Estrongiloidíase é a infecção parasitária causada pelo nematódeo Strongyloides stercoralis. Em indivíduos imunocompetentes cursa de forma crônica e assintomática na maioria dos casos; entretanto em imunocomprometidos pode assumir as formas graves como a hiperinfecção e/ou estrongiloidíase disseminada. Diante da baixa sensibilidade do diagnóstico parasitológico, vêm sendo realizadas pesquisas envolvendo o imunodiagnóstico, sobretudo voltadas para variações na obtenção das frações antigênicas heterólogas utilizando Strongyloides venezulensis como fonte de antígeno. O presente trabalho teve como objetivo aprimorar o diagnóstico sorológico da estrongiloidíase humana utilizando técnicas imunoproteômicas. Foram utilizadas amostras de fezes e soro de indivíduos imunocompetentes, divididos em três grupos de acordo com o resultado do exame parasitológico (P: positivos para S. stercoralis; OP: com outras parasitoses; N: negativos para quaisquer parasitos ou comensais) e imunocomprometidos divididos em dois grupos de acordo com o resultado do exame parasitológico (S+ID: positivos para S. stercoralis e S-ID: negativos para S. stercoralis ou qualquer parasito). As amostras de soro dos cinco grupos de pacientes foram submetidas a testes sorológicos a partir das frações antigênicas heterólogas de S. venezuelensis solúvel (TS) e de membrana (TM) utilizando as técnicas ELISA, DOT ELISA e Western-blotting. O ELISA apresentou 95% de sensibilidade em ambas frações antigênicas e 91,8% e 93,8% de especificidade para TS e TM, respectivamente. A técnica foi capaz de detectar anticorpos anti-Strongyloides em 34,4% e 28,1% em pacientes do grupo S+ID utilizando os antígenos TS e TM, respectivamente. O DOT ELISA foi executado somente com a fração TM e apresentou 95% de sensibilidade e 85,7% de especificidade detectando 28,1% de positividade no grupo S+ID. A banda de 40-35 KDa foi a única frequente em todos indivíduos do grupo P e a mais frequente em pacientes S+ID (62,5% para TS e 53,1% para TM). Géis réplicas foram realizados, excisados nas regiões de identificação proteica e submetidos a identificação de proteínas por espectrometria de massas. As proteínas mais abundantes nas frações antigênicas TS e TM foram actina e galectina. A fração TS foi purificada em coluna de afinidade para galectina, dada sua capacidade de modulação do sistema imunológico. As frações não ligada (NL) e galectina (Gal) foram utilizadas como antígenos na técnica ELISA apresentando 90% de sensibilidade e 89,8 e 75% de especificidade para NL e Gal, respectivamente. Foram positivos no ELISA 34,4% e 18,1% das amostras do grupo S+ID utilizando as frações NL e Gal, respectivamente. As técnicas sorológicas utilizando variações antigênicas constituem ferramentas alternativas importantes de diagnósticos que podem ser aplicadas à estrongiloidíase humana, sobretudo na população imunocomprometida / Strongyloidiasis is a parasitic infection caused by nematode Strongyloides stercoralis. In immunocompetent patients, it is chronic and asymptomatic in the most of cases. However, in immunocompromised patients, it can take on severe forms such as hyperinfection and / or disseminated strongyloidiasis. Due to the parasitological diagnosis\'s low sensitivity, research involving the immunoassay has been carried out, primarily focused on changes in the heterologous antigenic fractions using Strongyloides venezulensis as a source of antigen. The present work aimed to improve the serological diagnosis of human strongyloidiasis using immunoproteomic techniques. Feces and sera samples from immunocompetent patients were used and divided into three groups according to parasitological examination result (P: positive for S. stercoralis, OP: with other parasites, N: negative for any parasites or commensal). Samples from immunocompromised patients were divided into two groups according to the parasitological examination result (S+ID: positive for S. stercoralis and S-ID: negative for S. stercoralis or any parasite). Serum samples from the five groups of patients were submitted to serological tests by S. venezuelensis soluble (TS) and from membrane (TM) heterologous antigenic fractions by ELISA, DOT ELISA and Western blotting techniques. The ELISA test showed 95% sensitivity in both antigenic fractions and 91.8% and 93.8% specificity for TS and TM, respectively. This technique was able to detect anti-Strongyloides antibodies in 34.4% and 28.1% in S+ID group by using the TS and TM antigens, respectively. DOT ELISA was performed only with the TM fraction and showed 95% sensitivity and 85.7% specificity, detecting 28.1% positivity in S+ID group. The 40-35 KDa band was the only present in all P group individuals and the most frequent in S+ID patients (62.5% for TS and 53.1% for TM). Replicate gels were made, excised in the protein identification regions and submitted to protein identification by mass spectrometry. The most abundant proteins in TS and TM antigenic fractions were actin and galectin. TS antigen was purified on a galectin affinity column, due the immune modulation ability. The non-bound (NL) and galectin (Gal) fractions were used as antigens in the ELISA technique with 90% of sensitivity and 89.8 and 75% of specificity for NL and Gal, respectively. 