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Alkaptonuria (AKU) is a rare autosomal recessive disease, associated with deficiency of homogentisate 1,2-dioxygenase activity in the liver. This leads to an accumulation of homogentisic acid (HGA) and its oxidized derivatives in polymerized form in connective tissues especially in joints. Currently, AKU lacks an appropriate therapy. Hence, we propose a new treatment for AKU using the antioxidant N-acetylcysteine (NAC) administered in combinations with ascorbic acid (ASC) since it has been proven that NAC counteracts the side-effects of ASC. We established an in vitro cell model using human articular primary chondrocytes challenged with an excess of HGA (0.33 mM). We used this experimental model to undertake pre-clinical testing of potential antioxidative therapies for AKU, evaluating apoptosis, viability, proliferation, and metabolism of chondrocytes exposed to HGA and treated with NAC and ASC administered alone or in combination addition of both. NAC decreased apoptosis induced in chondrocytes by HGA, increased chondrocyte growth reduced by HGA, and partially restored proteoglycan release inhibited by HGA. A significantly improvement in efficacy was found with combined addition of the two antioxidants in comparison with NAC and ASC alone. Our novel in vitro AKU model allowed us to demonstrate the efficacy of the co-administration of NAC and ASC to counteract the negative effects of HGA for the treatment of ochronotic arthropathy.

Since 1992 extracorporeal shock wave therapy (ESWT) has been used in the treatment of pain, in various tendinopathies, including plantar fascitis, heel spur, epicondylitis, calcific and non-calcific tendonitis of the rotator cuff. The exact mechanism by which ESWT relieves tendon-associated pain is not known; however, numerous controlled and noncontrolled studies have demonstrated the efficacy of ESWT in the treatment of various tendinopathies, including plantar fascitis, heel spur and epicondylitis. Based on personal experiences and analysis of the literature, this review confirmed that ESWT is an effective therapeutic procedure with no side-effects in the treatment of calcific rotator cuff tendonitis, and which can be considered a valuable alternative to conventional treatments.

To study heel fat pad (HFP) inflammatory-oedematous lesions in selected patients with rheumatoid arthritis (RA) using ultrasonography (US) and power Doppler US (PDUS), to describe and compare US features of these lesions with those obtained with magnetic resonance imaging (MRI), and to describe changes in the lesions after a short-term follow-up with conventional or anti-tumour necrosis factor-alpha (TNFalpha) therapy.Twelve heels of eight RA outpatients with HFP inflammatory-oedematous lesions were studied by US, PDUS, and unenhanced MRI. All the patients were followed up and US was performed after 3 months. Five patients started on anti-TNFalpha therapy.HFP lesions appeared at US as a heterogeneous and hypoechoic subcalcaneal mass, with loss of normal lobular structure and increased thickness of HFP, because of focal rupture of fibrous septae with oedema and fluid. PDUS showed peripheral vascularization of HFP lesions in 9/12 heels. In 3/12 heels some vascular signals was also detectable inside the lesion, always along the residual echoic septa. No detectable flow was observed within the central fluid-filled spaces. MRI of the HFP lesions showed areas of mean intensity in T1-weighted sequences and high intensity in T2-weighted sequences, with poorly or well-defined margins. After 3 months, PDUS showed reduction in HFP lesion vascularity (associated with reduction in pain) in 10/12 heels, while poor regression of grey-scale US abnormalities was observed.Both US and MRI are capable of demonstrating structural abnormalities in the HFP. PDUS is useful to assess and monitor inflammatory vascularization of the HFP lesions.

Enthesitis is the inflammatory process marked by the insertion of tendons, ligaments and joint capsules on the bone and it is a cardinal feature (and diagnostic criteria) of spondyloarthropathies (SpA). Although it is usually revealed by clinical examination, recent studies using magnetic resonance imaging (MRI) and ultrasonography (US) have confirmed that enthesitis (as well as synovitis) can often be asymptomatic, both in the axial and peripheral skeleta. Therefore, a systematic US study of peripheral entheses could be useful in the diagnostic process of patients suspected with SpA, and peripheral enthesitis scoring systems have been proposed. Recently, power Doppler US (PDUS) has been proved to be useful for differentiating mechanical and inflammatory enthesitis and for monitoring the efficacy of therapy. This article reviews the main anatomical and histopathological aspects of enthesitis and describes the general US features of enthesis and the basic US features of enthesitis, in its various stages. The usefulness of US and PDUS in the diagnosis and assessment of SpA is discussed on the basis of the available literature and our experience.

OBJECTIVE: To assess the clinical characteristics and determinants of pain observed by general practitioners (GPs) in Italian patients with osteoarthritis (OA) of the hand, hip, and knee. METHODS: The 2764 GPs participating in the study were asked to enroll 10 consecutive patients with OA diagnosed according to the American College of Rheumatology (ACR) clinical criteria. To standardize the diagnosis, the GPs received ad hoc training from musculoskeletal system specialists. A questionnaire evaluating demographic data, the clinical characteristics of OA, and previous diagnostic and therapeutic interventions was administered by the GPs. RESULTS: 25,589 evaluable patients were enrolled during a mean period of 2.8 weeks by the GPs: 17,567 women (69%) and 7878 men (31%). The most painful OA joints were the knee in 12,827 patients (54%), the hip in 5645 patients (24%), and the hand in 5467 patients (23%)--percentages calculated on the 23,939 patients for whom this information was available. The weekly incidence of referrals to GPs for OA was higher for women and for knee OA. The median age of the patients was 70 years (range 50 to 104 years) and disease duration was 8.3 +/- 7.10 years. The most frequent comorbidities were hypertension (53%), obesity (22%), osteoporosis (21%), type II diabetes mellitus (15%), and chronic obstructive pulmonary disease (13%). The median pain visual analog scale (VAS) score was higher for women than for men, for hip OA, and for generalized OA (GOA) than for knee and hand OA (P < 0.0001). Intense pain, defined as VAS readings of >60 mm, was increased in women only in the knee (OR = 1.24; 95% CI 1.15 to 1.34) and in GOA (OR = 1.17; 95% CI 1.03 to 1.33). It was also significantly increased in patients older than 70 years (OR = 1.46; 95% CI 1.39 to 1.54), those with a low educational level (OR = 1.44; 95% CI 1.36 to 1.5), a BMI of > or =30 (OR = 1.52; 95% CI 1.42 to 1.61), a disease duration of more than 7 years (OR = 1.60; 95% CI 1.52 to 1.68), comorbidities (OR = 1.61; 95% CI 1.5 to 1.73), and GOA (OR = 2.05; 95% CI 1.91 to 2.19). Manual occupations were associated with highly intense pain only in men. CONCLUSIONS: The results of this study underscore the major impact of OA on care in general practice, the high frequency of OA-associated comorbidities, and the role of different risk factors in OA pain.

