Thesis (Ph. D.)--Cornell University, 1935. / Cover title. Reprinted from the Cornell veterinarian vol. XXV. no. 2, April, 1935. Conference number. Bibliography: p. 110.

Inaug.-Diss. (V.M.D.)--Hanover. Tierärztliche hochschule. / "Aus dem Staatlichen veterinär-untersuchungsamt Kassel." "Schrifttumsverzeichnis": p. 44-46.

Thesis (M.S.)--University of Wisconsin--Madison, 1941. / Typescript. eContent provider-neutral record in process. Description based on print version record.

Thesis (Ph. D.)--University of Wisconsin, 1942. / Typescript (carbon copy). eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves [36-40]).

Thesis (Ph. D.)--University of Wisconsin, 1938. / Typescript (carbon copy). eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (MMedVet (Gyn))--University of Pretoria, 2002. / Includes bibliographical references.

Thesis (Ph. D.)--University of Washington, 1999. / Vita. Includes bibliographical references (leaves 83-108).

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2014-11-17T19:30:12Z No. of bitstreams: 1 Alene Vanessa Azevedo dos Santos. Reacoes de oxido reducao como alvo de quimioterapia triconomicida.pdf: 13103565 bytes, checksum: ad5cc355a7ef3d05b87bd59e85b6f9e0 (MD5) / Made available in DSpace on 2014-11-17T19:30:12Z (GMT). No. of bitstreams: 1 Alene Vanessa Azevedo dos Santos. Reacoes de oxido reducao como alvo de quimioterapia triconomicida.pdf: 13103565 bytes, checksum: ad5cc355a7ef3d05b87bd59e85b6f9e0 (MD5) Previous issue date: 2008 / Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Trichomonas vaginalis e Tritrichomonas foetus são os agentes etiológicos da tricomoníase humana e bovina, respectivamente. A primeira é uma parasitose de grande importância em saúde pública, sendo a principal doença sexualmente transmissível não viral e a segunda ocasiona extensos prejuízos econômicos na pecuária. A tricomoníase humana pode promover a incidência de câncer de colo de útero e a transmissão do HIV. Na infestação humana a droga de escolha é o metronidazol, mas não há um composto eficiente para o tratamento de bovinos. Como este fármaco é usado para outras parasitoses e infecções bacterianas desde a década de 60, tem sido documentado o surgimento de casos refratários e cepas resistentes. Além disso, o fármaco pode ter efeitos mutagênicos e carcinogênicos. Assim sendo a busca por novos compostos tricomonicidas constitui uma demanda premente. No presente estudo foram avaliados os efeitos do dietilditiocarbamato de sódio (DETC) sobre os protozoários parasitas T. vaginalis e T. foetus. Observamos que a adição de DETC inibiu significativamente a proliferação de T. vaginalis, produzindo uma IC50 de 269,7nM, enquanto o metronidazol teve uma IC50 de 523,1nM. Em T. foetus os valores de IC50 foram semelhantes, sendo 497,8 e 459,7nM para o DETC e para o metronidazol, respectivamente. Estes compostos não afetaram significativamente a incorporação de [3H]timidina por esplenócitos murinos em concentrações de até 1000M. A atividade mitocondrial de macrófagos peritoneais murinos também não foi afetada por DETC, mesmo em 200μM, sugerindo seletividade no modo de ação. Objetivando determinar a existência de sinergismo entre DETC e metronidazol, foram determinados os valores de FIC (concentração inibitória fracionada) para os dois protozoários. Os valores de FIC foram 0,3 e 0,7 para T. vaginalis e T. foetus, respectivamente. Esta observação indica que ocorre a interação sinergística no protozoário parasita de humanos. A fim de determinar se a incubação com DETC poderia afetar a expressão de grupos sulfidrila dos parasitos empregamos a reação de Ellman para dosar os grupamentos tiol totais de T. vaginalis antes e após a adição do composto. Observamos que o tratamento com DETC por 24h significativamente (p<0,01) reduziu a concentração de grupos tióis do parasito. A detecção de tióis livres pela sonda fluorescente orto-phthalaldeído (OPA) em T. foetus sugere a participação de sulfidrilas no mecanismo de ação do DETC, uma vez que este reduz marcadamente a marcação dos parasitos pelo OPA, mas este efeito foi revertido pela pré-incubação com cisteína. Vale salientar que o aminoácido reverte, ainda, os efeitos do composto na proliferação parasitária. A mensuração de radicais livres por quimioluminescência amplificada pelo luminol indicou que a geração