34.4% and 18.1% of S+ID group were positive in the ELISA by NL and Gal fractions, respectively. Serological techniques using antigenic variations are an important alternative diagnostic tool and can be applied to human strongyloidiasis, especially in the immunocompromised population

A estrongiloidíase é a infecção parasitária causada pelo nematódeo Strongyloides stercoralis. O diagnóstico definitivo é realizado pela visualização de larvas, principalmente nas fezes. Porém as técnicas parasitológicas têm baixa sensibilidade. As técnicas sorológicas apresentam-se como importante alternativa diagnóstica. Pesquisas apontam para a utilização de antígenos heterólogos solúveis, principalmente de Strongyloides venezuelensis. A identificação e caracterização dos antígenos de membrana podem fornecer fonte alternativa de antígenos e assim auxiliar o desenvolvimento das técnicas imunológicas. O presente trabalho teve como objetivo a avaliação das técnicas ELISA e WB frente a diferentes frações antigênicas de larvas filarioides de S. venezuelensis. Foram utilizadas amostras de sangue e fezes de 92 indivíduos, 20 indivíduos com estrongiloidíase (grupo I), 32 indivíduos com outras parasitoses (grupo II) e 40 indivíduos negativos (grupo III) pelos métodos de Lutz, cultura em placa de ágar e Rugai. Para preparação dos antígenos foram utilizadas larvas infectantes obtidas a partir de ratos infectados experimentalmente com S. venezuelensis. Seis frações antigênicas foram preparadas: frações salinas solúveis e de membrana (PBS 0,01M pH 7,2 e SDS 1%, SS e MS; Tris-HCl 25mM pH 7,5 e CHAPS 1%, ST e MT, respectivamente) e frações alcalinas solúvel e de membrana (NaOH 0,15M e SDS 1%, SA e MA, respectivamente). Para a técnica ELISA foram utilizadas placas sensibilizadas com 10ug/mL de antígeno, soro dos indivíduos diluídos 1:200 em PBS 0,05% Tween 3% de leite (PBSTM) e o conjugado (anti IgG-humana peroxidase) em PBSTM. As amostras foram consideradas positivas quando o Índice ELISA foi maior que 1. Para a técnica de WB os soros foram diluídos 1:100 em Tris-HCl 5% de leite (TM) e o conjugado (anti IgG-humana peroxidase) em TM. Após as técnicas sorológicas foram determinadas os parâmetros de diagnóstico pela curva ROC como sensibilidade (SE), especificidade (ES), Likelihood ratio (LR) além da determinação da acurácia diagnóstica (AC) e do índice Kappa (k). A técnica ELISA destacou as frações de membrana com melhor desempenho em relação aos parêmetros diagnósticos estudados (SE 95%, ES 94,4%, AC 94,8%, LR 17,1, k 0,848). O WB revelou componentes antigênicos imunodominantes variando de 260-10kDa, mas destacam-se as frações de 40-35kDa mais frequentes em todas frações antigênicas. Pela técnica de WB, a fração ST apresentou melhor desempenho em relação aos parêmetros diagnósticos estudados (SE 100%, ES 93,1%, AC 94,5, LR 14,4,k 0,854). A utilização das frações de membrana no imunodiagnóstico da estrongiloidíase humana torna-se fonte acessível e eficaz em relação às frações purificadas, não necessitando de gastos complementares para sua obtenção / Strongyloidiasis is a parasitic infection caused by a nematode Strongyloides stercoralis. The definitive diagnosis is made by the larvae visualization in stool samples. However parasitological techniques have low sensitivity. Serological techniques became as suitable diagnostic alternative. Research indicates for the soluble heterologous antigen utilization, mainly Strongyloides venezuelensis. Identification and characterization of membrane antigen may constitute an alternative source of antigen and then assist the development of serological techniques. The aim of this study was evaluate ELISA and WB techniques behind different antigenic fractions of S. venezuelensis´ infective larvae. A total of 92 serum and stool samples was analyzed, 20 from individuals with strongyloidisis (group 1), 32 with other parasitic diseases (group 