Heel fat pad inflammation and degeneration have been frequently proved to cause talalgia. Painful heel fat pad is often confused with plantar fasciitis, and only magnetic resonance imaging (MRI) or ultrasonography (US) can differentiate these conditions. Scanty data are available about heel fat pad involvement in the course of chronic polyarthritis.To investigate with US the heel fat pad involvement in patients with rheumatoid arthritis (RA) and spondyloarthropathies (SpA); to describe and compare the clinical and sonographic features of this lesion in the two groups.The heels of 181 consecutive outpatients with RA and 160 with SpA were studied by US and radiography. A control group of 60 healthy subjects was examined by US.Two different patterns of involvement of the heel fat pad were observed. The inflammatory-oedematous pattern was more frequent in patients with RA (6.6%) than in those with SpA (1.8%), and was associated with talalgia--even if it was not associated with plantar fasciitis or enthesophyte (bony spur). The degenerative-atrophic pattern was less frequent (1.1% in RA, 1.9% in SpA), and was associated with plantar fasciitis and subcalcaneal enthesophyte.The inflammatory-oedematous lesion of the heel fat-pad is relatively frequent in RA and causes subcalcaneal pain. Degenerative-atrophic changes of the heel fat pad can be observed in RA and SpA, and seem to be associated with chronic abnormalities of the plantar fascia and of its enthesis.

Summary Objective: To investigate the efficacy and tolerability of a 3-month duration, twice a-year, intermittent treatment with oral chondroitin sulfate (CS) in knee osteoarthritis (OA) patients. Design: A total of 120 patients with symptomatic knee OA were randomized into two groups receiving either 800 mg CS or placebo (PBO) per day for two periods of 3 months during 1 year. Primary efficacy outcome was Lequesne’s algo-functional index (AFI); secondary outcome parameters included VAS, walking time, global judgment, and paracetamol consumption. Radiological progression was assessed by automatic measurement of medial femoro-tibial joint space width on weight-bearing X-rays of both knees. Clinical and biological tolerability was assessed. Results: One hundred and ten of 120 patients were included in the ITT analysis. AFI decreased significantly by 36% in the CS group after 1 year as compared to 23% in the PBO group. Similar results were found for the secondary outcomes parameters. Radiological progression at month 12 showed significantly decreased joint space width in the PBO group with no change in the CS group. Tolerability was good with only minor adverse events identically observed in both groups. Conclusion: This study provides evidences that oral CS decreased pain and improved knee function. The 3-month intermittent administration of 800 mg/day of oral CS twice a year does support the prolonged effect known with symptom-modifying agents for OA. The inhibitory effect of CS on the radiological progression of the medial femoro-tibial joint space narrowing could suggest further evidence of its structure-modifying properties in knee OA. © 2004 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.

Objectives To describe the clinical presentation and course of a relatively large group of Italian adult patients screened for mutation of the homogentisate dioxygenase gene causing alkaptonuria (AKU) and ochronosis, and to review typical and atypical facets of this condition. Methods We reviewed the medical records of 9 patients affected by ochronotic arthropathy who were observed in our institutions between 1979 and 2001. All patients were diagnosed as having AKU through a rapid urine test with alkali. Mutation screening was performed by single-strand conformation analysis of all homogentisate dioxygenase exons, followed by sequencing of altered conformers. Results Our 9 cases had similar clinical features and they reflected those described in the literature: a progressive degenerative arthropathy mainly affecting axial and weight-bearing joints associated with extraarticular manifestation. Musculoskeletal symptoms began in most of our patients around the age of 30 years with back pain and stiffness: involvement of the large peripheral joints usually occurred several years after spinal changes. Ochronotic peripheral arthropathy generally was degenerative, but joint inflammation was observed in some cases; this could be attributed to an inflammatory reaction of the ochronotic shard in the synovial membrane. Conclusions Ochronosis is a model of arthropathy with known etiologic factors. Over time, AKU, the genetically determined metabolic defect, leads to the accumulation of pigment and the development of this crippling condition. Most of the clinical findings may be explained by inhibition of collagen crosslinks, but some require additional interpretation. For example, inflammatory features of the ochronotic joint only occur in a minority of cases, and may be attributable to ochronotic shards. Further studies are needed to establish the genotype-phenotype correlation to identify mutations that are predictive of severe disease. For this purpose, the Italian Study Group on Alkaptonuria (www.dfc.unifi.it/aku) is enrolling affected patients in an on-line database to characterize the molecular defects and their relationship to clinical data.

Objective Newborns of mothers positive for anti-Ro/SSA autoantibodies may develop a series of electrocardiographic (EKG) disturbances. Prolongation of the corrected QT (QTc) interval was recently reported in a significant proportion of children with maternally acquired anti-Ro/SSA antibodies, with a concomitant disappearance of EKG abnormalities and acquired maternal autoantibodies during the first year, suggesting a direct, reversible electrophysiologic effect of anti-Ro/SSA antibodies on the ventricular repolarization. On this basis, we investigated whether these antibodies may also affect cardiac repolarization in anti-Ro/SSA–positive adult patients with connective tissue diseases. Methods Fifty-seven patients with connective tissue diseases were selected: 31 had anti-Ro/SSA antibodies and 26 did not (controls). In all subjects, we analyzed the QTc interval, heart rate variability, and signal-averaged high-resolution EKG recording. Results Anti-Ro/SSA–positive patients showed a significant prolongation of the mean QTc interval compared with the controls (mean ± SD 445 ± 21 versus 419 ± 17 msec; P = 0.000005). Eighteen of the 31 anti-Ro/SSA–positive patients (58%) and none of the 26 anti-Ro/SSA–negative patients had QTc values above the upper limit of normal (440 msec). Both groups had a reduction in heart rate variability, with a prevalence for the sympathetic nervous system and a high incidence of ventricular late potentials; these values were not significantly different between the 2 groups. Conclusion Adult patients with anti-Ro/SSA–positive connective tissue diseases showed a high prevalence of QTc interval prolongation. This feature, with the concomitant abnormalities in the autonomic tone and ventricular late excitability observed in all patients studied, suggests that anti-Ro/SSA–positive patients may have a particularly high risk of developing life-threatening arrhythmias.

