Anti-Ro/SSA autoantibody-mediated regulation of extracellular matrix fibulins in human epithelial cells of the salivary gland.

OBJECTIVES: The fibulins are a family of extracellular matrix (ECM) molecules that regulate the organ shape along with other growth factors and stromal cells and have recently been shown to be involved in a variety of cellular functions including proliferation, migration, differentiation, and survival. Important changes in acinar and ductal morphology and function, together with pronounced ECM remodelling, are detectable in the labial salivary glands (LSGs) of patients with Sjögren's syndrome (SS). Here we report the in vitro expression of the recently identified ECM proteins fibulin-6 and fibulin-7 by human salivary gland epithelial cells (SGECs). The ability of anti-Ro/SSA autoantibodies (Abs) to modulate fibulin-6 and fibulin-7 expression was investigated. METHODS: Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) and real-time PCR were used to analyse fibulin-6 and fibulin-7 mRNA expression. Confocal microscopy and fluorescence-activated cell sorting (FACS) were used to study expression of the proteins in primary human SGEC cultures, established from biopsies of minor LSGs, in both untreated control cells and anti-Ro/SSA Abs-treated cells. RESULTS: The methods used show the expression of fibulin-6 and fibulin-7 in SGECs. Treatment of cells with anti-Ro/SSA Abs results in a down-regulation of fibulin-6 mRNA expression whereas no significant differences were observed in fibulin-7 expression between untreated and treated cells. CONCLUSION: Dysregulation of fibulin expression in SGECs by anti-Ro/SSA Abs may contribute to disorganization of the ECM environment and thus cause injury to the salivary gland architecture and functionality observed in SS.

Hashimoto's thyroiditis in Melkersson-Rosenthal syndrome patient: casual association or related diseases?

Melkersson-Rosenthal syndrome (MRS) is a rare disorder of unknown etiology. MRS is classically defined as a triad of recurrent orofacial edema, relapsing paralysis of the facial nerve, and fissured tongue. The authors present the case of a 52-year-old woman with orofacial swelling and facial pain attacks. The patient reported to suffer of hypothyroidism and laboratory findings disclosed free triiodothyronine (FT(3)), free thyroxine (FT(4)), and thyrotropin (TSH) altered. Endocrinological consult led to the diagnosis of Hashimoto's thyroiditis. Antithyroper-oxidase antibodies (anti-TPO) were highly elevated and thyroid function tests had evidenced a clinically significant hypothyroidism. A link between MRS and immunological disorders such as sarcoidosis, Crohn's disease, unilateral anterior uveitis and multiple sclerosis was documented. The literature did not report any association between Hashimoto's thyroiditis and Melkersson-Rosenthal syndrome. The presence of the anti-TPO antibodies in the case reported here could suggest a possible correlation between immunological alteration characteristic of autoimmune thyroiditis and MRS.

[Hospital anxiety and depression]. / Ansia e depressione nelle ospedalizzazioni.

OBJECTIVE: The study investigate the presence of depressive disorders in patient who are taken in general hospital, to prevent and reduce the risk of developing a psychological pathology (anxious-depressive symptoms). METHODS: We used two tests: 1. General Health Questionnaire (GHQ-12) by Goldberg, a self-report questionnaire, consisting of 12 items, used to estimate the probability of detecting non-psychotic mental disorders and problems in every day's activity. 2. Personal Health Questionnaire (PHQ), a self-report questionnaire, consisting of 10 items. It is a new tool for detecting individuals with ICD-10 depressive disorders, used to estimate the probability to individuate the presence of major depression symptoms, and minor depression symptoms. Tests have been administered to 140 subjects (males' experimental group, 32 subjects; males' control group, 20 subjects; females' experimental group 58 subjects; females' control group, 30 subjects) from eight departments of university hospital, medical faculty, and social services. RESULTS: It can be noted that with GHQ-12, the presence of non-psychotic mental disorders does not come out: just the females' group, experimental and control, shows some problems with insomnia and stress. On the other hand, with PHQ, the presence of no great entity depressive symptoms comes out for all groups. In the experimental groups the quantity of depressive symptoms is greater than in the control groups. CONCLUSIONS: We have find the presence of a number of depressive symptoms into a hospitalized population. It is known that detecting such symptoms is important for protection and care of depressive disorders in hospitalized and non-hospitalized populations.

