Increased mRNA expression of collagen V gene in pulmonary fibrosis of systemic sclerosis.

BACKGROUND: Collagen V shows promise as an inducer of interstitial lung fibrosis in experimental systemic sclerosis (SSc). MATERIALS AND METHODS: Remodelling of the pulmonary interstitium was evaluated based on the clinical data and open lung biopsies from 15 patients with SSc. Normal lung tissues obtained from eight individuals who died of traumatic injuries were used as control group. Immunofluorescence, immunohistochemistry, morphometry, tri-dimensional reconstruction and a real-time polymerase chain reaction were used to evaluate the quantity, structure and molecular chains of collagen V. The impact of these markers was tested on clinical data. RESULTS: The main difference in collagen V content between SSc patients and the control group was an increased, abnormal and distorted fibre deposition in the alveolar septa and the pre-acinar artery wall. The lungs from SSc patients presented [alpha1(V)] and [alpha2(V)] mRNA chain expression increased, but [alpha2(V)] was proportionally increased compared with the control group. High levels of collagen V were inversely associated with vital capacity (r = -0.72; P = 0.002), forced vital capacity (r = -0.76; P < 0.001), forced expiratory volume in 1-s (r = -0.89; P < 0.001) and diffusing capacity for carbon monoxide (r = -0.62; P = 0.04). CONCLUSIONS: Abnormal collagen V fibres are overproduced in lungs from SSc patients and may play an important role in the pathogenesis of the disease as this molecule regulates tissue collagen assembly. The aberrant histoarchitecture observed in SSc can be related to the overexpression of the [alpha2(V)] gene of unknown origin.

Description of Lyme disease-like syndrome in Brazil. Is it a new tick borne disease or Lyme disease variation?

An emerging clinical entity that reproduces clinical manifestations similar to those observed in Lyme disease (LD) has been recently under discussion in Brazil. Due to etiological and laboratory particularities it is named LD-like syndrome or LD imitator syndrome. The condition is considered to be a zoonosis transmitted by ticks of the genus Amblyomma, possibly caused by interaction of multiple fastidious microorganisms originating a protean clinical picture, including neurological, osteoarticular and erythema migrans-like lesions. When peripheral blood of patients with LD-like syndrome is viewed under a dark-field microscope, mobile uncultivable spirochete-like bacteria are observed. PCR carried out with specific or conservative primers to recognize Borrelia burgdorferi sensu stricto or the genus Borrelia has been negative in ticks and in biological samples. Two different procedures, respectively involving hematoxylin and eosin staining of cerebrospinal fluid and electron microscopy analysis of blood, have revealed spirochetes not belonging to the genera Borrelia, Leptospira or Treponema. Surprisingly, co-infection with microorganisms resembling Mycoplasma and Chlamydia was observed on one occasion by electron microscopy analysis. We discuss here the possible existence of a new tick-borne disease in Brazil imitating LD, except for a higher frequency of recurrence episodes observed along prolonged clinical follow-up.

Description of Lyme disease-like syndrome in Brazil: is it a new tick borne disease or Lyme disease variation?

An emerging clinical entity that reproduces clinical manifestations similar to those observed in Lyme disease (LD) has been recently under discussion in Brazil. Due to etiological and laboratory particularities it is named LD-like syndrome or LD imitator syndrome. The condition is considered to be a zoonosis transmitted by ticks of the genus Amblyomma, possibly caused by interaction of multiple fastidious microorganisms originating a protean clinical picture, including neurological, osteoarticular and erythema migrans-like lesions. When peripheral blood of patients with LD-like syndrome is viewed under a dark-field microscope, mobile uncultivable spirochete-like bacteria are observed. PCR carried out with specific or conservative primers to recognize Borrelia burgdorferi sensu stricto or the genus Borrelia has been negative in ticks and in biological samples. Two different procedures, respectively involving hematoxylin and eosin staining of cerebrospinal fluid and electron microscopy analysis of blood, have revealed spirochetes not belonging to the genera Borrelia, Leptospira or Treponema. Surprisingly, co-infection with microorganisms resembling Mycoplasma and Chlamydia was observed on one occasion by electron microscopy analysis. We discuss here the possible existence of a new tick-borne disease in Brazil imitating LD, except for a higher frequency of recurrence episodes observed along prolonged clinical follow-up.

