Matrix metalloproteinase gene polymorphisms and susceptibility to systemic sclerosis.

The major pathological hallmark of the systemic sclerosis (SSc) is skin and internal organ fibrosis, which results from normal tissue architecture alterations and extracellular matrix (ECM) protein deposition. ECM components are degraded by matrix metalloproteinases (MMP). Promoter region polymorphisms in MMP genes may influence gene expression, resulting in an imbalance between ECM protein production and degradation. Here, we analyzed MMP1 -1607 1G/2G (rs1799750), MMP3 -1171 5A/6A (rs3025058), and MMP9 -1562 C/T (rs3918242) polymorphisms in relation to susceptibility to SSc and its clinical features. The patient group included 98 individuals with longstanding or recently diagnosed disease, meeting the American College of Rheumatology or LeRoy and Medsger criteria for SSc; the control group included 100 healthy blood donors. All participants were of European descent. Genotyping was performed by polymerase chain reaction followed by restriction digestion. Genotype and allele frequencies of MMP polymorphisms were similar between the two groups. In secondary analyses, significantly higher frequency of 1G/2G genotype from MMP1 polymorphism was observed for patients testing positive for antinuclear autoantibodies (P = 0.007), while 1G/1G genotype was associated with interstitial lung disease development (P = 0.018). The 6A/6A genotype from MMP3 polymorphism was absent in patients with calcinosis (P = 0.011), while the MMP3 5A/5A genotype correlated with the presence of anti-topoisomerase I antibodies (P = 0.009) and reduced diffusing capacity for carbon monoxide (P = 0.024). These results suggest that MMP polymorphisms are not associated with SSc susceptibility, although MMP1 and MMP3 variants are associated with specific SSc clinical and laboratory features.

Assessment of nailfold capillaroscopy in systemic sclerosis by different optical magnification methods.

BACKGROUND: Systemic sclerosis (SSc) is characterized by target-organ fibrosis and microvascular dysfunction, which can be assessed using nailfold capillaroscopy. Dermoscopy is a useful and easily performed method for diagnosing skin lesions. AIM: To compare conventional capillaroscopy, using the gold-standard method (conventional stereomicroscope nailfold capillaroscopy; SNFC), with polarized light noncontact dermoscopy (PNCD) and nonpolarized light contact dermoscopy (NPCD), and to evaluate their accuracy in diagnosing characteristic SSc-related alterations. METHODS: The study enrolled 45 patients with SSc. Capillaroscopy images and photographs were taken with three devices, SNFC, NPCD and PNCD, and these images were randomly analysed by a blinded observer. RESULTS: The scleroderma pattern was found in 83% of patients. PNCD and NPCD were highly sensitive in identifying the presence of focal capillary loss (96.4% and 100%, respectively), haemorrhage (96.2% and 92%, respectively), and scleroderma (91.9%, 94.6%), and showed high specificity for haemorrhage and enlarged loops. The intra-observer kappa values for detection of the scleroderma pattern by SNFC images, NPCD and PNCD were moderate to good: (κ = 0.71 (95% CI 0.44-0.95), κ = 0.60 (95% CI 0.35-0.83) and κ = 0.60 (95% CI 0.32-0.86), respectively. Evaluation of haemorrhage presence gave high kappa values for all methods: κ = 0.77 (95% CI 0.57-0.95), κ = 0.90 (95% CI 0.76-1.00) and κ = 0.95 (95% CI 0.85-1.00), respectively. CONCLUSIONS: Both polarized and nonpolarized dermoscopy are reliable methods for valuation of nailfold capillaroscopy in patients with SSc. They are easy to perform, with good rates of accuracy and results that are comparable with traditional capillaroscopy.

Interleukin-10 gene promoter and NFKB1 promoter insertion/deletion polymorphisms in systemic sclerosis.

