Дисертації з теми "Kinetoplasts"
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Dewar, Caroline E. "What do kinetoplastids need a kinetoplast for? : life cycle progression of Trypanosoma brucei in the presence and absence of mitochondrial DNA." Thesis, University of Edinburgh, 2016. http://hdl.handle.net/1842/17943.
Повний текст джерелаAlkhaldi, Abdulsalam Abdulhadi. "Drug development against kinetoplastid parasites." Thesis, University of Glasgow, 2012. http://theses.gla.ac.uk/3637/.
Повний текст джерелаBrewster, S. "Analysis of the kinetoplast DNA of Leishmania panamensis." Thesis, University of Cambridge, 1999. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.596899.
Повний текст джерелаTimm, Jennifer. "Structure-function studies of kinetoplastid proteins." Thesis, University of York, 2014. http://etheses.whiterose.ac.uk/7454/.
Повний текст джерелаRodriguez, Noris Marcela. "PCR diagnosis of Leishmania using nuclear and kinetoplast primers." Thesis, University of Cambridge, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.627544.
Повний текст джерелаTiesen, K. L. "Studies on monogenetic kinetoplastid flagellates of hemiptera." Thesis, University of Salford, 1986. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.376881.
Повний текст джерелаAli, Juma Ahmed Mohmed. "Pyrimidine salvage and metabolism in kinetoplastid parasites." Thesis, University of Glasgow, 2013. http://theses.gla.ac.uk/4664/.
Повний текст джерелаHitchcock, Robert Arthur. "Epigenetic control of the kinetoplastid spliced leader RNA." Diss., Restricted to subscribing institutions, 2009. http://proquest.umi.com/pqdweb?did=1998392041&sid=1&Fmt=2&clientId=1564&RQT=309&VName=PQD.
Повний текст джерелаBlom, Daniël. "Mechanism and evolution of RNA editing in kinetoplastids." [S.l. : Amsterdam : s.n.] ; Universiteit van Amsterdam [Host], 2000. http://dare.uva.nl/document/83225.
Повний текст джерелаGaston, Kirk W. "Editing and Modification of Threonyl-tRNAs in Kinetoplastids." The Ohio State University, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=osu1248965851.
Повний текст джерелаSmyth, Audra Jayne. "Sequence variation in Leishmania kinetoplast DNA minicircles and diagnosis of leishmaniasis." Thesis, University of Cambridge, 1992. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.386238.
Повний текст джерелаPalazzo, Setareh Seraji. "Kinetoplastid RNA editing ligases : functional analysis and editosome association /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/9303.
Повний текст джерелаIndranil, Mukherjee. "Ecology of kinetoplastid flagellates in freshwater deep lakes of Japan." 京都大学 (Kyoto University), 2016. http://hdl.handle.net/2433/217135.
Повний текст джерелаSample, Paul J. "Characterization of two unique pathways for wyosine biosynthesis in Kinetoplastids." The Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1397733440.
Повний текст джерелаCooper, Sinclair. "Complexity and dynamics of kinetoplast DNA in the sleeping sickness parasite Trypanosoma brucei." Thesis, University of Edinburgh, 2017. http://hdl.handle.net/1842/28819.
Повний текст джерелаWebster, Lauren. "Target identification and mechanism of action studies in folate metabolism in kinetoplastids." Thesis, University of Dundee, 2014. https://discovery.dundee.ac.uk/en/studentTheses/1b8c36a5-af4d-4085-99e1-3e09e0a9080a.
Повний текст джерелаRoberts, Matthew D. "Lipidomic investigations into the phospholipid content and metabolism of various kinetoplastids." Thesis, University of St Andrews, 2017. http://hdl.handle.net/10023/16983.
Повний текст джерелаBrito, Querido Jailson Fernando. "Structural study of mRNA translation in kinetoplastids by Cryo-electron microscopy." Thesis, Strasbourg, 2017. http://www.theses.fr/2017STRAJ108.
Повний текст джерелаKinetoplastid is a group of flagellated protozoans, which threatens more than 400 million people world-wide. They possess unusual large rRNA expansion segments (ES) in the 40S, such as ES6S, ES7S and ES9S and their location suggests an involvement in the initiation process. Furthermore, all mature mRNAs possess a conserved 5’ spliced-leader. Here, we purified from T. cruzi cell lysates native initiation complexes and native 40S subunits that we then analysed by cryo-EM. The structure of native initiation complexes reveals several kinetoplastid-specific aspects of translation, such as an intricate interaction network between eIF3 and ES6S and ES7S. Furthermore, it reveals the role of DDX60 in translation initiation in kinetoplastids. The structure of native 40S subunits reveals the existence of an uncharacterized factor (termed ηF) bound at platform of the 40S. The binding site of ηF suggests a role in translational control. Moreover, we reported a novel kinetoplastid-specific ribosomal (r-) protein (KSRP) bound to the 40S subunit. Our work represents the first structural characterization of kinetoplastids-specific aspects of translation initiation
Venkatesh, Divya. "Evolution of vesicular transport in kinetoplastids : dynamics and novel gene products." Thesis, University of Cambridge, 2016. https://www.repository.cam.ac.uk/handle/1810/269276.
Повний текст джерелаWang, Bingbing. "Kinetoplastid RNA editing : in vitro RNA editing and functional analysis of the editosome /." Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/9307.