Radionuclide imaging for the assessment of esophageal motility disorders in mixed connective tissue disease patients: relation to pulmonary impairment.

Esophageal functional abnormalities may lead to regurgitation, chronic esophagitis and life-threatening conditions such as aspiration pneumonia. In mixed connective tissue disease patients, previous reports showed that esophageal dysfunction varies according to the method employed for investigation. Our study was conceived to: (i) assess esophageal motility and mucosal aspects in patients with mixed connective tissue disease by endoscopy, cine-esophogram and scintigraphy focusing on the prevalence of each exam; and (ii) verify the association between pulmonary and esophageal dysfunctions. Twenty-four mixed connective tissue disease patients were enrolled for this study. Cine-esophogram and upper digestive endoscopy with mucosal biopsy were performed according to previous standardization. Radionuclide esophageal scintigraphy was performed with a semisolid meal with (99m)Tc. Eleven healthy individuals voluntarily submitted to scintigraphy as controls. Cine-esophogram showed esophageal delayed emptying in 90% of patients. At scintigraphy there was a significant delay in total esophageal transit time in the group of patients when compared to healthy controls (35.3 +/- 8.2 s. vs. 13.6 +/- 9.5 s.; P < 0.0001). The whole esophageal body showed dysmotility in 96% of patients. The cine-esophogram detected functional esophageal impairment similar to scintigraphic findings. Histopathologic examination found esophagitis in 95% of studied patients. Reduced lung volumes were associated with esophagitis and delayed esophageal clearance at scintigraphy, observed at the distal portion of the esophagus. Esophageal scintigraphy is easy to perform, with good acceptance by patients with low radiation exposition. It is a useful non-invasive test for follow-up and interventional studies concerning esophagus dysfunction.

Immune cell infiltration and broncovascular remodeling after nitric acid nasal instillation in a mouse bronchiolitis obliterans model.

Immune cell airway infiltration and the bronchovascular remodeling process have shown to be promising in the understanding of bronchiolitis obliterans (BO) pathogenesis. In this study we sought to validate the importance of immune cells, whether diffusely distributed or forming lymphoid follicles, collagen density, and vascular factors. Eight weeks after a single nitric acid (NA) nasal instillation, lung changes were characterized by lumen distortion, epithelial layer folding, reduction or total obliteration of terminal bronchiole (TB) lumen, and wall thickness increase. The morphologic changes in the TB and TA (terminal artery) lumen coincide with the measurement difference in the three groups. The TB diameter and lumen were significantly decreased in BO when compared with non-BO lungs (0.76 +/- 0.05 microm vs. 0.81 +/- 0.05 microm and 12,286.13 +/- 378.83 microm vs. 18,182.27 +/- 5,593.98 microm, p = 0.05 and p = 0.01, respectively). Equally significant was the increase in TB thickness in BO when compared with the non-BO group (201.72 +/- 35.75 microm vs. 149.75 +/- 40.61 microm, p = 0.007). The morphologic changes in immune cells seen in TB, TA, and bronchus-associated lymphoid tissue (BALT) also coincide with the quantification differences observed in the three groups. We concluded that immune cell infiltration and collagen/vascular remodeling are related to the spectrum of histologic changes in a BO nasal-induced model in mice and may be an appropriate target for prospective studies of human bronchiolitis.

Search for Borrelia sp. in ticks collected from potential reservoirs in an urban forest reserve in the State of Mato Grosso do Sul, Brazil: a short report.