Systemic sclerosis (SSc) is a connective tissue disease characterized by fibrotic, immunological and vascular abnormalities. Nuclear factor-kB (NFKB), as a key transcription factor involved in the regulation of immune responses, appears to be a good candidate for studies on the pathogenesis of autoimmune diseases, as well as the interleukin-10 (IL-10) polymorphism, which other studies have suggested an association with SSc. Our objective was to study the association of NFKB and IL-10 gene polymorphisms with SSc. One hundred and fifty-one SSc patients and 147 healthy bone marrow donors were enrolled in a case-control study. Blood was collected for DNA extraction; typing of IL-10 genes was made by polymerase chain reaction with sequence-specific primers (PCR-SSP), and NFKB gene typing was made by restriction fragment length polymorphism (RFLP). Patients underwent clinical evaluation, serology, Doppler echocardiography and chest high-resolution computed tomography. The frequency of IL-10 (-1082) GG genotype was found to be significantly higher in SSc patients (36.4%) as compared to healthy controls (22.4%) (P = 0.012). The frequency of heterozygous genotype GA was significantly lower (P = 0.004) in patients (38.4%) in comparison with control subjects (55.8%). A predominance of the high-producing IL-10 phenotype (GCC(+) /GCC(+) ) was observed in SSc patients compared with healthy controls (37.7% versus 24.5%, respectively; OR: 1.87, 95% CI: 1.10-3.19, P = 0.019). No significant difference was found in the allelic and genotype distribution of the NFKB promoter polymorphism between patients and controls. No statistically significant associations were found between IL-10 or NFKB polymorphisms clinical and laboratory features of SSc. Our results confirmed the association of the high-producing phenotype (GCC(+) /GCC(+) ) with increased risk for SSc, but found no correlation with NFKB polymorphisms.

Killer cell immunoglobulin-like receptor (KIR) genes in systemic sclerosis.

A previous study has suggested that the combination KIR2DS2(+)/KIR2DL2(-) was related to increased risk for systemic sclerosis (SSc), while others have failed to reproduce this finding. Our objective was to study this matter further and test the association of other KIR genes with SSc. One hundred and ten SSc patients and 115 healthy bone marrow donors were enrolled in a case-control study. Blood was collected for DNA extraction; typing of 15 KIR genes and human leucocyte antigen-C (HLA-C) was made by polymerase chain reaction with sequence specific primers (PCR-SSP), followed by electrophoresis on agarose gel. Patients underwent clinical evaluation, serology, Doppler echocardiography and chest high-resolution computed tomography. The frequency of the inhibitory KIR2DL2 was significantly lower in patients [29.1% versus 65.2% in controls, P < 0.0001; odds ratio (OR) = 0.22, 95% confidence interval 0.12-0.40]. When combinations of activating and inhibitory KIR genes were analysed, the presence of KIR2DS2 in the absence of KIR2DL2 (KIR2DS2(+)/KIR2DL2(-)) was more frequent in patients than in controls (25.5% versus 1.7%, respectively; P < 0.0001; OR = 19.29, 4.24-122.26). However, the presence of both KIR2DS2 and KIR2DL2 (KIR2DS2(+)/KIR2DL2(+)) was more frequent in controls (57.4%) than in patients (28.2%, P < 0.0001), suggesting a preponderant protective effect of KIR2DL2 over KIR2DS2. Stratification for HLA-C1 status did not change these results. No statistically significant associations were found between KIR phenotypes and clinical and laboratory features of SSc. Our results suggest a protective role of KIR2DL2(+) phenotype and confirmed the association of the combination KIR2DS2(+)/KIR2DL2(-) with increased risk for SSc.

Nailfold capillaroscopic findings in primary Sjögren's syndrome: clinical and serological correlations.

