Systemic sclerosis sine scleroderma: distinct features in a large Brazilian cohort.

OBJECTIVE: Systemic sclerosis sine scleroderma (ssSSc) is an infrequent SSc variant characterized by visceral and immunological manifestations of SSc in the absence of clinically detectable skin involvement. We sought to delineate the characteristics of ssSSc in a cohort of Brazilian patients and contrast them with those in the literature. METHODS: SSc patients seen at two academic medical centres in Brazil were retrospectively analysed. Patients were classified as ssSSc if they presented with RP, positive ANAs and at least one visceral involvement typical of SSc in the absence of skin thickening. Demographics, clinical and laboratory data were obtained by chart review. Literature review was performed by searching available original studies up until June 2012. RESULTS: Among the 947 consecutive patients with SSc, 79 (8.3%) were classified as ssSSc. Oesophagus was the most frequently affected organ (83.1%), followed by pulmonary involvement (63.2%). Compared with the limited cutaneous form of SSc, telangiectasia was the only variable significantly different after multivariate logistic regression analyses (odds ratio 0.46; 95% CI 0.27, 0.81). Compared with the diffuse cutaneous form of SSc, multivariate analyses revealed that ssSSc patients were less likely to be male (odds ratio 0.15; 95% CI 0.04, 0.57), have digital ulcers (odds ratio 0.26; 95% CI 0.13, 0.51) or anti-Scl70 antibodies (odds ratio 0.19; 95% CI 0.07, 0.55) and less frequently treated with CYC (odds ratio 0.23; 95% CI 0.12, 0.43). These features were comparable to those in the published literature. CONCLUSION: In this series, patients with ssSSc had a relatively mild disease with good prognosis.

Fluctuation of anti-endothelial cell antibody titers in " mixed connective tissue disease.

BACKGROUND: Antibodies directed against endothelial cell surface antigens have been described in many disorders and have been associated with disease activity. Since the most prominent histopathologic feature in mixed connective tissue disease (MCTD) is the widespread and unique proliferative vascular lesion, our aim was to evaluate the frequency of anti-endothelial cell antibodies (AECA) in this condition. OBJECTIVES: To evaluate the frequency of AECA in this disease and assess its clinical and laboratory associations. METHODS: Seventy-three sera from 35 patients with MCTD (Kasukawa's criteria), collected during a 7 year period, were tested for immunoglobulins G and M (IgG and IgM) AECA by cellular ELISA, using HUVEC (human umbilical vein endothelial cells). Sera from 37 patients with systemic lupus erythematosus (SLE), 22 with systemic sclerosis (SSc) and 36 sera from normal healthy individuals were used as controls. A cellular ELISA using HeLa cells was also performed as a laboratory control method. RESULTS: IgG-AECA was detected in 77% of MCTD patients, 54% of SLE patients, 36% of SSc patients and 6% of normal controls. In MCTD, IgG-AECA was associated with vasculitic manifestations, disease activity and lymphopenia, and was also a predictor of constant disease activity. Immunosuppressive drugs were shown to reduce IgG-AECA titers. Since antibodies directed to HeLa cell surface were negative, AECA was apparently unrelated to common epitopes present on epithelial cell lines. CONCLUSIONS: AECA are present in a large proportion of patients with MCTD and these antibodies decrease after immunosuppressive treatment.

Pathological pulmonary vascular remodeling is induced by type V collagen in a model of scleroderma.

OBJECTIVE: The pulmonary vascular remodeling in systemic sclerosis (SSc) is poorly understood and animal models are lacking. Type V collagen (COLV) is elevated in SSc and is implicated in the pathogenesis, and immunization with human COLV induces SSc-like skin and lung changes in rabbits and mice. Here we tested the hypothesis that COLV immunization will induce pathological and functional changes that phenocopy SSc-associated pulmonary vascular disease. METHODS: Pulmonary vascular changes in rabbits immunized with human COLV were extensively characterized by a combination of histology, electron microscopy and immunohistochemistry. Physiologic changes induced by COLV in explanted pulmonary artery rings were evaluated. The pattern of histopathologic alterations and gene expression induced in immunized rabbits were compared to those in SSc patients. RESULTS: COLV immunization was accompanied by striking pulmonary vascular abnormalities, characterized by reduced capillary density, perivascular inflammation, endothelial cell injury and collagen accumulation, that closely phenocopy changes seen in SSc patients. Moreover, pulmonary arteries from immunized rabbits showed impaired ex vivo vascular relaxation. Expression of COL5A2 was significantly increased in the lungs from immunized rabbits (p = 0.02), as well as in patients with SSc (P = 0.02). CONCLUSION: COLV immunity in rabbits is associated with marked vascular remodeling in the lung that phenocopies early-stage human SSc-associated pulmonary vascular disease. COLV immunization therefore represents a novel approach to model SSc pulmonary vascular pathology. Moreover, our findings suggest that COLV might represent a novel pathogenic autoantigen in SSc and future studies with the present model should be developed for possible association with PAH.

