<p>In South Africa, it is estimated that approximately 70% of the population frequently make use of traditional medicinal plants for their health care needs. The use of Lessertia frutescens by the&nbsp / various cultural groups in South Africa dates back to the earlier civilizations and continues to be used today to treat a multitude of ailments. To get the best results from a medicinal plant, one&nbsp / &nbsp / would need to ensure that the crude material is of good quality through interventions like being properly grown, well dried and correctly processed. This would add a measure of quality&nbsp / assurance, which will contribute towards the safety and efficacy aspect of herbal medicine. The aim of this study was to investigate what impact a particular season of harvest and the time in&nbsp / storage would have on the flavonoid and triterpenoid marker levels of Lessertia frutescens. To achieve this, the following was investigated: (1) storage variation of Lessertia frutescens leaves&nbsp / by comparing the results obtained from the High Performance Liquid Chromatography (HPLC) analysis of the flavonoids and triterpenoids, (2) seasonal variation of Lessertia frutescens&nbsp / leaves by comparing the results obtained from the HPLC analysis of the flavonoids and triterpenoids, (3) leaf and stem variation of Lessertia frutescens by comparing the results obtained from HPLC analysis of the flavonoids and triterpenoids. The hypotheses were: (1) the stored sample would indicate the same level of the biomarkers for the flavonoids and triterpenoids, as that of&nbsp / the freshly prepared sample, (2) the sample that was harvested during the summer season would indicate higher levels of the biomarkers of&nbsp / flavonoids and triterpenoids than the other three&nbsp / seasons, (3) the leaf sample would indicate the same level of the biomarkers for the flavonoids and triterpenoids, as that of the stem sample. An Agilent 1200 series HPLC was used for the&nbsp / determination of the flavonoids sutherlandin A and sutherlandin D as well as the triterpenoids sutherlandioside B and sutherlandioside D. Results show that for both sutherlandin A (summer:&nbsp / 3.67 &plusmn / 2.88 mg/ml / storage: 4.07 &plusmn / 2.88 mg/ml) and D (summer: 4.10 &plusmn / 1.06 mg/ml / storage: 4.25 &plusmn / 1.06 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the case of the storage&nbsp / amples. For both sutherlandioside B (summer: 3.01 &plusmn / 0.39 mg/ml / storage: 2.82 &plusmn / 0.39 mg/ml) and D (summer: 5.82 &plusmn / 0.42 mg/ml / storage: 4.66 &plusmn / 0.42 mg/ml) show significantly (P &lt / &nbsp / .0001)&nbsp / higher concentrations in the case of the fresh summer samples.For the seasonal comparison, results show that for sutherlandin A (summer: 3.67 &plusmn / 12.49 mg/ml / autumn: 4.75 &plusmn / &nbsp / 12.49 mg/ml / winter: 4.23 &plusmn / 12.49 mg/ml / spring: 6.56 &plusmn / 12.49 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the case of the spring sample. For sutherlandin D (summer: 4.10&nbsp / &nbsp / 10.32 mg/ml / autumn: 6.37 &plusmn / 10.32 mg/ml / winter: 5.25 &plusmn / 10.32 mg/ml / spring / 6.08 &plusmn / 10.32 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the case of the autumn sample. For both sutherlandioside B (summer: 3.01 &plusmn / 7.19 mg/ml / autumn: 2.15 &plusmn / 7.19 mg/ml / winter: 2.89 &plusmn / 7.19 mg/ml / spring: 1.47 &plusmn / 7.19 mg/ml) and D (summer: 5.82 &plusmn / 14.48 mg/ml / autumn: 3.33 &plusmn / 14.48 mg/ml / winter: 4.23 &plusmn / 14.48 mg/ml / spring: 2.50 &plusmn / 14.48 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the case of the autumn sample. For the summer&nbsp / leaf/stem comparison, results show that for sutherlandin A (leaf: 3.67 &plusmn / 8.18 mg/ml / stem: 4.67 &plusmn / 8.18 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the case of the stem&nbsp / sample. For the sutherlandin D (leaf: 4.10 &plusmn / 4.81 mg/ml / stem: 3.31 &plusmn / 4.81 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the case of the summer leaf sample. For both the&nbsp / sutherlandioside B (leaf: 3.01 &plusmn / 4.24 mg/ml / stem: 3.62 &plusmn / 4.24 mg/ml) and D (leaf: 5.82 &plusmn / 0.42 mg/ml / stem: 5.80 &plusmn / 0.42 mg/ml) show significantly (P &lt / 0.0001) higher concentrations in the&nbsp / case of the stem samples. Results demonstrate that the production of secondary metabolites are influenced by&nbsp / &nbsp / environmental factors like seasonal harvesting, as indicated by the variation in the chemical constituent composition of Lessertia frutescens depending on the season collected in. Moreover, the storage of Lessertia frutescens for a period of one year resulted in an&nbsp / increase of two of the four constituents being monitored. There was slight variations in the chemical constituents, depending on whether the leaf or stem material of Lessertia frutescens was being used. Finally, the type of chemical constituent being monitored was also important in the consideration of this study. Therefore, this study can be seen as a starting point to further&nbsp / &nbsp / investigations of these aspects, which are of clinical, pharmacological and economic</p>