A total of 128 ticks of the genus Amblyomma were recovered from 5 marsupials (Didelphis albiventris) - with 4 recaptures - and 17 rodents (16 Bolomys lasiurus and 1 Rattus norvegicus) captured in an urban forest reserve in Campo Grande, State of Mato Grosso do Sul, Brazil. Of the ticks collected, 95 (78.9%) were in larval form and 22 (21.1%) were nymphs; the only adult (0.8%) was identified as A. cajennense. Viewed under dark-field microscopy in the fourth month after seeding, 9 cultures prepared from spleens and livers of the rodents, blood of the marsupials, and macerates of Amblyomma sp. nymphs revealed spiral-shaped, spirochete-like structures resembling those of Borrelia sp. Some of them showed little motility, while others were non-motile. No such structures could be found either in positive Giemsa-stained culture smears or under electron microscopy. No PCR amplification of DNA from those cultures could be obtained by employing Leptospira sp., B. burgdorferi, and Borrelia sp. primers. These aspects suggest that the spirochete-like structures found in this study do not fit into the genera Borrelia or Leptospira, requiring instead to be isolated for proper identification.

Search for Borrelia sp. in ticks collected from potential reservoirs in an urban forest reserve in the State of Mato Grosso do Sul, Brazil: a short report

A total of 128 ticks of the genus Amblyomma were recovered from 5 marsupials (Didelphis albiventris) - with 4 recaptures - and 17 rodents (16 Bolomys lasiurus and 1 Rattus norvegicus) captured in an urban forest reserve in Campo Grande, State of Mato Grosso do Sul, Brazil. Of the ticks collected, 95 (78.9 percent) were in larval form and 22 (21.1 percent) were nymphs; the only adult (0.8 percent) was identified as A. cajennense. Viewed under dark-field microscopy in the fourth month after seeding, 9 cultures prepared from spleens and livers of the rodents, blood of the marsupials, and macerates of Amblyomma sp. nymphs revealed spiral-shaped, spirochete-like structures resembling those of Borrelia sp. Some of them showed little motility, while others were non-motile. No such structures could be found either in positive Giemsa-stained culture smears or under electron microscopy. No PCR amplification of DNA from those cultures could be obtained by employing Leptospira sp., B. burgdorferi, and Borrelia sp. primers. These aspects suggest that the spirochete-like structures found in this study do not fit into the genera Borrelia or Leptospira, requiring instead to be isolated for proper identification

Borrelia burgdorferi antibodies in dogs from Cotia county, São Paulo State, Brazil.

Dogs sera samples collected from Cotia County, São Paulo were tested using indirect immunoenzymatic test (ELISA) in order to study Lyme disease serology in dogs. ELISA method was standardized and G39/40 North American strain of Borrelia burgdorferi was used as antigen. Positive results were confirmed employing the Western blotting technique. Because of the possibility of cross-reactions, sera were also tested for different serological strains of Leptospira interrogans and L. biflexa using microscopic sera agglutination test. Twenty-three of 237 (9.7%) serum samples were positive in the ELISA; 20 of them (86.9%) were confirmed by the Western blotting, what suggests that Cotia may be a risk area for Lyme disease. Although 4 samples (1.7%) were positive for Lyme disease and leptospirosis, no correlation was found between the results (X(2) = 0.725; p = 0.394) what suggests absence of serological cross reactivity.

Oviposition and eclosion periods of Ixodes didelphidis Fonseca and Aragão, 1951 (Acari: Ixodidae) under laboratory conditions.

Oviposition and eclosion periods for Ixodes didelphidis were observed under two temperatures (25 degrees C and 27 degrees C) and 90-95% humidity. Although there was a significant increase in the eclosion period (p<0.05) and a tendency to increase the oviposition period at 25 degrees C, there was neither significant differences in the interval (days), until maximum peak of eclosion nor in the number of emerging larvae during the peak nor the total number of emerged larvae. These temperature values are not critical for embryological development of the species. Because at 27 degrees C and under high humidity the oviposition and eclosion periods are shorter, and the percentage of emerged larvae is higher, we consider this to be the ideal temperature for laboratory studies.

