Undergraduate honors paper--Mount Holyoke College, 2005. Dept. of Biological Sciences. / Includes bibliographical references (leaves 94-100).

No description available.

Monoclonal antibodies (MAbs) have been raised against $ beta$-tubulin of B. pahangi and A. suum. Anti-B. pahangi MAbs were used to investigate the heterogeneity of tubulins from nematodes and mammals. One-dimensional SDS-PAGE showed that MAbs P3D and 1B6 react with $ beta$-tubulin from a number of filarial and intestinal nematodes, but not with tubulin from protozoan and mammalian cells. Two-dimensional SDS-PAGE demonstrated that MAb P3D recognizes two isoforms of $ beta$-tubulin and 1B6 recognizes one. Limited proteolysis showed that MAb 1B6 reacted with the amino-terminal fragments and MAb P3D with the carboxyl-terminal fragments of $ beta$-tubulin. The effect of anti-B. pahangi MAbs on the viability of adult B. pahangi was assessed using MTT assay. It was found that MAbs P3D and 1B6 caused an 80% and 40% reduction respectively, in worm viability, whereas anti-chick MAb 357 or mebendazole drug had no effect. Immunogold labelling of B. pahangi demonstrated the presence of tubulin in the median and basal layers of the cuticle, hypodermal layer and somatic muscle blocks, as well as the uterus of B. pahangi. The reduction in the viability of worms may, therefore, be due to the disruption of microtubules in the body wall muscle of B. pahangi. The total MBZ binding was highest in the intestine followed by the body wall muscle and in the reproductive tract extracts of A. suum. Electron microscopy of A. suum tissues demonstrated that the tubulin content decreased from the intestine through the body wall muscle to the reproductive tract. One dimensional SDS-PAGE revealed the presence of $ alpha,$ $ beta sb1$ and $ beta sb2$ tubulin subunits in all tissues of A. suum. This data confirmed the reduction of tubulin from the intestine through the body wall muscle to the reproductive tract. Two dimensional SDS-PAGE followed by Western blotting demonstrated that $ alpha$ and $ beta$ tubulin isoform patterns are dissimilar in different tissues of A. suum. Body wall muscle, inte

Thesis (M.Sc.). / Written for the Institute of Parasitology. Title from title page of PDF (viewed 2009/08/07). Includes bibliographical references.

No description available.

Many parasites have evolved to release products that inhibit host defence mechanisms such as enzymes in the mammalian host, in order to promote and sustain their survival within the host. The human filarial nematode Brugia malayi produces larval microfilariae, which circulate in the blood stream. Their most abundant secreted product is a serine protease inhibitor Bm-SPN-2. Serine protease inhibitors (Serpins) are reported to be involved in how the nematodes avoid host immune defences, and in the case of Bm-SPN-2, the protein was found to specifically inhibit the enzymatic activity of human neutrophil elastase and cathepsin G in a dose-dependent manner. More recently, these two enzymes have been linked to the activation of a major innate cytokine IL-33, which is stored as a full-length 270-aa protein in the cell nucleus, and released as an active C-terminal domain upon stimulation. As full-length (FL) human and murine IL-33 are not commercially available, soluble murine and human FL-IL-33 were produced in transfected HEK 293T cells, following mutation of the nuclear binding motif. In this form, IL-33 is no longer retained in the nucleus and can be purified as a soluble protein. It was confirmed that once cleaved, recombinant human IL-33 was able to induce significant IL-6 secretion by mast cells. Bm-SPN-2 was then shown to block human full-length IL-33 cleavage by inhibiting human neutrophil cathepsin G in a dose dependent manner, supporting the hypothesis that Bm-SPN-2 may act in vivo to prevent IL-33 activation and the promotion of the TH2 immune response. However, in the in vivo setting, it was unexpectedly found that IL-33R (ST2) gene deficiency did not enhance the survival of B. pahangi microfilariae. Furthermore, in the absence of IL-33R, murine immune responses to microfilariae were not significantly altered compared to wild-type BALB/c mice, other than in a significant increase in IL-33 expression. Hence while Bm-SPN-2 can act in vitro to forestall one of the key events in TH2 induction, this has not yet been shown to be crucial to the immune response to the parasite in vivo.