Novel spotted fever group rickettsiosis, Brazil.

We report a clinical case of spotted fever group rickettsiosis acquired in Sao Paulo, Brazil. Definitive diagnosis was supported by seroconversion between acute-phase and convalescent-phase serum samples. Molecular analysis of skin samples indicated the agent was a novel spotted fever group strain closely related to Rickettsia africae, R. parkeri, and R. sibirica.

Collagen V-induced nasal tolerance downregulates pulmonary collagen mRNA gene and TGF-beta expression in experimental systemic sclerosis.

BACKGROUND: The purpose of this study was to evaluate collagen deposition, mRNA collagen synthesis and TGF-beta expression in the lung tissue in an experimental model of scleroderma after collagen V-induced nasal tolerance. METHODS: Female New Zealand rabbits (N = 12) were immunized with 1 mg/ml of collagen V in Freund's adjuvant (IM). After 150 days, six immunized animals were tolerated by nasal administration of collagen V (25 microg/day) (IM-TOL) daily for 60 days. The collagen content was determined by morphometry, and mRNA expressions of types I, III and V collagen were determined by Real-time PCR. The TGF-beta expression was evaluated by immunostaining and quantified by point counting methods. To statistic analysis ANOVA with Bonferroni test were employed for multiple comparison when appropriate and the level of significance was determined to be p < 0.05. RESULTS: IM-TOL, when compared to IM, showed significant reduction in total collagen content around the vessels (0.371 +/- 0.118 vs. 0.874 +/- 0.282, p < 0.001), bronchioles (0.294 +/- 0.139 vs. 0.646 +/- 0.172, p < 0.001) and in the septal interstitium (0.027 +/- 0.014 vs. 0.067 +/- 0.039, p = 0.026). The lung tissue of IM-TOL, when compared to IM, showed decreased immunostaining of types I, III and V collagen, reduced mRNA expression of types I (0.10 +/- 0.07 vs. 1.0 +/- 0.528, p = 0.002) and V (1.12 +/- 0.42 vs. 4.74 +/- 2.25, p = 0.009) collagen, in addition to decreased TGF-beta expression (p < 0.0001). CONCLUSIONS: Collagen V-induced nasal tolerance in the experimental model of SSc regulated the pulmonary remodeling process, inhibiting collagen deposition and collagen I and V mRNA synthesis. Additionally, it decreased TGF-beta expression, suggesting a promising therapeutic option for scleroderma treatment.

[Epidemiological characteristics of Lyme-like disease in children]. / Características clínico-epidemiológicas da doença Lyme-símile em crianças.