2) and 40 from individuals with negative coproparasitology (group 3) using Lutz, agar plate culture and Rugai methods. For the antigen preparation infective larvae of S. venezuelensis from experimental infected rats were employed. Six antigenic fractions were prepareted: saline soluble and from membrane fractions (0.01M PBS pH 7.2, and 1% SDS, SS and MS; 25mM Tris-HCl pH 7.5, 1% CHAPS, MT and ST, respectively) and alkaline soluble and membrane fractions (0.15 M NaOH and 1% SDS, SA and MA, respectively). For ELISA technique, plates were sensitized with 10 ug/mL of antigen, serum samples were diluted 1:200 in 0.05% Tween in PBS 3% milk (PBSTM) and conjugate (anti-human IgG peroxidase) in PBSTM. Positive samples were considered when ELISA index was greater than 1. To WB technique, serum samples were diluted 1:100 in Tris-HCl 5% milk (TM) and conjugate (anti-human IgG peroxidase) in the TM. After serological techniques diagnostics parameters were determined by ROC curve how sensitivity (SE), specificity (ES), Likelihood ratio (LR) and determination of diagnostic accuracy (AC) and Kappa (k) index. ELISA technique highlighted the membrane fractions with better performance compared to parameters diagnoses studied (95% SE, 94.4% ES, 94.8% AC, 17.1 LR, 0.848 k). The WB revealed immunodominant antigenic components ranging from 260-10kDa, but there are the fractions of 40-35kDa more frequent in all antigenic fractions. WB technique showed ST fraction better performance in relation to the diagnostic parameters (100% SE, 93.1% ES, 94.5% AC, 14.4 LR, 0.854 k). Membrane fractions in the immunodiagnosis of human strongyloidiasis become an accessible and effective source of antigens in relation to the purified fractions, requiring no additional expense to obtain it

Metazoan genomes contain thousands of protein-coding and non-coding RNA genes, most of which are differentially expressed, i.e., at different locations, at different times during development, or in response to environmental signals. Differential gene expression is achieved through complex regulatory networks that are controlled in part by two types of trans-regulators: transcription factors (TFs) and microRNAs (miRNAs). TFs bind to cis-regulatory DNA elements that are often located in or near their target genes, while microRNAs hybridize to cis-regulatory RNA elements mostly located in the 3’ untranslated region (3’UTR) of their target mRNAs. My work in the Walhout lab has centered on understanding how these trans-regulators interact with each other in the context of gene regulatory networks to coordinate gene expression at the genome-scale level. Our model organism is the free-living nematode Caenorahbditis elegans, which possess approximately 950 predicted TFs and more than 100 miRNAs Whereas much attention has focused on finding the protein-coding target genes of both miRNAs and TFs, the transcriptional networks that regulate miRNA expression remain largely unexplored. To this end, we have embarked in the task of mapping the first genome-scale miRNA regulatory network. This network contains experimentally mapped transcriptional TF=>miRNA interactions, as well as computationally predicted post-transcriptional miRNA=>TF interactions. The work presented here, along with data reported by other groups, have revealed the existence of reciprocal regulation between these two types of regulators, as well as extensive coordination in the regulation of shared target genes. Our studies have also identified common mechanisms by which miRNAs and TFs function to control gene expression and have suggested an inherent difference in the network properties of both types of regulators. Reverse genetic approaches have been extensively used to delineate the biological function of protein-coding genes. For instance, genome-wide RNAi screens have revealed critical roles for TFs in C. elegans development and physiology. However, reverse genetic approaches have not been very insightful in the case of non-coding genes: A single null mutation does not result in an easily detectable phenotype for most C. elegans miRNA genes. To help delineate the biological function of miRNAs we sought to determine when and where they are expressed. Specifically, we generated a collection of transgenic C. elegans strains, each containing a miRNA promoter::gfp (Pmir::gfp) fusion construct. The particular pattern of expression of each miRNA gene should help to identify potential genetic interactors that exhibit similar expression patterns, and to design experiments to test the phenotypes of miRNA mutants.