Повний текст джерелаRodrigues, Elisandra Márcia. "Estudos moleculares das enzimas envolvidas na biossíntese de selenocisteína em Trypanosoma brucei e Leishmania major." Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/76/76132/tde-05092008-090707/.
Повний текст джерелаOne of the main biological forms of the selenium incorporation is the amino acid form named selenocysteine (Sec, U), which is incorporated co-translationally at the emerging new polypeptide in the specific positions at the UGA codon, that is usually recognized as stop codon. The incorporation of the selenocysteine in E.coli is already solved with the involvement of the genes that codify to selenocysteine synthase (SELA), seryl tRNA synthetase (SerRS), a specific tRNASec (SELC), selenophosphate synthetase (SELD) and a selenocysteine-specific translation elongation factor (SELB). However, in eukarya there is no SELA homologue, but there are evidences about the requirement of the two enzymatic steps that replace the activity performed by SELA, the fosforilation of the serine followed by selenocysteylation through the phosphoseryl-tRNASec kinase (PSTK) and Sep-tRNA:Sec-tRNA synthase (SepSecS) enzymes, respectively. Currently, the selenocysteine synthesis and its incorporation is more studied in many organisms, but less explored in Kinetoplastid. Subsequently, the molecular studies were done with the enzymes involved in this pathway, especially in Trypanosoma brucei and Leishmania major. The SECIS element was identified in the region 3´ of the mRNA, that acts in the recognition of the UGA codon positioned within a gene\'s open reading frame on the insertion of the selenocysteine in Leishmania major and Leishmania infantum; the incorporation of 75Se into Leishmania proteins, the occurrence of selenocysteine-tRNASec in both Leishmania and Trypanosoma; in addition, the finding of all genes necessary for selenocysteine synthesis, such as: SELB, SELD, PSTK, and SECp43. Clones were obtained from the selB and selD genes in the pET28a(+) expression vector and the enzymes were expressed in Escherichia coli BL21 (DE3). The recombinant SELD protein was purified by affinity chromatography and its pI and molecular mass were determined using: isoeletrophocusing electrophoresis and native gel. The proteins SELB, SELD, SECp43, and sery-tRNA synhetase were immune located in the cytoplasm in T. brucei native cells. A new methodology \"PTP tagging\" was utilized for protein interaction studies by using target proteins SECp43, SELB and PSTK to search new tagged proteins in selenocysteine T. brucei synthesis. Future molecular and structural investigation of the enzymes involved in Kinetoplastida selenocysteine biosynthesis will provide relevant information for understanding of the synthesis of this amino acid as well as the development of the specific inhibitors, focusing the treatment of the disease caused by Trypanosoma brucei e Leishmania major parasites.
Evangelista, Jaqueline Pesciutti. "Estudos moleculares das enzimas Fosfoseril-tRNA sintease de Trypanosoma brucei e Leishmania major e Seril-tRNA sintease de Trypanosoma brucei." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/76/76132/tde-24072009-112648/.
Повний текст джерелаThe translation process study is central role in the cellular metabolism and attracts the interest of several groups, in particular, the study of the 21º amino acid, the selenocystein. The selenocystein incorporation pathway was described in Escherichia coli and recently in eukaryotes. The first step of this pathway is initiated by Seryl-tRNA Synthetase that aminoacilates the Ser-tRNASec (SELC) with serine. In E. coli, the second step is performed by the Sec-synthase (SELA) that removes hydroxyl group of the serine side chain, forming an aminoacrylil intermediary. This serves as an acceptor of seleno phosphate generating the selenocystein. In eukaryotes, the similar process is performed by PSTK and SepSecS, which phosphorylate serine and adds the selenium, respectively. Interested in this pathway, we performed initial molecular studies of the Phosphoseryl-tRNA synthetase of Trypanosoma brucei and Leishmania major and Seryl-tRNA synthetase of Trypanosoma brucei. The gene that encodes T. brucei Phosphoseryl-tRNA synthetase was obtained with several mutations. However, the gene encoding the T. brucei Phosphoseryl-tRNA synthetase was cloned into pET28 vector and the enzyme was expressed in E. coli cells, however at low amounts hampering the intended experiments. Therefore we initiated the investigation of the enzyme involved in the first step of this pathway, the Seryl-tRNA Synthetase from T. brucei. The enzyme was already cloned and expressing in the soluble fraction of E. coli. The recombinant protein was purified using 60% ammonium sulfate precipitation, hydrophobic and nickel affinity chromatography. Native gel experiments, DLS and anisotropy fluorescence was performed and allowed to conclude that, after purification, the enzyme remains stable and free of aggregation, with a hydrodynamic radius of 4.32 nm, molecular weight of 110kDa. Above 150nM protein its entirely in the dimeric form. Information about Ser-tRNASec binding can now be obtained from the technique of anisotropy seen that initial experiments with SerRS add Ser-tRNASec be shown to be promising.
Bachmaier, Sabine [Verfasser], and Michael [Akademischer Betreuer] Boshart. "Evolutionary repurposing of cAMP and PKA signaling pathways in kinetoplastids / Sabine Bachmaier ; Betreuer: Michael Boshart." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2015. http://d-nb.info/1177682095/34.