de espécies reativas de oxigênio foi significativamente (p <0,05) aumentada por DETC em T. vaginalis, mas não em T. foetus. O estresse oxidativo sobre os parasitas foi avaliado pela medida de substâncias reativas ao ácido tiobarbitúrico (TBARs). Observou-se que a combinação metronidazol-DETC significativamente aumenta a peroxidação lipídica em T. vaginalis (p <0,01) e T. foetus (p <0,05). A análise ultraestrutural por microscopia eletrônica de transmissão revelou que tanto DETC quanto o metronidazol produziram danos hidrogenossomais e desencadearam autofagia e esses efeitos foram mais acentuadas no parasitas incubados com a combinação de drogas. Tomados em conjunto, estes dados sugerem que a combinação metronidazol-DETC pode fornecer novas ferramentas para a efetiva quimioterapia da tricomoníase / Trichomonas vaginalis e Tritrichomonas foetus are the etiologic agents of the human and bovine trichomoniasis, respectively. The former is a parasitic disease of great relevance in public health – the major non-viral sexually-transmitted disease, whereas the latter causes remarkable losses in livestock productivity. Human trichomoniasis can promote the incidence of cervical cancer and HIV transmission. In human infestation the drug of choice is metronidazole, but there is no efficient compound for treating the cattle. Since the medication is also used for other parasitic and bacterial diseases, since the 60s, refractory cases and resistant strains have been documented. Besides, the drug may be mutagenic and carcinogenic. Therefore the search for new trichomonicidal compounds is required. In the present study we investigated the effects of sodium N,N-diethylthiolcarbamate (DETC) alone or combined with metronidazole upon the parasitic protozoa T. vaginalis and T. foetus. We notice that DETC significantly inhibited the T. vaginalis proliferation, producing an IC50 of 269.7nM, whereas metronidazole produced an IC50 of 523.1. In T. foetus the IC50 values were similar, being 497.8 and 459.7nM for DETC and metronidazole, respectively. These compounds did not significantly affect the incorporação de [3H]timidine incorporation by murine splenocytes in concentrations up to 1000μM. The mitochondrial activity of murine peritoneal macrophages was not affected by 200μM DETC, suggesting a selective mode of action. In order to determine whether there is synergism between DETC and metronidazole, we determined the fractional inhibitory concentrations (FIC) of these compounds upon both protozoa. The FIC values for T.vaginalis and T. foetus were 0.3 and 0.7, respectively. This observation indicates that synergistic interaction takes place only on the human pathogen. To determine whether the incubation with DETC could affect the expression of sulphydril groups of parasites we employed the Ellman’s reaction to measure the total thiol groups of T. vaginalis before and after the addition of the compound. We observed that treatment with DETC for 24h significantly (p <0.01) reduced the concentration of thiol groups of the parasite. The detection of free thiols by the fluorescent probe ortho- phthaldialdehyde (OPA) in T. foetus suggests the involvement of sulphydrils in the DETC mechanism of action, since it markedly reduces the OPA labeling of parasites, but this effect was reversed by cysteine preincubation with. It is noteworthy that the amino acid addition also reverts, the effects of the compound on the parasite proliferation. The measurement of free radicals by luminol-enhanced chemiluminescence indicated that reactive oxygen species generation was significantly (p <0.05) enhanced by DETC in T. vaginalis, but not T. foetus. The oxidative stress on the parasites was evaluated by measurement of the thiobarbituric acid-reactive substances (TBARs). We observed that the metronidazole-DETC combination significantly enhanced lipid peroxidation in T. vaginalis (p <0.01) and T. foetus (p <0.05). The ultrastructural analysis by transmission electron microscopy revealed that both DETC and metronidazole produced hydrogenosomal damage and triggered autophagy and these effects were more pronounced on the parasites incubated with the combined drugs. Taken together these data suggest that the metronidazole-DETC combination may furnish new tools in the effective chemotherapy of trichomoniasis.