Borrelia-like spirochetes recovered from ticks and small mammals collected in the Atlantic Forest Reserve, Cotia county, State of Sao Paulo, Brazil

Forty-four marsupials, 77 rodents and 161 ticks were captured in an Atlantic Forest Reserve in Cotia county, State of Sao Paulo, where human cases of Lyme disease (LD) simile were reported. Twenty-one borrelia-like spirochete isolates were recovered from the mammals' blood and rodent livers or spleens, and triturated ticks inoculated into BSK II medium. Our results suggest that the reservoirs and ticks collected may harbor borrelia-like spirochetes, some of which have an antigenic similarity with the unknown causative agent of LD simile in Brazil, and/or with North American Borrelia burgdorferi s.s.

Borrelia-like spirochetes recovered from ticks and small mammals collected in the Atlantic Forest Reserve, Cotia county, State of São Paulo, Brazil.

Forty-four marsupials, 77 rodents and 161 ticks were captured in an Atlantic Forest Reserve in Cotia county, State of São Paulo, where human cases of Lyme disease (LD) simile were reported. Twenty-one borrelia-like spirochete isolates were recovered from the mammals' blood and rodent livers or spleens, and triturated ticks inoculated into BSK II medium. Our results suggest that the reservoirs and ticks collected may harbor borrelia-like spirochetes, some of which have an antigenic similarity with the unknown causative agent of LD simile in Brazil, and/or with North American Borrelia burgdorferi s. s.

Parasitism by Ixodes didelphidis and I. loricatus (Acari: Ixodidae) on small wild mammals from an Atlatic Forest in the State of Sao Paulo, Brazil.

From January 1995 to June 1996, ticks were studied in a fragment of Atlantic Forest in a residential area in the city of Itapevi, State of Sao Paulo, Brazil. Cases of human Lyme disease-like illness were registered in this area during the spring of 1992. The monthly relative density of ticks was determined and the influence of seasonal climatic conditions was evaluated as well as the relationship between ticks and hosts. Ticks (n = 88) were collected from small mammals captured (n = 134) in monthly trappings where the traps were operated for five consecutive days. Immature ticks (n = 42, 47.7%) were identified as Ixodes spp. The adult ticks (n = 46, 52.3%) were identified as Ixodes didelphidis Fonseca and Aragão (n = 19, 21.6%), I. loricatus Neumann (n = 26, 29.5%) and Amblyomma cajennense (F.) (n = 1, 1.1%). The monthly correlations between tick numbers and meteorological data were not significant. The correlation between the different stages of ticks and the two seasons (cold-dry and warm-rainy) indicated that both immature and adult ticks showed a seasonal pattern of abundance. The highest numbers of immature ticks were observed on rodents, during the cold-dry season (from April to September) with a peak in August 1995. Adults ticks were predominant during the warm-rainy season (from October to March) with a peak in January 1995. Adult ticks were only collected on marsupials.

Anti-corpus luteum antibody: a novel serological marker for ovarian dysfunction in systemic lupus erythematosus?

OBJECTIVE: To investigate the presence of autoantibodies directed to corpus luteum (CoL) in systemic lupus erythematosus (SLE) sera and its correlation with menstrual disturbances. METHODS: We evaluated 87 female patients with SLE, < 40 years old, and 23 women with normal menses as controls. Anti-corpus luteum antibody was detected by immunoblot technique. RESULTS: Reactivity to a bovine CoL antigen was found in 22% of SLE sera. Characterization of the target antigen revealed a 67 kDa glycoprotein highly enriched in corpus luteum, but nearly absent in total ovary extract. Similarly, target antigen was also weakly detectable in tissues that produce or metabolize steroids, such as testis, adrenal cortex, and liver, and it was absent in adrenal medulla or HEp-2 cells. Anti-CoL antibody was easily distinguished from other frequent reactivities of SLE sera, including anti-RNP, anti-Sm, anti-Ro/La, anti-dsDNA, or anticardiolipin. The observation of anti-67 kDa reactivity to human CoL suggests a possible pathogenic role in gonadal dysfunction. Indeed, we observed an inverse association of anti-CoL antibody with the duration of hypergonadotropic amenorrhea. Supporting this hypothesis, in patients with normal or irregular menses, the presence of this antibody was associated with elevated serum level of follicle stimulating hormone, an early and specific sign of ovarian lesion. CONCLUSION: Anti-CoL antibody seems to be associated with early stages of ovarian dysfunction in SLE. Moreover, since similar association of antiovarian antibodies has been observed in an experimental model of autoimmune oophoritis, our findings raise the possibility of autoimmune ovarian lesion in patients with SLE.

