Hepatoprotective and anti-inflammatory effects of silibinin on experimental preeclampsia induced by 1-NAME in rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Genetic characterisation of Giardia duodenalis in dairy cattle in Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of mangiferin on the development of periodontal disease: Involvement of lipoxin A(4), anti-chemotaxic action in leukocyte rolling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Production of pro- and anti-inflammatory cytokines by monocytes from patients with paracoccidioidomycosis

Monocytes and macrophages can produce a large repertoire of cytokines and participate in the pathogenesis of granulomatous diseases. We investigated the production of pro- and anti-inflammatory cytokines by monocytes from patients with active paracoccidioidomycosis. Peripheral blood monocytes from 37 patients and 29 healthy controls were cultivated with or without 10 mug/ml of lipopolysaccharide (LPS) for 18 h at 37 degreesC, and the cytokine levels were determined in the culture supernatants by enzyme immunoassay. The results showed that the endogenous levels of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), IL-6, IL-8, IL-10 and transforming growth factor beta detected in the supernatant of patient monocytes cultivated without stimulus were significantly higher than those produced by healthy controls. These data demonstrated that monocytes from patients with active paracoccidioidomycosis produce high levels of cytokines with both inflammatory and anti-inflammatory activities. However, patient monocytes produced significantly lower TNF-alpha and IL-6 levels in response to LPS when compared to normal subjects, suggesting an impairment in their capacity to produce these cytokines after LPS stimulation. Concentrations of IL-1beta, IL-8 and IL-10 in cultures stimulated with LPS were higher in patients than in controls. These results suggest that an imbalance in the production of pro- and anti-inflammatory cytokines might be associated with the pathogenesis of paracoccidioidomycosis. (C) 2003 Editions scientifiques et medicales Elsevier SAS. All rights reserved.

High-fat diet associated with obesity induces impairment of mouse corpus cavernosum responses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização eletroforética e análise de subgrupo de rotavírus em rebanhos bovinos leiteiros do Estado de São Paulo

Foi realizado um estudo para determinar a ocorrência de infecção por rotavírus em rebanhos bovinos leiteiros. Foram analisadas 375 amostras de fezes de bezerros, na faixa etária 1 a 45 dias, provenientes de animais pertencentes a nove propriedades rurais, situadas em seis municípios da região nordeste do Estado de São Paulo. Destas, 193 pertenciam a animais com diarréia e 182 foram obtidas de animais clinicamente sadios. As técnicas utilizadas para a detecção de rotavírus foram o ensaio imunoenzimático (EIE) e a eletroforese em gel de poliacrilamida (EGPA). Por meio do EIE foram detectadas 11,2% (42/375) de amostras positivas, 15% delas (29/193) obtidas de animais com diarréia e 7,1% (13/182) colhidas de animais sem diarréia. A análise do perfil do genoma indicou a presença de seis eletroferótipos distintos, característicos de rotavírus do grupo A. Um único eletroferótipo foi detectado em três rebanhos, o qual permaneceu constante durante o período de amostragem. em dois rebanhos diferentes eletroferótipos foram detectados, embora com maior prevalência de um dado perfil. A caracterização das amostras positivas em subgrupos foi realizada por meio do EIE com duplo sanduíche, utilizando-se anticorpos monoclonais (MAb) específicos para antígenos de subgrupo (I e II). Foram caracterizadas como subgrupo I 52,4% (22/42) das amostras testadas, nenhuma reagiu com MAb de subgrupo II, enquanto as demais, 47,6% (20/42), não reagiram com nenhum dos dois subgrupos.

Absence of cross-reactivity to myeloperoxidase of anti-thyroid microsomal antibodies in patients with autoimmune thyroid diseases

Background: Thyroperoxidase is the major antigen of the thyroid microsomal antibodies (TMA) detected in autoimmune thyroid diseases. Its amino acid sequence has 44% homology with myeloperoxidase (MPO), an enzyme present in the primary granules of neutrophils and one of the major antineutrophil cytoplasmic antibodies (ANCA) antigens. The objective of the present study was to investigate the presence of cross-reactivity to MPO of TMA. Methods: We studied sera from 51 patients with autoimmune thyroid diseases, all of them TMA-positive. The presence of ANCA was investigated by indirect immunofluorescence and by capture enzyme-linked immunosorbent assay. Results: ANCA were positive in 3.9% of the TMA-positive sera and none of them reacted with MPO. In contrast, the ANCA-positive sera revealed antielastase activity. None of the ANCA-positive cases presented clinical signs of vasculitis. However, these 2 patients had been on prolonged treatment with propylthiouracil. Conclusions: We conclude that there is no cross-reactivity to MPO of TMA in patients with autoimmune thyroid diseases, possibly because of difference in the spatial configuration of the immunodominant region. The presence of ANCA in patients with autoimmune thyroid diseases without evidence of vasculitis might result from propylthiouracil-induced polyclonal activation.

