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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Imunomodula??o e a??o anti-Leishmania (Viannia) braziliensis por Ouratea cuspidata (Ochnaceae) / Immunomodulation and action anti-Leishmania (Viannia) braziliensis promastigotes by Ouratea cuspidata (Ochnaceae)

Ribeiro, Renata da Silva 27 February 2007 (has links)
Made available in DSpace on 2016-04-28T20:15:26Z (GMT). No. of bitstreams: 1 2007-Renata da Silva Ribeiro.pdf: 705558 bytes, checksum: e16373fa17f99c734d58ede2a54543c6 (MD5) Previous issue date: 2007-02-27 / Ouratea cuspidata (Ochnaceae) crude extracts were added to experimental systems containing either host macrophages, or Leishmania braziliensis promastigotes (L566). The biological effect by the use of the hexane (OCH), methanol (OCMeH) and ethyl-acetate (OCAcEt) extracts added at different concentrations: 2mg/mL; 4mg/mL; 8mg/mL, 16mg/mL e 32mg/mL. The OCMeOH was the only fraction to which the anti parasitic potentiality was achieved at doses rates that generated bio protective effects in the host cells. The parasiticide action, due by direct exposure of promastigotes to OCMeOH, as well as the parasitic growth, and morphological alterations intermediary metabolism alterations (mitochondrial activity) were concomitantly measured. Biological macrophagic functions were also evaluated using Cricetus cricetus peritoneal macrophages as a model (Mf). Bio-protective assays were carried out in order to determine the free radical generation by the extract constituents. The results were compared with non treated L566 and Mf. 78% of the 16 mg/106 Mf/mL treated Mf were viable but a significant decrease of their phagocytosis capability (92%) was detected. Such alterations were evident in 62% of the cells treated with 32mg/106 Mf/mL. At this concentration cells were 70% viable and presented 98% phagocytosis suppression. The macrophages enzymatic- mitochondrial activity was gradually diminished, with 87% activity at exposures to 8mg/106 Mf/mL; 66,3% at exposures to 16mg/106 Mf/mL, and 49% at exposures to 32mg/106 Mf/mL, respectively. The anti parasitic effects were not associated to the promastigotes lack of viability. Instead, a significant rising on the L566 cells counting was detected when compared to control non treated parasites, from the first 6 hours treatment until 24 hours exposure. Morphological changes were detected in 62% of the 8mg/106 L566/mL parasite treated cells; 78% after 16mg/106 L566/mL treatment, and 98% of the cells presented morphological changes after 32mg/106 L566/mL treatment. Cells were either round-shaped, showing incomplete mitosis, or presenting double flagella, suggesting that the present extract containing substances that might interfere in the parasitic topoisomerase function. The parasitary mitochondrial activity evidenced the occurrence of a metabolic acceleration due by the OCMeOH treatment. The mitochondrial enzymatic activity was of 220% at exposures to 8mg/106 L566/mL; 389% at exposures to 16mg/106 L566/mL and of 480% at exposures to 32mg/106 L566/mL, respectively, when compared to the non treated parasites (100%). The anti parasitic potential (Therapeutic Index) was considered to be positive (acceptable) (TI=4,0) when estimated in function of the morphological changes observed at the extract concentration of 32mg/106 Mf/mL (LD62%), and at 8mg/106 L566/mL (ED62%), respectively. Since bi-flavonoids are the main constituents present in the OCMeOH, results were suggestive the anti parasitic effect was due to this group of secondary metabolites. As such biologically active molecules are