BACKGROUND: To determine the prevalence, age distribution, seasonality and clinical characteristics of Lyme-simile disease in Brazilians less than 15 years of age. METHODS. From July, 1998 to November, 2000, a cross-sectional study was conducted in 333 patients with skin rash and fever. Paired blood samples were collected for identification of the pathogens. Only 193 samples which were negative for other pathogens (Parvovirus B19 Human, Herpesvirus 6 Human, Measles, Rubella, Dengue, Scarlet fever and Enterovirus), were tested for borreliosis by Enzyme-Linked Immunosorbent Assay and Western-blotting. Other clinical, socioeconomic, demographic and climatic variables were studied. RESULTS: Prevalence of the disease was 6.2%(12/193). Of the variables studied, there was predominance in: <6 years old (83.2%); females (66.7%); being from the city of Franco da Rocha (58.3 %); and a summer/fall seasonality. The duration of care was 4 days. Signs and symptoms with statistical significance were itching; absence of lip notch and ocular pain; irritability and good clinical condition. Other clinical data presented were: pruritus (90%), irritability (80%) and fever (38 masculineC) (58.3%) with a duration of 1 to 3 days. Erythema was maculo-papular (40%), urticaria-like (25%) and scarlatiniform (16.7%), occurring predominately on the trunk (60%). There were no primary clinical evidences of Lyme-simile disease in the patients under study. The sensitivity and specificity of the clinical diagnosis as opposed to the laboratory diagnosis was zero. There was no initial clinical suspicion of the disease in the 10 cases studied and followed up for two years that showed no evidence of cardiologic or neurological complications. This is the first study of Lyme-simile in Brazilian children. CONCLUSION: Prevalence of Lyme-simile disease was low, and it was not remembered at the initial diagnosis of those with skin rash. However, practical knowledge is necessary, demanding increased medical attention.

Sudden deafness and Lyme disease.

OBJECTIVES/HYPOTHESIS: Otolaryngological manifestations are common in Lyme disease, affecting up to 75% of patients. One of these symptoms is sudden deafness. Hearing loss has been frequently described in Lyme disease; on the other hand, titers seropositive for, the causal agent of this disease, have been found in almost 20% of cases of sudden deafness. No consensual information exists on the outcome of Borrelia-seropositive patients or on the importance of determining Borrelia antibody titers. The present study aimed to determine the prevalence of seropositivity for Borrelia in sudden deafness, describing clinical characteristics and outcomes. STUDY DESIGN: This was a prospective observational study. METHODS: Forty-seven consecutive patients with sudden deafness were enrolled in the study. Demographic data, the presence of tinnitus and vertigo, and low- and high-frequency pure-tone averages were recorded. The percentage of hearing recovery was determined. Data obtained from Borrelia-seropositive patients were described and compared with those from the seronegative group. RESULTS: Titers positive for antibodies were present in 21.3% of the cases. Seropositive and seronegative groups of patients were homogeneous concerning age, sex distribution, the presence of tinnitus and vertigo, and high- and low-frequency hearing thresholds. Hearing outcome was not significantly different between the groups of patients. CONCLUSIONS: No distinctive clinical characteristic was found between seropositive and seronegative subjects. The hearing outcome of treated Borrelia-seropositive patients was similar to that of the seronegative group.

Architectural remodelling in lungs of rabbits induced by type V collagen immunization: a preliminary morphologic model to study diffuse connective tissue diseases.

The pathogenesis of diffuse connective tissue diseases (DCTD) is still unknown and has been extensively studied regarding its autoimmunity aspects related to extracellular matrix (ECM) remodelling, with an emphasis on the collagens at the inflammatory site. The present paper describes the pulmonary architectural and repair/remodelling responses to injury after immunization of rabbits with human type V collagen. The animal model consisted of rabbits immunized with collagen mixed with Freund's adjuvant and sacrificed 7, 15, 30, 75, and 120 days after the first of four doses of antigen. Pulmonary architecture remodelling response was evaluated by histology, morphometry, and the immunofluorescence method, according to compartments of reference (parenchyma and interstitium) and injury: 1 inflammation (polymorphonuclear and mononuclear cells); 2-repair (fibrosis) and 3-ECM remodelling (collagen system). The results showed an intense inflammatory involvement of the pulmonary vascular and bronchiolar parenchyma, characterized by increased wall thickness in small arteries, infiltrations by pseudoeosinophils, and mononuclear cells. Progressive remodelling of the pulmonary ECM was characterized by collagen deposition in the septal and bronchovascular interstitium, especially in rabbits sacrifices at 75 and 120 days. The ECM remodelling process was not reproduced when rabbits were inoculated with collagen types I and III. We conclude that the model reproduces morphologic changes similar to those observed in many DCTD, encouraging realization of other experiments to gain a better understanding of the pathogenesis of these diseases.

Synovial remodeling process induced by type V collagen immunization in rabbits.

