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Auranofin Targets Thioredoxin Reductases in Trichomonas vaginalisJauregui, Jose 01 January 2017 (has links)
Trichomonas vaginalis is an anaerobic, parasitic protozoan, responsible for trichomoniasis, the world’s most common, non-viral sexually transmitted infection. Lacking many of the defenses present in other organisms to combat oxidative stress, Trichomonas vaginalis relies extensively on the thioredoxin system—NADPH, thioredoxin reductase, and thioredoxin—as a means to protect against exposure to excess oxygen. Current trichomoniasis treatment relies exclusively on the 5-nitroimidazole drugs, but fear of drug-resistant strains and allergic reactions to 5-nitroimidazole treatment necessitate the discovery of a new treatment method for trichomoniasis. Previous research has shown that auranofin, an FDA-approved drug, was effective at inhibiting activity of one of Trichomonas vaginalis’ isoforms of thioredoxin reductase (of which the organism has five total). Our research showed that only two of the isoforms were transcribed and expressed at high levels, and that both of these isoforms were susceptible to auranofin treatment. Not only that, these two isoforms were also shown to be susceptible to various auranofin analogs, having comparable or lower IC50 values. Further tests on these analogs might show that they are actually better treatment candidates if they exhibit less symptoms than auranofin. Experiments examining how mRNA and protein levels were modulated in response to two different concentrations of auranofin treatment showed that while some isoforms show increased levels, no one isoform experienced any drastic changes. Together, this data suggests that further studies should focus on these two most highly expressed isoforms of thioredoxin reductase.
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Cytosolická hydrogenáza prvoka Trichomonas vaginalis / Cytosolic hydrogenase in Trichomonas vaginalisDohnálková, Alena January 2015 (has links)
Trichomonas vaginalis is a flagellated microaerophilic protozoan from the group Excavata that cause trichomoniasis, the most common nonviral sexually transmitted disease in the world. This thesis deals with the study of hydrogenases, enzymes catalyzing reversible conversion of protons and electrons to molecular hydrogen. In T. vaginalis, hydrogenases have been identified so far only in hydrogenosomes, modified anaerobic mitochondia that are involved in energy metabolism. We proved the presence of this enzyme also in the cytosol of T. vaginalis. Among several hydrogenase paralogues present in the genome, we selected an appropriate gene for the putative cytosolic hydrogenase (C-Hyd) and verified its cytosolic localization in the cells with overexpressed C-Hyd protein. Based on the determination of hydrogenase activities in different cell compartments and fractions obtained by affinity chromatography, we demonstrated the hydrogenase activity of C-Hyd protein, which means that C-Hyd is a functional hydrogenase. Identification of hydrogenase in T. vaginalis cytosol changes our understanding of trichomonad core metabolism and opens the door for the research of unexplored metabolic capabilities of this parasite.
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Incidence of Trichomonas gallinae in mourning dove, Zenaidura macroura, populations of ArizonaStraus, Milo August, 1942- January 1966 (has links)
No description available.
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Effects of fecal contaminant Eschericia [sic] coli on the bovine venereal pathogen Tritrichomonas foetus in cultureMadden, Tanya D. January 2007 (has links)
Thesis (M.S.)--University of Wyoming, 2007. / Title from PDF title page (viewed on Nov. 7, 2008). Includes bibliographical references (p. 30-35).
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Biological characterization of Tritrichomonas foetus of bovine and feline originStockdale, Heather Dawn, Blagburn, Byron L., January 2008 (has links) (PDF)
Thesis (Ph. D.)--Auburn University, 2008. / Abstract. Vita. Includes bibliographical references (p. 83-89).