Siögren's syndrome: anti-Ro and anti-La autoantibodies trigger apoptotic mechanism in the human salivary gland cell line, A-253.

AIM: The Sjögren's syndrome (SS) is an autoimmune rheumatic disease that targets salivary and lacrimal glands, characterized by a high concentration of autoantibodies in the serum. The anti-Ro and anti-La autoantibodies are present in approximately 70-90% of the patients with primary SS and this presence is correlated to extraglandular manifestations. The objective of this work was to explore the cellular apoptotic pathway triggered by binding and penetration of anti-Ro and anti-La autoantibodies, isolated from the total IgG fraction of patients with primary SS, in the human salivary gland cell line A-253. METHODS: The sera were obtained from 13 healthy volunteers and 13 patients with primary SS. The IgG was obtained from sera through precipitation with ammonium sulfate and the anti-La and anti-Ro autoantibodies were purified using Sepharose 4B-Ro and Sepharose 4B-La affinity columns. The methods used to evaluate the apoptosis were: DNA fragmentation, immunofluorescence and immunoenzymatic tests. RESULTS: In the salivary gland cells, the anti-Ro and anti-La autoantibodies: 1) are able to penetrate; 2) induce DNA fragmentation and cleavage and activation of the effector caspase-3. In the same experimental condition, IgG purified from healthy sera did not have any apoptotic effect on the human salivary gland cell line. CONCLUSION: Anti-Ro and anti-La autoantibodies mediate the apoptosis the human salivary gland cells A-253 in a caspase-3 dependent manner.

Anti-Ro and anti-La autoantibodies induce TNF-alpha production by human salivary gland cells: an in vitro study.

In this report, we demonstrate that both TNFR1 and the TNFR2 are expressed on the salivary gland cell line A-253 cell membrane. Furthermore, cell treatment with anti-Ro and anti-La autoantibodies from Sjögren IgG determined TNF-alpha production, clarifying which could be the inducer of the extrinsic pathway of apoptosis in salivary gland cells.

Autoantibodies from Sjögren's syndrome induce activation of both the intrinsic and extrinsic apoptotic pathways in human salivary gland cell line A-253.

Sjögren's syndrome (SS) is an autoimmune rheumatic disease that targets salivary and lachrymal glands, characterized by a high concentration of serum autoantibodies directed against nuclear and cytoplasmic antigens. It is known that autoantibodies can enter viable cells and this phenomenon has functional consequences including activation of apoptotic process. The objective of this work was to explore whether autoantibodies contained in IgG purified from Sjögren sera trigger apoptotic process in an experimental model represented by the human salivary gland cell line A-253. To define if the intrinsic or extrinsic pathways are activated, we examined which caspases are critical for inducing cell death. The results have demonstrated that morphological changes and DNA laddering, consistent with apoptotic cell death, occurred in A-253 cells treated with IgG from Sjögren sera. Sjögren IgG induced cleavage and activation of the effector caspase-3 and degradation of the caspase-3 substrate poly(ADP-ribose)polymerase. Both the intrinsic and extrinsic apoptotic pathways were activated, since both caspase-8 and caspase-9 cleavages occurred. In conclusion, autoantibodies contained in IgG purified from Sjögren sera mediate apoptosis of the A-253 cell line in a caspase-dependent manner.

Study on the effectiveness of a nifedipine gel for treatment of Raynaud?s phenomenon.

The aim of this study is to evaluate the efficacy of a nifedipine gel in patients with primary or secondary Raynaud?s phenomenon. Photopletismography was the instrumental examination test used to evaluate recovery time (time necessary for recuperation of normal capillary circulation) in 17 patients with primary or secondary Raynaud?s phenomenon before and after the application of the gel. It emerged that of the 17 patients who used the gel, in 3 cases the recovery time was reduced, in 9 cases the recovery time was cancelled (no spasm occurred), in 5 cases the recovery time was not modified. Therefore, in more than 70 percent of patients the drug had a positive effect. Besides, 50 percent of the patients referred an improvement of the subjective symptomatology with reduction of cooling, torpidity, ache and paresthesias of the fingers. The results obtained, even if related to a restricted number of patients and to a brief interval of time, show the effectiveness of this drug in patients with primary or secondary Raynaud?s phenomenon. We believe that these results, presented here for the first time, are important for investigators involved in the study of Raynaud?s disease.

[The strange case of a patient affected by acromegaly with osteoporomalacia without hypogonadism]. / Lo strano caso di un paziente affetto da acromegalia con osteoporomalacia senza ipogonadismo.