Abnormalities in the biochemical markers of bone turnover in children with juvenile chronic arthritis.

OBJECTIVE: To study the biochemical markers of bone turnover in children with juvenile chronic arthritis (JCA) and to compare these parameters with those in healthy children in order to evaluate the relationships between age, disease activity and biochemical variables. METHODS: Sixty-two children with JCA and 157 healthy children were studied. Serum samples were analyzed for their concentrations of minerals, osteocalcin (OC) and bone-specific alkaline phosphatase (BAP). Urine samples were examined to monitor the excretion of hydroxyproline (HYP) and deoxypyridinoline crosslinks (DPD). RESULTS: OC, BAP, HYP/Cr, DPD/Cr values were decreased in healthy girls more than 12 years of age and in healthy boys more than 14 years of age compared to younger children from the same population. Lower levels of OC and BAP were observed in younger children with JCA (girls < or = 12 yrs.; boys < or = 14 yrs.) compared to healthy children of the same age. Older girls with JCA (> or = 13 yrs.) were found to have increased HYP/Cr and DPD/Cr values compared to older healthy children. CONCLUSION: These results indicate that abnormalities of bone metabolism occur in an age-related fashion in JCA. This was demonstrated by a reduction in the markers of bone formation in younger JCA patients. Moreover, in older girls the markers of bone resorption were found to be elevated. Taken together, these findings suggest that bone formation is reduced from early childhood to mid-puberty, while resorption levels increase in children with JCA after this time.

Eye disease in a patient with rheumatoid arthritis.

We report the case of a 40-year-old woman with diffuse uveitis, sensorineural hearing loss and cerebrospinal fluid pleocytosis as features of Vogt-Koyanagi-Harada syndrome who developed symmetric polyarthritis and stiffness of small and large joints, in addition to rheumatoid arthritis. Although their target tissues are distinct, both diseases have a possible autoimmune origin strongly associated with HLA-DRB4.

Bone marrow findings in systemic lupus erythematosus patients with peripheral cytopenias.

We studied 21 bone marrow specimens from 21 patients with systemic lupus erythematosus (SLE) and peripheral cytopenias: anaemia (Hb < 10 g/dl), and/or leucopenia (white blood cell count < 4 x 10(9)/l), and/or thrombocytopenia (platelets < 150 x 10(9)/l). None of the patients had used immunosuppressive drugs in the 2 months before the study, and 11 (52.4%) had never used these drugs. The global and specific series cellularity, degree of fibrosis and necrosis were evaluated by bone marrow trephine; morphological abnormalities and iron stores were evaluated by cytological smears. The most important abnormalities viewed in bone marrow biopsies were: global hypocellularity (47.6%), increased reticulin proliferation (76.2%) with myelofibrosis in one patient, and necrosis (19.0%). The marrow aspirates were difficult to obtain in four patients, who showed an increased reticulin proliferation on histological analysis. Plasmocytosis was present in 26.7% of cases and in one there was a serum monoclonal component (IgG kappa). Iron stores were normal or increased in 26.7% of specimens and decreased or absent in 73.3%. The most frequent peripheral abnormality was leucopenia in 90.4% (19/21) and granulocytic hypoplasia was observed in 47.3% (9/19) of these patients. We conclude that the bone marrow may be a target organ in SLE with cytopenias.