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Estudo da hidrólise extracelular de nucleotídeos em Trichomonas gallinaeBorges, Fernanda Pires January 2006 (has links)
Trichomonas gallinae é um protozoário flagelado que parasita o trato digestivo superior de várias aves, incluindo pombos domésticos, frangos e perus. O estudo dos mecanismos de patogenicidade deste parasito é de fundamental importância, uma vez que a infecção causada pelo mesmo tem envolvido grandes perdas econômicas. Sabe-se que além da função energética, o ATP desempenha inúmeras funções fisiológicas, como sinalização extracelular e mecanismos citolíticos. As concentrações dos nucleotídeos, tais como ATP e ADP, no meio extracelular são controladas por um grupo de enzimas denominadas ectonuncleotidases. Fazem parte deste grupo as NTPDases (nucleosídeo trifosfato difosfoidrolases) e a ecto-5´- nucleotidase, as quais participam do controle dos níveis extracelulares dos nucleotídeos e nucleosídeos. A presença dessas atividades enzimáticas pode estar associada com a virulência e evasão dos parasitos, servindo como um mecanismo de escape dos efeitos citolíticos do ATP. Portanto, este estudo teve como objetivo caracterizar as atividades da NTPDase e 5´-nucleotidase, envolvidas na degradação dos nucleotídeos extracelulares em T. gallinae. A hidrólise de ATP, ADP e AMP foi ativada na presença de cátions divalentes e a adição de quelantes de cátions no meio de incubação reduziu significativamente a atividade específica da NTPDase e 5´-nucleotidase. As enzimas apresentaram ampla especificidade de substrato, observada através da hidrólise de outros nucleotídeos trifosfatados, difosfatados e monofosfatados, quando adicionados ao meio como substrato. O KM para o ATP foi de 65,62 ± 15,55 μM e para o ADP foi de 122,66 ± 3,51 μM. A Vmax para o ATP e o ADP foi de 0,20 ± 0,03 e 0,70 ± 0,09 nmolPi/min/106 trofozoítos, respectivamente. Para o AMP, o KM foi 466 ± 57 μM, com uma Vmax de 3,70 ± 0,59 nmolPi/min/106 trofozoítos. A influência de outras enzimas que hidrolisam nucleotídeos extracelulares foi descartada através do uso de inibidores específicos. A hidrólise do ATP, ADP e AMP indicou a presença de uma cadeia enzimática na superfície do parasito, composta por uma NTPDase e uma ecto-5’-nucleotidase. A presença de atividades enzimáticas capazes de hidrolisar nucleotídeos extracelulares pode representar um mecanismo de sobrevivência dos parasitos nos seus ambientes naturais. A compreensão dos processos bioquímicos extracelulares destes parasitos pode ampliar o conhecimento a respeito dos mecanismos envolvidos no parasitismo. / Trichomonas gallinae is a flagellated protozoan which parasitizes a variety of birds all over the world, including domestic pigeons, chickens and turkeys. The study of patogenicity mechanisms is relevant, since the infection caused by this parasite involves significant economic loss. Besides the energetic function, extracellular ATP plays several physiological functions, such as extracellular signaling and cytolysis. Extracellular nucleotide levels are controlled by group of enzymes named ectonucleotidases. This group of enzymes includes the NTPDases (nucleoside triphosphate diphosphohydrolases) and an ecto-5´-nucleotidase, which participates in the control of extracellular nucleotide and nucleoside levels. The presence of these enzyme activities could be associated with the virulence and evasion of the parasites, escaping from the cytolytic effects of ATP. Therefore, the aim of this study was to characterize the NTPDase and 5´-nucleotidase activities, involved in extracellular nucleotide degradation in T. gallinae. ATP, ADP and AMP hydrolysis were activated in the presence of divalent cations and the addition of cation chelating agents in the incubation medium significantly decreased the specific activity of NTPDase and 5´-nucleotidase. The enzymes presented broad substrate specificity because others triphosphate, diphosphate and monophosphate nucleotides were also hydrolysed when they were added to the mixture as substrates. The KM value for ATP was 65.62 ± 15.55 μM and for ADP was 122.66 ± 3.51 μM. The Vmax values for ATP and ADP were 0.20 ± 0.03 and 0.70 ± 0.09 nmolPi/min/106 trichomonads, respectively. For AMP, KM was 466 ± 57 μM, with the Vmax value of 3.70 ± 0.59 nmolPi/min/106 trichomonads. The influence of other enzymes able to hydrolyze extracellular nucleotides was tested through the use of specific inhibitors. ATP, ADP and AMP hydrolysis indicated the presence of an enzyme chain in the surface of the parasite, composed by an NTPDase and an ecto-5´-nucleotidase. The presence of enzyme activities able to hydrolyze extracellular nucleotides can represent a survival mechanism of the parasites in their natural environments. The study of extracellular biochemical processes of these parasites can improve the knowledge related to the mechanisms involved in the parasitism.