Made available in DSpace on 2015-03-26T13:28:10Z (GMT). No. of bitstreams: 1 texto completo.pdf: 3335039 bytes, checksum: 432a8a4734747e35f11922a30da30b08 (MD5) Previous issue date: 2012-02-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This study aimed at evaluating the disinfection process of sewage sludge from a UASB reactor by greenhouse solar drying. Seven lots of sewage sludge were monitored between February 2010 and May 2011, each lot referring to an event of sludge discharge from the UASB reactor. The monitoring period of time of the different sludge lots varied from 50 to 130 days, with sampling every week. The following variables were monitored: pH, moisture, total solids, total coliforms, Escherichia coli, total and viable helminth eggs. In addition, somatic coliphages spiking experiments were conducted in sludge samples dried in the same field greenhouse, as well as in samples dried in a laboratory incubator kept under controlled temperature. A high sludge dewatering during the drying treatment in the greenhouse was observed, with moisture and total solids contents reaching levels, respectively, below 10% and above 90% when the sludge was subjected to periods of treatment longer than 75 days. Microbial decay during sludge drying was variable, but effective. Exponential decay was quite clear for total coliforms and Escherichia coli, but not so much for total and viable helminth eggs. Regarding the bacterial removal, an influence of the temperature was noticed, with higher decreasing of total coliforms and E.coli in the warmer periods. E.coli concentrations below 10³ MPN/gTS (the CONAMA Resolution 375/2006 standard for Class A biosolids) were obtained with more than 60 days of treatment in the greenhouse. Calculated values of Kb 20 for E.coli varied widely, between 0.004 and 0.142 d-1. Wide variations in the decay of helminth eggs were also observed, with calculated values of Koh 20 for total eggs ranging between 0.008 and 0.035 d-1. Concentrations of viable helminth eggs were always low, requiring only 15 days to reach counts below 0.25 eggs/gTS (the CONAMA Resolution 375/2006 standard for Class A biosolids). The results of the spiking experiments conducted in the greenhouse also showed intense exponential decay of somatic coliphages, with high Kcs 20 values ranging between 0.187 and 0.248 d-1. In contrast, in the spiking experiments performed in laboratory under conditions of controlled temperature (30ºC) the decay was much less intense, with Kcs 20 values between 0.028 and 0.078 d-1. Finally, this work presents a model for estimating the microbiological quality of biosolids treated by greenhouse drying based on the microbiological quality of raw sewage. Using stochastic modeling (taking into account variations on the input variables) it was estimated that, with 60 days of sludge drying in the greenhouse, the probability of reaching the bacteriological quality established in COMANA Resolution 375/2006 for biosolids classes A and B (103 and 106 E.coli/gST, respectively) was 66.2% and 97% respectively. In the case of helminth eggs these chances were respectively 64.3% and 99.8%. That is, the results suggest high stability and reliability of the greenhouse drying treatment process of the UASB sewage