No description available.

Brugia malayi is one of the causative agents of the disabling and disfiguring disease known as Lymphatic Filariasis (LF). This infection is a well-established ailment in tropical and subtropical countries and recently the drug ivermectin has been introduced for the LF control programs. Ivermectin (IVM) is an excellent microfilaricide, but is not markedly macrofilaricidal. However, it causes a long-lasting reduction in the production of new larvae by female worms, suggesting that adult stages are affected. However, the mechanism by which IVM produces such effect in the adult worm is not well understood. One major reason is our incomplete understanding about the biological effect of IVM on adult stages. The present study was carried out to examine the in vitro effects of IVM on B. malayi adult worms using Brugia-gerbil animal model. And also to have some leads in understanding the drug-uptake and location of probable targets in the worm body by using fluorescent labeled IVM and confocal microscopy. / The antifilarial effects of IVM were examined using three parameters: mf release by female worms, and motility, and viability in both male and female worms. The results reported in this study demonstrate that although IVM did not kill the adult worm, but showed significant antifilarial effects on B. malayi adult stages when examined in an in vitro system. Confocal microscopy images of the worms incubated in bodipy FITC-IVM showed strong specific localization signal in the anterior cephalic region of both male and female worms. These observations suggest the early/initial interactions of the drug with its probable receptors that could be located specifically in the head region.

Made available in DSpace on 2016-06-28T12:15:32Z (GMT). No. of bitstreams: 2 320.pdf: 1676340 bytes, checksum: 711128ac80440a81a6bb4dc9ff8ef57b (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2012 / Made available in DSpace on 2016-07-05T22:00:06Z (GMT). No. of bitstreams: 3 320.pdf.txt: 125592 bytes, checksum: 94d4fa2436a2862cb43733491b5ab4ce (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 320.pdf: 1676340 bytes, checksum: 711128ac80440a81a6bb4dc9ff8ef57b (MD5) Previous issue date: 2012 / Fundação Oswaldo Cruz. Centro de Pesquisas Aggeu Magalhães. Recife, PE, Brasil / A Filariose Linfática (FL) no Brasil é causada pela espécie Wuchereria bancrofti e consiste em um problema de saúde pública. O principal foco ativo de transmissão atualmente no país é a Região Metropolitana do Recife - PE, que desde 2003 iniciou o Programa de Controle/Eliminação da FL, tendo como estratégia principal o Tratamento Coletivo (TC) com Dietilcarbamanzina (DEC). Este trabalho, aprovado pelo Comitê de Ética em Pesquisa do Centro de Pesquisas Aggeu Magalhães, analisou o TC nessas áreas, acompanhando 30 moradores, no período de 2003 a 2009. Para essa análise além das ferramentas tradicionais da pesquisa filarial - Filtração (MF/mL de sangue) e Antígeno Circulante Filarial (Og4C3) - também foi utilizada a pesquisa de anticorpos através de um antígeno recombinante (Bm14). Essa nova metodologia desenvolvida é recomendada para ser empregada como uma forma de avaliar o progresso dos programas de controle e eliminação da FL nas áreas sob intervenção. Os resultados obtidos indicam redução na positividade para a FL pelas três metodologias: o Bm14 reduziu de 90 por cento para 80,00 por cento, o Og4C3 de 100 por cento para 60,00 por cento e a microfilaremia (MF) de 100 por cento para 0 por cento. A análise da densidade de MF/mL de sangue e a positividade para o Bm14 revelou que o grupo com maior densidade de MF/mL no sangue (= 57 MF/mL) apresentou maior percentual de redução na positividade para o anticorpo do que o grupo de menor densidade ( 57 MF/mL) em 2009. A taxa de anticorpos-positivos apresentou um percentual de redução de 11,11 por cento no último ano. A diminuição nas taxas de positividade apresentadas pelo Bm14 e o padrão de decaimento observado na análise das Densidades Óticas média e mediana do anticorpo durante os seis anos da pesquisa indicam que o monitoramento dos anticorpos com o antígeno recombinante Bm14 foi capaz de reconhecer indivíduos infectados e também de identificar redução dos níveis de anticorpos produzidos por eles após exposição aos parasitos filariais. Sugerindo que o TC com DEC teria surtido efeito na eliminação dos vermes adultos e conseqüente desaparecimento das microfilárias da circulação sanguínea