Made available in DSpace on 2014-07-29T15:30:36Z (GMT). No. of bitstreams: 1 dissertacaopatriciabreu.pdf: 1725826 bytes, checksum: d77d7164c903bff7a0df515bcc455897 (MD5) Previous issue date: 2008-12-17 / This study evaluated the frequency of Trichomonas vaginalis infection in human immunodeficiency virus positive (HIV+) and negative (HIV-) women in Goiania, Goiás, Brazil, comparing the presence of the parasite in the two groups and correlating it with the conditions of immunodeficiency present in these women. The diagnostic techniques used, wet mount microscopy, culture and cytology, were also evaluated, and the principal inflammatory alterations in the two groups were assessed. The HIV+ samples (test group) were collected at the Hospital of Tropical Diseases and in the Maternal and Child Healthcare Hospital, whereas the HIVnegative samples (control group) were collected at the Maternity Hospital. Swabs were used for wet mount microscopy (saline solution) and for culture (Diamond s medium), and Ayre s spatula and brush were used for the cytology smears, which were fixed using a commercial fixative. A total of 237 samples were analyzed, 125 HIV-positive test samples and 112 HIV-negative controls. The overall frequency of T. vaginalis was 13.9%, 18.4% in the HIV+ and 8.9% in the HIV- group. This difference was statistically significant (p<0.05); however, the infection was not associated with immunodeficiency according to CD4, viral count and lymphocytes. There was a significant difference in the prevalence of the parasite between HIV+ and HIVpregnant women (22.6% versus 12.5%). Culture identified a frequency of T. vaginalis of 13.9%, while cytology identified a rate of 13.5% and wet mount microscopy 11.4%. Perinuclear halos were the most frequent inflammatory alteration; however, there was no difference between the groups / O estudo avaliou a freqüência da infecção por Trichomonas vaginalis em mulheres HIV+ (vírus imunodeficiência humana) e HIV- em Goiânia-GO, comparando a presença do parasito e correlacionando com as condições de imunodeficiência. Avaliou também as técnicas de diagnóstico: exame a fresco, cultura e citologia, e apontou as principais alterações inflamatórias nos dois grupos. As amostras de HIV+ (grupo teste) foram coletadas no Hospital de Doenças Tropicais e no Hospital Materno Infantil e as de HIV negativas (grupo controle) na Maternidade Nascer Cidadão. Foram utilizados swabs para os exames a fresco (salina) e para a cultura (meio Diamond), espátula de Ayre e escovinha para os esfregaços citológicos que foram submetidos a fixadores comerciais. Foram examinadas 237 amostras: 125 do teste e 112 do controle. A freqüência por T. vaginalis foi 13,9%, sendo 18,4% nas HIV+ e 8,9% nas HIV-. O resultado foi estatisticamente significativo (p<0,05), porém a infecção não foi associada à imunodeficiência (CD4, carga viral e linfócitos). Houve diferença significativa entre grávidas HIV+ e HIV- (22,6% vs 12,5%). A Cultura obteve 13,9% da presença de T.vaginalis, a Citologia 13,5% e o exame a fresco 11,4%. Halos perinucleares predominaram na avaliação das alterações inflamatórias, porém não houve diferença entre os grupos.

Trichomonas vaginalis and Tritrichomonas foetus are the etiologic agents of human and bovine trichomoniasis, respectively. As microaerophilic protozoans; both share a wide array of clinical manifestations ranging from vaginitis to abnormal pregnancies. Human trichomoniasis receives minimal public health attention despite of its worldwide high prevalence rate. Emerging evidence of metronidazole-resistant T vaginalis strains facilitates a concern to understand this protozoan. Cysteine proteases have been implicated as important virulence factors produced by T vagina/is. This study explores the expression of one particular legumain-like cysteine protease known as Tv AE 1. Furthermore, it highlights the relationship between inhibitory effects of trichomonal cells caused by sanguinarine and chelerythrine. A system for obtaining legumains by expressing it in methylotrophic yeast, Pichia pastor is, has been described. The recombinant legumains were produced and processed by the yeast to their inactive and mature forms. Secondly, T foetus cells were transfected with TvAEl construct. Localization and enzymatic studies on legumains will provide evidence into the pathogenicity ofT vagina/is. This study revealed the vesicularization of recombinantly unprocessed TvAEl proteins. Thirdly, plant derived compounds, sanguinarine (SA) and chelerythrine (CHE) were assessed in vitro for their inhibitory effects against T vagina/is and T. foetus. Treatment of SA and CHE for 24 h led to a significant inhibitory growth of in vitro cultures for all three trichomonal strains, G3, Tl and Dl, compared to untreated cells. For these bovine and human trichomonal strains, SA was slightly more effective inhibitor than CHE. With IC5o values between 3 - 8 micromolar for the alkaloids, CHE had less inhibitory effect compared to SA. These findings are significant considering the association between cysteine pro teases and trichomoniasis. Further elucidation of the exact anti protozoal mechanism of both compounds toward legumains may lead to the development of these potent agents against trichomonads.