sludge. / O presente trabalho teve como objetivo o acompanhamento da higienização de lodo de esgoto proveniente de reator UASB por secagem em estufa. Foram monitorados sete lotes de lodo, entre fevereiro de 2010 e maio de 2011, sendo cada lote referente a um evento de descarte de lodo do reator UASB. O tempo de acompanhamento dos vários lotes de lodo variou de 50 a 130 dias, com realização de coletas semanais. Foram monitoradas as seguintes variáveis: pH, umidade, sólidos totais, coliformes totais, Escherichia coli e ovos totais e viáveis de helmintos. Foram ainda realizados experimentos de inoculação de colifagos somáticos em lodos submetidos à secagem na mesma estufa de campo e também sob condições de temperatura controlada (estufa laboratorial). Foi observada elevada desidratação do lodo durante a secagem em estufa, sendo obtidos teores de umidade menores que 10% e de sólidos totais acima de 90% quando os lotes foram submetidos a períodos de tratamento superiores a 75 dias. O decaimento microbiano durante a secagem do lodo se mostrou variável, mas eficiente. Decaimento exponencial foi bastante nítido para coliformes totais e Escherichia coli, porém essa tendência não foi tão clara para os dados de ovos viáveis e totais de helmintos. Para a remoção das bactérias, foi possível perceber influência da temperatura (sazonalidade), com quedas das concentrações de coliformes totais e E.coli mais acentuada nos períodos mais quentes. Valores de E.coli abaixo de 10³ NMP/gST, limite estabelecido pela Resolução CONAMA 375/2006 para biossólido Classe A, foram obtidos com tempos de secagem acima de 60 dias. Os valores de Kb 20 calculados para E.coli foram bastante variáveis, entre 0,004 e 0,142 d-1. Variações amplas também foram observadas no decaimento de ovos de helmintos, com valores de Koh 20 calculados para ovos totais entre 0,008 e 0,035 d-1. As concentrações de ovos viáveis de helmintos foram sempre baixas, sendo necessários apenas 15 dias para se obter contagens abaixo de 0,25 ovos/gST (biossólido Classe A de acordo com a Resolução CONAMA 375/2006). Os resultados dos experimentos de campo de inoculação de colifagos somáticos também mostraram intenso decaimento exponencial para esse bacteriófago em estufa, com valores de Kcs 20 elevados e variando entre 0,187 e 0,248 d-1. Em contrapartida, nos experimentos de inoculação realizados em laboratório sob condições temperatura controlada (30ºC) o decaimento foi bem menos intenso, com valores de Kcs 20 entre 0,028 e 0,078 d-1. Por fim, o trabalho apresenta um modelo de estimativa da qualidade microbiológica de biossólidos tratados por secagem em estufa a partir da qualidade microbiológica do esgoto bruto. Por meio de modelagem estocástica (que leva em consideração variações em torno das variáveis de entrada) estimou-se que, com 60 dias de secagem em estufa, a probabilidade de alcançar a qualidade bacteriológica preconizada para as Classes A e B na Resolução CONAMA 375/2006 (103 e 106 E.coli/gST, respectivamente) seria de 66,2% e 97%, respectivamente. No caso dos ovos de helmintos essas chances seriam, respectivamente, de 64,3% e 99,8%. Ou seja, os resultados sugerem estabilidade e confiabilidade consideráveis do processo de tratamento por secagem em estufa do lodo de reator.

