-8.7 years) were evaluated. Body composition (fat and lean mass) was assessed by dual-energy X-ray absorptiometry (DXA). Two regions of interest were determined for regional fat analysis. Serum leptin and insulinemia were measured by radioimmunoassay, lipoproteins with colorimetric methods and apolipoproteins nephelometrically. RESULTS: Plasma leptin levels are strongly related to total fat mass, in grams (r=0.73, p<0.001) or as a percentage of soft tissue (r=0.75, p<0.001), and to adiposity, calculated as ratio between lean and fat mass (r=0.76, p<0.001). A significant correlation was also found between serum leptin and central fat distribution (r=0.29, p<0.01). As concerns biochemical markers, serum leptin was significantly related to fasting insulin (r=0.38, p<0.001), total cholesterol (r=0.29, p<0.01), Apolipoprotein-B (r=0.35, p<0.001), and triglycerides (r=0.22, p<0.05). When corrected for total fat mass, the partial correlation coefficients remain significant for percentage of total body fat (r=0.27, p<0.01), adiposity (r=0.23, p<0.01), and fat proportion in android region (r=0.18, p<0.05). CONCLUSIONS: These data indicate that leptin levels are related to adiposity and fasting insulin levels; indeed fast insulin mantains significant correlation with leptin (r=0.23, p<0.01) after controlling for fat mass. Android distribution of fat mass in elderly women is associated with leptin concentration.

Influence of insulin-like growth factor-1 and leptin on bone mass in healthy postmenopausal women.

This study examines the influence of circulating insulin-like growth factor-1 (IGF-1) and serum leptin on bone mass as well as modulation of bone mass during skeletal development. Moreover, an inverse relationship between IGF-1 and leptin is reported. To evaluate the effects of serum IGF-1 and serum leptin on bone mass in healthy postmenopausal women, and the possible role of IGF-1 in leptin production, we studied a population of 123 women, aged 39-82 years. Bone mineral density (BMD) was determined by whole-body dual-energy X ray absorptiometry, which also enables measurement of body composition. Bone metabolism was assessed by measuring serum total alkaline phosphatase (TAP) and urinary hydroxyproline/creatinine (HP/Cr) excretion. IGF-1 correlated significantly with age (r = -0.28, p < 0.01) and years since menopause (r = -0.24, p < 0.01). A negative correlation was also found with weight and body mass index (r = -0.15, p < 0.05 and r = -0.19, p < 0.05, respectively). Leptin values were strongly correlated with weight (r = 0.7, p < 0.01), BMI (r = 0.7, p < 0.01), fat mass (r = 0.77, p < 0.01), and lean mass (r = 0.39, p < 0.01); a significant correlation was found with total body BMD (r = 0.29, p < 0.01), TAP (r = 0.15, p < 0.05), and HP/Cr (r = 0.18, p < 0.05). After adjustment for BMI, the significance of these relationships disappeared, demonstrating the lack of effect of serum leptin on BMD and bone turnover independent of body weight. On the other hand, the relationship between BMD and fat mass remained statistically significant after adjusting for serum leptin (r = 0.15, p < 0.05). Controlling for BMI eliminated the significant inverse correlation between IGF-1 and leptin; significant differences in leptin levels were found among women in the lower and higher quartile of IGF-1, suggesting that leptin production may be inhibited only at high values of serum IGF-1. We conclude that serum IGF-1 and serum leptin have no direct effect on bone mass and bone turnover.

[The efficacy and safety of glucosamine sulfate in the treatment of gonarthritis]. / Efficacia e tollerabilità della glucosamina solfato nel trattamento della gonartrosi.

This study was performed to evaluate the therapeutic efficacy and tolerability of glucosamine sulfate in patients with gonarthritis. During the 12-month study period, the signs and symptoms of the disease were evaluated, as well as the dosage of the urinary pyridinoline. In this trial, we demonstrated that glucosamine sulfate has a chondroprotective activity, which was significant after the first 3 months of therapy. Moreover, this study showed that the side effects due to glucosamine sulfate were mild to moderate and did not require discontinuation of the drug.