To establish by ultrasonography (US) the frequency of calcaneal entheses involvement in erosive osteoarthritis (EOA), nodal osteoarthritis (NOA), RA and PsA, and to compare these results in order to aid clinicians in the differential diagnosis among these diseases. A comparison between US results and radiography was also made.The heels of 56 consecutive outpatients with EOA, 209 with NOA, 158 with RA and 125 with PsA were studied by US and radiography. A control group of 50 subjects was examined by US.US showed no significant difference in inferior calcaneal enthesophytosis among the four diseases. The frequency of posterioinferior enthesophytosis was lower in RA (34%) in comparison with the other diseases (57% in EOA, 47% in NOA, 49% in PsA). Achilles enthesitis was found in 8% of PsA and in 2% of RA. Retrocalcaneal bursitis was found in 18% of RA and in 6% of PsA. Posterior erosions were present in 12% of RA and 5% of PsA. Inferior erosions were present in 6% of RA and in 1% of PsA. Plantar fasciitis was found in 26% of RA, in 37% of PsA, and in 15% of NOA and 12% of EOA. Subcalcaneal panniculitis was observed in 10% of RA and in 1% of PsA. In the control group, only posterioinferior and inferior enthesophytosis (22% and 18% respectively) were found. Kappa statistics show excellent agreement between US and radiography in detecting posterioinferior (kappa = 0.89) and inferior enthesophytosis (kappa = 0.83), and entheseal erosions (kappa = 0.86).The calcaneal lesions that could be found in EOA are similar to those observed in NOA. The frequency of calcaneal enthesophytosis is similar in EOA, NOA, and PsA, but inflammatory lesions of calcaneal entheses and of the adjacent bursae are more frequent in RA and in PsA. In terms of heel involvement, EOA seems to be similar to NOA. US shows an excellent concordance with radiography in detecting entheseal cortical bone abnormalities.

The objective of this study was to determine the HLA class II associations of the anticardiolipin (aCL) and anti-beta(2)GPI (a beta(2)GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). A cohort of 577 European SLE patients was enrolled. aCL and a beta(2)GPI were measured by ELISA methods. Molecular typing of HLA-DRB1, DRB3, DRB4, DRB5, DQA1 and DQB1 loci was performed by the polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP) method. aCL of IgG, IgM and IgA isotypes were detected in 22.8%, 14% and 13.9% of patients, respectively. IgG and IgM a beta(2)GPI were detected in 20% of patients, aCL showed positive association with HLA DRB1*04, DRB1*0402, DRB1*0403, DRBI*07, DRB3*0301, DQA1*0201, DQA1*0301, DQB1*0302, and negative association with DQA1*0501, DRB3*0202. a beta(2)GPI showed positive association with DRB1*0402, DRB1*0403, DQB1*0302. DRB1*0402 carried the highest relative risk for the presence of both aCL (RR = 8.1) and a beta(2)GPI (RR = 4.6). Our results confirm the already described associations of aCL with HLA DR4 and DR7, but also demonstrate that, among the alleles at the DRB1*04 locus; the *0402 was most represented both in aCL and in a beta(2)GPI positive patients. In addition, HLA class II associations of a beta(2)GPI are for the first time extensively examined in a large cohort of European SLE patients.

Linear scleroderma (LS) is characterized by scleroatrophic lesions affecting limbs and legs, unilaterally. Neurological involvement may be associated with ipsilateral facial and skull involvement in disorders referred to clinically as LS 'en coup de sabre', and Parry-Romberg syndrome. We report a child with LS presenting with a severe neurological disorder characterized by epilepsy, progressive mental deterioration and a rapid process of atrophy involving the ipsilateral cerebral hemisphere, but not associated with an overlying facial structure involvement. Functional brain studies showed a reduction in the diameter of the left internal carotid and of the left middle cerebral artery. Our observations suggest that neuroimaging studies should be considered in all patients with linear scleroderma, and such studies become necessary when neurological symptoms occur.

In this study, we evaluated pain sensitivity in patients with fibromyalgia or other types of chronic, diffuse musculoskeletal pain to establish whether fibromyalgia represents the end of a continuum of dysfunction in the nociceptive system. One hundred and forty five patients and 22 healthy subjects (HS) completed an epidemiological questionnaire to provide information about fatigue, stiffness, sleep, the intensity of pain (VAS 0-100) and its extent both at onset and at present. Algometry was performed at all American College of Rheumatology (ACR) tender points and at ten control points. Patients were divided into five main groups: fibromyalgia (FS) patients, secondary-concomitant fibromyalgia (SCFS) patients, patients with widespread pain (WP) but not reaching the ACR criterion of 11 tender points, patients with diffuse multiregional pain (MP) not reaching the ACR criteria (widespread pain, tender point counts), and patients with multiregional pain associated with at least 11 tender points (MPTE). von Frey monofilaments were used to assess superficial punctate pressure pain thresholds. Heat and cold pain thresholds were determined with a thermal stimulator. Ischemic pain was assessed by the cold pressure test and the submaximal effort tourniquet test. The scores for stiffness and present pain intensity gradually increased concomitantly with the increase in tender point count and pain extent. The pressure pain thresholds for positive tender and positive control points were significantly lower in the SCFS, FS and MPTE groups than in HS, MP and WP groups, the latter three groups displaying similar values. In all groups, there were no differences in pain thresholds between positive tender and positive control points. The heat pain threshold and the pain threshold in the cold pressure test were lower in the FS and SCFS groups than in HS. The cold pressure tolerance was lower in patients with widespread pain than in HS. In the von Frey test, all patient groups except MP had similar values, which were significantly lower than in HS. Finally, all patient groups displayed lower tourniquet tolerance than HS. In each psychophysical test, patients with widespread pain and patients with multiregional pain showed similar thresholds; however, the thresholds in the MP or MPTE groups differed from those in the FS and SCFS groups. In the FS group, pain thresholds and pain tolerance did not differ according to the presence of ongoing pain at the stimulated site and were not correlated to ongoing pain. The results indicate that dysfunction in the nociceptive system is already present in patients with multiregional pain with a low tender point count; it becomes more and more severe as the positive tender point count and pain extent increase and it is maximal in fibromyalgia patients.

Enthesitis is an inflammatory lesion of the tendon, ligament and capsular insertions into the bone, and it is a fundamental element in the diagnosis of spondyloarthropathies. Sonography is the method of choice for studying periarticular soft tissues because it is capable of detecting both the early (oedema, thickening) and the late alterations (erosions and enthesophytes); it is also an inexpensive, biologically harmless and easily repeatable technique. The aim of this study was to compare the prevalence of quadricipital enthesitis in psoriatic arthritis (PsA) and rheumatoid arthritis (RA) patients, and to document any clinical and echostructural differences in this lesion between the two diseases. The results show that enthesitis is more frequent in PsA patients, more than half of whom are asymptomatic. Knee inflammation was found in the PsA patients with enthesitis regardless of the concomitant presence of joint effusion; none of the RA patients suffered from enthesitis alone. Quadricipital enthesitis is more frequent in male patients. There was no significant correlation between the presence of peripatellar psoriatic lesions and enthesitis. Sonographic examinations of patients with enthesitis revealed that those with RA had dominantly inflammatory lesions, whereas PsA patients also showed major new bone deposition.