Les Wolbachia sont des alpha-proteobactéries présentes chez de nombreux arthropodes et nématodes filaires. Ces bactéries héritées maternellement induisent chez leurs hôtes des phénotypes allant du parasitisme au mutualisme, avec le long de ce continuum des phénotypes tels que la féminisation (F), l'incompatibilité cytoplasmique (IC) ou la mort des mâles. Wolbachia est ainsi un modèle particulièrement intéressant pour étudier les différents types de relations symbiotiques.Chez Brugia malayi, comme pour les autres nématodes filaires, Wolbachia vit en symbiose obligatoire avec son hôte. L'élimination de la bactérie par des traitements antibiotiques entraîne une perte de fertilité voire la mort du nématode. Chez l'isopode terrestre Armadillidium vulgare, Wolbachia induit la féminisation des mâles génétiques en femelles fonctionnelles entraînant des biais de sex-ratio vers les femelles dans la descendance.Pour comprendre les mécanismes impliqués dans ces deux symbioses, nous avons mis au point une nouvelle méthode de capture pour isoler l'ADN de Wolbachia et séquencer 8 souches de Wolbachia d'isopodes (F et IC). Une étude de génomique comparative a permis d'établir un premier pan-génome des bactéries du genre Wolbachia et d'identifier 2, 5 et 3 gènes présents seulement chez les souches mutualistes, féminisantes ou induisant la mort des mâles. L'expression des gènes potentiellement impliqués dans la féminisation ou le mutualisme a été étudiée au cours du développement de l'hôte. L'étude de l'interactome protéique bactérie-hôte a ensuite été initiée en utilisant comme appât des protéines bactériennes à domaines eucaryotes en vue d'identifier les cibles de Wolbachia chez l'hôte. / Bacteria of the genus Wolbachia are gram-negative alpha-proteobacteria present in many arthropods and filarial nematodes. These obligate intracellular bacteria are maternally inherited and induce a large number of phenotypes across the symbiosis continuum from mutualism to parasitism, including feminization (F), cytoplasmic incompatibility (CI) or male killing. Studying Wolbachia symbioses is therefore of particular interest in the investigation of symbiotic relationships.In Brugia malayi and other filarial nematodes, they are obligate leading to a loss of worm fertility, and eventual death upon their depletion with antibiotic. In arthropods, they rather are parasitic. In the isopod crustacean Armadillidium vulgare they cause feminization when present: genetic males develop as functional female leading to female biased sex-ratio progenies.In order to understand the molecular mechanisms of these two symbioses, we set up a new capture procedure to catch Wolbachia DNA and performed whole-genome sequencing on 8 Wolbachia strains, symbionts of isopods (F & CI). Comparative genomics led to the establishment of the Wolbachia pan-genome as well as the identification of phenotype related gene patterns. We identified 2, 5 and 3 genes that are only found in mutualist, feminizing and male killing strains, respectively. Expression of genes potentially involved in feminization and mutualism were also analyzed throughout host post-embryonic development. Host-symbiont interactome approach was then initiated by protein-protein interaction studies using bacterial proteins with eukaryote like motifs as bait in order to identify Wolbachia host targets involved in symbiosis.