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Estudo da hidrólise extracelular de nucleotídeos em Trichomonas gallinaeBorges, Fernanda Pires January 2006 (has links)
Trichomonas gallinae é um protozoário flagelado que parasita o trato digestivo superior de várias aves, incluindo pombos domésticos, frangos e perus. O estudo dos mecanismos de patogenicidade deste parasito é de fundamental importância, uma vez que a infecção causada pelo mesmo tem envolvido grandes perdas econômicas. Sabe-se que além da função energética, o ATP desempenha inúmeras funções fisiológicas, como sinalização extracelular e mecanismos citolíticos. As concentrações dos nucleotídeos, tais como ATP e ADP, no meio extracelular são controladas por um grupo de enzimas denominadas ectonuncleotidases. Fazem parte deste grupo as NTPDases (nucleosídeo trifosfato difosfoidrolases) e a ecto-5´- nucleotidase, as quais participam do controle dos níveis extracelulares dos nucleotídeos e nucleosídeos. A presença dessas atividades enzimáticas pode estar associada com a virulência e evasão dos parasitos, servindo como um mecanismo de escape dos efeitos citolíticos do ATP. Portanto, este estudo teve como objetivo caracterizar as atividades da NTPDase e 5´-nucleotidase, envolvidas na degradação dos nucleotídeos extracelulares em T. gallinae. A hidrólise de ATP, ADP e AMP foi ativada na presença de cátions divalentes e a adição de quelantes de cátions no meio de incubação reduziu significativamente a atividade específica da NTPDase e 5´-nucleotidase. As enzimas apresentaram ampla especificidade de substrato, observada através da hidrólise de outros nucleotídeos trifosfatados, difosfatados e monofosfatados, quando adicionados ao meio como substrato. O KM para o ATP foi de 65,62 ± 15,55 μM e para o ADP foi de 122,66 ± 3,51 μM. A Vmax para o ATP e o ADP foi de 0,20 ± 0,03 e 0,70 ± 0,09 nmolPi/min/106 trofozoítos, respectivamente. Para o AMP, o KM foi 466 ± 57 μM, com uma Vmax de 3,70 ± 0,59 nmolPi/min/106 trofozoítos. A influência de outras enzimas que hidrolisam nucleotídeos extracelulares foi descartada através do uso de inibidores específicos. A hidrólise do ATP, ADP e AMP indicou a presença de uma cadeia enzimática na superfície do parasito, composta por uma NTPDase e uma ecto-5’-nucleotidase. A presença de atividades enzimáticas capazes de hidrolisar nucleotídeos extracelulares pode representar um mecanismo de sobrevivência dos parasitos nos seus ambientes naturais. A compreensão dos processos bioquímicos extracelulares destes parasitos pode ampliar o conhecimento a respeito dos mecanismos envolvidos no parasitismo. / Trichomonas gallinae is a flagellated protozoan which parasitizes a variety of birds all over the world, including domestic pigeons, chickens and turkeys. The study of patogenicity mechanisms is relevant, since the infection caused by this parasite involves significant economic loss. Besides the energetic function, extracellular ATP plays several physiological functions, such as extracellular signaling and cytolysis. Extracellular nucleotide levels are controlled by group of enzymes named ectonucleotidases. This group of enzymes includes the NTPDases (nucleoside triphosphate diphosphohydrolases) and an ecto-5´-nucleotidase, which participates in the control of extracellular nucleotide and nucleoside levels. The presence of these enzyme activities could be associated with the virulence and evasion of the parasites, escaping from the cytolytic effects of ATP. Therefore, the aim of this study was to characterize the NTPDase and 5´-nucleotidase activities, involved in extracellular nucleotide degradation in T. gallinae. ATP, ADP and AMP hydrolysis were activated in the presence of divalent cations and the addition of cation chelating agents in the incubation medium significantly decreased the specific activity of NTPDase and 5´-nucleotidase. The enzymes presented broad