known COX2 selective inhibitors, its internal use should be avoided. Otherwise, bi- flavonoids are good candidate substances to be applied as topical medicine to treat the American Tegumentar Leishmaniasis. / Os efeitos dos extratos de Ouratea cuspidata (Ochnaceae) foram avaliadas a partir da exposi??o de c?lulas hospedeiras e de promastigotas de Leishmania (Viannia) braziliensis (L566) ?s fra??es oriundas da sua parti??o hex?nica (OCH), metan?lica (OCMeOH) e acetato de et?lica (OCAcEt). Dentre estas, a fra??o OCMeOH foi a ?nica que apresentou potencial antiparasit?rio em doses que geraram bioprote??o aos sistemas hospedeiros. Assim, procederam-se an?lises de a??o parasiticida direta do extrato OCMeOH sobre formas promastigotas, onde, tamb?m, foram avaliados o crescimento, altera??es morfol?gicas e fun??es do metabolismo mitocondriais. Do mesmo modo, foram realizadas observa??es semelhantes em macr?fagos peritoneais de hamsters Cricetus cricetus (Mf) afim de se determinar a toxicidade relativa para o hospedeiro. Finalmente, foram realizados ensaios complementares de bioprote??o e gera??o de radicais livres, com a finalidade de confirmar a poss?vel aplica??o terap?utica dos constituintes presentes no extrato. Em todas as etapas experimentais, foram utilizadas cinco concentra??es de OCMeOH (2mg/mL; 4mg/mL; 8mg/mL; 16mg/mL e 32mg/mL), sendo que os resultados obtidos foram comparativamente avaliados em rela??o a sistemas de controle (L566 e Mf n?o tratados). Os (Mf) tratados com 16 mg/106 Mf/mL do extrato sofreram uma diminui??o significativa (92%) de sua capacidade fagocit?ria, mas permaneceram vi?veis, em sua maioria (78%). As altera??es morfol?gicas mostraram-se mais evidentes em 62% das c?lulas tratadas com 32mg/106 Mf/mL, com 70% de viabilidade e 98% de inibi??o da fagocitose. A atividade enzim?tica-mitocondrial macrof?gica apresentou diminui??o gradativa, com a preserva??o de 87% das fun??es enzim?ticas nas c?lulas tratadas com 8mg/106 Mf/mL; 66,3% daquelas tratadas com 16mg/106 Mf/mL e 49% das tratadas com 32mg/106 Mf/mL. As L566 n?o apresentaram a perda da viabilidade ap?s exposi??o de 24horas ?s diferentes concentra??es de OCMeOH. Ao contr?rio, houve um aumento significativo no n?mero de promastigotas que, a partir de seis horas de cultivo, mostraram-se sempre superiores ? quantidade de parasitos n?o tratados. Foram detectadas altera??es morfol?gicas em 62% das L566 tratadas com 8mg/106 L566/mL; 78% daquelas tratadas com 16mg/106 L566/mL e 98% das tratadas com 32mg/106 L566/mL. As promastigotas tratadas apresentaram-se arredondadas, com mitose incompleta, ou apresentando dois flagelos, sugerindo que as subst?ncias presentes no extrato podem interferir nas topoisomerases parasit?rias. A atividade mitocond rial parasit?ria evidenciou a ocorr?ncia de acelera??o metab?lica induzida pelo tratamento com OCMeOH. A atividade enzim?ticamitocondrial foi de 220% no tratamento com 8mg/106 L566/mL; de 389% com 16mg/106 L566/mL e 480% com 32mg/106 L566/mL quando comparadas ao controle (100%). O potencial antiparasit?rio (Indice Terap?utico) foi considerado positivo (IT=4,0) em fun??o das altera??es morfol?gicas observadas nas concentra??es de 32mg/106 Mf/mL (LD62%), e 8mg/106 L566/mL (ED62%). Sendo biflavon?ides os constituintes presentes em maior propor??o, atribuiu-se a este grupo de subst?ncias a a??o antiparasit?ria e imunot?xica observadas para OCMeOH. Os bi-flavon?ides presentes no extrato metan?lico de Ouratea cuspidata est?o entre os inibidores seletivos de COX2, sugerindo o desenvolvimento de pesquisas que visem sua utiliza??o t?pica em animais naturalmente infectados.
2

