Estudo pareado da cardiopatia chagásica no Rio Grande do Sul, Brasil

São estudados 666 pares de indivíduos, sendo 344 masculinos e 322 femininos, com sorologia positiva/negativa para doença de Chagas, obtidos em inquérito sorológico-eletrocardiográfico entre populações rurais não selecionadas de 17 municípios do sul Rio Grande do Sul, Brasil. Na análise dos eletrocardiogramas foram consideradas só as alterações sugestivas de cardiopatia chagásica: bloqueio átrio-ventricular de 1º, e 2º e 3º, BCRD isolado ou associado ao HBAE, HBAE isolado, extrasístoles ventriculares freqüentes e/ou polifocais e/ou bigeminadas e trigeminadas; alterações de ST e T; zonas eletricamente inativas. Com este critério apresentaram eletrocardiogramas alterados 201 pessoas soropositivas (30,2%) e 66 soronegativas (9,9%). O gradiente de 20,3%, sendo 21,5% nos homens e 18,9% nas mulheres, mostrou-se significativo ao nível de p < 0,001.

Fatores que podem afetar a criação e manutenção de caramujos infectados e a produção de cercárias de Schistosoma masoni

Foram estudados fatores que afetam a produção em massa de cercárias do Schistosoma mansoni. Rotíferos e ostracodos causaram diminuição de produção em nossa colônia. Os rotíferos foram erradicados facilmente lavando-se os aquários e a alface com ácido acético diluído. Já o ostracodo foi de difícil erradicação, produzindo mortalidade nos caramujos infectados, que alcançou níveis de 50-60% por mês. Moluscos infectados mantidos na incubadora a temperatura constante e em total escuridão apresentaram maior eliminação quando iluminados, a aproximadamente 30ºC. A eliminação de cercárias foi praticamente a mesma entre os pHs 5-7. Traços de metais pesados adicionados à água destilada afetaram negativamente a eliminação. Finalmente, a manutenção em incubadora, embora apresenta-se maior mortalidade de moluscos, foi mais simples e eficiente para a produção em massa de cercárias que o sistema convencional com água corrente.

Estudo pareado da cardiopatia chagásica no Rio Grande do Sul, Brasil: comportamento das alterações eletrocardiográficas em função da cor

O trabalho analisa o comportamento das alterações eletrocardiográficas em 150 indivíduos de cor preta, naturais da área rural do sul do Rio Grande do Sul, pareados com outros tantos brancos do mesmo sexo, idade e procedência. O grupo tinha idade entre 9 e 78 anos, com média de 39,2 e era composto de 57 homens e 93 mulheres. Com referência à sorologia, 84 eram positivos e 66 negativos para a infecção pelo Trypanosoma cruzi. A prevalência de alterações eletrocardiográficas foi significativamente maior entre os pretos, tanto soropositivos quanto soronegativos. As alterações eletrocardiográficas consideradas sugestivas de miocardiopatia chagásica, predominaram significativamente entre os positivos pretos. E mesmo entre os pretos negativos as alterações foram mais numerosas e de maior gravidade com respeito aos negativos brancos. Na opinião dos autores a maior prevalência e severidade das alterações eletrocardiográficas entre os pretos, podem ser atribuídos a vários fatores entre os quais se destacariam: a pobreza e marginalidade das pessoas de cor que implicam trabalhos mais pesados e desgastantes; habitações triatomínica; alimentação mais pobre quantitativa e qualitativamente. Outro fator negativo seriam os valores mais elevados de pressão arterial encontrados entre os pretos que, associados a possíveis influências de ordem racial, podem contribuir e/ou criar condições de patologia miocárdica.