Tolerability of nabumetone: a pilot in vitro study showing absence of injury to articular cartilage.

Nabumetone, a compound of the naphthylalkanone class, has shown considerable anti-inflammatory, analgesic and antipyretic effects, together with high systemic and organ-specific tolerability. Its reputation for tolerability has been increased by an in-vitro study evaluating the mature collagen metabolism's markers under nabumetone treatment, which shows that, nabumetone does not interfere with collagen synthesis. Therefore, unlike some nonsteroidal anti-inflammatory drugs, nabumetone might not be injurious to articular cartilage. This feature makes nabumetone even safer for long-term treatment of rheumatic and orthopaedic conditions.

Serum osteocalcin radioimmunoassay in bone diseases.

Osteocalcin (or bone Gla protein, BGP) is a non-collagenous vitamin K-dependent protein accounting for 1-2% of the total bone proteins. It represents a specific index of osteoblastic activity and directly reflects the bone turnover. Serum levels of osteocalcin were measured by a radioimmunoassay method. In 40 postmenopausal women with osteoporosis, mean serum BGP levels were lower than the normal range (3.69 +/- 1.35 ng/ml), whereas they significantly increased in 7 patients with osteomalacia (10.48 +/- 3.05 ng/ml), in 12 patients with secondary hyperparathyroidism (11.1 +/- 4.9 ng/ml) and in 41 patients with Paget's disease (12.09 +/- 6.5 ng/ml). Four patients with primary hyperparathyroidism showed very high BGP levels (64.0 +/- 32.3 ng/ml), which strikingly fell after the surgical removal of a parathyroid adenoma. These results confirm that the quantitation of serum osteocalcin is a specific and sensitive method in the diagnosis of bone disease, represents a useful index of bone turnover and is particularly helpful in the follow-up of patients with treated bone disease.

Calcitonin levels in normal subjects according to age and sex.

To evaluate the effect of age and sex on plasma calcitonin in human beings, the concentrations of the hormone were measured in 63 normal subjects aged 13-87 years of both sexes. In another study 30 healthy women were studied, 14 of them were pre-menopausal, 16 were post-menopausal. Plasma calcitonin was determined by means of a radioimmunoassay, using delayed tracer addition for increasing sensitivity. The antibody was produced in rabbits against pure synthetic human calcitonin and was specific for the aminoacid sequence 17 to 32 in the calcitonin molecule. A second antibody was used as a precipitating agent. A mean plasma calcitonin level of 71.3 +/- 37.0 SD pg/ml was observed. Women were found to have lower levels than men, the mean values being 63.4 +/- 34.7 pg/ml and 92.6 +/- 35.4 pg/ml respectively. The difference was statistically significant (P less than 0.005). No significant correlation was found between calcitonin levels and age of subjects. Premenopausal women, however, showed higher levels of plasma calcitonin than post-menopausal women, the mean values being 88.5 +/- 38.4 pg/ml and 54.0 +/- 33.6 pg/ml, respectively. This difference was also significant (P less than 0.01). Possible implications of these data are discussed.

[Behavior of plasma and urinary cAMP after intravenous fructose load]. / Comportamento dell'AMPc plasmatico e urinario dopo carico endovenoso di fruttoso.

Increased excretion of uric acid and cAMP has been observed in many pathological conditions in man, but interrelation between behaviour of uric acid and cAMP is not well known. To investigate these aspects of purine metabolism the authors have studied behaviour of plasma and urinary cAMP in 8 subjects after intravenous rapid load of fructose (0.5 g/Kg b.w.) compared with the uric acid one. The results have shown that urinary excretion of cAMP after load of fructose is increased, but levels of plasma cAMP remain unchanged. These data confirm consensual relation between urinary uric acid and cAMP excretion, but the exact mechanism of this relation remains unexplained.

[Asu(1,7)E-CT, an analog of eel calcitonin. A comparative study in man with reference to other synthetic calcitonins]. / La (Asu) E-CT, un analogo della calcitonina di anguilla. Studio comparativo nell'uomo rispetto ad altre calcitonine sintetiche.