Utilidade dos métodos imuno-histoquímicos para o diagnóstico anátomo-patológico.

In order to determine the value of immunohistochemical staining methods for the morphologic diagnosis, we studied 949 histologic specimens sent for consultation to the Immunohistochemistry Laboratory of Department of Pathology of the Medical School of Botucatu in the period 1984-1989. All case were submitted to the immunoperoxidase staining with the methods PAP or ABC. Immunohistochemical stains confirmed the original morphologic diagnosis in 468 cases (49.3%); made the definitive diagnosis from a list of differential diagnostic possibilities in 244 cases (25.7%); provided contributory information in 74 cases (7.8%); were non-contributory in 114 cases (12%) and rendered an unsuspected diagnosis in 49 cases (5.2%). In some cases with non-contributory information the differences in methods of fixation might have led to suboptimal preservation of tissue antigens. The immunohistochemical staining may provide important and sometimes essential informations for definitive diagnosis. This technique was particularly useful for differential diagnosis between carcinoma, lymphoma and melanoma.

Contribuição do estudo imuno-histoquímico para o diagnóstico anátomo-patológico.

In order to determine the value of immunohistochemical staining methods for morphologic diagnosis specimens of 949 cases received at the Immunohistochemistry Laboratory of the Department of Pathology of the Medical School of Botucatu, in the period 1984-1989 were reviewed. All of them were submitted to the immunoperoxidase staining (PAP or ABC). The main morphologic diagnosis was confirmed in 468 cases (49.3%); the definitive diagnosis was made in 244 cases (25.7%) that had only differential diagnosis, and contributory information was provided in 74 cases (7.8%); the immunohistochemical staining was non-contributory in 114 cases (12%). It rendered an unsuspected diagnosis in 49 cases (5.2%). The analysis of these cases shows that immunohistochemical methods may provide important and sometimes essential informations for definitive diagnosis. This technique is particularly useful for distinguishing between carcinoma, lymphoma and melanoma.

Peripheral giant cell granuloma. An immunohistochemical and ultrastructural study.

OBJECTIVE: To study the nature of multinucleated and mononuclear cells from peripheral giant cell granuloma (PGCG). MATERIALS AND METHODS: Formalin-fixed, paraffin-embedded sections of 40 cases of PGCG were immunohistochemically stained for vimentin, alpha I-antichymotrypsin, CD68, S-100 protein, lysozyme, leucocyte common antigen (LCA), factor VIII-related antigen and muscle cell actin. Six cases of PGCG were also studied by transmission electron microscopy. RESULTS: Vimentin, alpha I-antichymotrypsin and CD68 were expressed in both the mononuclear and multinucleated giant cells. Dendritic mononuclear cells, positive for S-100 protein, were noted in 67.5% of the lesions, whereas lysozyme and leucocyte common antigen were detected in occasional mononuclear cells. Ultrastructural examination showed mononuclear cells with signs of phagocytosis and sometimes interdigitations with similar cells. Others presented non-specific characteristics and the third type exhibited cytoplasmic processes and occasional Birbeck granules. Some multinucleated giant cells showed oval nuclei, abundant mitochondria and granular endoplasmic reticulum whereas others presented with irregular nuclei and a great number of cytoplasmic vacuoles. CONCLUSIONS: Immunohistochemical and ultrastructural results suggest that PGCGs of the jaws are composed mainly of cells of the mononuclear phagocyte system and that Langerhans cells are present in two thirds of the lesions.