This study aimed to craft production in copper stills, the spirits of honey of different flowers, juice of sugar cane, honey bees, sugar cane and grape mixed (juice of cane sugar and honey from different flowers) and subsequent aging in oak barrels.The tests were conductedby mixing 75% juice with 25% inoculum (30L and 10L, respectively). The grape juice were cane sugar, honey bees-of-seven different flowers and mixed (where the ratio of 50% by volume for each party).Samples were taken must and wine, quantifying pH, sulfuric acid, ART, Brix, and also ethanol content in wine. In conducting fetched, three fractions were taken: head (8%), heart (84%) and tail (8%), quantifying pH, density, alcohol content, dry extract and acetic acid.The operating temperatures of the still were split head, with initial and final temperatures respectively equal to 89.3 and 93.0 o C; fraction of heart, 93.0 and 96.0 ° C and split tail, 96.0 and 97 , 0 ° C. The duration of each step was about 25 minutes, 70 minutes and 23 minutes respectively, and volumes were obtained 500mL, 500mL and5380mL in the same order. During aging, were measured monthly, alcoholic, acetic acid, pH, solids, density, evaporation rate, visual appearance and color.The results indicated that it is possible to produce brandy honey from different flowers, with appropriate physical and chemical characteristics.The production of grape brandy using resulted in mixed drink with different physical and chemical characteristics in relation to the sugar cane and similar to those obtained with the respective pure honey.This mixture reduces the production costs of brandy,when compared with that produced from pure honey. When we worked with the honey wine, which showed better fermentation efficiencywas honey G (mixed flora, from the state of Ceara).However, when working with mixed wine, the best performance was obtained with Mel C (mixed flora, native of the City of Pao de Acucar - AL). / Fundação de Amparo a Pesquisa do Estado de Alagoas / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Este estudo objetivou a producao artesanal, em alambique de cobre, de aguardentes de mel de abelhas de diferentes floradas, de caldo de cana-de-acucar, de mel de abelhas de cana-de-acucar e de mostos mistos (caldo de cana-de-acucar e mel de abelhas de diferentes floradas) e posterior envelhecimento em barris de carvalho. Os ensaios foram conduzidos misturando-se 75% de mosto com 25% de inoculo (30L e 10L, respectivamente). Os mostos foram de caldo de cana-de-acucar, de mel-de-abelhas de 7 diferentes floradas e mistos (sempre na proporcao de 50%, em volume, para cada uma das partes). Foram retiradas amostras de mosto e de vinho, quantificando-se pH, acidez sulfurica, ART, Brix e, tambem, teor de etanol no vinho. Na conducao das alambicadas, foram retiradas 3 fracoes: de cabeca (8%), de coracao (84%) e de cauda (8%), quantificando-se pH, densidade, teor alcoolico, extrato seco e acidez acetica. As temperaturas de operacao do alambique foram: fracao de cabeca, com temperaturas inicial e final, respectivamente iguais a 89,3 e 93,0oC; fracao de coracao, 93,0 e 96,0oC e fracao de cauda, 96,0 e 97,0oC. A duracao de cada etapa foi de cerca de 25 minutos, 70 minutos e 23 minutos, respectivamente, e os volumes obtidos foram 500mL, 5380mL e 500mL, na mesma ordem. Durante o envelhecimento, foram quantificados, mensalmente, teor alcoolico, acidez acetica, pH, extrato seco, densidade, taxa de evaporacao, aspecto visual e cor. Os resultados obtidos indicaram que e possivel a producao de aguardente de mel de abelhas de diferentes floradas, com caracteristicas fisico-quimicas adequadas. A producao de aguardente utilizando-se mostos mistos resultou em bebida com caracteristicas fisico-quimicas diferenciadas em relacao a da cana-deacucar e semelhantes as obtidas com os respectivos meis puros. Esta mistura reduz os custos de producao da aguardente, quando se compara com a produzida a partir de meis puros. Quando se trabalhou com mosto de mel de abelhas, o que apresentou melhor eficiencia de fermentacao foi o mel G (florada mista, oriundo do estado do Ceara).Porem, quando se trabalhou com mosto misto, o melhor desempenho foi obtido com o mel C (florada mista, oriundo do Municipio de Pão-de-Acucar – AL).