The pathogenesis of diffuse connective tissue diseases is still unknown despite studies of the autoimmunity aspects related to extracellular matrix elements, mainly the collagens. Articulations are frequently affected by the synovitis process in these diseases. The objective of the present study was to verify the morphologic aspects of the synovial membrane of rabbits immunized with type V collagen, which has some particular characteristics 75 days after the first antigen inoculation and when compared to control animals. The synovial membrane of the animals sacrificed after 75 days of immunization presented an intense remodeling phenomenon along the connective tissue screen and interlobular septa of the adipose-muscle tissue screen compartment. The remodeling process determined type I and III collagen fiber depositions in the vascular and connective tissue compartments of the synovial membrane. The nutrient vessels of the adipose-muscle compartment showed a similar remodeling process, which resulted in small vessel occlusion. Few residual inflammatory foci consisting of monocytes and eosinophils were observed. Thus, our experimental model reproduces morphologic changes in different tissues, characterized by an extracellular matrix remodeling process similar to those observed in many diffuse connective tissue diseases such as systemic lupus erytematosus and scleroderma. Therefore, this model could be useful in understanding the pathogenesis and the treatment of these diseases.

Cross-cultural adaptation and validation of the Brazilian version of the Scleroderma Health Assessment Questionnaire (SHAQ).

The Scleroderma Health Assessment Questionnaire (SHAQ) is a feasible multisystem specific tool that has been extensively used as an additional assessment for systemic sclerosis (SSc). The aim of this study is to cross-culturally adapt and validate the Brazilian version of the SHAQ. Construct validity was assessed based on the correlations between SHAQ and both the Medical Outcomes Survey Short Form 36 version 2 (SF-36v2™) and the Health Assessment Questionnaire Disability Index (HAQ-DI). The correlation between the SHAQ and disease severity was assessed by Spearman's correlation coefficient. The reproducibility of the SHAQ was evaluated by the intraclass correlation coefficient (ICC). Among the 151 consecutive outpatients evaluated, 59 % had limited SSc subtype. The overall disease severity visual analog scale (VAS) of the SHAQ was statistically significantly correlated to HAQ-DI, pain VAS, and the SF-36v2™ physical component summary score (r = 0.595, r = 0.612, and r = -0.582, respectively; p < 0.001). Further analysis of all SF-36v2™ components revealed statistically significant correlations between overall disease severity VAS and bodily pain (r = -0.621, p < 0.001), vitality (r = -0.544, p < 0.001), physical function (r = -0.510, p < 0.001), and role limitation-physical dimensions (r = -0.505, p < 0.001). Moreover, digestive, pulmonary, and overall disease severity VASs were statistically significantly correlated to the number of organs involved (r = 0.178, p = 0.029; r = 0.214, p = 0.008; r = 0.282, p < 0.001). We also demonstrated high reproducibility for SHAQ (ICC = 0.757, 95 % confidence interval = 0.636-0.842). The Brazilian version of the SHAQ demonstrated both construct and discriminant validities as well as good reproducibility.

Amplification of the flgE gene provides evidence for the existence of a Brazilian borreliosis.

INTRODUCTION: The symptoms of Brazilian borreliosis resemble the clinical manifestations of Lyme disease (LD). However, there are differences between the two in terms of epidemiological and laboratory findings. Primers usually employed to diagnose LD have failed to detect Borrelia strains in Brazil. OBJECTIVE: We aimed to identify the Brazilian Borrelia using a conserved gene that synthesizes the flagellar hook (flgE) of Borrelia burgdorferi sensu lato. METHOD: Three patients presenting with erythema migrans and positive epidemiological histories were recruited for the study. Blood samples were collected, and the DNA was extracted by commercial kits. RESULTS: The gene flgE was amplified from DNA of all selected patients. Upon sequencing, these positive samples revealed 99% homology to B. burgdorferi flgE. CONCLUSION: These results support the existence of borreliosis in Brazil. However, it is unclear whether this borreliosis is caused by a genetically modified B. burgdorferi sensu stricto or by a new species of Borrelia spp.

Survival, causes of death, and prognostic factors in systemic sclerosis: analysis of 947 Brazilian patients.