Повний текст джерелаKable, Moffett Lee. "Kinetoplastid RNA editing : analysis of the mechanism of guide RNA directed uridylate insertion into precursor messenger RNA /." Thesis, Connect to this title online; UW restricted, 1998. http://hdl.handle.net/1773/9295.
Повний текст джерелаSantos, Elisangela Madureira dos. "Avaliação da associação entre expressão da proteína “Kinetoplastid Membrane Protein-11” (KMP-11) e virulência de Leishmania amazonensis." reponame:Repositório Institucional da FIOCRUZ, 2011. https://www.arca.fiocruz.br/handle/icict/4246.
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Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.
As leishmanioses formam um grupo de doenças que afetam 350 milhões de pessoas atualmente, atingindo 88 países em todo o mundo com uma estimativa de 1-2 milhões de novos casos por ano. O desfecho da infecção causada por Leishmania depende tanto de fatores do patógeno como do hospedeiro, embora a virulência de Leishmania possa ser modulada por fatores ambientais e genéticos relacionados aos hospedeiros mamíferos e vetores, os determinantes moleculares são elementos-chave no estabelecimento da infecção, ou seja, são os fatores que determinam a virulência. A KMP-11 é uma glicoproteína que está presente em todos os cinetoplastideos. O presente estudo avaliou a expressão do gene de KMP-11 de L. amazonensis ao nível de RNA e ao nível de proteína, durante passagens sucessivas de promastigotas de fase estacionária através de cultivo in vitro, investigando se há associação entre a sua expressão e a virulência dos parasitos. A avaliação da virulência dos parasitos mantidos em cultura foi realizada através do acompanhamento da evolução da infecção experimental murina (modelo in vivo) durante um período de dez semanas, juntamente com a observação do surgimento de ulceração. A quantificação da carga parasitária foi realizada nos linfonodos drenantes das lesões, através da através da técnica de diluição limitante A avaliação também foi realizada pela infecção de macrófagos murinos (modelo in vitro). Os resultados de medição de pata foram analisados pelo teste não paramétrico ANOVA 2 fatores, seguido do pós-teste de Bonferroni. Além disso, foi realizada a determinação da proporção de metacíclicas nos promastigotas de fase estacionária mantidos em cultura, através da técnica de lise pelo complemento. Nossos resultados mostraram que há um decréscimo da virulência dos promastigotas de fase estacionária ao longo do número de passagens, pois os camundongos infectados com as passagens iniciais desenvolveram lesões maiores do que aqueles com os promastigotas mantidos em cultura por mais tempo. Quanto ao surgimento de ulcerações, na 10a semana pós-infecção, todos os animais infectados com promastigotas de 1a passagem apresentavam lesões ulceradas, enquanto que nenhum dos camundongos infectados com promastigotas de 20a passagem apresentava lesão ulcerada. Na infecção in vitro, a carga parasitária nos macrófagos testados diminuiu em função do número das subculturas, o que foi demonstrado através do decréscimo da porcentagem média de macrófagos infectados e pela quantidade média de amastigotas a cada 100 macrófagos infectados. A quantificação da carga parasitária foi realizada nos linfonodos drenantes da lesão dos camundongos infectados, confirmando a diminuição da virulência dos promastigotas. A quantificação da proporção de promastigotas metacíclicas demonstrou que a porcentagem diminui ao longo do tempo de subcultivo. A avaliação da expressão de KMP-11 na superfície de promastigotas por citometria de fluxo demonstrou um decréscimo na expressão da proteína proporcional ao número de subculturas. Verificou-se, portanto, uma associação entre a expressão da proteína KMP-11 e a virulência de promastigotas de L. amazonensis. Os resultados dos ensaios de PCR em tempo real demonstraram que não há diferença estatisticamente significativa na quantidade de transcritos do gene da proteína KMP-11 entre as passagens analisadas. Entretanto, a perda da virulência associada com a diminuição da expressão da proteína KMP-11 indica que esta molécula possua uma função na infectividade dos promastigotas de Leishmania amazonensis, atuando possivelmente como um fator de virulência.