OBJECTIVE: To describe the capillaroscopic abnormalities observed in patients with primary Sjögren's Syndrome (pSS), associating them with clinical and serologic features, and comparing these findings to those observed in normal controls. METHODS: Sixty-one consecutive patients with pSS were studied by clinical evaluation, serology, and nailfold capillary microscopy (NCM). Twenty-one normal controls were also examined. Capillaroscopic findings were recorded in a standardized way by a blinded observer Capillary loss on NCM was evaluated using a deletion score. RESULTS: NCM was normal in 59.0% of pSS patients; 29.5% had non-specific abnormalities, and 11.5% presented a SD-like pattern. Patients presented a higher deletion score than controls (p < 0.001). Other capillaroscopic parameters (number of dilated, bizarre, and meandering capillaries; capillary hemorrhages; venous plexus visibility) did not differ significantly between patients and controls. Among patients, the deletion score was higher in those with systemic manifestations (p = 0.022) and Raynaud's phenomenon (p = 0.050). No association between the presence of antinuclear antibodies, rheumatoid factor, anti-SSA/Ro and anti-SSB/La with qualitative or quantitative NCM findings was found. Among the 7 patients with SD-like pattern on NCM, 6 had Raynaud's phenomenon, but only 2 presented autoantibodies related to systemic sclerosis (1 with anticentromere and 1 with low titer antitopoisomerase I). None of these patients met the ACR criteria for SSc. CONCLUSIONS: SD-like pattern on NCM is observed in a small but significant proportion of pSS patients. The association of systemic involvement with a higher deletion score may be related to the hypothesis that these manifestations represent clinical expressions of systemic vasculitis.

Alcohol intake and blood pressure: the importance of time elapsed since last drink.

BACKGROUND: A positive association of chronic exposure to alcoholic beverages with blood pressure and the prevalence of hypertension has been described in epidemiological surveys, but the influence of time elapsed since last ingestion in this setting was not demonstrated. DESIGN: A cross-sectional, population-based survey. METHODS: In total 1089 adults from Porto Alegre, randomly selected from a population-based, multi-stage probability sample, were interviewed at home. The average daily alcohol intake of each subject was calculated taking into account the concentration of ethanol in the beverages (distilled or fermented beverages), and the time elapsed between the last ingestion of ethanol and the moment of blood pressure determination. Standardized sitting blood pressure and anthropometric parameters were collected. The magnitude and shape of the associations were analyzed considering blood pressure as a continuous variable and the prevalence of arbitrarily defined hypertension. Simple and multiple linear regression models, including models to identify nonlinear associations, with quadratic and cubic terms of the amount of alcohol consumed, were employed. Blood pressure means were compared by analysis of variance and analysis of covariance. The association between hypertension and exposure to ethanol was analyzed through logistic regression models, controlling for various potential confounders. RESULTS: Positive nonlinear associations of the amount of alcohol consumed with blood pressure and the prevalence of hypertension (> or = 160/95 mmHg) were found, independent of age, years of education, smoking, and use of oral contraceptive and antihypertensive drugs. The consumption of 30 g/day ethanol was associated with increases of 1.5 and 2.3 mmHg in diastolic and systolic blood pressures, respectively, for men, and 2.1 and 3.2 mmHg, respectively, for women. The prevalence of hypertension was higher among those ingesting more than 30 g/day (odds ratio = 2.9, P < 0.01). The time elapsed between the last ingestion and blood pressure measurement was independently associated with the prevalence of hypertension. Men with last consumption of alcohol 13-23 h prior to measurement had odds of being hypertensive 2.6 (confidence interval 1.3-5.0) greater than did subjects who had consumed alcoholic beverages 24 h and more before the blood pressure determination. For men, systolic and diastolic blood pressures were lower during the first 3 h after ingestion and increased afterward. Frequency of consumption and type of beverage consumed were not independently associated with level of blood pressure. CONCLUSION: A time-dependent association between alcohol consumption and effects on blood pressure, demonstrated in experimental studies, was found for free-living individuals selected at random.

Study of the usefulness of optic fundi examination of patients with hypertension in a clinical setting.

