Roberto Marcolongo

Alkaptonuria, ochronosis, and ochronotic arthropathy.

OBJECTIVESnTo 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.nnnMETHODSnWe 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.nnnRESULTSnOur 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.nnnCONCLUSIONSnOchronosis 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.

Anticardiolipin and anti-β2GPI antibodies in a large series of European patients with systemic lupus erythematosus

Objective. To test the prevalences and the clinical associations of anticardiolipin (aCL) and anti-β2GPI (aβ2GPI) antibodies in a large series of European patients with systemic lupus erythematosus (SLE). Methods. 574 SLE patients from 7 European countries were tested for aCL and aβ2GPI by ELISA methods. Results. aCL of IgG, IgM, and IgA isotypes were detected in 22.8%, 14%, and 13.9% of the patients, respectively. IgG and IgM aβ2GPI were detected in 20% of the patients. The presence of aCL was highly associated with the presence of aβ2GPI. Medium-high titer IgG aCL and aβ2GPI were associated with thrombosis, with similar sensitivity, specificity, and positive predictive value. When present at mediumhigh titer, IgG aCL were associated with thrombocytopenia, IgM aCL with hemolytic anemia, and cerebrovascular accidents, IgA aCL with livedo reticularis and Raynauds phenomenon. Conclusion. aCL, when present at medium-high titer, are as important as ab2GPI, as a risk factor for thrombosis. Medium-high titer a...

Analysis of HLA DP, DQ, and DR allesles in adult Italian rheumatoid arthritis patients

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.

Polysorbate 80 and Helicobacter pylori: a microbiological and ultrastructural study

BackgroundThe frequent occurrence of chemoresistant strains reduces the chances of eradication of H. pylori infection and prompted the investigation of non-antibiotic substances active against this organism. Some surfactants enhance the effectiveness of antibiotics for their permeabilizing properties towards bacteria. We examined the antimicrobial activity to H. pylori of the surfactant polysorbate 80, used alone and in association with amoxicillin, clarithromycin, metronidazole, levofloxacin and tetracycline. We also aimed to study the ultrastructural alterations caused upon H. pylori by polysorbate 80, alone and in combination with antibiotics. Twenty-two H. pylori strains were tested using the broth dilution method. After incubation, broth from each dilution was subcultured onto agar enriched with foetal bovine serum to determine the minimum bactericidal concentration (MBC). Synergistic effect of polysorbate 80 with antibiotics was investigated by the broth dilution and disc diffusion techniques. Ultrastructural alterations of organisms treated with polysorbate 80, alone and in association with antibiotics were analyzed by transmission electron microscopy.ResultsMBCs of polysorbate 80 ranged from 2.6 (1.1) μg/ml to 32 (0) μg/ml. Polysorbate 80 exerted a synergistic effect when associated with metronidazole and clarithromycin: polysorbate 80 and metronidazole MBCs decreased byu2009≥u20094 fold; clarithromycin MBCs for two resistant strains decreased by 20 and 1000 times. The principal alteration caused by polysorbate 80 consisted in the detachment of the outer membrane of bacteria.ConclusionsThe bactericidal activity of polysorbate 80 and the synergistic effect of the association with metronidazole and clarithromycin could be useful in the treatment of H. pylori infection.

The Role of Ultrasonography of Peripheral Entheses in the Diagnosis and Assessment of Spondyloarthropathies

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.

Extracorporeal Shock Wave Therapy for Calcific Tendonitis of the Rotator Cuff: A Review

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.

THE EXCRETION OF OXALIC ACID IN GOUTY PATIENTS: 121

Utilizing the very satisfactory HPLC procedure recently reported by Hughes et al. (1), we have determined the urinary excretion of oxalic acid in control subjects (aged 20 to 70 years) and in gouty patients (from 35 to 73 years), before and after allopurinol treatment. The urinary excretion in normal subjects was 27.5±1.89 mg/24h, in gouty patients 31.3±3.08 mg/24h before allopurinol, and 43.22±4.66 mg/24h after treatment. It is evident that there is no substantial difference between normal subjects and gouty patients, while allopurinol significantly increases (p<0.05) the urinary excretion of oxalic acid. Since allopurinol inhibits the incorporation of glycine into purine ribonucleotides, this increase suggests that the glycine might be channeled toward the formation of glyoxylate and oxalic acid in gouty patients (2).Results suggest that the behavior of urinary oxalic acid should be under constant control during allopurinol treatment.(1) Hughes H. et al. (1982) Anal. Biochem. 119, 1-3(2) Dean D.M. et al. (1968) Clin. Sci. 35, 325-335

SEX STEROIDS INFLUENCE ON URIC ACID BINDING TO HUMAN PLASMA PROTEINS: 136

Uric acid binding to human plasma proteins has been studied in various physiological and paraphysiological conditions,including the following groups:I-a group of children aged 4-10 years;2-a group of healthy males;3-a group of subjects treated with estrogen preparations for prostatic cancer;4-a group of regularly menstruating females with normal cycle;5-a group of post-menopausal females;6-a group of pregnant females;7-a group of females treated with oral contraceptives;8-a group of females treated with estrogen preparations for post menopausal symptoms.The results obtained demonstrate a significant correlation between the increase of plasma estrogen and the increase of the uric acid binding to human plasma proteins.

THE |[ldquo]|SWITCH-OFF|[rdquo]| MECHANISM OF SPONTANEOUS RESOLUTION OF ACUTE GOUT ATTACK: 119

In order to investigate the intriguing problem of the spontaneous resolution of acute gout attack,we examined the hypothesis that occurrence of urate crystals size changes or dissolution by oxygen radicals and/or lysosomal enzymes released by stimulated PMNs represents the more likely switch-off mechanism. The absorption spectra and the concentrations of uric acid,allantoin and urea were determined before and after in vitro exposition of MSU crystals to O2 flow generated from photoreduction of riboflavin and O2.The results showed a progressive dissolution of MSU crystals that started after 5 hrs of incubation under O2 flow and was completed after 20 hrs.A progressive decrease of uric acid and an increase of allantoin and urea concentration were simultaneously observed.These results were also confirmed by polarizing and electron microscopy and microcalorimetric techniques. Taking into account these observations,the spontaneous resolution of acute gout attack could be attributed to the dissolving effect on urate crystals by oxygen radicals released extracellularly by phagocytising PMNs.