Objective: to study the frequence of deltoideal proximal insertion enthesitis (DPIE) in patients affected with spondyloarthritis (SpA) and to evaluate its clinical, sonographic and radiological characteristics. Methods: a retrospective study of clinical, sonographic and radiological examinations of the shoulders of 100 symptomatic consecutive outpatients with SpA, compared to 4 groups of control patients: 100 with Rheumatoid Arthritis, 100 with Osteoarthritis, 100 with Painful Shoulder, and 50 with shoulders undamaged by local pathological processes. Results: the frequence of DPIE in the course of SpA was 9%. DPIE appears most frequently in Psoriatic Arthritis (PsA) (17%, 7/41). DPIE does not appear to be related to the sex or the age of the patient. The clinical signs and symptoms are similar to those of an impingement syndrome. Sonography reveals thickening and hypoechogenicity of the enthesis, associated or not to the subchondral osseous rearrangement and enthesophytosis. Radiology only reveals the ent...

Objective—To determine the dissolving ability (DA) of linear pentasodium tripolyphosphate (PSTP), cyclic trisodium metaphosphate (TSMP), polymeric sodium metaphosphate (SMP) on synthetic crystals of calcium pyrophosphate dihydrate (CPPD) and on crystalline aggregates of menisci from patients with chondrocalcinosis (CC). Methods—Synthetic CPPD crystals were mixed with phosphate buVered saline (PBS), which contained the diVerent polyphosphates, for one hour at 37°C. The calcified menisci were obtained from the knees of four female patients with CPPD disease who underwent total arthroscopic meniscectomy for degenerative meniscal lesions. Meniscal cryosections and fragments were incubated in SMP (15 mg/ml PBS) at 37°C for one hour and 24 hours, respectively. Histological evaluation on meniscal samples after polyphosphate incubation was carried out by ordinary transmitted light microscopy and polarised light microscopy. The dissolution of CPPD crystals by polyphosphates was assessed by atomic absorption spectroscopy, which determined the amount of calcium liberated from synthetic crystals and meniscal fragments. Cytotoxicity of SMP was evaluated by tetrazolium salt assay and by an ultrastructural study on cultured chondrocytes. Results—SMP and PSTP showed higher DA on CPPD crystals than TSMP. Analysis of the DA values at increasing concentrations of SMP showed that a concentration of 15 mg/ml completely dissolved 2.0 mg CPPD crystals.The solution of meniscal CPPD crystals showed a significant increase of calcium concentration after three hours and 24 hours of SMP incubation (p=0.0001; Kruskal-Wallis analysis of variance) compared with fragments incubated in PBS control solution. Macroscopic and microscopic evaluation of meniscal specimens showed a notable reduction of CPPD deposits. A 50% inhibitory dose on cultured chondrocytes was reached at the maximum concentration of SMP used in this work (15 mg/ml); ultrastructural analysis did not show morphological alterations in the treated cells. Conclusion—The results of this study indicate that linear polyphosphates are eVective in dissolving both synthetic and ex vivo CPPD crystal aggregates. This suggests a potential therapeutic use for these molecules in the treatment of symptomatic CC. (Ann Rheum Dis 2001;60:962‐967)

Objective This study investigated the in vitro effects of chondroitin sulfate (CS) on human articular chondrocytes cultivated in the presence or in the absence of interleukin-1beta (IL-1beta) during 10 days of culture with and without pressurization cycles. Design The effects of CS (10 and 100 microg/ml) with and without IL-1beta were assessed in the culture medium of cells exposed to pressurization cycles in the form of synusoidal waves (minimum pressure 1 Mpa, maximum pressure 5 Mpa) and a frequency of 0.25 Hz for 3 h by immunoenzymatic method on microplates for the quantitative measurement of human proteoglycans (PG). On the 4th and 10th day of culture the cells were used for morphological analysis by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Results The presence of IL-1beta determines a significant decrease in PG concentration measured in the culture medium. When the cells are cultured in the presence of IL-1beta and CS, a statistically significant restoration of PG levels is observed. Under pressurization conditions, we observed that PG concentration in the medium of cells presents a significant increase at baseline conditions, in the presence of IL-1beta+CS10 and IL-1beta+CS100, but not with IL-1beta alone. The results concerning metabolic evaluation are confirmed by the morphologic findings obtained by TEM and SEM. Conclusions These in vitro studies confirm the protective role of CS, which counteracts the IL-1beta induced effects and they confirm the importance of pressure on chondrocyte metabolism and morphology.

The pharmacokinetics of diclofenac sodium 150 mg in a new prolonged-release formulation (Dicloreum® CR [CR]) and a commercially available formulation (Dealgic® 75 [D]) were compared in 12 healthy volunteers. In this randomized, crossover study with a Latin-square design, diclofenac was administered as a single dose to subjects after eating. Blood samples were obtained before dosing and at 1, 2, 4, 6, 8, 14, 24, and 36 hours after dosing, and the plasma concentrations of diclofenac were determined by high-performance liquid chromatography. Possible adverse events were monitored during the study. After log-transformation of the comparable variables, no statistically significant differences were found between the two formulations with respect to the time between dosing and the appearance of the drug in serum, time to reach maximum concentration, elimination half-life, or mean residence time. The extent of absorption was assessed by estimating the area under the plasma concentration-time curve (AUC). AUC 0–24 was statistically significantly higher for CR than for D (2992.8 ± 280.2 and 2527.6 ± 109.9, respectively). No significant differences were found in AUC 0–36 , AUC 24–36 , or AUC 0–∞ . CR showed a statistically significantly higher maximum plasma concentration than D (516.9 ± 74.3 ng/mL and 382.9 ± 46.9 ng/mL for CR and D, respectively). The results of the present study suggest that CR is a useful, prolonged-release formulation for continuous, once-daily therapy.