substrate specificity because others triphosphate, diphosphate and monophosphate nucleotides were also hydrolysed when they were added to the mixture as substrates. The KM value for ATP was 65.62 ± 15.55 μM and for ADP was 122.66 ± 3.51 μM. The Vmax values for ATP and ADP were 0.20 ± 0.03 and 0.70 ± 0.09 nmolPi/min/106 trichomonads, respectively. For AMP, KM was 466 ± 57 μM, with the Vmax value of 3.70 ± 0.59 nmolPi/min/106 trichomonads. The influence of other enzymes able to hydrolyze extracellular nucleotides was tested through the use of specific inhibitors. ATP, ADP and AMP hydrolysis indicated the presence of an enzyme chain in the surface of the parasite, composed by an NTPDase and an ecto-5´-nucleotidase. The presence of enzyme activities able to hydrolyze extracellular nucleotides can represent a survival mechanism of the parasites in their natural environments. The study of extracellular biochemical processes of these parasites can improve the knowledge related to the mechanisms involved in the parasitism.
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Estudo da campilobacteriose e tricomonose genitais bovina no Distrito Federal e GoiásLeal, Diogo Ramos 25 April 2012 (has links)
Dissertação (mestrado)—Universidade de Brasília, Faculdade de Agronomia e Medicina Veterinária, Programa de Pós-Graduação em
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2012_DiogoRamosLeal.pdf: 470837 bytes, checksum: d797fb6fe6116d5c430378002bffff11 (MD5) / A Campilobacteriose Genital Bovina (CGB) e a Tricomonose Genital Bovina (TGB) são doenças de transmissão venérea que têm como algumas de suas características a ausência de sinais clínicos nos touros e a condição de portadores assintomáticos. Nas fêmeas, as principais manifestações são retorno ao estro em intervalos irregulares e aumento do intervalo de partos, esses sinais podem fazer com que essas doenças não sejam detectadas de imediato e permaneçam no rebanho trazendo prejuízos. A CGB e a TGB são responsáveis por perdas econômicas devido principalmente ao descarte e necessidade de reposição de animais subférteis, à queda na produção de bezerros e ao aumento no intervalo de partos. Exames negativos para as duas doenças são exigidos para o ingresso de um animal em centrais de coleta e processamento de sêmen, bem como em transações comerciais internacionais. Devido à importância dessas doenças, o objetivo deste trabalho foi estudar a prevalência da CGB e da TGB na região do Distrito Federal e entorno, bem como em criatórios da raça Curraleiro Pé- Duro em fazendas localizadas nos estados do Distrito Federal e Goiás. Para detecção da CGB foi utilizado o exame de imunofluorescencia direta (IFD) e para TGB o cultivo em meio de Diamond. Os estudos demonstraram a presença da CGB em alguns animais abatidos em frigoríficos do Distrito Federal e entorno e em alguns rebanhos de Curraleiro Pé-Duro, determinando a necessidade de diagnóstico dessa enfermidade em rebanhos que apresentem baixos índices reprodutivos, bem como em rebanhos que comercializem reprodutores. O cultivo para TGB não apresentou resultados positivos, o que não significa a ausência do agente nesses rebanhos, mas levanta a necessidade de estudos mais amplos com outros métodos mais sensíveis de diagnóstico. Medidas de prevenção e controle da CGB e da TGB, bem como algumas práticas de manejo foram avaliadas e apresentadas neste trabalho. _______________________________________________________________________________________ ABSTRACT / The bovine genital campylobacteriosis (BGC) and the bovine genital trichomoniasis (BGT) are venereal transmitted diseases which absence of clinical signs in bulls and the male carrier state are some of their characteristics. In females, the main manifestations are return to estrus (repeated breeding) at irregular intervals and elongation of the calving interval, these signals can be not detected and