Anti-parasitic and anti-viral immune responses in insects

Terenius, Olle January 2004 (has links)
<p>Insects encounter many microorganisms in nature and to survive they have developed counter measures against the invading pathogens. In <i>Drosophila melanogaster</i> research on insect immunity has mainly been focused on infections by bacteria and fungi. We have explored the immune response against natural infections of the parasite <i>Octosporea muscaedomesticae</i> and the <i>Drosophila</i> C virus as compared to natural infections of bacteria and fungi. By using Affymetrix <i>Drosophila</i> GeneChips, we were able to obtain 48 genes uniquely induced after parasitic infection. It was also clearly shown that natural infections led to different results than when injecting the pathogens. </p><p>In order to search for the ultimate role of the lepidopteran protein hemolin, we used RNA interference (RNAi). We could show that injection of double stranded RNA (dsRNA) of <i>Hemolin</i> in pupae of <i>Hyalophora cecropia</i> led to embryonic malformation and lethality and that there was a sex specific difference. We continued the RNAi investigation of hemolin in another lepidopteran species, <i>Antheraea pernyi</i>, and discovered that hemolin was induced by dsRNA<i> per se</i>. A similar induction of hemolin was seen after infection with baculovirus and we therefore performed <i>in vivo</i> experiments on baculovirus infected pupae. We could show that a low dose of ds<i>Hemolin</i> prolonged the period before the <i>A. pernyi</i> pupae showed any symptoms of infection, while a high dose led to a more rapid onset of symptoms. By performing <i>in silico</i> analysis of the hemolin sequence from <i>A. pernyi</i> in comparison with other<i> Hemolin</i> sequences, it was possible to select a number of sites that either by being strongly conserved or variable could be important targets for future studies of hemolin function.</p>
3

Bioassay-guided phytochemical study of indigenous medicinal plants of Ethiopia

Gutu, Ketema Tolossa January 2018 (has links)
In many developing countries, farmers and pastoralists still rely on their indigenous knowledge, practices and locally available plants to control nematode parasitic infections, both in livestock and humans. The overall aim of my thesis was to undertake bioassay-guided phyto-chemical study of extracts and their constituents from Ethiopian anti-parasitic plants used by healers to control gastrointestinal nematode parasites in livestock to validate their ethno-medicinal use and to characterise and identify their active ingredients. As a first experiment (Chapter Three), four types of crude extracts (water, 70% methyl-alcohol, absolute methanol and acetone) of four indigenous Ethiopian medicinal plants (Adenia species, Cissus ruspolii, Ipomoea eriocarpa and Euphorbia thymifolia) were screened against Teladorsagia circumcincta egg hatching in vitro, not only as a first step to validate the traditional healers claim but also to choose the most promising plant extract(s) for further phyto-chemical studies. The egg hatching inhibition (EHI) test results revealed that the anti-parasitic properties of these plants depended on plant species, dose, and solvent polarity. The water extracts of both C. ruspolii and Adenia sp. exhibited largest, up to 100% EHI but also larval migration inhibition activities, and were selected for further studies. The second experiment (Chapter Four) assessed the nature of active constituents in these extracts by physico-chemical methods. It was observed that the major constituents of both plant extracts responsible for the EHI activities are likely highly polar, water-soluble, small and moderately heat-labile molecules. The third and fourth experiments (Chapters Five and Six) consisted of separating Cissus ruspolii and Adenia sp. water extracts into discrete fractions by gel-permeation chromatography, EHI tests of Bio-Gel P-2 fractions followed by thin layer chromatography (TLC) profiling of these fractions to detect separated spots (in day light, under UV-light or after staining with various staining reagents) and also to see how elution patterns of separated spots affected by column parameters. The EHI tests on the fractions obtained revealed that the active constituents of C. ruspolii and Adenia sp. water crude extracts were eluted into few fractions based on their molecular sizes. The TLC profilings of these fractions identified spot patterns of active and inactive fractions, which allowed pooling of active constituents based on their EHI and TLC profiling into three pools for each plant. The fifth experiment (Chapter Seven) was to isolate and purify compounds from these pools using various preparative planar and column chromatographic methods. Sequential applications of column chromatography followed by preparative thin layer chromatography isolated and purified five active compounds from C. ruspolii and two active compounds from Adenia sp. The sixth experiment (Chapter Eight) was to characterize and propose/elucidate structures of compounds from the active fractions using chromatographic, analytical and spectroscopic methods. In this regard, the structures of two oleanane type triterpenoid saponins isolated from one of active fractions of Adenia sp. were proposed based on their mass spectrometry (MS) and nuclear magnetic resonance (NMR) data with support of compounds property, TLC and literature. Similar outcomes for C. ruspolii were not achieved due to lack of sufficient sample to run 13C-nuclear magnetic resonance spectroscopy and distortionless enhancement by polarization transfer (DEPT), contamination of some purified compounds with ill-characterised substance from the preparative TLC matrix and in some cases mass spectrometry (MS) and nuclear magnetic resonance (NMR) data did not support each other. The last experiment (Chapter Nine) was to assess anthelmintic efficacy and safety of C. ruspolii and Adenia sp. crude water extracts in Heligmosomoides bakeri infected mice. This in vivo test revealed that both plant extracts exhibited significant reduction in worm burdens and worm egg excretion, with moderate effects on haematology and organ weights at tolerated dosages. In conclusion, both in vitro and in vivo data revealed that Adenia sp. and C. ruspolii have anthelmintic properties, thus validating traditional healer claims and supporting ethno-medicinal use. The bioassay-guided phytochemical study resulted in the isolation of a number of active compounds from these plants, for some of which a structure has been proposed.
4

