Analysis of the chemical components of hydatid fluid from Echinococcus granulosus

Introduction The aim of this study was to explore the environment of Echinococcus granulosus (E. granulosus) protoscolices and their relationship with their host. Methods Proteins from the hydatid-cyst fluid (HCF) from E. granulosus were identified by proteomics. An inductively coupled plasma atomic emission spectrometer (ICP-AES) was used to determine the elements, an automatic biochemical analyzer was used to detect the types and levels of biochemical indices, and an automatic amino acid analyzer was used to detect the types and levels of amino acids in the E. granulosus HCF. Results I) Approximately 30 protein spots and 21 peptide mass fingerprints (PMF) were acquired in the two-dimensional gel electrophoresis (2-DE) pattern of hydatid fluid; II) We detected 10 chemical elements in the cyst fluid, including sodium, potassium, calcium, magnesium, copper, and zinc; III) We measured 19 biochemical metabolites in the cyst fluid, and the amount of most of these metabolites was lower than that in normal human serum; IV) We detected 17 free amino acids and measured some of these, including alanine, glycine, and valine. Conclusions We identified and measured many chemical components of the cyst fluid, providing a theoretical basis for developing new drugs to prevent and treat hydatid disease by inhibiting or blocking nutrition, metabolism, and other functions of the pathogen. .

Enzyme markers to identify and characterize Opisthorchis viverrini in Thailand and Lao PDR.

We conducted an allozyme electrophoretic study to explore potential enzyme markers to distinguish Opisthorchis viverrini in Thailand and Lao PDR. Twenty-eight enzymes encoding presumptive 32 loci were established. The enzymes glucose-6-phosphate dehydrogenase and pyruvate kinase were diagnostic between two geographically separate isolates from Thailand. Twelve enzymes, ie, aconitate hydratase, aldolase, creatine kinase, enolase, esterases, fumarate hydratase, aspartate aminotransferase, glucose-phosphate isomerase, alanine aminotransferase, isocitrate dehydrogenase, malic enzyme, and pyruvate kinase, also provided diagnostic markers for these two isolates from Thailand and one isolate from Lao PDR.

Estudo da resposta imune humoral (IgG específica) para antígenos de larvas infectantes (L3) de Wuchereria bancrofti, entre portadores e não portadores de filariose bancroftiana no Município de Olinda-PE / Study of the immune humoral response (IgG specific) for antigens of infective larvae (L3) of Wuchereria bancrofti, between bearers and not bearers of elephantiasis filarial in the city of Olinda-PE

A filariose linfática bancroftiana é uma doença parasitária humana de grande complexidade em sua dinâmica de infecção, necessitando ainda de maiores esclarecimentos, principalmente, em aspectos relacionados à tolerância e imunopatologia. A existência daimunidade protetora em comunidades endêmicas de filariose permanece como objeto de intenso debate e o grupo denominado “endêmicos normais”, tem sido alvo de interesse para elucidar muitas questões referentes à imunologia da doença. O presente trabalho tem como objetivo verificar através de um estudo do tipo caso-controle, as diferenças entre portadorese não portadores de filariose linfática bancroftiana pelo reconhecimento humoral de bandas protéicas de extrato secretório-excretório de larvas infectantes de Wuchereria bancrofti. Osquatro setores censitários de Olinda-PE, com maior prevalência de filariose, foram escolhidos como área de estudo. Consideramos “grupo controle” portadores da filariose bancroftiana e “grupo de casos” os de endêmicos normais. O extrato antigênico foi obtidopor incubação de larvas infectantes em meio Hanks por 72h em 5% de CO2. As bandas protéicas foram separadas por eletroforese (SDS-PAGE) e por western-blot, transferidas e incubadas com os soros humanos. Foram considerados portadores da doença os positivospela técnica da gota espessa ou filtração e endêmicos normais os residentes de área endêmica negativos pelo teste de detecção parasitária e captura de antígenos (Og4C3). Participaram do estudo 325 indivíduos, 130 considerados endêmicos normais e 195portadores de filariose bancroftiana. As bandas protéicas da composição do extrato de L3 foram as de 200, 175, 138, 105, 100, 76, 55, 49, 42, 39, 38, 32, 28, 14 kDa. Na comparaçãodo reconhecimento das proteínas entre os grupos estudados, apenas as proteínas 175 e 105 kDa não diferiram significativamente, os portadores de filariose bancroftiana não reconheceram a proteína 175 e no grupo...