One hundred and twenty postmenopausal women with osteoporosis were enrolled into an open, controlled study of 24 months’ duration, with the aim of evaluating the metabolic action and the effect on bone mineral density (BMD) of calcitriol (0.5 µg/day), alendronate (10 mg/day) and combined treatment with alendronate (10 mg/day) + calcitriol (0.5 µg/day). We also studied 30 patients treated with placebo (calcium 500 mg/day). BMD was measured by total body dual-energy x-ray absorptiometry (DPX-Lunar) as total body BMD and at different sites of interest including the spine, trunk, arms, legs and pelvis. BMD appeared to be significantly higher in the group of patients receiving combined therapy compared with the group treated with alendronate or calcitriol alone. In the placebo group, BMD decreased significantly. Depending on the skeletal regions in which BMD was measured (total body or selected areas), 15 to 23% of patients treated with calcitriol, 37 to 48% of patients treated with alendronate, and 69 to 78% of patients treated with alendronate + calcitriol, respectively, had a BMD increase greater than the lowest significant densitometric difference. In patients treated with alendronate, 24-hour urinary calcium and hydroxyproline levels fell significantly after 3 months, and alkaline phosphatase levels fell significantly after 6 months; these parameters remained unchanged in all patients on combined treatment. We concluded that combined treatment with calcitriol + alendronate was more effective than therapy with alendronate alone.

To determine the prevalence of morphologic bowel lesions in patients with inflammatory rheumatic diseases and to better define the interactions between intestinal and articular pathology, 177 patients [39 with reactive arthritis (ReA), 40 with psoriatic arthritis (PsA), 23 with ankylosing spondylitis (AS), 21 with undifferentiated spondyloarthropathy (USpA) and 54 with rheumatoid arthritis (RA)] underwent ileocolonoscopy followed by multiple biopsies of the large bowel and the ileum and ileocecal valve. Biopsies were then examined with light and electron microscopy. During the endoscopic examination various degrees of gut inflammation were observed in 13% of ReA, 5% of PsA, 26% of AS, 14% of USpA and 11% of RA patients. At the histological examination those percentages were respectively 51%, 45%, 48%, 38%, and 15%, and at the electron microscopic examination 76%, 53%, 90%, 60%, and 50%. Our results show that an involvement of the gut is a factor in a large percentage of patients with spondyloarthropathy and, to a lesser extent, with RA. The involvement of the intestine in RA manifests itself mainly in ultrastructural lesions, thus this involvement is not so obvious as in the spondyloarthropathies; however, it could nonetheless play an important role in the etiopathogenesis of this disease.

Twenty outpatients suffering from rheumatoid arthritis and 20 outpatients with scapulohumeral periarthritis were treated for 30 days with oral diclofenac sodium, either using one 150-mg prolonged-release capsule per day (10 patients with each diagnosis) or one 50-mg, enteric-coated tablet 3 times a day (10 patients per diagnosis). During treatment, visual analog scale scores for pain intensity, the severity of symptoms and signs, and results of laboratory tests (erythrocyte sedimentation rate, C-reactive protein, and rheumatoid factor) were monitored, as well as the local tolerability of the administered formulations. Systemic tolerability was also monitored by means of routine blood and urine laboratory tests. The single daily administration of 150 mg as a prolonged-release capsule dosage form yielded almost identical and very good anti-inflammatory and analgesic activity as the multiple daily administration of the 50-mg tablet: 16 cases of good activity and 4 of no efficacy with the prolonged-release formulation versus 15 cases of good efficacy and 5 cases of no efficacy with the enteric-coated tablet. As predictable, based on the different characteristics of the diseases, some patients with rheumatoid arthritis responded only partially to diclofenac treatment (60% had positive results), while in nearly all patients with scapulohumeral periarthritis, treatment with this nonsteroidal anti-inflammatory drug yielded the best clinical outcome (95% had positive results). Drug tolerability, both at the gastrointestinal and systemic levels, was excellent; no adverse events were reported. The prolonged-release formulation, which enables once-daily dosing, appears to be an optimal means to treat articular rheumatic diseases.

Objective. To evaluate the ability of low-dose cyclosporin A (CsA) to control radiologic disease progression, and to assess the clinical efficacy and tolerability of CsA, compared with conventional disease-modifying antirheumatic drugs (DMARDs), in patients with early active rheumatoid arthritis (RA). Methods. In this long-term, multicenter, prospective, open, blinded end point, randomized trial, 361 consenting patients with early (< 4 years since diagnosis) active RA were enrolled. Of the eligible patients, 167 were treated with CsA at 3 mg/kg/day, and 173 with DMARDs. The decision to use conventional antirheumatic drugs as controls was based on the fact that joint erosion could be expected to occur after 1 year regardless of the type of DMARD being used. The possibility of switching therapies in both groups was intended to keep the largest possible number of patients in the study. Results. Blinded evaluation of hand and foot radiographs after 12 months of treatment showed that CsA led to a significant (P < 0.001) delay in the mean ± SD progression in the eroded joint count (1.3 ± 3.1 versus 2.4 ± 3.0 for the control group) and in the joint damage score (3.6 ± 8.9 versus 6.9 ± 9.1 for the control group), both measured by the Larsen-Dale method. When only the patients without erosion at baseline were considered (37 in the CsA-treated group and 54 in the control group), erosion appeared in only 10.8% of the CsA-treated patients, but in 51.8% of the controls (P = 0.00005). Low-dose CsA was as effective as traditional DMARDs in controlling clinical symptoms. Maintenance on the initially prescribed treatment regimen (“survival on treatment”) was also better at 12 months with CsA than with DMARDs (89.2% versus 77.5%; P = 0.002). The tolerability of CsA was acceptable. Conclusion. These 12-month results suggest that low-dose CsA decreases the rate of further joint damage in previously involved joints as well as the rate of new joint involvement in previously uninvolved joints, in patients with early RA.

Whether a genetic predisposition to develop the antiphospholipid syndrome (APS) and to produce anticardiolipin antibodies (aCL) and lupus anticoagulant (LAC) exists has been addressed by family studies and by population studies on primary APS and on aCL in diseases other than primary APS. Various studies suggest a familial occurrence of aCL and LAC, with or without clinical evidence of APS. This familial tendency could be genetically determined, because APS, aCL, and LAC occur in families carrying haplotypes which contain HLA-DR4, -DR7, and -DRw53. Population studies on primary APS also indicate that HLA genes have a role in conferring susceptibility to develop primary APS. Again, DR4, DR7, and DRw53 are the relevant loci. Population studies on aCL in diseases other than primary APS indicate that aCL are associated with DR4, DR7, and DRw53, at least when they are found in patients with systemic lupus erythematosus. Because HLA-DR4, -DR7, and -DRw53 are in linkage disequilibrium, the genetic association of aCL could be with DRw53 and, depending on the regional frequency of DR4 or DR7, it could be linked with either DR4 or DR7. HLA-DR4 seems to be more important in Anglo-Saxons, whereas DR7 emerges in populations of Latin origin. In this report we review our studies and the pertinent literature in this field.