Acromegaly is a rare disease that, in the majority of cases, is due to the presence of a benign growth hormone (GH)-producing tumor of the pituitary. Growth hormone has profound effects on linear bone growth, bone metabolism, and bone mass. In acromegaly, the skeletal effects of chronic GH excess have been mainly addressed by evaluating bone mineral density (BMD). Most data were obtained in patients with active acromegaly, and apparently high or normal BMD was observed in the absence of hypogonadism. The Autors describe a case of patient affected by acromegaly without hypogonadism with serious osteoporosis and biological signs of osteomalacia.

Osteoporosis in adult with Marfan syndrome: casuality or causality?

Marfan syndrome (MFS) is an inherited connective tissue disorder transmitted as an autosomal dominant trait. Recent studies indicate that decreased bone mineral density (BMD) occurs in the spine, femoral necks and greater trochanters of some adults and children with MFS. Since there is uncertainty regarding the BMD status of patients with MFS, the authors present a case report on a female patient with MFS and alteration of phosphocalcic metabolism.

The "submerged" connective tissue diseases.

BACKGROUND: Many patients present with laboratory abnormalities suggestive of connective tissue diseases (CTD), but with a few clinical symptoms, therefore not fulfilling the criteria for a specific diagnosis. This fact led us to suppose the existence of a changeable long "preclinical" phase in CTD development. In the past, we have already dealt with this subject, but the existence of a few reports underlining this concept led us to describe some "paradigmatic" clinical cases, and we suggest the definition of "submerged connective tissue diseases (Sub-CTD)". METHODS: During the last 5 years, we observed 25 patients (23 females and 2 males), with a mean age of 32 years, who were enrolled in a prospective study, with a mean follow-up of 23 months. All the patients were ANA-positive and went to our observation presenting only 1 clinical symptom and other laboratory abnormalities suggestive of CTD. RESULTS: The following development of the clinical picture allowed us to make a specific CTD diagnosis in 22 cases. The follow-up study shows the correlation between the starting symptoms, the serological markers (detected at the first examination) and the definitive diagnosis. CONCLUSIONS: From this point of view, we emphasize the importance of the detection of very sensitive and specific serological markers, which are useful for an early diagnosis, particularly in this "submerged" phase of the disease.

[Biochemical markers of bone turnover and YKL 40 in ankylosing spondylitis. Relation to disease activity]. / Marcatori biochimici di turnover osseo e YKL 40 nella spondilite anchilosante. Rapporto con l'attività di malattia.

BACKGROUND: YKL-40 is a glycoprotein produced by chondrocytes and synovial cells. The plasmatic levels of this metabolite increase in some pathologies such as rheumatoid arthritis and osteoarthrosis, so much so that it can be considered an effective marker of disease activity and in the therapeutic monitoring of these diseases. It has been interesting to dose a group of both male and female subjects affected by seronegative spondylarthritis, comparing this parameter with the disease activity indexes and with the bone turnover markers. METHODS: The study has been carried out on 48 subjects (26 males and 22 females) between 17 and 68 years affected by spondylarthritis, diagnosed in conformity with ARA standards. None of the patients carried out basic treatment or by glycocorticoids, and 22 patients took FANS when required. In these subjects the disease activity markers (VES, PCR, fibrinogen, mucoprotein) and some of the classic bone remodelling markers (blood calcium and phosphates, calciuria, phosphaturia, Ca++, Ntx, osteocalcine, bone isoenzyme of alkaline phosphatase, hydroxyproline, procollagen and YKL-40) were dosed. RESULTS: The comparison between different parameters pointed out that the highest values are obtained in subjects of most advanced age with the highest phlogosis indexes, without any correlation with sex. The quite interesting comparison shows a correlation between the bone remodelling indexes and YKL-40, being particularly remarkable when the disease is more aggressive or during relapse. CONCLUSIONS: It is then possible to confirm that, though preliminary, these data may suggest evaluations on wider case histories to research YKL-40 as a surgical monitoring marker in seronegative poliarthritis.

[Sex hormones and male osteoporosis. A physiologic prospective for prevention and therapy]. / Ormoni sessuali ed osteoporosi maschile. Una prospettiva fisiologica di prevenzione e terapia.