Pesquisa de anticorpos contra Brucella ovis em ovinos do Estado de São Paulo

Eight hundred and fifty sheep sera from 18 herds, located in 15 municipalities of the State of São Paulo, were examined by complement fixation test, indirect enzyme immunoassay and gel diffusion test for antibodies to Brucella ovis. The combination of the serological test results and the clinical and epidemiological data suggest that none of the examined animals was infected. The complement fixation and the gel diffusion tests showed good specificity, but the indirect enzyme immunoassay showed false positive results in nine of the sera.

Detection of circulating Paracoccidioides brasiliensis antigen in urine of paracoccidioidomycosis patients before and during treatment

For the diagnosis and follow-up of paracoccidioidomycosis patients undergoing therapy, we evaluated two methods (immunoblotting and competition enzyme immunoassay) for the detection of circulating antigen in urine samples. A complex pattern of reactivity was observed in the immunoblot test. Bands of 70 and 43 kDa were detected more often in urine samples from patients before treatment. The immunoblot method detected gp43 and gp70 separately or concurrently in 11 (91.7%) of 12 patients, whereas the competition enzyme immunoassay detected antigenuria in 9 (75%) of 12 patients. Both tests appeared to be highly specific (100%), considering that neither fraction detectable by immunoblotting was present in urine samples from the control group. gp43 remained present in the urine samples collected during the treatment period, with a significant decrease in reactivity in samples collected during clinical recovery and increased reactivity in samples collected during relapses. Reactivity of some bands was also detected in urine specimens from patients with 'apparent cure.' The detection of Paracoccidioides brasiliensis antigens in urine appears to be a promising method for diagnosing infection, for evaluating the efficacy of treatment, and for detecting relapse.

Quantitative cell-cycle protein expression in oral cancer assessed by computer-assisted system

The knowledge of cell-cycle control has shown that the capacity of malignant growth is acquired by the stepwise accumulation of defects in specific genes regulating cell growth. Histologic diagnosis might be improved by a quantitative evaluation of more specific diagnosis biomarkers, which could help to precisely identify pre-malignant and malignant oral lesions. The aim of the present study is to evaluate whether computer-based quantitative assessment of p53, PCNA and Ki-67 immunohistochemical expression, could be used clinically to foresee the risk of oral malignant transformation. This retrospective study was carried out in ninety-five oral biopsies, 27 were classified as fibrous inflammatory hyperplasia, 40 as leukoplakia and 28 as oral squamous cell carcinoma. Sixteen out of the 40 leukoplakia were diagnosed as non-dysplastic leukoplakia, the other 24 being dysplastic leukoplakia, of which 50.0% were classified as moderate to severe dysplasia. Comparison of the four groups of oral tissues showed significant rises in p53 and Ki-67 positivity index, which increased steadily in the order benign, pre-malignant, and malignant. In contrast, it was not possible to relate higher PCNA levels with pre-malignant and malignant oral lesions. We therefore conclude that PCNA immunohistochemistry expression is probably an inappropriate marker to identify oral carcinogenesis, whereas joint quantitative evaluation of p53 and Ki-67, appears to be useful as a tumor marker, providing a pre-diagnostic estimate of the potential for cell-cycle deregulation of the oral proliferate status.

Morphological, cytochemical, and ultrastructural study of thrombocytes and leukocytes in neotropical fish, Brycon orbignyanus Valenciennes, 1850 (Characidae, Bryconinae)

Morphological, cytochemical and ultrastructural studies are important to demonstrate the function of the blood cells, which is very little understood in teleosts. In peripheral blood of piracanjuba' Brycon orbignyanus, thrombocytes, lymphocytes, monocytes, neutrophils and heterophils were studied and characterized. Thrombocytes had a fusiform or oval shape with PAS-positive granules. Lymphocytes presented small size with sparse basophilic cytoplasm. Monocytes were large in size, presented basophilic cytoplasm that may be foamy or vacuolated, with non-specific esterase staining. The neutrophils presented lightly neutrophilic granule cytoplasm, with positivity for PAS and peroxidase. The heterophils were large in size, with eosinophilic and basophilic granules cytoplasm and PAS-positive. Transmission electron microscopy study demonstrated that the thrombocytes, lymphocytes and monocytes features were similar to other teleosts. In ultrastructural study only one type of neutrophils was observed. Cytochemical findings indicated that neutrophils and monocytes of B. orbignyanus may be involved in phagocytosis, and neutrophils play an important microbicidal role.