Idiopathic hypereosinophilic syndrome (IHES) is a rare condition of uncertain etiology characterized by marked peripheral blood eosinophilia and organ system dysfunction that cannot be explained by any factor other than the presence of eosinophils or their potentially toxic products. Diagnostic criteria include 1) a sustained eosinophilia greater than 1500/mm3 for longer than 6 months, 2) absence of other causes of eosinophilia, including parasitic infections and allergic diseases, and 3) multiorgan involvement (ie, lungs, heart, central nervous system, skin, liver, joints). Steroids represent the initial therapeutic approach, although for those patients unresponsive to steroids, cytotoxic chemotherapy should be considered. We describe a case of IHES characterized by polyarticular inflammatory joint involvement, hypereosinophilia, and urticarioid skin manifestations without visceral involvement. Synovial fluid smears as well as pathology of skin lesions and knee synovial membrane confirmed the presence of numerous eosinophils. The patient was successfully treated with a combination therapy of a cysteinyl leukotriene receptor antagonist (montelukast) and ketotifen. Hypereosinophilic syndrome like in this patient is a rare cause of polyarthritis.

The efficacy and tolerability of aceclofenac and naproxen sodium in the treatment of ankylosing spondylitis (AS) were evaluated and compared in this double-blind, multicenter, controlled study. Of the 130 patients who entered the study, 126 patients met the inclusion criteria. Efficacy was evaluated at baseline, 15 days, and 1, 2, and 3 months using a visual analog scale for spontaneous pain, a zero to three-point scale for pain on movement and pain at rest, and measurements of chest expansion, hand-to-floor distance, Schober's test, and normal daily activities. No significant between-group differences were seen for any of the variables at baseline, except hand-to-floor distance. Both drugs provided effective analgesia and a corresponding improvement in functional activity. Overall efficacy assessment made by the physician and patients was not statistically significant between the two treatment groups. The overall incidence of adverse effects was higher in the naproxen group than the aceclofenac group, 22 versus 15 patients, respectively. All adverse effects resolved. However, the overall assessment of tolerability given by the physician and patients was significantly (P

The recommendations for the management of osteoarthritis (OA) of the knee firstly proposed by the EULAR in 2000, have been updated in 2003. One of the most important objectives of the expert charged to provide these recommendations was their dissemination. Thus, the information generated may be used by each individual country to produce their own set of management guidelines and algorithms for treatment in primary care. The Italian Society of Rheumatology (SIR) and the Italian League against Rheumatism (LIMAR) have organised a Consensus on the EULAR recommendations 2003 with the aim to analyse their acceptability and the applicability according to our own experience and local situations in the Italy. The results of this Consensus have demonstrated that a large majority of the EULAR recommendations are endorsed by the Italian experts. Furthermore, the final document of the Italian Consensus clearly indicated the need that the specialists involved in the management of knee OA strongly encourage the dissemination of the EULAR 2003 recommendations also in Italy.

We describe a male patient who developed systemic sclerosis following orchiectomy, radiotherapy and anti-androgenic treatment for prostatic adenocarcinoma. This case appears interesting as it further supports the possibility of a relationship between neoplasia and systemic sclerosis. The concurrence of scleroderma and iatrogenic hypoandrogenism suggests that hormonal influences may also play a role in the pathogenesis of this connective tissue disorder.

Osteoarthritis of the hands is a very common disease that can present a large number of different clinic pictures, such as nodal (NOA) and erosive (EOA) forms. EOA in particular is a rare subset of hand osteoarthritis characterised by faster destructive changes involving the distal interphalangeal (DIP) and proximal interphalangeal (PIP) joints. In the early stages of the disease the differential diagnosis from other arthritides, such as rheumatoid or seronegative arthritis, may pose a challenge. Nailfold capillaroscopy is a non-invasive technique that allows the in vivo study of the microvascular environment. In this study the authors have compared the capillaroscopic microvascular patterns in 56 patients with EOA, in 46 patients with NOA, and in 50 normal controls. The abnormalities that could be found in EOA patients were similar to those described by some authors in patients with psoriatic arthritis. The authors discuss the significance of these abnormalities and the possible relationship between EOA and psoriatic arthritis.

Alkaptonuria (AKU) is a rare genetic disease associated with deficient homogentisate 1,2-dioxygenase activity in the liver. This leads to the accumulation of homogentisic acid (HGA) and its oxidized/polymerized products in connective tissues, which in turn become characterized by the presence of melanin-like pigments (ochronosis). Since at present, further studies are necessary to support the use of drugs for the treatment of AKU, we investigated the effects of various anti-oxidants in counteracting melanin-like pigmentation and oxidative stress related to HGA and its metabolites.We set up an in vitro model using human serum treated with 0.33 mM HGA and tested the anti-oxidants ascorbic acid, N-acetylcysteine, phytic acid (PHY), taurine (TAU), ferulic acid (FER) and lipoic acid (LIP) for their ability to prevent or delay the production of melanin-like pigments, as well as to reduce oxidative post-translational modifications of proteins. Monitoring of intrinsic fluorescence of HGA-induced melanin-like pigments was used to evaluate the efficacy of compounds.Our model allowed us to prove efficacy especially for PHY, TAU, LIP and FER in counteracting the production of HGA-induced melanin-like pigments and protein oxidation induced by HGA and its metabolites.Our model allows the opening of new anti-oxidant therapeutic strategies to treat alkaptonuric ochronosis.

We analyzed the distribution of DRB1, DQA1, DQB1, and DPB1 allelic variants in 48 rheumatoid arthritis (RA) patients, compared with 109 Italian random controls, using PCR amplification and hybridization with specific oligonucleotides. We confirm the previously reported increase of DR4 specificity, in comparison with healthy Italian individuals. In particular, we find a statistically significant positive association of DRB1 ∗ 0401 and DRB1 ∗ 0404 alleles with RA. However, when we compare the DR4 + groups, none of the DRB1 ∗ 04 alleles is increased in the RA group. By sequence analysis, performed on 10 patients, we demonstrate that the DRB1 ∗ 04 genes of RA show no difference from the DRB1 ∗ 04 sequences previously published. From the molecular analysis of the other DRB1 polymorphic variants, we find a trend of positive association of DRB1 ∗ 0101 in DR4-negative patients versus DR4-negative healthy controls and, in the group of DR4-negative and/or DR1-negative patients, a similar increase of DRB1 ∗ 06. Also, we observe in RA patients a statistically significant increase of DQA1 ∗ 0301 and DQB1 ∗ 0302 accompanied by a significant decrease of DQA1 ∗ 0201, DQA1 ∗ 0501 and DQB1 ∗ 0201. Finally, from the analysis of DPB1 gene, it can be assessed that the distribution of DPB1 alleles does not differ significantly between RA patients and healthy controls.