The leishmaniases are a group of diseases that currently, affects 350 million people reaching 88 countries throughout world with an estimated incidence of 1-2 million new cases per year. The outcome of the infection caused by Leishmania depends on factors from the pathogen and from the host. Although Leishmania virulence can be modulated by environmental and genetic factors related to mammalian hosts and vectors, molecular determinants are key elements in the establishment of infection and for the determination, of virulence. KMP-11 is a glycoprotein which is present in all kinetoplastids. This study evaluated the gene expression of KMP-11 in L. amazonensis at RNA level and at protein level, during successive passages of in vitro culture, investigating a possible correlation between KMP-11 expression and virulence of parasites. The evaluation of the virulence of cultured parasites was performed by monitoring of the progression of the lesions in experimental murine infection (in vivo model) during ten weeks, along with the emergence of cutaneous ulcers. The quantification of parasite load was performed on draining lymph nodes using the limiting dilution analysis. The assessment of parasite virulence was also performed by the infection of murine macrophages (in vitro model). The paw measurement results were analyzed by nonparametric test ANOVA 2 way, followed by Bonferroni post-test. Furthermore, the metacyclic promastigotes proportion in stationary growth phase from cultures with different numbers of passages was evaluated by complement lysis. Our results showed a decrease in promastigotes of stationary phase virulence that correlated with the increase of the number of passages, as mice infected with the early passages developed larger lesions than those infected with promastigotes cultured for longer periods and higher numbers of passages. Concerning the development of ulcers, at 10th week post-infection, all animals infected with promastigotes of first passage presented ulcerated lesions, whereas none of the mice infected mice with promastigotes of the 20th passage showed an ulcerated lesion. Analyzing the in vitro infection, the parasite burden in macrophages decreased with the number of subcultures, as demonstrated by the decrease in the percentage of infected macrophages and in the number of amastigotes per 100 infected macrophages. The quantification of parasites in draining lymph nodes of the infected mice confirmed the decrease in the virulence of promastigotes from cultures with more passages. The estimation of the metacyclic promastigote proportions showed that the percentages decline through the time of subculture. The evaluation of KMP-11 expression on the surface of promastigotes by flow cytometry showed a decrease in protein expression proportional to the number of subcultures. Therefore, there was an association between the expression of KMP-11 protein and the virulence of L. amazonensis promastigotes. The results of real-time PCR assays showed that there is no statistically significant difference in the amount of gene transcripts of KMP-11 protein in the analyzed passages. However, the loss of virulence associated with decreased protein expression of KMP-11 indicates that this molecule may have a role in promastigotes infectivity of Leishmania amazonensis, possibly acting as a virulence factor.
Perez, Arina Marina 1982. "Caracterização funcional da proteína LaRbp38 nos telômeros e no cinetoplasto de Leishmania spp." [s.n.], 2013. http://repositorio.unicamp.br/jspui/handle/REPOSIP/317094.
Повний текст джерелаTese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia
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Resumo: LaRbp38 e uma proteína exclusiva de protozoários tripanosomatideos, entre os quais está os agentes etiológicos da leishmaniose, uma doença endêmica presente em diversas regiões do Brasil. LaRbp38 e codificada por um gene nuclear, que parece exercer diferentes funções nas maquinarias de replicação nuclear e mitocondrial. Foi primeiramente descrita como proteína estabilizadora de RNA mitocondrial e parece estar envolvida com a replicação de DNA mitocondrial. Em Leishmania, LaRbp38 também interage in vivo com DNA mitocondrial, com sequencias ricas em GT e com DNA telomerico simples e dupla fita. Nesta tese mostramos estudos que nos levaram a caracterizar novas propriedades estruturais e biológicas desta proteína. Na primeira parte da tese mostramos, que a LaRbp38 inteira e mutantes truncados da proteína são capazes de interagir com diferentes tipos de DNAs: DNA simples e dupla fitas telomericos e kDNA, porem com diferentes afinidades. Desta forma, foi possível mapear a vizinhança de um domínio de interação desta proteína aos diferentes tipos de DNA (DBD). Como este domínio não compartilha similaridade estrutural com nenhum domínio descrito em outras proteínas, isto sugere que este pode ser um novo domínio presente exclusivamente em tripanosomatideos. Estes resultados estão compilados no artigo entitulado: "Mapping the boundaries of the DNA-binding domain of Leishmania amzonensis Rbp38 (LaRbp38)". Na segunda parte da tese, mostramos a localização subcelular da proteína e como ela e capaz de translocar por diferentes compartimentos celulares utilizando um sinal de localização mitocondrial presente no N-terminal e um sinal de localização nuclear, presente no Cterminal da proteína. Embora a proteína esteja presente de forma mais abundante no cinetoplasto, e possível visualizá-la no núcleo quando o ciclo celular do parasita e sincronizado ou quando este e submetido a um estresse genotoxico. Baseado nesses achados também foram realizados ensaios de interação proteina-proteina, onde foi possível determinar a interação entre LaRbp38 e a proteína importina ?, uma proteína que esta diretamente ligada ao transporte de proteínas ao núcleo via NLS. Estes resultados também foram compilados em um artigo, que esta em fase de preparação, entitulado: "The protein LaRbp38 