Granulomatous hypersensitivity to Schistosoma mansoni egg antigens in human Schistosomiasis: I. Granuloma formation and modulation around polyacrylamide antigen-conjugated beads

We have developed an in vitro model of granuloma formation for the purpose of studying the immunological components of delayed type hypersensitivity granuloma formation in patients infected with Schistosoma mansoni. Our data show that 1) granulomatous hypersensitivity can be studied by examining the cellular reactivity manifested as multiple cell layers surrounding the antigen conjugated beads; 2) this reactivity is a CD4 cell dependent, macrophage dependent, B cell independent response and 3) the in vitro granuloma response is antigenically specific for parasite egg antigens. Studies designed to investigate the immune regulation of granulomatous hypersensitivity using purified populations of either CD4 or CD8 T cells have demonstrated the complexity of cellular interactions in the suppression of granulomatous hypersensitivity. The anti-S. mansoni egg immune responses of individual patients with chronic intestinal schistosomiasis can be classified either as soluble egg antigen (SEA) hypersensitive with maximal granulomatous hypersensitivity or SEA suppressive with activation of the T cell suppressor pathway with effective SEA granuloma modulation. Our data suggest that T cell network interactions are active in the generation of effective granuloma modulation in chronic intestinal schistosomiasis patients.

Analysis of toxoplasma gondii proteins after Triton X-114 solubilization and hidropholic chromotograhy

The distribution of the surface proteins of toxoplasma gondii radiodinated were studied using the phase separation technique and ability of binding in the phenyl-Sepharose column. Eight polypeptides with Mr 22 to 180 distributed exclusively in the detergent rich-phase, while six polypeptides with mol. wt. 15,000 to 76,000 distributed exclusively in the detergent poor-phase. Twopolypeptides with 15,000 and 70,000 distributed on both phase. All the polypeptides present in the detergent rich-phase binding in the phenyl-Sepharose column, and can be isolated in two peak according with their relative hydrophobicities.two polypeptides hydrophobic with Mr 60 and 66 recognized by human serum were isolated by the association of the two technique. Our result showed that the surface proteins of t. gondii present different degrees of hydrophobicity and that the use of hydrophobic interaction chromatography after Triton X-114 extraction may be an important isolation method of membrane proteins.

Characterization of plasma menbrane polypeptides of trypanosoma from bats

Cell surface proteins of Trypanosoma dionisii, Trypanosoma vespertilionis and Trypanosoma sp. (M238) were radiodinated and their distribution both in the detergent-poor (DPP) and dertergent-enriched phase (DRP) was studied using a phase separation technique in Triton X-114 as well as polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS-PAGE). Significant differences were observed in the proteins present in the DRP when the three species of trypanosoma were compared. Two major bands with 88 and 70 KDa were observed in T. sp. (M238) but were not detectable in T. dionisii and T. vespertilionis. Three polypeptides whith 96, 77 and 60 KDa were identified in the DRP of T. vespertilionis. Three major bands with 84, 72 and 60 KDa were observed in the DRP of T. dionisii. Two polypeptides with 34-36 KDa present in the DPP, were observed in the three Trypanosome species analyzed. Our observations show that T. sp. (M238) has characteristic surface polypeptides not found in T. vespertilionis.

Identification and characterization of sex-linked proteins of Schistosoma mansoni

The proteins of adults worms (male and female) of two isolates (BH and RJ) of Shistosoma mansoni were extracted using Triton X-114 phase separation. The SDS-polyacrilamide gel electrophoresis profiles of the three phases (detergent, aqueous and insoluble proteins) obtained were compared after Coomassie blue and silver staining, surface radioiodination and Western blotting. No major differences were detected between the 2 isolates. Of the 25 or more proteins which partitioned into the detergent phase, only about 8 proteins could be surface radiodinated on live adult worms. A comparison was also made between the profiles of mael and females worms, isolated from bisexually infected mice. Two major female-specific and one male-specific band were detected by silver and/or Coomassie staining. The female bands, 32 KDa and 18 KDa, partitioned into the detergent and aqueous phase, respectively. The male-specific band of 42 KDa remained in the insoluble phase. Antigenic differences between male and females protins were detected by Western vlotting using a sera from infected Nectomys squamipes.