Perfil protéico e reconhecimento antigênico de extratos de larvas infectantes (L3) de Wuchereria bancrofti / Proteic profile and antigenic recognition of extracts from Wuchereria bancrofti L3 infective larvae

A caracterização protéica dos extratos de larvas infectantes (L3) de Wuchereria bancrofti foi realizada por eletroforese em gel de poliacrilamida, em presença de dodecil sulfato de sódio (SDS-PAGE) e o reconhecimento antigênico das proteínas por Western-blot. O maior número de bandas protéicas reconhecidas foi evidenciado nos extratos AgSE (105, 100, 76, 55, 49, 39 e 32 kDa) e AgS (100, 76, 55, e 49 kDa) na presença de soros de indivíduos endêmicos normais. As bandas de 49 e 55 kDa foram reconhecidas pelos soros dos microfilarêmicos, endêmicos normais (residentes de área endêmica livres de infecção filarial) e portadores da forma crônica da doença. As larvas infectantes foram obtidas pela dissecção de mosquitos Culex quinquefasciatus infectados com sangue microfilarêmico de voluntários portadores de microfilaremia, residentes do Município de Olinda-PE. Nos 792 indivíduos investigados pela técnica da gota espessa mensurada (60æl de sangue) 87 foram positivos (11 por cento). A diferenca da positividade entre homens e mulheres não foi significativa e a faixa etária de 11 a 19 anos foi a de maior prevalência.

A new method for concentration of proteins in the calcareous corpuscles separated from the spargana of Spirometra erinacei

Calcareous corpuscles are a characteristic structure found in larval and adult stage cestodes. These corpuscles are known to contain several protein components and to possess protein-binding activity. However, the proteins bound to calcareous corpuscles in situ have not been studied. The present study was undertaken to identify the proteins on calcareous corpuscles. Calcareous corpuscles were purified from the plerocercoids (= spargana) of Spirometra erinacei, and serially dissolved using 0.1 M sulfamic acid solution. Collected supernatants were examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining. The results showed that only the fraction remaining after the 19th dissolved fraction contained proteins. A total of 20 protein molecules were detected in gel, with major bands at 56, 53, 46, 40, 35, 29, 28, 24.5, 21, 19, 16, 13, 10 and 8 kDa. In particular, the proteins corresponding to the 21 and 16 kDa bands were most abundant. Our results demonstrated for the first time the protein contents of the calcareous corpuscles of spargana. Further studies on the functions of these proteins are required.

Preliminary analysis of Sm14 in distinct fractions of Schistosoma mansoni adult worm extract

In previous studies it was shown that the recombinant molecule, r-Sm14, induces high levels of protection against Schistosoma mansoni infection in two outbred animal models and immune crossprotection against infection by Fasciola hepatica in Swiss outbred mice. r-Sm14 was derived from a living worm extract, called SE, and is being developed as the molecular basis of an anti-helminth bivalent vaccine against the two parasites, for medical and veterinary application. Present data refer to SDS-PAGE and Western Blotting analysis of four different preparations of S. mansoni adult worms focusing Sm14 identification. The extracts correspond to the initial fraction of the SE extraction process, containing products released by living worms (SEi); SE2, reextraction of adult worms in PBS; and SE of separated male and female adult worms. In all extracts it was possible to detect the component of 14 kDa, that was recognized by specific anti-rSm14 antibody raised in rabbits

Gnathostoma spinigerum: analysis of protein patterns by two dimensional gel electrophoresis.

The protein extracts from male (MS) and female (FS) adults and advanced third-stage larvae (LS) of Gnathostoma spinigerum were separated by high resolution two-dimensional gel electrophoresis (2-DE). The polypeptide spots, as detected by silver staining, were subsequently identified. The spot patterns of LS, MS and FS were highly complex and consisted of more than 75, 44, 52 prominent spots, respectively. In addition, the stage-specific protein patterns were identified. This 2-DE database should provide an important reference for future biological and biochemical studies of G. spinigerum.

Hydrolytic activity of an onchocercoma

The onchocercoma or nodule produced by the nematode Onchocerca volvulus (Filarioidea) in the skin of patients suffering from onchocerciasis has not been examined by histochemical techniques. In this work we have used histochemical techniques to evaluate 5 hydrolytic enzymes, namely phosphatases, esterases and beta-glucuronidase. The results show increased enzymatic activity at the sites of major metabolic activity and within reactive cells including macrophages (mc) and giant cells (gc) of the onchocercoma