remain in the herd resulting in economic loss. The BGC and the BGT are responsible for economic losses due to the need for disposal and replacement of infected animals, the reduction in calving rate and the increase in calving interval. Negative tests are required for a bull to enter in an artificial insemination centre, as well as in international trades. The aim was to study the prevalence of BGC and BGT in the region of the Federal District and the surrounding areas, as well as herds of Curraleiro Pé-Duro breed on farms located in the states of the Federal District and Goiás. To detect BGC, it was used the direct immunofluorescence (DIF) survey and to BGT cultivation on Diamond`s medium. The results have shown the presence of BGC in animals at slaughterhouses in the Federal District and surrounding areas and in some herds of Curraleiro Pé-Duro, exposing the need for the diagnosis of disease in herds that have low reproductive rates, as well as trading in livestock breeding. The cultivation for BGT did not show positive results, which does not mean the absence of the agent in these herds, but it alerts to the necessity in further studies with others diagnostic methods. Measures of prevention and control of BGC and BGT and some management practices have been reviewed and are presented in this study.
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Estudo da hidrólise extracelular de nucleotídeos em Trichomonas gallinaeBorges, Fernanda Pires January 2006 (has links)
Trichomonas gallinae é um protozoário flagelado que parasita o trato digestivo superior de várias aves, incluindo pombos domésticos, frangos e perus. O estudo dos mecanismos de patogenicidade deste parasito é de fundamental importância, uma vez que a infecção causada pelo mesmo tem envolvido grandes perdas econômicas. Sabe-se que além da função energética, o ATP desempenha inúmeras funções fisiológicas, como sinalização extracelular e mecanismos citolíticos. As concentrações dos nucleotídeos, tais como ATP e ADP, no meio extracelular são controladas por um grupo de enzimas denominadas ectonuncleotidases. Fazem parte deste grupo as NTPDases (nucleosídeo trifosfato difosfoidrolases) e a ecto-5´- nucleotidase, as quais participam do controle dos níveis extracelulares dos nucleotídeos e nucleosídeos. A presença dessas atividades enzimáticas pode estar associada com a virulência e evasão dos parasitos, servindo como um mecanismo de escape dos efeitos citolíticos do ATP. Portanto, este estudo teve como objetivo caracterizar as atividades da NTPDase e 5´-nucleotidase, envolvidas na degradação dos nucleotídeos extracelulares em T. gallinae. A hidrólise de ATP, ADP e AMP foi ativada na presença de cátions divalentes e a adição de quelantes de cátions no meio de incubação reduziu significativamente a atividade específica da NTPDase e 5´-nucleotidase. As enzimas apresentaram ampla especificidade de substrato, observada através da hidrólise de outros nucleotídeos trifosfatados, difosfatados e monofosfatados, quando adicionados ao meio como substrato. O KM para o ATP foi de 65,62 ± 15,55 μM e para o ADP foi de 122,66 ± 3,51 μM. A Vmax para o ATP e o ADP foi de 0,20 ± 0,03 e 0,70 ± 0,09 nmolPi/min/106 trofozoítos, respectivamente. Para o AMP, o KM foi 466 ± 57 μM, com uma Vmax de 3,70 ± 0,59 nmolPi/min/106 trofozoítos. A influência de outras enzimas que hidrolisam nucleotídeos extracelulares foi descartada através do uso de inibidores específicos. A hidrólise do ATP, ADP e AMP indicou a presença de uma cadeia enzimática na superfície do parasito, composta por uma NTPDase e uma ecto-5’-nucleotidase. A presença de atividades enzimáticas capazes de hidrolisar nucleotídeos extracelulares pode representar um mecanismo de sobrevivência dos parasitos nos seus ambientes naturais. A compreensão dos processos bioquímicos extracelulares destes parasitos pode ampliar o conhecimento a respeito dos