Immunological profiles of patients from endemic areas infected with Schistosoma mansoni

Crude extracts of eggs (SEA) adult worms (SWAP) or cercariae (Cerc) have been used to stimulate Peripheral Blood Mononuclear cells (PBMC) and have provided rather distinct profiles of responses in different types of patients. In genenral it is clear that patients with early infections respond strongly to SEA while response to SWAP are developed more slowly. As infection progresses into the more chronic phases, a general pattern is seen whic leads to lower anti-SEA proliferative responses in the face of higher responses to SWAP and variable anti-cerc responsiveness. Cured not re-exposed patients express very high levels of anti-SEA proliferation. It has recently been seen that those individuals who live in endemic areas and have continued water contact, but are reapeatedly stool-negative (who are presumed to have self-cured or be putatively resistant; endemic normals) are strongly responsive to antigenic extracts, particularly to SEA. Furthermore, our results show that endemic normal individuals have significantly higher IFN gamma production upon PBMC stimulation with schistosome antigens than infected individuals. With the emergence of more studies it is becoming apparent that both the intensity and the prevalence of a given area may influence or shape the general responsiveness of the population under study.

Human schistosomiasis mansoni: studies on in vitro granuloma modulation

Infection with Schistosoma mansoni induces humoral and T cell mediated responses and leads to delayed hipersensitivity that results in granulomatous inflamatory disease around the parasite eggs. Regulation of these responses resulting in a reduction in this anti-egg inflamatory disease is appsrently determined by idiotypic repertoires of the patient, associated with genetic background and multiple external factors. We have previously reported on idiotype/anti-idiotype-receptor transactions in clinical human schistosomiasis. These findings support a hypothesis that anti-SEA cross-reactive idiotypes develop in some patients during the course of a chronic infection and participate in regulation of anti-SEA cellular immune responses. We repport here on experiments wich extend those observations to the regulation of granulomatous hypersensitivity measured by an in vitro granuloma model. T cells from chronic intestinal schistosomiasis patients were stimulated in vitro with anti-SEA idiotypes and assayed in an autologous in vitro granuloma assay for modulation of granuloma formation. These anti-SEA idiotype reactive T cells were capable of regulating autologous in vitro granuloma formation. This regulatory activity, initiated with stimulatory anti-SEA idiotypic antibodies, was antigenically specific and was dependent on the present of intact (F(ab')2 immunoglobulin molecules. The ability to elicit this regulatory activity appears to be dose dependent and is more easily demonstrated in chronically infected intestinal patients or SEA sensitized individuals. These data support the hypothesis that anti-SEA cross reactive idiotypes are important in regulating granulomatous hypersensitivy in chronic intestinal schistosomiasis patients and these cross-reactive idiotypes appear to play a major role in cell-cell interactions which result in the regulation of anti-SEA cellular immune responses.

Partial isolation and some properties of enterotoxin produced by Bacillus cereus strains

Extracellular proteins produced by Bacillus cereus AL-42 and AL-15 were fractioned by chromatography on QAE-Sephadex and Sephadex G75. This last chromatographic process resulted in three peaks. The major peak showed vascular permeability activity to rabbits, lethality to mice, and cytotoxicity to Vero and Hela cells. The analysis by SDS-PAGE after ultrafiltration confirm recent findings that the enterotoxin is a compound with molecular mass > 30.000.

N-terminal amino acid sequences of the major outer membrane proteins from a Neisseria meningitidis group B strain isolated in Brazil

The four dominant outer membrane proteins (46, 38, 33 and 28 kDa) were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in a semi-purified preparation of vesicle membranes of a Neisseria meningitidis (N44/89, B:4:P1.15:P5.5,7) strain isolated in Brazil. The N-terminal amino acid sequence for the 46 kDa and 28 kDa proteins matched that reported by others for class 1 and 5 proteins respectively, whereas the sequence (25 amino acids) for the 38 kDa (class 3) protein was similar to class 1 meningococcal proteins. The sequence for the 33 kDa (class 4) was unique and not homologous to any known protein.