Orientador: Silmara Marques Allegretti / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-12T15:42:28Z (GMT). No. of bitstreams: 1 Oliveira_ClaudineideNascimentoFernandesde_M.pdf: 7853084 bytes, checksum: e21dff7dd2d35d9545d8717dc30fdb79 (MD5) Previous issue date: 2008 / Resumo: A esquistossomose mansonica e uma doenca cronica e debilitante. Estima-se que mais de 200 milhoes de pessoas estejam infectadas no mundo pela doenca, e no Brasil estima-se que existe de 8 a 10 milhoes de pessoas infectadas. O tratamento da esquistossomose e baseado na quimioterapia com o praziquantel, principalmente devido a sua atividade contra todas as especies de Schistosoma patogenicas ao homem. Infelizmente, o uso extensivo e inapropriado desse farmaco pelo mundo, culminou com o aparecimento de esquistossomas tolerantes, gerando uma preocupacao sobre a selecao da resistencia a esse medicamento. Entao, para se controlar a esquistossomose, ha necessidade de se desenvolver novas opções de farmacos, como alternativa ao praziquantel. As plantas medicinais vem sendo aplicadas e testadas como novas alternativas medicamentosas para o tratamento de parasitoses. Este estudo teve por objetivo avaliar a atividade esquistossomicida da planta Phyllanthus amarus L. sobre Schsistosoma mansoni, linhagem BH em camundongos Swiss. Foram testados os extratos hexanico e etanolico nas concentracoes 100, 150 e 250mg/Kg, e a fracao de Lignanas 50 e 100mg/Kg, administrados em dose unica por tubagem esofagica. Os animais foram divididos em dois grupos de acordo com o periodo de tratamento (30 ou 45 dias apos a infeccao para avaliar a acao dos extratos nos vermes jovens e adultos, respectivamente). Foram analisados os seguintes parametros: quantidade/ porcentagem de vermes adultos nas veias mesentericas, porta e nas visceras; proporcao entre machos e femeas; reducao no numero de vermes; reducao no numero de ovos eliminados para o ambiente externo e retidos no tecido intestinal; alteracao no oograma; aspecto visual dos granulomas nas visceras; alteracao no tamanho dos granulomas encontrados no figado. De acordo com os resultados obtidos, o tratamento feito com o extrato etanolico 250mg/Kg no 30° dia de infeccao, foi o que apresentou maior reducao do numero de vermes (63%), e o grupo tratado com o extrato hexanico 100mg/Kg nesse mesmo periodo foi o que apresentou maior quantidade de vermes nas visceras (n=3,1). Ja os tratamentos feitos com o extrato etanolico 100mg/Kg e a fracao de Lignanas 50mg/Kg apos 30 dias de infeccao, conseguiram cessar a postura de ovos, o que fez com que os orgaos desses grupos fossem pouco lesados. Ocorreu ausencia de granulomas nos figados observados histologicamente para o grupo tratado com o extrato etanolico 100mg/Kg. No 45° dia apos a infeccao o grupo tratado com o extrato hexanico 150mg/Kg foi o que apresentou resultado mais significativo, uma vez que ele conseguiu reduzir o numero de ovos imaturos e aumentar o de ovos maduros, indicando que este tratamento alterou a oviposicao do verme. Os resultados obtidos para os extratos hexanico e etanolico e fracao de Lignanas demonstraram potencial atividade nos diferentes parametros avaliados, evidenciando que a planta P. amarus possui efeito contra o S. mansoni linhagem BH. / Abstract: The schistosomiasis is a chronic and debilitating disease. It is estimated more than 200 million people worldwide are infected by the disease, and in Brazil is estimated 8 to 10 million people infected. The treatment of schistosomiasis is based on praziquantel chemotherapy, mainly due to its activity against all species of pathogenic Schistosoma to humans. The inappropriate and extensive use of this drug in the world culminated in the appearance of tolerant worms, generating a concern about the selection of resistance this drug. So, to control schistosomiasis, there is need to develop new options for drugs, as an alternative to the praziquantel. Medicinal plants have been implemented and tested as a new alternative drug for the treatment of parasitic. The objective of this study was to assess the presence of antichistosomal activity of the plant Phyllanthus amarus L. using Schistosoma mansoni infected mice of BH strain. The hexane and ethanolic extracts with 100, 150 and 250mg/Kg, concentrations and the Lignans fraction 50 and 100mg/kg, were administered with single oral dose by esophageal intubation. The animals were divided into two groups according to the treatment period (30 or 45 days after infection to evaluate the effect of the extracts in young and adult worms respectively). Analyzes performed consists in a set of parameters: quantity / percentage of adult worms in the veins mesenteric, port and the viscera; proportion of male and female worms, worms reduction; eggs reduction eliminated environment and retained in the intestinal tissue; change in oograma; visual aspect of granulomas in the viscera; change