Evaluation by dual X-ray absorptiometry (DXA) of bone mineral density in children with juvenile chronic arthritis.

OBJECTIVE: To study the relationship between bone mineral loss and disease subtype, disease duration and corticosteroid use in children with juvenile chronic arthritis (JCA). METHODS: Bone mineral density (BMD) was evaluated by dual X-ray absorptiometry (DXA), using a Hologic QDR 1000 densitometer. Sixty-two children with JCA and 157 healthy children, aged 5-18 years, were studied. Bone mass was measured in the lumbar spine at the L1-L4 level (LS), in the femoral neck (FN) and in the distal one-tenth radius (DR). RESULTS: A decrease in bone mineral density was observed in 50-60% of the JCA patients in the three regions studied. Those patients who had undergone corticosteroid treatment showed significant bone loss in the DR and LS (trabecular bone), but not in the FN (cortical bone). Bone mass loss was seen for all three disease subtypes, being higher in the patients with polyarticular JCA (particularly in the DR), although this different was not significant. There was a significant difference in disease duration between the children with decreased BMD and those with no BMD decrease in the same regions. CONCLUSION: A decrease in bone mineral density was found in 50-60% of all the JCA patients in this series, regardless of the disease subtype. Corticosteroid use apparently had an effect on the BMD in the trabecular bone. The data also show a correlation between the loss of BMD in both cortical and trabecular bone and a long disease duration.

[Hormonal and biochemical parameters in postmenopausal osteoporosis]. / Parâmetros bioquímios e hormonais na osteoporose pós-menopausa.

In the recent years, many authors had studied the relationship between the calciotropic and gonadotropic hormones actions on postmenopausal bone loss, named calcium negative balance, with different results (Riggs et al 1983; Prince et al, 1995). We evaluated 187 female patients, aged 40 to 80 with the following distribution: 24 normals, 49 patients with osteopenia and 114 patients with osteoporosis, according to WHO classification. The aim of this study was to analyse the relationship between biochemical parameters (seric and urinary calcium), gonadotropic (seric FSH-foliculi stimulant-hormone and seric oestradiol) and calciotropic hormone (PTH) and postmenopausal bone loss. The results had shown the diminution on bone mineral density was related with elevated levels of FSH (p < 0.00001), lower levels of oestradiol (p < 0.00001) and, however, no differences on seric and urinary calcium (respectively, p > 0.70 and p > 0.52) or PTH (p > 0.70) were demonstrated.

[Anti-beta-2-microglobulin antibodies in systemic lupus erythematosus]. / Estudo da antibeta-2-microglobulina sérica em portadores de lúpus eritematoso sistêmico.

The aim of this work was the study of 88 patients with SLE for clinical, laboratorial and demographical parameters. The patients sera were assayed for the presence of anti-beta-2-microglobulin antibodies by ELISA. These spectrofotometric results were converted in a standard-deviation unity in relation to the average of 29 control individuals values. It was established a cut-off of 2.5 standard-deviation in order to determine the limit of normality as well as the cut-off of 10 standard deviation was chosen in order to defined the higher values. Several statistical significant associations were observed between the anti-beta-2-microgobulin and the clinical and laboratorial parameters. The clinical applications of those associations were determined by the predictive value. It was only considered the values greater then 75%. The negative predictive values found in the lower cut-off were 92% to ANA (homogeneous-peripheral pattern), 89% to high levels CIC, 81% to mucocutaneous vasculitis and 80% to depressed CH50 levels. The positive predictive values found in 88% to high level of mucoprotein, 83% to mucocutaneous vasculitis, 83% to the onset-disease-time lower six years and 78% to limphopenia. Special attention may be paid to the high sensibility and specificity of the anti-beta-2-microglobulin test to some of the parameters in the evaluation of the disease activity, mainly in mucocutaneous vasculitis.