OBJECTIVE: To analyze survival, prognostic factors, and causes of death in a large cohort of patients with systemic sclerosis (SSc). METHODS: From 1991 to 2010, 947 patients with SSc were treated at 2 referral university centers in Brazil. Causes of death were considered SSc-related and non-SSc-related. Multiple logistic regression analysis was used to identify prognostic factors. Survival at 5 and 10 years was estimated using the Kaplan-Meier method. RESULTS: One hundred sixty-eight patients died during the followup. Among the 110 deaths considered related to SSc, there was predominance of lung (48.1%) and heart (24.5%) involvement. Most of the 58 deaths not related to SSc were caused by infection, cardiovascular or cerebrovascular disease, and cancer. Male sex, modified Rodnan skin score (mRSS) > 20, osteoarticular involvement, lung involvement, and renal crisis were the main prognostic factors associated to death. Overall survival rate was 90% for 5 years and 84% for 10 years. Patients presented worse prognosis if they had diffuse SSc (85% vs 92% at 5 yrs, respectively, and 77% vs 87% at 10 yrs, compared to limited SSc), male sex (77% vs 90% at 5 yrs and 64% vs 86% at 10 yrs, compared to female sex), and mRSS > 20 (83% vs 90% at 5 yrs and 66% vs 86% at 10 yrs, compared to mRSS < 20). CONCLUSION: Survival was worse in male patients with diffuse SSc, and lung and heart involvement represented the main causes of death in this South American series of patients with SSc.

Abnormal collagen V deposition in dermis correlates with skin thickening and disease activity in systemic sclerosis.

OBJECTIVE: The physiological and mechanical properties of the skin, the primary tissue affected by systemic sclerosis, depend on the assembly of collagen types I, III and V, which form heterotypic fibers. Collagen V (COLV) regulates heterotypic fiber diameter, and the maintenance of its properties is important for maintaining normal tissue architecture and function. Based on a COLV-induced experimental SSc model, in which overexpression of abnormal COLV was a prominent feature, we assumed that this abnormality could be present in SSc patients and could be correlated to disease duration, skin thickening and disease activity. METHODS: Skin biopsies from 18 patients (6 early-stage and 12 late-stage) and 10 healthy controls were studied. Skin thickening assessment was performed with the Modified Rodnan Skin Score (MRSS), and activity was calculated using the Valentini Disease Activity Index. Morphology, morphometry of COLV deposition in dermis, as well as, quantitative RT-PCR and 3D-reconstruction of the dermal fibroblast culture were performed. RESULTS: Structurally abnormal COLV was overexpressed in SSc skin, mainly in the early stages of the disease, when compared to normal controls and late-stage. A positive correlation between COLV expression and MRSS and disease activity was observed. Collagen V alpha-1 and alpha-2 mRNA expression levels were higher in SSc. Tridimensional reconstruction of SSc dermal heterotypic fibers confirmed the presence of atypical COLV. CONCLUSION: Increased synthesis of abnormal COLV and its correlation with disease stage, activity and MRSS suggest that this collagen can be a possible trigger involved in the pathogenesis of SSc.

Experimental diabetes modulates collagen remodelling of joints in rats.

UNLABELLED: The aim of this study was to evaluate extracellular matrix components in articular cartilage, ligaments and synovia in an experimental model of diabetes. Young Wistar rats were divided into a streptozotocin-induced (STZ; 35 mg/kg) diabetic group (DG; n=15) and a control group (CG; n=15). Weight, blood glucose and plasma anti-carboxymethyllysine were measured 70 days after STZ infusions. Knee joints, patellar ligaments, and lateral and medial collateral ligaments were isolated and stained with hematoxylin-eosin and Picrosirius. The total collagen content was determined by morphometry. Immunofluorescence was employed to evaluate types I, III, and V collagen in ligaments and synovial tissues and types II and XI collagen in cartilage. RESULTS: Higher blood glucose levels and plasma anti-carboxymethyllysine were observed in DG rats when compared to those in CG rats. The final weight was significantly lower in the DG rats than in the CG rats. Histomorphometric evaluation depicted a small quantity of collagen fibers in ligaments and articular cartilage in DG rats, as well as increased collagen in synovial tissue. There was a decrease in cartilage proteoglycans in DG rats when compared with CG rats. Immunofluorescence staining revealed an increase of collagen III and V in ligaments, collagen XI in cartilage, and collagen I in synovial tissue of DG rats compared with CG rats. CONCLUSION: The ligaments, cartilage and synovia are highly affected following STZ-induced diabetes in rats, due the remodeling of collagen types in these tissues. This process may promote the degradation of the extracellular matrix, thus compromising joint function. Our data may help to better understand the pathogenesis of joint involvement related to diabetes.