translocates between the nucleus and the kinetoplast in Leishmania (L.) amazonensis promastigotes". Outro estudo que realizamos para compreender a função da LaRbp38, o qual também esta na forma de um artigo: "LaRbp38 can form part of a shelterin-like complex in L.amazonensis telomeres", mostramos evidencias sobre a interação entre as proteínas LaRbp38 e a LaTRF de L.amazonensis. Aqui, uma analise in silico pela busca de motivos conservados em LaRbp38, nos levou a descobrir que esta proteína contem um motivo do tipo TRFH docking encontrado em proteínas telomericas que interagem com as proteínas da família das TRFs no complexo shelterina de vertebrados e mamíferos (ex: TIN2, PINX1 e APOLLO). Juntas, as TRFs e suas interatoras tem a função na manutenção dos telomeros. Sendo assim, utilizando ensaios de interação proteina-proteina, conseguimos mostrar que LaRbp38 interage fisicamente com a LaTRF, usando um motivo TRFH docking diferente daquele que foi primeiramente encontrado in silico. Nossos resultados mostram que a LaRbp38 interage com a LaTRF usando o motivo ALKTL, que compartilha similaridade de sequencia, com motivos descritos em proteínas interatoras de TRFs e bastante conservado entre as Rbp38 de tripanosomatideos. Estes resultados podem indicar que a LaRbp38 cumpre função análoga a uma das proteínas de vertebrados descritas como interatoras de TRFs, a proteína TIN2, que a exemplo de LaRbp38, também tem função nas mitocôndrias
Abstract: LaRbp38 is a trypanosomatid protein found exclusively in these protozoa, among which are the etiological agents of leishmaniasis, an endemic disease present in several regions of Brazil. LaRbp38 is a protein encoded by a nuclear gene, which probably plays different roles in both mitochondrial and nuclear replication machineries. It was first described as a mitochondrial RNA stabilizing protein involved in the replication of mitochondrial DNA. In Leishmania, LaRbp38 also interacts in vivo with mitochondrial DNA, GT-rich sequences and single- and double-stranded telomeric DNA. Here we show the results that led us to characterize some new biological and structural features of this protein. In the first part of the thesis we show that the entire LaRbp38 and its truncated mutants are able to interacts with different GT-rich DNAs and were possible to map the boundearies of a DNA-binding domain (DBD). This domain doesn't share any sequence or structural similarities with the domains described in other proteins suggesting that it could be a new domain present exclusively in trypanosomatids. These results are compiled in the article entitled: "Mapping the boundaries of the DNA-binding domain of Leishmania amzonensis Rbp38 (LaRbp38)." The second part of the thesis shows the subcellular localization of the protein and how it is able to translocate to different cellular compartments using an N-terminal mitochondrial localization signal (MLS) and a nuclear localization signal (NLS) present in the C-terminus of the protein. Although the protein is seem more abundantly in the mitochondria associated with kinetoplast DNA, its nuclear localization seems to be cell cycle dependent and enhanced at the end of S phase or when parasites are subjected to genotoxic stress. In order to confirm that the protein is able to translocate to the nucleus, we used different in silico approaches. The results strongly suggest the existence of a non-classical and also non-bipartite NLS at the C-terminus of LaRbp38. Based on these findings we did protein-protein interaction assays and verified that LaRbp38 can associate in vitro with importin ?, which is directly linked to protein transport to the nucleus via a NLS. These results were also compiled in an article, which is in preparation, entitled: The LaRbp38 protein translocates between the nucleus and the kinetoplast in Leishmania amazonensis promastigotes. Another study that was carried out and present in the third part of the thesis shows evidence about the interaction between LaRbp38 and the telomeric L.amazonensis LaTRF protein. These results are presented as an article entitled: "LaRbp38 can form part of a shelterina-like complex in L.amazonensis telomeres," Here, an in silico analysis search for conserved motifs in LaRbp38, showed that this protein contains a motif, the TRFH-docking-like typically found in proteins that associate with the TRF paralogue proteins (TRF1 and TRF2) in the shelterin complex of vertebrates and mammallian telomeres (eg.TIN2, PINX1 and APOLLO). TRFs and their interactors work together to regulate the dynamics of telomeric chromatin and telomere length maintenance. By using protein-protein interaction assays we show that LaRbp38 physically interacts with LaTRF. This interaction, however, seems to occurs via a new TRFH docking motif, which is different from the conserved core motif [FY]xLxP. This new TRFH-docking-like motif (ALKTL) aligns and share similarities with the TRH-docking motif described in the shelterin protein TIN2. This motif is also very conserved among the Rbp38 orthologues of other trypanosomatids. Curiously, TIN2 is a telomeric protein that shows nuclear and mitochondrial localization
Doutorado
Genetica de Microorganismos
Doutora em Genética e Biologia Molecular
Panethymitaki, Chrysoula. "Structure-function studies of kinetoplastid myristoylCoA : protein N-myristoyltransferase and two substrates, the 'Leishmania' vaccine antigen candidates, HASPA and HASPB." Thesis, Imperial College London, 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.413682.
Повний текст джерелаSánchez, Arcila Juan Camilo. "Estudo de determinantes antigênicos para respostas imunes de células humanas em KMP-11 (Kinetoplastid membrane protein – 11) de Leishmania Amazonensis." reponame:Repositório Institucional da FIOCRUZ, 2010. https://www.arca.fiocruz.br/handle/icict/4103.
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CNPq e PEC-PG
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Rio de Janeiro, RJ, Brasil.