The aim of the present study was to demonstrate the mitogenic and differentiating properties of platelet-rich plasma releasates (PRPr) on human chondrocytes in mono- and three-dimensional cultures. In order to assess if PRPr supplementation could maintain the chondrocyte phenotype or at least inhibit the cell de-differentiation even after several days in culture, we performed a proteomic study on several cell cultures independently grown, for different periods of time, in culture medium with FCS, human serum (HS), and releasates obtained from PRP and platelet-poor plasma (PPP). We found that PRP treatment actually induced in chondrocytes the expression of proteins (some of which novel) involved in differentiation.

We report a case of subcutaneous panniculitis-like T-cell lymphoma (SPTCL), associated with macrophage activation syndrome, mimicking a lupus erythematosus panniculitis (LEP). A 29-year-old woman presented with high fever, general malaise, nausea, vomiting, and subcutaneous nodules and ulcerating lesions located on the lower extremities. The histopathology showed an infiltration of the panniculus, mostly involving fat, and periadnexial and perivascular structures consistent with lymphocytic lobular panniculitis (LLP). LLP is a shared feature of LEP and SPTCL. The immunophenotyping of the cell infiltrate was crucial for a correct diagnosis.

According to the criteria proposed by Ryan and McCarty,1 the diagnosis of calcium pyrophosphate dihydrate (CPPD) deposition disease has been based on radiological evidence of the characteristic calcifications and on verification of the synovial liquid of CPPD crystals. Joint ultrasonography is an innocuous diagnostic technique that is well tolerated by patients, and is the elected method for observing calcified deposits in soft tissues.2 We carried out a longitudinal study, enrolling patients affected with ultrasonographic chondrocalcinosis according to previously proposed criteria3 from a sample of consecutive patients that came to our joint ultrasonography department for gonalgia (fig. 1). A total of 47 patients were identified, of which 14 had joint effusion. Figure 1 Hyperechoic deposits. Deposits (arrows) are shown that are compatible with calcium pyrophosphate dihydrate (CPPD) calcifications in the context of the synovial membrane (1), hyaline cartilage of the …

The use of unidimensional scales to measure pain intensity has been criticised because of the multidimensional nature of pain. We conducted multiple linear regression analyses to determine which dimensions of pain--sensory versus affective--predicted scores on unidimensional scales measuring pain intensity and emotions in 109 Italian women suffering from chronic, non-malignant musculoskeletal pain. We then compared the results with earlier findings in two groups of cancer patients suffering from acute post-operative pain and chronic cancer-related pain, respectively. Age, physical capacity and scores on the multidimensional affect and pain survey (MAPS) were used to predict patients' ratings on one visual analogue scale (VAS) and three numerical rating scales (NRS) measuring pain intensity, anxiety and depressed mood. Unidimensional pain intensity ratings were predicted better from sensory than from affective pain predictors, and the affective predictors made no unique contribution (NRS), or only a very small one (VAS). Both sensory and emotional pain aspects were unique predictors of NRS anxiety and depression. Therefore, in contrast to earlier findings in two different types of cancer patients, in subjects affected by chronic non-malignant musculoskeletal pain, the scores on unidimensional pain intensity scales mainly reflect sensory pain dimensions, supporting the discriminant validity of the NRS and VAS used. However, the patients had some difficulty in distinguishing between sensory and emotional information. For this reason, several unidimensional scales to rate pain intensity and emotions separately should be used to obtain a complete picture of the status and needs of any given patient.

Changes in expression profiles for 17 proteins were ascertained in human mature osteoblasts compared to pre-osteoblasts (differentiation markers). A differential approach was used to highlight proteomic changes between human osteosarcoma cells and mature osteoblasts, showing a relative over-expression of 8 proteins (proliferation and tumor indicators), as well as under-expression of proteins also found down-regulated in pre-osteoblasts (specific markers of osteoblast differentiation). Our findings confirmed the differences between cell lines and primary human cell cultures and suggested caution on the use of osteosarcoma to study anti-osteoporotic drugs in humans.

Two poly(amido-amine)s (oligoPAM and oligoNER) containing bis-phosphonate residues were obtained by a Michael-type polyaddition of pamidronate and neridronate to 1,4-bis(acryloyl)piperazine. The SEC (size-exclusion chromatography) and the MALDI-TOF (matrix assisted laser desorption ionization) analyses were consistent with the presence of oligomeric species (2-3 kDa) and with a narrow polydispersity index. The thermodynamic results (log Ks, -DeltaH(o) , and DeltaS(o) obtained at 25 degrees C in 0.15 M NaCl) of both the oligomers and the corresponding low molecular weight precursors were in line with a cluster structure formed during the protonation of the basic nitrogen in the pamidronate. The solubility of the oligoNER with a longer aliphatic chain was improved at high pHs, allowing the evaluation of their solution properties. Preliminary biological results show that both the oligomers do not negatively affect the in vitro viability, proliferation, and cellular activity of either normal animal or human osteoblasts.

Objective: To investigate by high frequency ultrasonography the appearance of calcium pyrophosphate dihydrate (CPPD) calcifications, in the most commonly affected sites in CPPD disease, and the relationship between ultrasonographic CPPD deposits and the presence of CPPD crystals in synovial fluid. Methods: Three ultrasonographic patterns of CPPD calcification were identified and 11 patients enrolled. A control group comprised 13 patients with no evidence of CPPD deposits. Synovial fluid was aspirated from all patients and controls and examined for identification of crystals. All patients underwent a standard radiography examination at the same sites investigated by ultrasound. Results: In all patients with ultrasonographically defined CPPD deposits, CPPD crystals were found in the synovial fluid. In two cases, standard radiographic examination did not show evidence of the calcific deposits that were identified by ultrasonography. CPPD crystals were not found in the synovial fluid of controls. In four control group patients, ultrasonography identified calcifications defined as deposits of another nature. Conclusions: The ultrasonographic pattern used in this study for the diagnosis of CPPD disease demonstrated a very high correlation with the presence of CPPD crystals in synovial fluid. Ultrasonography demonstrated a sensitivity and specificity at least equal to that of radiography in identifying CPPD crystal calcifications.