Made available in DSpace on 2014-12-17T15:03:17Z (GMT). No. of bitstreams: 1 FernandaLC.pdf: 619525 bytes, checksum: b172a729547f262e355961911646e833 (MD5) Previous issue date: 2007-03-19 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The technology of anaerobic reactors for sanitary wastewater treatment has been extensively developed in Brazil, and today it is practically consolidated. They present several advantages, such as low construction and operating costs, and low sludge production, the anaerobic reactors are an attractive alternative to minimize problematic lack of basic sanitation in urban areas, and also of the rural areas. The anaerobic filters have been widely used in Brazil. It produces an effluent with low concentration of organic matter and solids suspended, besides conserving the nutrients, therefore, it is good for use in irrigation, but the practice must be associated with knowledge of the pathogens presence. The main objective of this study was to evaluate the efficiency of anaerobic filters in removal faecal coliforms and helminth eggs, and to verify if the effluent can be used for agricultural purposes, according to the World Organization of Health (WHO, 1989). The protocol used to enumerate helminths eggs was the modified Bailenger method, (Ayres and Mara, 1996) recommended by WHO for evaluation of raw effluent and treated effluent. The membrane filtration method was utilized to determine the concentrations of faecal coliforms. Three different systems of sewer treatment composed by anaerobic filters were analyzed. The results, in a general analysis, showed that all the researched systems reached a larger removal than 93% to helminth eggs, resulting in an effluent with smaller average than 1 egg/L. One of these systems, Sistema RN, reached a larger removal than 99%, confirming the good performance of the anaerobic filters in removal helminths eggs. Even with low concentrations of eggs in the influent, the filters were capable to remove this parameter efficiently. About faecal coliforms, it was observed for all the researched systems an effluent with 106 CFU/100mL. The high concentrations to faecal coliforms in the effluent just allow reuse for restricted irrigation, in agreement with the guidelines of WHO. Although the researched systems have not removed faecal coliforms efficiently, the results indicated a good efficiency of the anaerobic filters in removal helminth eggs / A tecnologia de reatores anaer?bios para o tratamento de esgoto sanit?rio vem sendo extensivamente desenvolvida no Brasil, e hoje encontra-se praticamente consolidada. Apresentando diversas vantagens, como baixos custos de constru??o e opera??o, e baixa produ??o de lodo, os reatores anaer?bios s?o uma alternativa bastante atrativa para a mitiga??o dos problemas de saneamento b?sico urbano, e tamb?m das ?reas rurais. Os filtros anaer?bios v?m sendo bastante aplicados no Brasil. Sua utiliza??o produz um efluente com baixa concentra??o de mat?ria org?nica e s?lidos suspensos, al?m de conservar os nutrientes, sendo por isso muito bom para irriga??o com fins produtivos, desde que sejam resguardados os cuidados com a presen?a de organismos patog?nicos. Este trabalho teve por objetivo avaliar a efici?ncia de filtros anaer?bios na remo??o de coliformes fecais e ovos de helmintos, e verificar se os mesmos atendem ?s recomenda??es sanit?rias para reuso em irriga??o, segundo a Organiza??o Mundial de Sa?de - OMS (WHO, 1989). Para enumera??o dos ovos de helmintos foi utilizada a t?cnica de Bailenger modificada (Ayres e Mara, 1996), que deu origem ? metodologia atualmente recomendada pela OMS para avalia??o de ?guas residu?rias brutas e tratadas. Para determina??o de coliformes fecais foi utilizado o m?todo da membrana filtrante. Foram analisados tr?s diferentes sistemas de tratamento de esgoto compostos por filtros anaer?bios. Numa an?lise geral dos resultados, observou-se que todos os sistemas pesquisados atingiram efici?ncia maior que 93% para remo??o de ovos de helmintos, resultando em um efluente final com valor m?dio menor que 1 ovo/L. Um dos sistemas, o Sistema RN, alcan?ou uma remo??o maior que 99%, confirmando o bom desempenho dos filtros anaer?bios na remo??o de ovos de helmintos. Mesmo com baixas concentra??es de ovos no afluente, os filtros foram capazes de remover eficientemente este par?metro. Em rela??o ? contagem de coliformes fecais, foi observado, para todos os sistemas pesquisados um efluente final com cerca de 106 UFC/100mL. As altas concentra??es de coliformes fecais no efluente dos filtros permitem a reutiliza??o apenas para irriga??o restrita, de acordo com as diretrizes da OMS. Apesar dos sistemas pesquisados n?o removerem eficazmente coliformes fecais, os resultados encontrados no presente estudo indicaram uma boa efici?ncia dos filtros anaer?bios na remo??o de ovos de helmintos

No description available.