The purpose of this study was to evaluate the usefulness of direct opthalmoscopy by non-opthalmologists in patients with hypertension. In a cross-sectional survey, we analysed the association between optic fundi abnormalities, individually and according to the criteria of Keith and Wagener (KW), with blood pressure and duration of known hypertension in 400 non-diabetic hypertensive patients. The optic fundi abnormalities were more frequent in patients with diastolic blood pressure (DBP) > 105 mm Hg (P = 0.002), SBP > 180 mm Hg (P < 0.0001) and with a duration of known hypertension > 3 years (P = 0.002). The severity of hypertension did not vary in parallel with the KW classes I and II: 34.5% of patients classified as KW I had a diastolic pressure of > 105 mm Hg compared with only 25.3% of those classified as KW II. Class III abnormalities were infrequent (2.5% of the whole cohort). In a logistic regression model, diffuse arteriolar narrowing was associated with DBP (P = 0.002) and age (P < 0.001). Abnormalities of the arteriovenous crossings were associated with SBP (P = 0.001) and duration of disease (P = 0.008). The positive predictive value of any fundoscopic abnormality to estimate the severity of hypertension was 59% and the negative value was 60%. The results of this study demonstrate that optic fundi examination by internists and cardiologists does not give an accurate assessment of the severity of hypertension in most patients, and that the Keith-Wagener classification of retinopathy has a limited applicability.(ABSTRACT TRUNCATED AT 250 WORDS)

Alcoholic beverage consumption and associated factors in Porto Alegre, a southern Brazilian city: a population-based survey.

OBJECTIVE: To describe the pattern of alcoholic beverage consumption and the prevalence of at risk drinking behaviors, as well as their association with demographic and socioeconomic factors in the adult population of Porto Alegre, a southern Brazilian city. METHOD: In a cross-sectional, population-based, multistage random sampling study, 1,091 (600 female) individuals (92% of those eligible) were selected and interviewed at home. Exposure to alcohol was measured by the CAGE questionnaire and by inquiring about the type, quantity and frequency of alcoholic beverage consumption. An average consumption of 30 g per day or more, a level of exposure associated with health risks, was considered as heavy drinking. Two positive answers to the GAGE questionnaire represented the cutoff for indicating dependence. RESULTS: The prevalences were: 9.3% (95% CI: 7.6 to 11.0) for dependence, 15.5% (13.4 to 17.7) for heavy drinking and 12.3% (10.4 to 14.2) for daily drinking; 24.1% (21.7 to 26.6) were abstinent. Women consumed alcoholic beverages in lower frequency and amounts than men. The most widely consumed beverages were beer, wine and "cachaça," a Brazilian sugarcane spirit. In a logistic regression model, increasing age, lower education and income, and nonwhite race were associated with heavy drinking and dependence. Households with 3-4 persons were associated with the lowest risk of heavy drinking, but the prevalence of dependence was higher in crowded households. The presence of another heavy drinker or dependent in the household was associated with heavy drinking but not with dependence. CONCLUSIONS: The study characterized a detailed pattern of alcoholic beverage use and indicated that at risk drinking is an important public health problem in a developing country. The risk factors for heavy drinking and dependence were the same, with the exception of age at starting to drink, heavy drinking or dependence-positive household members.

[Prevalence of systemic arterial hypertension and associated risk factors in the Porto Alegre metropolitan area. Populational-based study]. / Prevalência de hipertensão arterial sistêmica e fatores associados na região urbana de Porto Alegre. Estudo de base populacional.

PURPOSE: To evaluate the contemporaneous prevalence of hypertension in Porto Alegre, RS, and its association with biological, socioeconomic and environmental factors. METHODS: It was done an observational and analytical study with a cross-sectional design, of a representative sample of the adults of the urban region. The study was planned with a power to describe the main estimates with 0.5% confidence limits of +/- 2%. One thousand and ninety one individuals, selected at random in conglomerates and in a multiple stage process, were interviewed. The data were obtained in the domiciles through standardized questionnaires and physical examination. RESULTS: The prevalence of hypertension, defined by blood pressure (BP) > or = 160/95 mmHg, was 12.6% (CI = 10.6 to 14.6). Considering as hypertensives the individuals with BP < 160/95 mmHg under drug treatment, the prevalence increased to 19.2% (CI = 16.9 to 21.5). The corresponding figures for the 140/90 mmHg criteria were 25.8% (CI = 23.2 to 28.4) and 29.8% (CI = 27.1 to 32.5). Among those using anti-hypertensive drugs (11%), 58.9 had BP < 160/95 mmHg and 35.5% < 140/90 mmHg; 57.7% of the hypertensives (160/95 criteria) were aware of diagnosis; 28% had body mass index above 27kg/m2, 15.5% consumed more than 30g per day of ethanol, 35.1% were smokers, and 17.8% ex-smokers. The prevalence increased with age and was higher in individuals with obesity, strong family history of hypertension, low education and in those which abused from alcoholic beverages. In a logistic regression model, these putative risk factors showed to be independent of others. CONCLUSION: The contemporaneous prevalence of hypertension in Porto Alegre demonstrates that the prevalence rates have not decreased in the last 15 years. It was also shown an inadequate BP control in almost 50% of those under drug treatment, and finally, the association of hypertension with well-known risk factors.