Survey for tick-borne zoonoses in the state of Espirito Santo, southeastern Brazil.

Blood samples collected from 201 humans, 92 dogs, and 27 horses in the state of Espirito Santo, Brazil, were tested by polymerase chain reaction, indirect immunofluorescence assays, and indirect enzyme-linked immunosorbent assay for tick-borne diseases (rickettsiosis, ehrlichiosis, anaplasmosis, borreliosis, babesiosis). Our results indicated that the surveyed counties are endemic for spotted fever group rickettsiosis because sera from 70 (34.8%) humans, 7 (7.6%) dogs, and 7 (25.9%) horses were reactive to at least one of the six Rickettsia species tested. Although there was evidence of ehrlichiosis (Ehrlichia canis) and babesiosis (Babesia canis vogeli, Theileria equi) in domestic animals, no human was positive for babesiosis and only four individuals were serologically positive for E. canis. Borrelia burgdorferi-serologic reactive sera were rare among humans and horses, but encompassed 51% of the canine samples, suggesting that dogs and their ticks can be part of the epidemiological cycle of the causative agent of the Brazilian zoonosis, named Baggio-Yoshinari Syndrome.

Hepatozoon canis infecting dogs in the State of Espírito Santo, southeastern Brazil.

From May 2007 to March 2008, blood samples were collected from 92 healthy dogs living in 21 households (17 farms in rural area, and 4 homes in urban area) in 6 counties of the State of Espírito Santo, southeastern Brazil. In addition, ticks were collected from these dogs. A mean of 4.4+/-3.0 dogs (range: 1-12) were sampled per household; 78 and 14 dogs were from rural and urban areas, respectively. Polymerase chain reaction (PCR) designed to amplify fragments of the 18S rDNA gene of Babesia spp or Hepatozoon spp revealed amplicons of the expected size in 20 (21.7%) dogs for Babesia, and 54 (58.7%) dogs for Hepatozoon. All Babesia-positive dogs were also Hepatozoon-positive. Among the 21 households, 15 (71.4%) from 3 counties had at least one PCR-positive dog, including 13 farms (rural area) and 2 homes (urban area). A total of 40 PCR products from the Hepatozoon-PCR, and 19 products from the Babesia-PCR were submitted to DNA sequencing. All generated sequences from Hepatozoon-PCR were identical to each other, and to corresponding 18S rDNA sequences of H. canis in GenBank. Surprisingly, all generated sequences from the Babesia PCR were also identical to corresponding 18S rDNA sequences of H. canis in GenBank. Dogs from 10 rural and 2 urban households were found infested by Rhipicephalus sanguineus ticks. Immature of Amblyomma cajennense ticks were found in dogs from only 4 rural households (also infested by R. sanguineus). All but one household with R. sanguineus-infested dogs had at least one Hepatozoon-infected dog. Statistical analysis showed that the presence of ticks (i.e. R. sanguineus) infesting dogs in the households was significantly (P<0.05) associated with at least one PCR-positive dog. There was no significant association (P>0.05) between PCR-positive dogs and urban or rural households. Canine hepatozoonosis caused by H. canis is a high frequent infection in Espírito Santo, Brazil, where it is possibly vectored by R. sanguineus. Since all infected dogs were found apparently healthy, the pathogenicity of H. canis for dogs in Espírito Santo is yet to be elucidated.

[Occurrence of ticks (Acari: Ixodidae) in the municipality of Goiatins, Tocantins]. / Ocorrência de carrapatos (Acari: Ixodidae) no município de Goiatins, Tocantins.

