Production and characterization of monoclonal antibodies against Campylobacter fetus subsp. venerealis / Produção e caracterização de anticorpos monoespecíficos contra Campylobacter fetus subsp. venerealis

Myeloma cells Sp2/0-Ag14 and spleen cells from BALB/c mouse immunized with sonicated Campylobacter fetus subsp. venerealis NCTC 10354 were fused with polyethylene glycol (PEG) for the selection of clones producing antibodies. Clones were obtained by limiting dilution and screened for the production of specific antibodies to C. fetus subsp. venerealis NCTC 10354 by indirect ELISA and western blot against a panel of bacteria: C. fetus subsp. venerealis NCTC 10354, C. fetus subsp fetus ADRI 1812, C. sputorum biovar sputorum LMG 6647, C. lari NCTC 11352, and Arcobacter skirrowii LMG 6621 for the ELISA and C. fetus subsp. venerealis NCTC 10354 and C. sputorum biovar sputorum LMG 6647 for the western blotting. Fifteen clones producing monoclonal antibodies (MAbs) anti-C. fetus subsp. venerealis of the IgM (1) and IgG (14) classes were further screened for species-specificity. Four clones of the 15 obtained were producers of species-specific monoclonal antibodies (MAbs): two were specific for C. fetus subsp. venerealis and two were specific for C. fetus subsp. fetus. None of the clones were reactive against C. sputorum biovar sputorum LMG 6647. All clones recognized a protein with molecular mass of approximately 148 kDa from lysed C. fetus subsp. venerealis NCTC 10354.

Para a produção de anticorpos monoclonais contra Campylobacter fetus subsp. venerealis foram utilizadas as linhagens de células de mieloma Sp2/0-Ag14 e células de baço de camundongos BALB/c imunizados com sonicado de C. fetus subsp. venerealis NCTC 10354. A detecção dos anticorpos monoclonais foi realizada por ELISA indireto utilizando antígeno sonicado de C. fetus subsp. venerealis NCTC 10354. A clonagem foi realizada por diluição limitante e os clones foram caracterizados por ELISA indireto utilizando um painel de bactérias escolhidas em função da prevalência e habitats: C. fetus subsp. venerealis NCTC 10354, C. fetus subsp. fetus ADRI 1812, C. sputorum biovar sputorum LMG 6647, C. lari NCTC 11352 e Arcobacter skirrowii LMG 6621; e no "western blotting" utilizando antígenos sonicados de C. fetus subsp. venerealis NCTC 10354 e C. sputorum biovar sputorum LMG 6647. Foram obtidos 15 clones produtores de anticorpos anti- C. fetus subsp. venerealis das classes IgM (1) e IgG (14). Quatro clones dentre os 15 clones obtidos foram produtores de anticorpos monoclonais espécie-específicos: dois clones reagiram com maior especificidade contra C. fetus subsp. venerealis NCTC 10354 e dois clones reagiram com maior especificidade contra C. fetus subsp. fetus ADRI 1812. Nenhum dos clones reagiu contra C. sputorum biovar sputorum LMG 6647, comprovando a especificidade dos anticorpos monoclonais testados. Todos os clones reconheceram uma proteína de massa molecular de aproximadamente 148 kDa no sonicado de C. fetus subsp. venerealis NCTC 10354.

Correlação entre a Visual Gait Assessment Scale, Edinburgh Visual Gait Scale e Escala Observacional da Marcha em crianças com paralisia cerebral diparética espástica / Correlation among the Visual Gait Assessment Scale, Edinburgh Visual Gait Scale and Observational Gait Scale in children with spastic diplegic cerebral palsy

CONTEXTUALIZAÇÃO: Desordens da marcha são comuns em crianças com paralisia cerebral (PC) diparética espástica. Com o intuito de aprimorar a marcha dessas crianças e quantificar os desfechos de tal intervenção, torna-se necessário que se faça uma análise instrumentada pré e pós-intervenção. OBJETIVOS: Correlacionar a Edinburgh Visual Gait Scale (EVGS), a Visual Gait Assessment Scale (VGAS) e a Escala Observacional de Marcha (EOM). MÉTODOS: Estudo transversal de análise da marcha por meio das escalas EVGS, VGAS e EOM, envolvendo oito crianças com PC diparética espástica, nível I ou II do Gross Motor Function Classification System (GMFCS), avaliadas por três examinadores. O estudo foi aprovado pelo Comitê de Ética em Pesquisa da Universidade Estadual de Campinas - UNICAMP. Os dados foram analisados pelo índice Kappa ponderado, considerando um nível de significância de 5%. RESULTADOS: O estudo intra-avaliadores mostrou que a concordância entre os métodos na classificação dos sujeitos foi de moderada a excelente (k=0,41, 1,00), sendo a comparação entre a VGAS e a EVGS a de maior índice de concordância, enquanto a EOM obteve grande discordância em comparação com as outras escalas. A concordância interavaliadores se mostrou predominantemente alta. CONCLUSÃO: Os resultados fornecem evidências de que a VGAS e a EVGS são mais adequados para avaliação da marcha de crianças com PC diparética quando comparadas à EOM.