Glu298Asp eNOS polymorphism is not associated with SLE.

To examine potential associations of the Glu298Asp polymorphism in the coding region of the endothelial nitric oxide synthase (eNOS) gene with susceptibility to and clinical expression of systemic lupus erythematosus (SLE). One hundred thirteen consecutive patients of European ancestry with diagnosis of SLE, satisfying the American College of Rheumatology criteria, from the outpatient clinic of the Serviço de Reumatologia of the Hospital de Clínicas de Porto Alegre, and 206 healthy controls from the same geographic area were genotyped by polymerase chain reaction for the Glu298Asp polymorphism in the coding region of the eNOS gene. Clinical, demographic, and laboratorial data were collected. Clinical manifestations of SLE and related diseases were evaluated for the association with specific genotypes. The allelic and genotypic distribution of the Glu298Asp did not differ significantly between SLE patients and controls. We found no association of the polymorphism with lupus nephritis, antiphospholipid syndrome, cardiovascular disease (CVD), and risk factors to CVD. The presented results in this study do not provide support for a major role of eNOS Glu298Asp neither in the susceptibility for SLE or clinical manifestations, although there was low statistical power for the latter evaluation.

The chemokine receptor CCR5 genetic polymorphism and expression in rheumatoid arthritis patients.

OBJECTIVES: To identify the genetic polymorphism of the chemokine receptor CCR5 (the Delta32 allelic variant) in patients with rheumatoid arthritis (RA) and compare the findings with healthy controls. To compare the CCR5 phenotypic expression in T cells and monocytes isolated from the peripheral blood and synovial fluid in a subgroup of RA patients. METHODS: CCR5 genes of 92 RA patients and 160 healthy controls were genotyped using specific primers flanking the region of deletion. The ethnic distribution was similar between the groups. Flow cytometric analysis was used for immunophenotyping the T cells and monocytes isolated from the peripheral blood and synovial fluid of eight RA patients. The isolated cells were triple stained with CD4 or CD8, CD25 and CCR5 monoclonal antibodies. RESULTS: There was no difference in the CCR5Delta32 genotypic frequency between the RA patients and the control group (0.055 and 0.063, respectively, p = 0.989). No homozygote for the CCR5Delta32 allele was seen in either group. Five heterozygotes were identified in the RA patient group, whose disease was shown to be aggressive. A significant enrichment of activated CCR5+ monocytes was seen in the synovial fluid of the RA patients subjected to arthrocentesis, who were all homozygotes for the CCR5 wild-type genotype. CONCLUSION: A protective role for the CCR5 allelic variant in RA development was not observed. Disease severity in the heterozygotes suggests that other proinflammatory mechanisms might overcome this mutation in vivo. The activated CCR5+ monocyte enrichment in the rheumatoid synovial fluid might indicate that this cell population has an important role in the pathogenesis of the disease.

Raised serum S100B protein levels in neuropsychiatric lupus.