mecanismos envolvidos no parasitismo. / Trichomonas gallinae is a flagellated protozoan which parasitizes a variety of birds all over the world, including domestic pigeons, chickens and turkeys. The study of patogenicity mechanisms is relevant, since the infection caused by this parasite involves significant economic loss. Besides the energetic function, extracellular ATP plays several physiological functions, such as extracellular signaling and cytolysis. Extracellular nucleotide levels are controlled by group of enzymes named ectonucleotidases. This group of enzymes includes the NTPDases (nucleoside triphosphate diphosphohydrolases) and an ecto-5´-nucleotidase, which participates in the control of extracellular nucleotide and nucleoside levels. The presence of these enzyme activities could be associated with the virulence and evasion of the parasites, escaping from the cytolytic effects of ATP. Therefore, the aim of this study was to characterize the NTPDase and 5´-nucleotidase activities, involved in extracellular nucleotide degradation in T. gallinae. ATP, ADP and AMP hydrolysis were activated in the presence of divalent cations and the addition of cation chelating agents in the incubation medium significantly decreased the specific activity of NTPDase and 5´-nucleotidase. The enzymes presented broad substrate specificity because others triphosphate, diphosphate and monophosphate nucleotides were also hydrolysed when they were added to the mixture as substrates. The KM value for ATP was 65.62 ± 15.55 μM and for ADP was 122.66 ± 3.51 μM. The Vmax values for ATP and ADP were 0.20 ± 0.03 and 0.70 ± 0.09 nmolPi/min/106 trichomonads, respectively. For AMP, KM was 466 ± 57 μM, with the Vmax value of 3.70 ± 0.59 nmolPi/min/106 trichomonads. The influence of other enzymes able to hydrolyze extracellular nucleotides was tested through the use of specific inhibitors. ATP, ADP and AMP hydrolysis indicated the presence of an enzyme chain in the surface of the parasite, composed by an NTPDase and an ecto-5´-nucleotidase. The presence of enzyme activities able to hydrolyze extracellular nucleotides can represent a survival mechanism of the parasites in their natural environments. The study of extracellular biochemical processes of these parasites can improve the knowledge related to the mechanisms involved in the parasitism.
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Analýza lysosomů Trichomonas vaginalis / Analysis of lysosomes of Trichomonas vaginalisZimmann, Nadine January 2021 (has links)
Lysosomes represent the central degradative compartment of eukaryote cells. Harboring a variety of acid hydrolases at acidic pH, this organelle is designed for the degradation and recycling of material for cellular homeostasis and sustenance. Studies on mammalian lysosomes have been extensive and revealed a long list of lysosomal proteins. While the function of most of these remains elusive, it is not surprising that a large subset have been found to be hydrolases. However, little is known about the biogenesis and function of this organelle in parasitic protists, and even less about its role in secretion. This work aimed to shed light on the (phago-)lysosomal proteome of the human parasite Trichomonas vaginalis, its protein targeting, and involvement in hydrolase secretion. Our studies revealed a lysosomal proteome of 462 proteins in 21 functional classes. Hydrolases represented the largest functional class and included proteases, lipases, phosphatases, and glycosidases. The identification of a large set of proteins involved in vesicular trafficking and cytoskeleton rearrangement indicates a dynamic phagolysosomal compartment. Our research, as well as the research of others, have identified several hydrolases also in the secretome, including the cysteine protease TvCP2. However, previously the mode...
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