BACKGROUND: Male osteoporosis is often an underestimate and non-acknowledged pathology because it is less frequent then post-menopausal women osteoporosis. The causes of osteoporosis in males were reviewed, considering the importance of the assessment of sexual hormones, even without symptoms of hypogonadism. METHODS: Sixty-two patients ranging in age from 45 to 75 years, males, were studied. None of them had assumed steroids nor other drugs causing osteoporosis. These patients have been subjected to bone mineral analysis (BMA) together with the following sexual hormones: LH, FSH, total testosterone, free testosterone, SHBG, estrone, estradiol. Two methods of BMA have been employed (Norland 2780 and UBA 575 Walker and Sonix). RESULTS: When osteoporosis or osteopenia were present, always there were modifications of sex hormones values. Statistic evaluation (linear regression, Pearson coefficient, Multiple regression, Backward Stepwise Multiple Regression), showed that there was a significant association between total testosterone and osteoporosis. CONCLUSIONS: Total testosterone resulted the most predictive sex hormone for the loss of bone mass; therefore, it is important to evaluate sex hormones in males with osteoporosis, for a correct and "physiological" therapy.

Controlled study with a new sustained-release formulation of nifedipine in essential hypertensive patients.

The authors studied the antihypertensive effect and tolerability of a new sustained-release formulation of nifedipine 50 mg once a day, in comparison with nifedipine retard 20 mg twice a day in patients with mild or moderate primary arterial hypertension. Both treatments significantly lowered blood pressure with no difference in daily blood pressure profile. At steady state, the two drugs determined comparable plasma levels of nifedipine as measured immediately before the morning dose. After a 12-month treatment, the new formulation of nifedipine still displayed satisfactory blood pressure control in both supine and standing positions, with no change in tolerability throughout the study. In conclusion, this new sustained-release formulation of nifedipine has similar efficacy and tolerability to conventional treatment with nifedipine retard 20 mg twice a day.

[Significance of anticardiolipin antibodies in connective tissue diseases]. / Significato degli anticorpi anticardiolipina nelle connecttiviti.

Recent attention has focused on the possibility that the presence of Anticardiolipin Antibodies (ACA) in patients with connective tissue diseases, particularly with Systemic Lupus Erythematosus (SLE), may be associated with clinical and serological symptoms that identify a particular subset of patients. This group is characterized by recurrent arterial and/or venous thrombosis, multiple abortions and or intrauterine fetal death, presence of Lupus AntiCoagulant (LAC), false positive VDRL, thrombocytopenia and neuro-psychiatric diseases. These clinical features may identify the so-called "Anticardiolipin Syndrome". In this work, we have measured ACA, by ELISA test, in 194 serum samples: 97 SLE patients, 5 Mixed Connective Tissue Disease (MCTD), 8 Progressive Systemic Sclerosis (PSS), 7 Dermato/PolyMyositis (D PM), 3 Sjögren Syndrome (SS), 3 Unclassifiable Connective Tissue Disease (UCTD), 9 Rheumatoid Arthritis (RA), 1 Idiopathic Anticardiolipin Syndrome, 19 cases of Miscellanea, 42 healthy controls. The Optical Density (O.D.) was greater than 0 (higher than 0) in 89 serum samples (out of the 194): 43 SLE, 4 MCTD, 4 PSS, 3 D PM, 1 SS, 7 RA, 10 cases of Miscellanea, the Idiopathic Anticardiolipin Syndrome and 16 healthy controls. The O.D. was greater than m (greater than mean healthy controls level) + 3 Standard Deviations (S.D.) in a restricted number of cases (25 out of the 194): 14 SLE, 2 MCTD, 1 PSS, 1 D/PM, 1 SS, 1 RA, 3 cases of Miscellanea and 2 healthy controls. Therefore, we have noticed, first of all, the lack of specificity of positive results obtained: all the groups of patients, healthy controls included, had low as well as high levels of ACA. Moreover, we have examined the relationship between the presence of ACA and typical clinical features of the so-called. "Anticardiolipin Syndrome"; there was not difference of clinical symptoms between patients with low or high ACA levels. We have also clinically examined ACA negative patients; most of them had one or several clinical features of the "Syndrome", until almost complete clinical picture. Therefore, no correlation was found between clinical picture and immunological features of the so-called "Anti-cardiolipin Syndrome"; we would not exclude the existence of the "clinical" subset of patients, but of the "immunological" subset.

[Connective tissue diseases and HEp-2 antinuclear antibodies]. / Connettiviti e anticorpi antinucleari HEp-2.