OBJECTIVE: To test serum S100B protein levels in patients with and without neuropsychiatric systemic lupus erythematosus (NPSLE) and controls. METHODS: 87 patients with SLE, 23 with and 64 without neuropsychiatric involvement, and 25 control subjects were prospectively evaluated. NPSLE diagnosis was made according to the American College of Rheumatology nomenclature and case definitions for neuropsychiatric lupus syndromes. Serum S100B protein levels were determined with a luminescence immunoassay. Statistical analysis was performed using Mann-Whitney and Kruskal-Wallis tests. RESULTS: Among the patients with NPSLE, 9 presented psychosis; 4, cranial neuropathy; 3, cerebrovascular disease; 1, seizures; 1, chorea; 1, peripheral polyneuropathy; 1, multiplex mononeuropathy; 3, dementia. Serum concentrations of S100B protein were significantly higher in patients with NPSLE (median 0.164 ng/ml, interquartile range 0.113-0.332) than in non-NPSLE patients (0.062 ng/ml, 0.026-0.109) and controls (0.088 ng/ml, 0.013-0.124) (p<0.001). Patients with anti-dsDNA antibodies had higher S100B protein levels (p = 0.001). No significant associations were found of lupus activity (among non-NPSLE cases), antiphospholipid antibodies, and reduced complement levels with S100B concentration. CONCLUSIONS: Serum S100B protein level is raised in NPSLE, reflecting continuing neurological damage. The association of anti-dsDNA antibodies with higher S100B protein concentration deserves further study.

Oesophageal dysmotility in systemic sclerosis: comparison of HRCT and scintigraphy.

The aim of this study was to compare oesophageal abnormalities observed in high-resolution CT with radionuclide transit in patients with systemic sclerosis. 76 patients with systemic sclerosis were evaluated by high-resolution CT and oesophageal transit scintigraphy. Residual activity > or =20% (in relation to peak activity) at 15 s after the beginning of the swallow of the labelled liquid (in supine position) was considered indicative of oesophageal dysfunction. Supra-aortic and infra-aortic oesophageal coronal diameters were measured in high-resolution CT. Oesophageal dilatation was deemed present when the diameters exceeded 10 mm. 19 patients (25%) had supra-aortic oesophageal dilatation and 48 patients (63.1%) had infra-aortic dilatation. The prevalence of radionuclide transit delay was 77.6%. All patients (19/19) with supra-aortic dilatation had oesophageal dysfunction, compared with 70.2% (40/57) of the patients with no supra-aortic dilatation (p = 0.004). Oesophageal dysfunction was present in 97.9% (47/48) of patients with infra-aortic dilatation, compared with 42.9% (12/28) in patients without it (p < 0.001). Receiver operating characteristic (ROC) curves have demonstrated that the supra-aortic and infra-aortic diameters had good discriminatory capacity for oesophageal dysfunction in systemic sclerosis (area under the curve, 95% confidence interval: 0.80, 0.70-0.89 and 0.92, 0.86-0.98, respectively). There is a clinically significant association between oesophageal dysmotility and high-resolution CT findings of oesophageal coronal dilatation. The evaluation of infra-aortic oesophageal coronal diameter can provide additional useful information about the functional and anatomic conditions of the oesophagus in systemic sclerosis.

A study of the association between epistaxis and the severity of hypertension.

Hypertension (HTN) has frequently been cited as a general risk factor for epistaxis. However, studies dealing with this association have yielded equivocal results. In this study, a sample of 121 hypertensives (blood pressure > or = 140/90 mmHg) was selected to evaluate the association between the severity of HTN and a previous history of epistaxis. Patients with an average blood pressure > or = 160/100 mmHg were classified as suffering from a more severe form of HTN and were compared with those with a less severe form of the disease (160/100 mm Hg < or = blood pressure > or = 140/90 mm Hg). The frequency of epistaxis did not differ among patients categorized by the severity of HTN. Users of aspirin were found to be twice as likely to have a history of epistaxis. In addition, there was a statistical tendency for an association between a history of epistaxis and the duration of hypertension. We conclude that the severity of HTN and a history of epistaxis were not associated in a cohort of hypertensive patients. The identification of other risk factors for epistaxis, including the duration of HTN, deserves further study.