Summary Objective This study investigated the in vitro effects of hyaluronic acid (HA) of molecular weight (MW) 500–730kDa on human articular chondrocytes cultivated for 48h in the presence of interleukin-1β (IL-1β) with and without hydrostatic cyclical pressure. Design The effects of 10 and 100μg/ml HA with and without IL-1β were assessed in the culture medium of cells exposed to pressurization cycles in the form of sinusoidal waves (minimum pressure 1MPa, maximum pressure 5MPa) at a frequency of 0.25Hz for 3h, by the immunoenzymatic method on microplates for the quantitative measurement of human proteoglycans (PG) and by the Griess method for nitrites (NO). Morphological analyses were performed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Results The presence of IL-1β determines a significant decrease in PG and a significant increase in NO concentrations measured in the culture medium. When the cells are cultured in the presence of IL-1β and HA at the two concentrations, a statistically significant restoration of PG and a decrease in NO levels are observed. Under pressurization conditions, we observed that the PG concentration in the medium of cells presented a very significant increase in all the conditions used in the study, except for IL-1β alone. NO production decreased very significantly in the presence of IL-1β+HA 10 and IL-1β+HA 100. The results of metabolic evaluation are confirmed by morphological findings obtained by TEM and SEM. Conclusions These in vitro studies confirm both the protective role of HA (MW 500–730kDa), which counteracts the IL-1β-induced effects, and the importance of pressure on chondrocyte metabolism and morphology.

OBJECTIVE The main objective of the AMICA project was to photograph the Italian scenario of osteoarthritis (OA) and its treatment in general and specialty practice. The study was designed to evaluate their prescription modalities to determine whether they matched the recently proposed treatment guidelines for OA (ACR 2000; EULAR 2000; APS 2002). METHODS The study involved 2764 general practitioners (GPs) and 316 specialists who enrolled a total of 25,589 patients with OA of the hand, knee, and hip. RESULTS Pharmacological treatment alone was prescribed to 55% of the patients seen by GPs, 25% of those seen by rheumatologists, 8% of those seen by orthopedic surgeons, and 17% of those seen by physical medicine specialists (GPs versus specialists, P < 0.001). Specialists often prescribed a combined pharmacological and nonpharmacological approach (rheumatologists 51%, orthopedic surgeons 66%, physical medicine specialists 76%). Concomitant comorbidities and their treatment do not seem to influence OA prescription modalities except for peptic ulcer and anticoagulant therapy. The presence of peptic ulcer was associated with a reduction in NSAID prescriptions (OR 0.61, CI 0.53 to 0.69) and more frequent use of Coxibs (OR 1.15, CI 1.03 to 1.28) and simple analgesics (OR 1.42; CI 1.26 to 1.61), as well as physical therapy. NSAIDs and Coxibs also were less frequently prescribed if patients were receiving anticoagulant therapy (NSAIDs OR 0.86, CI 0.70 to 1.06; Coxibs: OR 0.77; CI 0.64 to 0.93). Gastroprotective therapy was more frequently used in patients treated with NSAIDs, Coxibs, and analgesics. There was no significant difference in therapies prescribed for patients with hypertension or cardiac disease (myocardial infarction and/or angina pectoris). CONCLUSIONS The published guidelines appear to be properly used by most of the physicians in terms of the pharmacological approach; however, the increased use of Coxibs has not reduced the amount of prescribed gastroprotection. No specific precautions were observed in the treatment of patients with hypertension or cardiac problems. Nonpharmacological treatments are mainly used in conjunction with medications and did not take into account the findings of evidence-based medicine. Continuing education of GPs and specialists caring for OA patients is essential.

We present the case of a women diagnosed with Wegener's disease at the age of 26 years, refractory to corticosteroid and cyclophosphamide therapy. Treatment with intravenous immunoglobulin (IVIg) was started, leading to partial clinical remission of disease. During IVIg treatment she became pregnant. IVIg therapy was continued, the disease went into remission, and after 40 weeks the patient delivered a healthy boy. After 6 months from the delivery, the patient became pregnant again. Now she is at the 22nd week of pregnancy and she is doing very well. This case supports the beneficial effect of IVIg in Wegener's granulomatosis and illustrates its safety and efficacy during pregnancy.

Bisphosphonates (BPs) are widely used in the treatment of a variety of bone-related diseases, particularly where the bone turnover is skewed in favor of osteolysis. The mechanisms by which BPs reduce bone resorption directly acting on osteoclasts are now largely clarified even at molecular level. Researches concerning the BP's effects on osteoblast have instead shown variable results. Many in vitro studies have reported positive effects on osteoblasts proliferation and mineralization for several BPs; however, the observed effects differ, depending on the variety of different model system that has been used.We have investigated if neridronate, an aminobisphosphonate suitable for pulsatory parenteral administration, could have an effect on human osteoblastic proliferation and differentiation in vitro.We have investigated whether prolonged addition of neridronate (from 10(-3) to 10(-11) M) to different human osteoblasts cultures, obtained from 14 different bone specimens, could affect the cells number, the endogenous cellular alkaline phosphatase (ALKP) activity, and the formation of mineralized nodules.Our results show that neridronate does not negatively affect in vitro the viability, proliferation, and cellular activity of normal human osteoblasts even after a long period addition of the drug (20 days) at concentrations equal or lower than 10(-5) mol/l (therapeutic dose). In addition, neridronate seems to enhance the differentiation of cultured osteoblasts in mature bone-forming cells. A maximum increase of alkaline phosphatase activity (+50% after 10 days; P0.01) and mineralized nodules (+48% after 20 days; P0.05) was observed in cultures treated with neridronate 10(-8) M.These results encourage the use of neridronate in long-term therapy of demineralizing metabolic bone disorders.

The discovery of extracellular nucleic acids in the circulation was firstly reported in 1948. In the last few years it has been demonstrated that the entire spectrum of genetic changes seen in primary tumors could also be detected in the serum of patients with solid tumors. This observation has also opened up exciting possibilities for tumor detection and monitoring. More recently investigators started looking for other forms of non-host DNA in the plasma/serum so that in 1997 the presence of fetal DNA in the plasma/serum of pregnant women was demonstrated. This finding suggested that maternal plasma fetal DNA would be a very valuable material for noninvasive prenatal diagnosis and monitoring. It has been also postulated that the presence of the two-way trafficking of nucleated cells and free DNA between the mother and fetus may have potential implications for the development of certain autoimmune diseases. Concerning autoimmune disorders, Tan was the first author to describe the presence of high levels of circulating DNA in patients with systemic lupus erythematosus (SLE) in 1986. Later on different authors demonstrated that elevated levels of serum DNA was also present in patients with other diseases including rheumatoid arthritis. We have analyzed both circulating free DNA and DNA extracted from nucleated blood cells in scleroderma and in lupus patients but, by using gel electrophoresis, we were able to define the pattern of the DNA, instead of simply dosing its amount in the circulation. We have found that SLE and SSc have anomalous patterns of DNA both in serum and in the Buffy-coat and that these patterns are typical for each disorder. It is possible that understanding the biological significance of the diversity in DNA pattern exhibition in white blood cells may give new insights into the pathophysiology of autoimmune disorders. It is also conceivable that circulating and immune-competent cellular DNA markers might offer the promise of precise quantitative analysis useful for diagnostic purposes, without the need to establish difficult cutoffs as is necessary for protein markers.