Due to a suspected human case of Brazilian Lyme-like disease in the city of Goiatins, Tocantins State, an epidemiological survey was carried out in eight counties in this region during September 2007 and February 2008, where 1,890 ticks were collected from domestic animals and from the environment. A total of eight tick species were identified: Rhipicephalus sanguineus, Rhipicephalus (Boophilus) microplus, Dermacentor nitens, Amblyomma cajennense, Amblyomma oblongoguttatum, Amblyomma ovale, Amblyomma parvum and Amblyomma tigrinum. The last four species were described for the first time in this region. Although human parasitism by ticks is frequently described in Goiatins, no ticks collected from humans were analyzed. The study of ixodids in this region contributes with the survey of Brazilian ticks, as well as the elucidation of the possible transmission of the agent that caused the Brazilian Lyme-like disease case in Goiatins.

Amplification of the flgE gene provides evidence for the existence of a Brazilian borreliosis / Amplificação do gene flgE fornece evidência para a existência de uma borreliose brasileira

INTRODUCTION: The symptoms of Brazilian borreliosis resemble the clinical manifestations of Lyme disease (LD). However, there are differences between the two in terms of epidemiological and laboratory findings. Primers usually employed to diagnose LD have failed to detect Borrelia strains in Brazil. OBJECTIVE: We aimed to identify the Brazilian Borrelia using a conserved gene that synthesizes the flagellar hook (flgE) of Borrelia burgdorferi sensu lato. METHOD: Three patients presenting with erythema migrans and positive epidemiological histories were recruited for the study. Blood samples were collected, and the DNA was extracted by commercial kits. RESULTS: The gene flgE was amplified from DNA of all selected patients. Upon sequencing, these positive samples revealed 99% homology to B. burgdorferi flgE. CONCLUSION: These results support the existence of borreliosis in Brazil. However, it is unclear whether this borreliosis is caused by a genetically modified B. burgdorferi sensu stricto or by a new species of Borrelia spp.

INTRODUÇÃO: Os sintomas da borreliose brasileira se assemelham às manifestações clínicas da Doença de Lyme (DL), porém, existem diferenças epidemiológicas e laboratoriais entre essas enfermidades. Primers normalmente utilizados para diagnosticar a DL não conseguiram detectar cepas de borrelia no Brasil. OBJETIVO: O objetivo desse trabalho foi identificar a borrelia brasileira usando um gene conservado que sintetiza o gancho flagelar (flgE) da Borrelia burgdorferi sensu lato. MÉTODO: Três pacientes com eritema migratório e epidemiologia positiva foram recrutados para o estudo. Amostras de sangue foram coletadas, e o DNA foi extraído por kits comerciais. RESULTADOS: O gene flgE foi amplificado a partir do DNA de todos os pacientes selecionados. Após o sequenciamento, essas amostras positivas revelaram homologia de 99% para B. burgdorferi. CONCLUSÃO: Estes resultados reforçam a existência de borreliose no Brasil. No entanto, não está claro se esta borreliose é causada por uma variante geneticamente modificada da B. burgdorferi sensu stricto ou por uma nova espécie de Borrelia spp.

Autoantibody profile in the experimental model of scleroderma induced by type V human collagen.

UNLABELLED: The aim of this study is to evaluate the humoral autoimmune response in the experimental model of systemic sclerosis (SSc) induced by human type V collagen (huCol V). New Zealand rabbits were immunized with huCol V in Freund's complete adjuvant (FCA) and boosted twice with 15 days intervals with huCol V in Freund's incomplete adjuvant. Control groups included animals injected only with FCA or bovine serum albumin. Bleeding was done at days 0, 30, 75 and 120. Tissue specimens were obtained for histopathological investigation. Serological analysis included detection of antibodies against huCol V and anti-topoisomerase I (Anti-Scl70) by enzyme-linked immunosorbent assay, antinuclear antibodies (ANA) by indirect immunofluorescence, and rheumatoid factor (RF) by a latex agglutination test. Target antigens were characterized by immunoblot. Histological analysis revealed extracellular matrix remodeling with fibrosis and vasculitis. Anti-Scl70 and ANA were detected as early as 30 days in all huCol V animals. The universal ANA staining pattern was Golgi-like. This serum reactivity was not abolished by previous absorption with huCol V. Characterization of the target antigen by immunoblot revealed two major protein fractions of 175,000 and 220,000 MW. Similarly to ANA, there was a gradual increase of reactivity throughout the immunization and also it was not abolished by preincubation of serum samples with huCol V. RF testing was negative in hyperimmune sera. CONCLUSION: The production of autoantibodies, including anti-Scl70, a serological marker for SSc associated with histopathological alterations, validates huCol V induced-experimental model and brings out its potential for understanding the pathophysiology of SSc.