As leishmanioses formam um grupo de doenças antropozoonóticas, endêmicas em 88 países e presentes em quase todos os estados brasileiros. O desenvolvimento de uma vacina contra das leishmanioses é altamente desejável já que a terapia e as características biológicas e ecoepidemiólogicas dos parasitos e seus vetores associados não facilitam o controle da doença. Kinetoplastid Membrane Protein-11 (KMP-11) é uma molécula candidata a vacina contra as leishmanioses. Utilizando ferramentas in vitro e in silico, foi avaliada a antigenicidade de 13 peptídeos sintéticos abrangendo a sequência inteira de KMP-11. Para os estudos in vitro foram usadas células mononucleares de sangue periférico (PBMC) de pacientes com leishmaniose cutânea (LC) do estado do Rio de Janeiro. Estas células foram empregadas para testar a antigenicidade dos 13 peptídeos individualmente e da proteína integral KMP-11 recombinante, através de ensaios de ELISA e ELISPOT. Na dosagem de citocinas por ELISA observamos que a proteína KMP-11 recombinante estimulou respostas de citocinas quase sempre superiores às induzidas pelos peptídeos isolados. No que se refere a IFN-, dois dos 13 peptídeos (P9 e P10) estimularam níveis desta citocina significativamente (p<0,05) mais baixos do que os observados com a proteína inteira. Dez peptídeos (P4, P5, P6, P7, P8, P9, P10, P11, P12 e P13) apresentaram níveis de IL-10 e TNF-α significativamente inferiores aos observados com a proteína inteira. KMP-11 mostrou-se um potente indutor de IL-10 em PBMC de pacientes com LC, confirmando resultados anteriormente publicados, mas também foi capaz de induzir a produção de IFN-γ e altos níveis de TNF-α, em níveis superiores aos dos peptídeos estudados. Na avaliação da razão IFN-γ/IL-10 observou-se um acentuado contraste entre a maioria dos peptídeos e a proteína KMP-11. As respostas a 11 dos 13 peptídeos mostraram um claro viés de resposta de tipo 1 (IFN-γ>IL-10), a exceção dos peptídeos P1 e P10 (IFN-γ
Jagu, Elodie. "Design, synthesis and biological evaluation of new polyamine derivatives as antikinetoplastid agents." Thesis, Université Paris-Saclay (ComUE), 2016. http://www.theses.fr/2016SACLS589/document.
Повний текст джерелаThis project is at the interface of chemistry and biology and relies on the expertise of two different teams. This thesis involves the design and development of inhibitors directed against Kinetoplastids. It is urgent to develop new therapeutic strategies to respond to drug resistance and toxicity of currently used drugs against these parasites. Polyamine metabolism and transporter have been demonstrated as essential for parasite growth. Therefore, these systems are potential drug targets for development of antikinetoplastid compounds. We chose to synthesize polyamine derivatives and evaluate their biological activity against Kinetoplatids. Fifty-four compounds, divided into three chemical series, have been synthesized and evaluated. Many have shown a micromolar biological activity in vitro against parasite. In vivo evaluation is foreseen for the most promising derivative
Salem, Hemida Manar Mahfouz. "Identification of antikinetoplastid compounds from Psorothamnus polydenius and P. arborescens." Columbus, Ohio : Ohio State University, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1127103915.
Повний текст джерелаBoudot, Clotilde. "Recherche de nouvelles molécules trypanocides." Thesis, Limoges, 2019. http://www.theses.fr/2019LIMO0078.
Повний текст джерелаKinetoplastid diseases are vectorial parasitoses caused by flagellated blood protozoa. Among these, African Trypanosomiasis, due to a parasite of the genus Trypanosoma, affects both humans and animals. In humans, this disease, known as sleeping sickness, progresses classically in 2 stages: the hemolymphatic stage characterized by multiplication of the parasite in blood and lymph and the nervous stage characterized by the presence of the parasite in the brain. In the absence of appropriate therapy, death is inevitable. Currently, the treatments proposed in human and veterinary medicine are old, toxic and at the origin of cases of resistance. The search for new molecules is therefore essential to control this pathology. It is in this context that we studied two families of molecules which recognize parasitic sites: (i) Nitroimidazoles that interact with nitroreductases to generate toxic intermediates, and (ii) Phenanthroline derivatives targeting telomerases to disrupt trypanosome DNA synthesis. Our thesis research evaluated the trypanocidal power of different molecules from these two families both by in vitro tests and in a mouse model infected with a strain of Trypanosoma brucei brucei. The purpose of this work was to identify new drug candidates. The results obtained have made it possible to identify compounds of interest that open up new pathways of research to control this parasite, as well as all kinetoplastidae
Iyengar, Preethi Ranganathan. "MYSTERIES OF THE TRYPANOSOMATID MAXICIRCLES: CHARACTERIZATION OF THE MAXICIRCLE GENOMES AND THE EVOLUTION OF RNA EDITING IN THE ORDER KINETOPLASTIDA." VCU Scholars Compass, 2015. http://scholarscompass.vcu.edu/etd/4010.
Повний текст джерелаPortman, Neil. "Deconstructing the trypanosome cytoskeleton : from structures to functions via components and complexes." Thesis, University of Oxford, 2011. http://ora.ox.ac.uk/objects/uuid:e04fef74-b111-4992-aad6-ddb3169ff95b.
Повний текст джерелаFleming, Ian Murray Cameron. "Studies on RNA Modification and Editing in Trypanosoma brucei." The Ohio State University, 2016. http://rave.ohiolink.edu/etdc/view?acc_num=osu1452245560.
Повний текст джерелаYakovich, Adam J. "Old targets and new beginnings a multifaceted approach to combating Leishmaniasis, a neglected tropical disease /." Columbus, Ohio : Ohio State University, 2007. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1193247442.
Повний текст джерелаBoni, Sara Macente. "Avaliação de método diagnóstico não invasivo para leishmaniose tegumentar americana através da reação em cadeia da polimerase." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/5/5134/tde-06012017-113026/.