Some characteristics of antinuclear antibodies that might be of use for diagnostic and/or prognostic purposed were studied using indirect immunofluorescence on HEp-2 cells in 55 cases of various types of connective tissue disease. For each nuclear (homogeneous, speckled, granular, dotted, pulverulent and centromeric) and nucleolar fluorescence pattern (homogeneous, conglutinated and dotted), the following parameters were observed; C3 fixing capacity, degree of antibody affinity and sensitivity to RNAse, DNAse and trypsin. The results were very interesting, especially in relation to the diagnosis of progressive systemic sclerosis and of the related subsets, but were insufficient for reliable, conclusive prognostic evaluation.

[Clinical significance of antinuclear antibodies in progressive systemic sclerosis]. / Significato clinico degli anticorpi antinucleari nella sclerosi sistemica progressiva.

Indirect immunofluorescence (IIF) was used to detect antinuclear antibodies (ANA) in 42 clinical cases. In each case cryostatic rat kidney slices and cultivated HEp-2 cells were used as substrates. Clinical diagnoses were as follow: Progressive Systemic Sclerosis (PSS) 25 cases, of which 8 were acrosclerotic, 8 diffuse, 5 CREST syndrome, 1 overlap PSS + Systemic Lupus Erythematosus (SLE) and 3 PSS + myopathy; Localised scleroderma (morphea): 3 cases; Mixed Connective Tissue Disease (MCTD): 3 cases; "Idiopathic" Raynaud's Disease (RD): 4 cases; Dermatomyositis (DM): 2 cases (1 paraneoplastic); SLE: 1 case; Unclassifiable Connective Tissue Disease (UCTD): 4 cases. The ANA-positive cases identified by the traditional technique were divided according to pattern into 4 categories: homogeneous, peripheral, speckled, nucleolar. In contrast those identified using HEp-2 cells were divided into 9 pattern groups: (nuclear type) centromere, fine speckled, coarse speckled, diffusely grainy, homogeneous: (nucleolar type) speckled, clumpy, homogeneous. The results demonstrated a higher general incidence of positivity with HEp-2 cells and confirmed the close connection between Anticentromere ANA and CREST syndrome. A similarly close connection was noted between MCTD and both nuclear diffusely grainy and nucleolar speckled patterns. A fairly clear connection was also noted between acrosclerotic or diffuse SSP and a fine speckled nuclear pattern. It is felt that ANA tests using IFI on HEp-2 cells should lead to significant progress in the field of diagnosis and prognosis and the study of PSS subsets.

[Capillaroscopic studies in connective tissue inflammations]. / Studi capillaroscopici nelle connettiviti.

Capillarioscopy has been unjustifiably neglected in the study of connective tissue diseases, where examination of the microcirculation is clearly important. A study of 80 cases is reported. 12 systemic lupus, 11 progressive systemic sclerosis (PSS), 20 rheumatoid arthritis (including 3 juvenile and 2 Still's disease); 9 Raynaud's disease (of which 3 idiopathic, 4 with rheumatoid arthritis and 2 with UCTD); 1 dermatomyositis; 11 other CTD (2 overlap syndrome--1 lupus + dermatomyositis; 1 lupus + PSS--3 Sjögren syndromes with rheumatoid arthritis, 1 MCTD, 2 primary mixed cryoglobulinaemia, 1 systemic vasculitis, 1 Behçet syndrome, and 1 UCTD); 9 miscellaneous forms (3 psoriatic arthropathy, 1 rheumatic pelvispondylitis, 1 allergic dermatitis, 1 pulmonary TB, 1 ulcerative colitis; 1 scapulohumeral periarthritis, 1 unclassifiable rheumatism; 7 healthy subjects). During capillarioscopy, from one to nine slides were prepared for each subject. These were interpreted separately by three persons who were unaware of the respective diagnosis. Calibre, tortuosity, length and number of capillaries were recorded, plus the visibility of the subpapillar plexus, height and number of the termal, subungual and/or ungual vallum haemorrhage, plugging. It was found that PSS, dermatomyositis, MCTD, and overlap-PSS revealed a very typical common pattern, possibly pathognomonic, namely marked reduction in the number of capillaries + megacapillaries. The other forms presented less evocative diagnostic patterns, though they were fairly indicative in some instances. Clinical correlations of particular significance with respect to prognosis, however, were not observed.