BACKGROUND: Gait disorders are very common in children with spastic diplegia cerebral palsy (CP). In order to improve the CP children's gait and to quantify the outcomes of this intervention it becomes essential to perform an instrumented analysis before and after the intervention. OBJECTIVES: To analyze the correlation among the Edinburgh Visual Gait Scale (EVGS), the Visual Gait Assessment Scale (VGAS) and the Observational Gait Scale (OGS). METHODS: Cross sectional study aiming to analyze the gait of 8 children with spastic diplegia CP with level I or II in the Gross Motor Function Classification System (GMFCS) through the EVGS, VGAS and OGS scales performed by 3 examiners. This study was approved by the Research Ethics Committee of the Universidade Estadual de Campinas (UNICAMP). Weighted Kappa scores were used to analyze the data considering a significance level of 5%. RESULTS: The intra-rater analyses showed a moderate to excellent agreement (k=0.41, 1.00) among the methods of the children's classification, being the comparison between VGAS and the EVGS scales presented the highest level of agreement, while the OGS scale presented a considerable disagreement in comparison with other scales. The inter-rater agreement showed to be predominantly high. CONCLUSIONS: The results provide evidence that the VGAS and the EVGS scales are more suitable for children's spastic diplegia CP gait assessment when compared to OGS.

Primary culture of Rhodnius prolixus (Hemiptera: Reduviidae) salivary gland cells

In the present paper, we developed a primary culture of Rhodnius prolixus salivary gland and main salivary canal cells. Cells remained viable in culture for 30 days. Three types of cells were indentified in the salivary gland cultures, with binuclear cells being the most abundant. The supernatants of salivary cultures contained mainly 16-24 kDa proteins and presented anticoagulant and apyrase activities. Secretion vesicles were observed budding from the cellular monolayer of the main salivary canal cells. These results indicate that R. prolixus salivary proteins may be produced in vitro and suggest that the main salivary canal may have a possible secretory role.

Host nutritional status as a contributory factor to the remodeling of schistosomal hepatic fibrosis

Weaning Swiss mice were percutaneously infected with 30 cercariae of Schistosoma mansoni and submitted to a shifting either from a deficient to a balanced diet or vice-versa, for 24 weeks. The nutritional status was weekly evaluated by measurements of growth curves and food intake. Hepatic fibrosis and periovular granulomas were studied by histological, morphometric and biochemical methods. All mice fed on a deficient diet failed to develop periportal "pipestem" fibrosis after chronic infection. An unexpected finding was the absence of pipestem fibrosis in mice on normal diet, probably related to the sample size. The lower values for nutritional parameters were mainly due to the deficient diet, rather than to infection. Liver/body weight ratio was higher in "early undernutrition" group, after shifting to the balanced diet. Volume density and numerical density of egg granulomas reached lowest values in undernourished animals. The amount of collagen was reduced in undernourished mice, attaining higher concentrations in well-fed controls and in "late undernutrition" (balanced diet shifted to a deficient one), where collagen deposition appeared increased in granulomas. That finding suggested interference with collagen degradation and resorption in "late" undernourished animals. Thus, host nutritional status plays a role in connective tissue changes of hepatic schistosomiasis in mice.

Chemotherapeutic effects on larval stages of Schistosoma mansoni during infection and re-infection of mice

The sensitivity of the larval stages of Schistosoma mansoni to chemotherapy with praziquantel and oxamniquine was tested in mice during primary and secondary infections and after different intervals from cercarial exposure. Worm recovery by perfusion of the porto-mesenteric system, followed by counting and a morphometric study of the parasite, allowed the conclusion that the relative resistance of the larval stages of S. mansoni to schistosomicide drugs, demonstrated in primary infections, also persists when the host is already infected. This indicates that a therapeutic failure may result when an infected host is treated some time after being re-infected, because of the presence of migrating, drug-resistant, immature forms of the parasite