Повний текст джерелаIntroduction: Etiologic diagnosis of tegumentary leishmaniasis is based on the detection of the parasite in injury samples collected by invasive method. DNA detection of the parasite by PCR, could be a more sensible alternative, but is not available in routine practice and it was standardized in clinical samples by invasive methods such as scrapes, aspirate or biopsy of the lesion. A proposal for the diagnosis of tegumentary leishmaniasis lesions would obtaining material (cutaneous or mucosal) through less invasive collection methods, and it was possible to detect parasite DNA from small quantities of clinical specimen. In this study we evaluate the effectiveness of the PCR in samples collected by non-invasive method (swab injury) as a tool to be used in the diagnosis of leishmaniasis (mucosal and localized cutaneous), as well as for early detection of Leishmania in mucosa from patients with cutaneous lesions active and as an evaluation tool of therapeutic response in mucosal leishmaniasis. Methodology: Between August 2013 to July 2015 were selected 57 patients from the Leishmaniasis out clinic from the Institute of Infectious Diseases Emilio Ribas, which samples of cutaneous lesion or mucosa were collected by swab and biopsy of the lesions. In parallel, routine laboratory was carried out for the diagnosis of leishmaniasis in patients with active lesions (histopathology, search and Leishmania culture, serology and Montenegro skin test antigen). The samples taken by biopsy or swab were assessed by polymerase chain reaction having as targets the minicircle kinetoplast DNA (kDNA) of Leishmania for conventional PCR and gene heat shock protein 70kDa (Hsp70) for conventional PCR and real-time PCR. Results: Leishmania DNA detection in samples taken by swab of active lesions was similar to the samples taken by biopsy from the same lesion. When compared to the methods commonly used in the diagnosis of tegumentary leishmaniasis, PCR material collected by swab showed superior performance. It was shown that using the primers for the target kDNA obtained more effectively compared with the target Hsp70, or by conventional PCR and by real-time PCR (sensitivity 94.1%, 42.4% and 39.4%, respectively). When analyzing samples from patients already treated for mucosal leishmaniasis observed positivity of 86% to kDNA and 22% for Hsp70. Samples of nasal mucosa and active cutaneous leishmaniasis, collected by swab for early detection of disease, it obtained 92.9% positivity with kDNA and 28.6% with Hsp70. Conclusions: Our results suggest that the biological sample collection method using the swab for the molecular diagnosis of tegumentary leishmaniasis had compared efficacy with biopsy collection method. The DNA detection collected by swab samples allows to analyze the presence of DNA of the parasite in tissue without damage and can detect the presence of Leishmania even before clinical changes are present. The monitoring of therapeutic response mucosal leishmaniasis can be made by Leishmania DNA detection in samples per swab
Andrino, Marcos Luiz Alves. "Padronização e validação de dois sistemas de amplificação quantitativa para a detecção do DNA mitocondrial e nuclear de Trypanosoma cruzi, em amostras sanguíneas e teciduais de camundongos Swiss infectados." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/99/99131/tde-23022017-112204/.
Повний текст джерелаSerological techniques are the gold standards for the diagnosis of Chagas\' disease, but are not very effective in evaluating the response to treatment, since seronegativation may take many years. Serology is also used to identify reactivation episodes in patients with some degree of immunodeficiency, for example those co-infected with HIV. Hemoculture and xenodiagnosis have high specificity and low sensitivity, requiring 30 to 120 days for releasing a final result, and they can generate false-negative results especially in the chronic phase of infection. Therefore, real-time PCR (qPCR), a technique with high sensitivity and specificity, could be used to detect and quantify the parasite load, allowing the diagnosis of reactivation episodes and the monitoring of patients undergoing treatment. The choice of amplification targets and qPCR primers is challenging since there is as yet no consensus in the literature about the best amplification target sequence and the best primers. In the present study, primers from the nuclear (F2/B3) and mitochondrial, kDNA (32F/148R) T. cruzi sequences were designed. Samples were obtained from the blood, brain, heart, lung, liver, spleen, kidney, intestine, adrenal glands, adipose tissue and skeletal muscle tissue of 24 adult Swiss mice, infected intraperitoneally with 103 trypomastigote forms of the Y strain of Trypanosoma cruzi. The samples were collected at the 13th, 26th and 61st post-infection days, corresponding to different parasite load levels (low, medium and high), and were analyzed by qPCR with SYBR Green. The results showed that the nuclear and mitochondrial DNA primers detected T. cruzi DNA in a specific way. The nuclear primers detected higher parasite load levels than the kDNA ones, although the kDNA primers presented higher analytical sensitivity (0.002 and 0.0002 of a single parasite, respectively). The two qPCRs showed adequate reproducibility and repeatability indexes, i.e., below 25%. The efficiency parameters, (90% - 110%) and linearity (R2 >=0.98) of the two qPCRs showed adequate values according to the established literature. The comparison of the threshold cycle (CT) of the two qPCRs found no statistical difference. Regarding the parasite load, it was possible to detect the parasite DNA in all blood and tissue samples, with universal distribution, however heterogeneous, and at all stages of infection. The animal model used in this study was adequate to validate the two qPCRs for the detection and quantification of the parasite load. According to established parameters, the two qPCRs, with nuclear and mitochondrial primers, were successfully standardized and validated, being able to quantify all types of samples (blood and organs), in the acute, subacute and chronic phases of the disease, signaling positively to the use of both molecular assays in the diagnosis of T. cruzi infections.