Anti-parasitic and anti-viral immune responses in insects

Terenius, Olle January 2004 (has links)
Insects encounter many microorganisms in nature and to survive they have developed counter measures against the invading pathogens. In Drosophila melanogaster research on insect immunity has mainly been focused on infections by bacteria and fungi. We have explored the immune response against natural infections of the parasite Octosporea muscaedomesticae and the Drosophila C virus as compared to natural infections of bacteria and fungi. By using Affymetrix Drosophila GeneChips, we were able to obtain 48 genes uniquely induced after parasitic infection. It was also clearly shown that natural infections led to different results than when injecting the pathogens. In order to search for the ultimate role of the lepidopteran protein hemolin, we used RNA interference (RNAi). We could show that injection of double stranded RNA (dsRNA) of Hemolin in pupae of Hyalophora cecropia led to embryonic malformation and lethality and that there was a sex specific difference. We continued the RNAi investigation of hemolin in another lepidopteran species, Antheraea pernyi, and discovered that hemolin was induced by dsRNA per se. A similar induction of hemolin was seen after infection with baculovirus and we therefore performed in vivo experiments on baculovirus infected pupae. We could show that a low dose of dsHemolin prolonged the period before the A. pernyi pupae showed any symptoms of infection, while a high dose led to a more rapid onset of symptoms. By performing in silico analysis of the hemolin sequence from A. pernyi in comparison with other Hemolin sequences, it was possible to select a number of sites that either by being strongly conserved or variable could be important targets for future studies of hemolin function.
5

I. Total synthesis of [plus or minus] ovalicin and its analogues II. Bio-based polymers from vegetable oil III. New synthetic methods of diacetylene fatty acids

Zhao, Huiping January 1900 (has links)
Doctor of Philosophy / Department of Chemistry / Duy H. Hua / I. Ovalicin is a natural product isolated from the culture of fungus Pseudorotium ovalis Stolk, it selectively inhibit type 2 methionine amino-peptidase (MetAP 2), which related to many physiological activities such as angiogenesis. Total synthesis of [plus or minus] ovalicin, its C4(S*), C4(S*)C5(S*) stereo-isomers, and C5 regio-isomer were synthesized via an intramolecular Heck reaction of (Z)-3-(t-butyldimethyl silyloxy)-1-iodo-1,6-heptadiene utilizing a catalytic amount of palladium acetate. Subsequent epoxidation, dihydroxylation, methylation (or stereochemistry inversion before or after methylation) and oxidation led to a variety of ketones, key intermediates for synthesis of ovalicin and its analogues. Introduction of side-chain to ketones by lithium (Z)-6-methylhepta-2,5-dien-2-ide and following functional group transformation led to ovalicin and its analogues. Anti-trypanosomal activities of various ovalicin analogues and synthetic intermediates were evaluated. II. Bio-based polymers from vegetable oils are renewable and environment-friendly materials. Dihydroxylated, trihydroxylate, tetrahydroxylated and hexahydroxylated triglycerides, triamino and triisopropylamino glycerides were synthesized from model triglyceride glyceryl trioleoate. These monomers were cross-linked with 1, 4-phenylene diisocynate to make polyurethanes and polyureas. The physical properties of these polymers were examined by gel content and swelling value measurements, thermodynamic and viscoelastic properties were studied from TGA, DSC and DMA measurements. The structure-property relationship was discussed based on these measurements. III. Diacetylenic fatty acids were widely applied in material science to regulate alignment on surface and stabilize self-assembled nanomaterials. A novel synthetic method of diacetylenic fatty acids from vegetable oils was developed. Its self-assembling properties on alumina surface were measured and discussed.

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