(Asu) E-CT is a deaminodicarba-analog of the synthetic eel-calcitonin (E-CT) that shows specific activity and the potency reasonably high in comparison with that of the most potent natural hormone. The structure of its molecule indicates that the disulphide bond in calcitonins is not essential for the biological activity but only for the maintenance of the specific conformation by forming an intramolecular bridge. The instability of calcitonins should mainly be attributed to the presence of the disulfide bond and (Asu)E-CT proved to be more stable "in vitro" than native calcitonins. The more prolonged hypocalcemic effect of E-CT and its aminosuberic analog (Asu)E-CT has been accounted for to a greater stability of and persistence at the receptor site. (Asu) E-CT has been largely studied in Japan on experimental animals and successfully used in the treatment of hypercalcemia in man. On the contrary investigations on human administration of this analog are very scarce. The present paper reports studies carried out in normal subjects and Paget's disease patients to investigate the effects of (Asu)E-CT in man in comparison with the effects of synthetic human calcitonin (H-CT) and synthetic salmon calcitonin (S-CT). Two different experimental procedures have been used: 1) rapid intravenous injection of (Asu)E-CT (80 MRC. U.) or respectively of H-CT and S-CT (100 MRC. U.) in 15 subjects (7 normals and 8 with Paget's disease); 2) slow 7 days continuous subcutaneous infusion of similar daily amounts of (Asu)E-CT, H-CT and S-CT administered by a microjet pump device in 21 subjects (7 normals and 14 with Paget's disease). The intravenous administration of (Asu)E-CT induced a rapid and persistent decrease in total plasma calcium, ionized calcium and plasma phosphate that was more evident in Paget's disease patients than in normal subjects. No clearly cut differences have been observed with the hypocalcemic and hypophosphatemic effect of H-CT and S-CT administered intravenously; nevertheless the hypocalcemic effect proved to be more persistent in Paget's disease patients treated with (Asu)E-CT. After intravenous infusion of (Asu)E-CT the plasma level of cAMP rose more evidently in pagetic than in normal subjects but the rise was lower than in H-CT and S-CT treated subjects.(ABSTRACT TRUNCATED AT 400 WORDS)

[Behavior of urinary excretion of cyclic AMP in post-menopausal osteoporosis during therapy with salmon calcitonin]. / Comportamento dell'escrezione urinaria di AMP ciclico nella osteoporosi post-menopausale in corso di terapia con calcitonina di salmone.

Salmon calcitonin has been administered at a dose of 100 U.M.R.C. for 6 months in 10 patients suffering from post-menopausal osteoporosis. During treatment, calcium plasma levels fluctuated but tended to fall, while urinary excretion of cyclic AMP rose, this pointing to an enhancement of parathyroid function. After 6 months an increase in intestinal calcium absorption and a decrease in bone turnover were also observed, the bone mineral content evaluated by bone densitometry showed a slight quantitative increase but proved to be substantially unchanged. Salmon calcitonin thus proved capable of interfering with bone turnover by reducing the bone resorption. By fostering an increase in parathyroid hormone production, it also made it possible to exploit that anabolic effect which, in low doses, the parathormone exerts on the bone tissue.

[Metabolic changes induced with salmon calcitonin in post-menopausal osteoporosis]. / Modificazioni metaboliche indotte dalla calcitonina di salmone nella osteoporosi post-menopausale.

10 patients suffering from post-menopausal osteoporosis were subjected to venous infusion of salmon calcitonine at a dose of 100 U.M.R.C. Essential inital biochemical data were hypocalcaemia associated with plasma increase in AMPc and, in the late phase, an increase in urinary excretion of phosphate and AMPc. In addition to conserving its physiological properties in osteoporotic disease also, the hormone is thus able to provoke a secondary secretion of parathormone. The use of CT in the treatment of post-menopausal osteoporosis is therefore recommended not only because of its evident action on bone reabsorption processes, which usually predominate in this dysmetabolic disease of the skeleton, but also in the indirect secretion of parathyroid hormone which is capable of positively affecting synthesis of 1-25 (OH)2 D3 and intestinal calcic absorption.