Migration within the body of an infected host is one of the most important parts in the life cycle of flukes, including schistosomes. Migration of avian and mammalian visceral schistosomes has been a quite well studied topic (Haas a Haeberlein, 2009), which became more attractive after the discovery of T. regenti, an avian schistosome which is able to migrate through the nervous tissues of infected birds and mammals as well. Migration of T. regenti and T. szidati schistosomula within the definitive (duck) and the accidental (mouse) hosts is the main topic of the diploma thesis. This work continues with the research of K. Blažová (Faculty of Science, Charles University in Prague) who studied migration of T. regenti within the definitive hosts infected perorally with cercariae or hepatopancreases of the infected intermediate snail, Radix lagotis (unpblished). She proved that T. regenti schistosomula are able to use the central nervous system for migration to the nasal mucosa of infected birds. In our work, we focused on the early phase of migration within the perorally infected birds and mice. Invasion of esophagus by T. regenti cercariae in vitro is not conditioned by secretion of glandular products, including cathepsin B2 of T. regenti (TrCB2). Activity of TrCB2 against mucins, the main components...

In this study, we examined the immunomodulatory effect of excretory/secretory products, crude adult extracts and crude larvae extracts from Hymenolepis diminuta on the intestinal epithelilal cell line from a rat. For determination of the immunomodulation effect of all H. diminuta extracts was used relative gene expression of TNFa, IL-17re and IL-33 from epithelial cells and it was tested using real-time PCR. Our result showed that excretory/secretory products had the strongest antiinflammatory effect on the epithelial cells. We assume that crude adult extracts play an important role in increase of gene expression of IL-33 and also in the immunomodulatory ability of H. diminuta in the host organism.

In a 4-cell C. elegans embryo the ventral blastomere EMS requires polarity signaling from its posterior sister cell, P2. This signaling event enables EMS to orient its division spindle along the anterior-posterior (A/P) axis and to specify the endoderm fate of its posterior daughter cell, E. Wnt pathway components have been implicated in mediating P2/EMS signaling. However, no single mutants or various mutant combinations of the Wnt pathway components disrupt EMS polarity completely. Here we describe the identification of a pathway that is defined by two tyrosine kinase related proteins, SRC-1 and MES-1, which function in parallel with Wnt signaling to specify endoderm and to orient the division axis of EMS. We show that SRC-1, a C. elegans homolog of c-Src, functions downstream of MES-1 to specifically enhance phosphotyrosine accumulation at the P2/EMS junction in order to control cell fate and mitotic spindle orientation in both the P2 and EMS cells. In the canonical Wnt pathway, GSK-3 is conserved across species and acts as a negative regulator. However, in C. elegans we find that GSK-3 functions in a positive manner and in parallel with other components in the Wnt pathway to specify endoderm during embryogenesis. In addition, we also show that GSK-3 regulates C. elegans germline development, a function of GSK-3 that is not associated with Wnt signaling. It is required for the differentiation of somatic gonadal cells as well as the regulation of meiotic cell cycle in germ cells. Our results indicate that GSK-3 modulates multiple signaling pathways to regulate both embryogenesis and germline development in C. elegans.

Pathogens and parasites are increasingly recognized as important components within host populations, communities, and ecosystems. Parasite contributions to ecosystem function most likely manifest as density-mediated impacts of parasites on their hosts, the direct contributions of parasite biomass to a system, and via parasite-induced changes in host behavior and physiology (trait-mediated impacts). Here, a framework was constructed that can be used to conceptualize parasite contributions to ecosystem function (Chapter 1). Then the influence of parasite attack on host movement was explored to further evince the mechanistic underpinnings of trait-mediated parasite impacts (Chapter 2). Additionally, mesocosms were created across a gradient of parasitism to examine how these mechanisms are likely to unfold at larger biological scales (Chapter 3). Lastly, a series of differential equations was created to model host-parasite-ecosystem interactions and generate theoretical predictions about how and when parasites are likely to influence ecosystem processes (Chapter 4). Parasites have many characteristics of ecosystem engineers, but their role has historically been ignored. These studies begin to explore the role that parasitism may have as one of the drivers of ecosystem processes.