Fersing, Cyril. "Synthèse et étude des relations structure-activité de nouvelles 3-nitroimidazo (1,2-a) pyridines anti-kinétoplastidés." Thesis, Aix-Marseille, 2018. http://www.theses.fr/2018AIXM0275/document.
Повний текст джерелаThe kinetoplastids of the Leishmania and Trypanosoma genus are the causative agents of neglected tropical diseases that threaten nearly half a billion people in the intertropical zone, resulting in 50 000 deaths per year. Among the molecules in clinical development to treat these pathologies, fexinidazole is a prodrug belonging to the 5-nitroimidazoles family, which exerts its anti-infectious action via a bioactivation step catalyzed by parasitic nitroreductases (NTR), enzymes whose cofactor is a flavin. In order to identify novel nitroheterocycles as parasitic NTR substrates, a small chemical library of imidazo[1,2-a]pyridines synthesized by our laboratory was screened in vitro, leading to the identification of a Hit molecule active both on Leishmania donovani and Trypanosoma brucei brucei. This compound served as a starting point for a pharmacomodulation work, initially in position 8 of the imidazo[1,2-a]pyridine ring: the introduction of various chemical groups using the pallado-catalyzed coupling reactions of Suzuki-Miyaura, Sonogashira and Buchwald-Hartwig, or SNAr reactions, highlighted several "lead" compounds with a significantly improved biological profile. In a second step, the pharmacomodulation work was extended to positions 2, 3 and 6 of the imidazo[1,2-a]pyridine ring in order to complete the structure-activity relationship data, to study in particular the impact of the redox potential and to optimize the physicochemical and in vitro pharmacokinetic parameters of the best compounds in order to initiate the study of their in vivo activity on a trypanosomiasis mouse model
Abraham, Rebecca Jane. "Repurposing of robenidine and characterization of novel analogues for treatment of infectious diseases." Thesis, 2017. https://hdl.handle.net/2440/131986.
Повний текст джерелаThesis (Ph.D.) -- University of Adelaide, School of Animal and Veterinary Sciences, 2018
DAVID, Vojtěch. "High-throughput analysis of uridine insertion and deletion RNA editing in \kur{Perkinsela}." Master's thesis, 2015. http://www.nusl.cz/ntk/nusl-187804.
Повний текст джерелаFLEGONTOVA, Olga. "Diversity and biogeography of diplonemid and kinetoplastid protists in global marine plankton." Doctoral thesis, 2017. http://www.nusl.cz/ntk/nusl-367441.
Повний текст джерелаKALTENBRUNNER, Sabine. "Characterization of TbPH1, a kinetoplastid-specific pleckstrin homology domain containing kinesin-like protein." Master's thesis, 2017. http://www.nusl.cz/ntk/nusl-317938.
Повний текст джерелаConcepcón-Acevedo, Jeniffer. "Analysis of the Spatiotemporal Localization of Mitochondrial DNA Polymerases of Trypanosoma brucei." 2013. https://scholarworks.umass.edu/open_access_dissertations/716.
Повний текст джерелаTÝČ, Jiří. "Kinetoplastids biology, from the group phylogeny and evolution into the secrets of the mitochondrion of one representative: \kur{Trypanosoma brucei}, the model organism in which new roles of the evolutionary conserved genes can be explored." Doctoral thesis, 2015. http://www.nusl.cz/ntk/nusl-187471.
Повний текст джерелаBruhn, David F. "Mitochondrial DNA polymerase IB: Functional characterization of a putative drug target for African sleeping sickness." 2011. https://scholarworks.umass.edu/dissertations/AAI3461989.
Повний текст джерелаYan, Yifei. "Study of cox1 trans-splicing in Diplonema papillatum mitochondria." Thèse, 2011. http://hdl.handle.net/1866/5423.
Повний текст джерелаDiplonema papillatum is a single cellular organism that lives in the ocean. Its mitochondrial genome possesses a special feature: all genes are fragmented in multiple pieces that are called modules and each module is encoded by a different chromosome. Expression of a gene requires trans-splicing that successfully assemble a full-length mRNA from all modules of the gene. It was previously shown that the cox1 gene is encoded in nine modules that are all located on different chromosomes; moreover, a stretch of six non-encoded Us exist between Module 4 and 5 in the mature mRNA [1]. No consensus sequence of known splicing sites was identified near the modules. We speculate that trans-splicing of the cox1 gene is directed by guide RNAs (gRNAs) via a mechanism that is similar to U-insertion/deletion editing in kinetoplastids, the sister group of diplonemids. We have detected populations of small RNA molecules that could come from mitochondrial. We found that the six Us were added to the 3’ end of Module 4 in a similar way to the Us added by the TUTase in kinetoplastid U-insertional editing. Sequence profiles of possible trans-splicing gRNAs were constructed in regular expressions based on our knowledge of known gRNAs in kinetoplastid RNA editing. According to the complementarity between the gRNA and the two adjacent modules, primers were designed for RT-PCR that aims to detect gRNA sequences. Among the results, we identified sequences that match or partially match the gRNA profiles. A pilot in vitro assay did not reconstitute trans-splicing of module 3, 4 and 5, suggesting that further technical improvements are needed.
Gowri, V. S. "Analysis Of Protein Evolution And Its Implications In Remote Homology Detection And Function Recognition." Thesis, 2007. http://hdl.handle.net/2005/568.
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