Características clínico-epidemiológicas da doença Lyme-símile em crianças / Epidemiological characteristics of Lyme-like disease in children

OBJETIVO: Determinar a prevalência, distribuição etária, sazonalidade, características clínicas da doença Lyme-símile em menores de 15 anos. MÉTODOS: De julho/1998 a dezembro/2000 foi conduzido um estudo transversal em 333 pacientes, com exantema e febre. Foram coletadas amostras pareadas de sangue para a identificação de patógenos. Somente em 193 amostras, negativas aos outros patógenos (Parvovirus B19, Herpesvírus 6 humano, Sarampo, Rubéola, Dengue, Escarlatina e Enterovírus), foram realizadas a pesquisa da borreliose pelos métodos de Enzyme-Linked Immunosorbent Assay e Western-blotting. Outras variáveis clínicas, socioeconômicas, demográficas e climáticas foram estudadas. RESULTADOS: A prevalência da doença foi de 6,2 por cento(12/193). Das variáveis estudadas, houve predomínio em <6anos(83,2 por cento); sexo feminino (66,7 por cento); procedência da cidade de Franco da Rocha (58,3 por cento); com sazonalidade no outono-verão. O intervalo de atendimento foi de quatro dias. Sinais e sintomas com significância estatística: prurido, ausência da fissura labial e bom estado clínico. Outros dados presentes foram: irritabilidade (80 por cento); febre (?38ºC) (58,3 por cento) com duração de um a três dias. O exantema foi do tipo máculo-papular (33,3 por cento), urticariforme (25 por cento) e escarlatiniforme (16,7 por cento); predominando em tronco (60 por cento). Não houve apresentação clínica característica para diagnóstico da doença de Lyme-símile nestes pacientes. A sensibilidade e especificidade para o diagnóstico clínico contraposta com o diagnóstico laboratorial foi zero. O acompanhamento de 10 casos durante dois anos não evidenciou complicações cardiológicas ou neurológicas. Este é o primeiro estudo desta doença em crianças brasileiras. CONCLUSÃO: A prevalência da doença Lyme-símile foi baixa, não tendo sido lembrada no diagnóstico inicial dos exantemas, mas seu conhecimento é necessário, necessitando maior atenção médica.

BACKGROUND: To determine the prevalence, age distribution, seasonality and clinical characteristics of Lyme-simile disease in Brazilians less than 15 years of age. METHODS. From July, 1998 to November, 2000, a cross-sectional study was conducted in 333 patients with skin rash and fever. Paired blood samples were collected for identification of the pathogens. Only 193 samples which were negative for other pathogens (Parvovirus B19 Human, Herpesvirus 6 Human, Measles, Rubella, Dengue, Scarlet fever and Enterovirus), were tested for borreliosis by Enzyme-Linked Immunosorbent Assay and Western-blotting. Other clinical, socioeconomic, demographic and climatic variables were studied. RESULTS: Prevalence of the disease was 6.2 percent(12/193). Of the variables studied, there was predominance in: <6 years old (83.2 percent); females (66.7 percent); being from the city of Franco da Rocha (58.3 percent); and a summer/fall seasonality. The duration of care was 4 days. Signs and symptoms with statistical significance were itching; absence of lip notch and ocular pain; irritability and good clinical condition. Other clinical data presented were: pruritus (90 percent), irritability (80 percent) and fever (?38ºC) (58.3 percent) with a duration of 1 to 3 days. Erythema was maculo-papular (40 percent), urticaria-like (25 percent) and scarlatiniform (16.7 percent), occurring predominately on the trunk (60 percent). There were no primary clinical evidences of Lyme-simile disease in the patients under study. The sensitivity and specificity of the clinical diagnosis as opposed to the laboratory diagnosis was zero. There was no initial clinical suspicion of the disease in the 10 cases studied and followed up for two years that showed no evidence of cardiologic or neurological complications. This is the first study of Lyme-simile in Brazilian children. CONCLUSION: Prevalence of Lyme-simile disease was low, and it was not remembered at the initial diagnosis ...