Experimental Neuroschistosomiasis: inadequacy of the murine model

Neuroschistosomiasis is rarely observed in human pathology, but it is of considerable importance. To investigate its pathogenesis, consequences and response to treatment, an experimental model would be desirable, but is not yet available, in spite of a few indications of a suitable mouse model in the literature. Severe, recent and late Schistosoma mansoni infections in outbred and inbred strains of mice revealed widespread distribution of parasite eggs in several organs, but only exceptionally did eggs reach the encephalus, thus revealing the inadequacy of the mouse as an experimental model for neuroschistosomiasis

Significance of schistosomal granuloma modulation

Hepatic Schistosoma mansoni periovular granulomas undergo changes in size, cellular composition and appearance with time. This phenomenom, known as "immunological modulation", has been thought to reflect host immunological status. However, as modulation has not been observed outside the liver, participation of local factors, hitherto little considered, seems crucial. Components of the extracellular matrix of periovular granulomas of the mouse were particularly studied in three different organs (liver, lung and intestine) and during three periods of infection time (acute, intermediate and chronic) by means of histological, biochemical and imunofluorescence techniques, while quantitative data were evaluated by computerized morphometry, in order to investigate participation of local factors in granuloma modulation. Results confirmed modulation as a exclusively hepatic phenomenom, since pulmonary and intestinal granulomas, formed around mature eggs, did not change size and appearance with time. The matricial components which were investigated (Type I, III and IV collagens, fibronectin, laminin, proteoglycans and elastin) were found in all granulomas and in all organs examined. However, their presence was much more prominent in the liver. Elastin was only found in hepatic granulomas of chronic infection. The large amount of extracellular matrix components found in hepatic granulomas was the main change responsible for the morphological aspects of modulation. Therefore, the peculiar environment of the liver ultimately determines the changes identified in schistosomal granuloma as "modulation".

An experimental approach to the pathogenesis of pipestem fibrosis (Symmers' fibrosis of the liver)

Pathogenesis of schistosomal hepatic fibrosis (pipestem fibrosis of the liver) was investigated by means of the murine model. Although worm load appears as the main pathogenetic factor, alone it is not sufficient to produce that characteristic lesion. By comparing the findings in animals with heavy and prolonged Schistosoma mansoni infection, which developed or not pipestem fibrosis, it was observed that the lesion was more frequent in intact animals than in the splenectomized one. However, the size of the spleen, the number of recovered worms, the number of eggs per gram of liver tissue, the level of serum idiotype and anti-idiotype antibodies, the size and volume of periovular granulomas formed in the liver, all that failed to show statistically significant differences between the two groups. After analysing all these data, other factors, that apparently have been hitherto negleted, rested to explain the findings. Among them, the timing and sequence of the egg-induced intrahepatic vascular changes seemed crucial. The sequential development of intrahepatic portal vein obstruction, followed by the opening of periportal collateral veins and the continous arrival of schistosome eggs going to be lodged into the latter, appeared as essential steps in the pathogenesis of pipestem fibrosis.

Immunostimulation as adjuvant for the chemotheraphy of experimental schistosomiasis

Animais imunodeprimidos respondem mal ao tratamento anti-esquistossomotico. A capacidade para responder adequadamente pode ser restaurada pela administracao de soro imune. Nao se sabe se a estimulacao de animais normais pode aumentar a eficacia da droga. Camundongos infectados com 50 cercarias do Schistosoma mansoni e tratados 2 meses depois com adjuvante de Freund responderam da mesma forma que os controles nao estimulados ao tratamento com 25, 50 e 100 mg/kg pc de oxamniquine. Nao houve correlacao, entre niveis de anticorpos sericos e a eficacia da droga...

AIDS numa comunidade de Londrina: populaçäo consciente ou desinformada? / Acquired immunodeficiency syndrome in a Londrina's comunity: a conscient population or a bad-informed population?

Considerando que a AIDS tornou-se um dos mais importantes problemas de saúde pública dos tempos atuais em face do grande aumento dos índices mundiais relativos a esta doença, foi avaliado o nível de conscientizaçäo da populaçäo de Londrina, mais especificamente na área de abrangência da Unidade Básica de Saúde "Dr. Newton L. Câmara - Vila Casoni", visando verificar nesta regiäo, o nível de conscientizaçäo da populaçäo situada na faixa etária dos 13-40 anos, além de levantar conhecimentos naquela comunidade, com relaçäo à prevençäo da AIDS. Os resultados obtidos revelam que existe preocupaçäo por parte da populaçäo, sendo satisfatórios os níveis de conscientizaçäo e prevençäo relativos à AIDS