in the size of granulomas found in the liver. According to the results, the treatment with ethanolic extract 250mg/Kg in 30th days of infection, presented the greatest reduction in the number of worms (63%), and the group treated with the hexane extract 100mg/kg in same period presented the highest number of worms in the viscera (n=3.1). The treatments made with ethanolic extract 100mg/kg and the Lignans fraction 50mg/kg after 30 days of infection managed to stop the egg laying. So that the organs of these groups were slightly injured. There was absence of granulomas in the liver observed histological for the group treated with ethanolic extract 100mg/kg. In the 45th days after infection the group treated with the hexane extract 150mg/Kg presented the most significant result, as to reduced the number of immature eggs, and increased of mature eggs, indicating that this treatment has changed the worm oviposition. The results for the ethanol, hexane extracts and fraction of Lignans demonstrated potential activity in the various parameters measured, suggesting that the plant P. amarus has effect against S.mansoni strain BH. / Mestrado / Mestre em Parasitologia

>Magister Scientiae - MSc / In South Africa, it is estimated that approximately 70% of the population frequently make use of traditional medicinal plants for their health care needs. The use of Lessertia frutescens by the various cultural groups in South Africa dates back to the earlier civilizations and continues to be used today to treat a multitude of ailments. To get the best results from a medicinal plant, one would need to ensure that the crude material is of good quality through interventions like being properly grown, well dried and correctly processed. This would add a measure of quality assurance, which will contribute towards the safety and efficacy aspect of herbal medicine. The aim of this study was to investigate what impact a particular season of harvest and the time in storage would have on the flavonoid and triterpenoid marker levels of Lessertia frutescens. To achieve this, the following was investigated: (1) storage variation of Lessertia frutescens leaves by comparing the results obtained from the High Performance Liquid Chromatography (HPLC) analysis of the flavonoids and triterpenoids, (2) seasonal variation of Lessertia frutescens leaves by comparing the results obtained from the HPLC analysis of the flavonoids and triterpenoids, (3) leaf and stem variation of Lessertia frutescens by comparing the results obtained from HPLC analysis of the flavonoids and triterpenoids. The hypotheses were: (1) the stored sample would indicate the same level of the biomarkers for the flavonoids and triterpenoids, as that of the freshly prepared sample, (2) the sample that was harvested during the summer season would indicate higher levels of the biomarkers of flavonoids and triterpenoids than the other three seasons, (3) the leaf sample would indicate the same level of the biomarkers for the flavonoids and triterpenoids, as that of the stem sample. An Agilent 1200 series HPLC was used for the determination of the flavonoids sutherlandin A and sutherlandin D as well as the triterpenoids sutherlandioside B and sutherlandioside D. Results show that for both sutherlandin A (summer: 3.67 ± 2.88 mg/ml; storage: 4.07 ± 2.88 mg/ml) and D (summer: 4.10 ± 1.06 mg/ml; storage: 4.25 ± 1.06 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the storage samples. For both sutherlandioside B (summer: 3.01 ± 0.39 mg/ml; storage: 2.82 ± 0.39 mg/ml) and D (summer: 5.82 ± 0.42 mg/ml; storage: 4.66 ± 0.42 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the fresh summer samples. For the seasonal comparison, results show that for sutherlandin A (summer: 3.67 ± 12.49 mg/ml; autumn: 4.75 ± 12.49 mg/ml; winter: 4.23 ± 12.49 mg/ml; spring: 6.56 ± 12.49 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the spring sample. For sutherlandin D (summer: 4.10 ± 10.32 mg/ml; autumn: 6.37 ± 10.32 mg/ml; winter: 5.25 ± 10.32 mg/ml; spring; 6.08 ± 10.32 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the autumn sample. For both sutherlandioside B (summer: 3.01 ± 7.19 mg/ml; autumn: 2.15 ± 7.19 mg/ml; winter: 2.89 ± 7.19 mg/ml; spring: 1.47 ± 7.19 mg/ml) and D (summer: 5.82 ± 14.48 mg/ml; autumn: 3.33 ± 14.48 mg/ml; winter: 4.23 ± 14.48 mg/ml; spring: 2.50 ± 14.48 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the autumn sample. For the summer leaf/stem comparison, results show that for sutherlandin A (leaf: 3.67 ± 8.18 mg/ml; stem: 4.67 ± 8.18 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the stem sample. For the sutherlandin D (leaf: 4.10 ± 4.81 mg/ml; stem: 3.31 ± 4.81 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the summer leaf sample. For both the sutherlandioside B (leaf: 3.01 ± 4.24 mg/ml; stem: 3.62 ± 4.24 mg/ml) and D (leaf: 5.82 ± 0.42 mg/ml; stem: 5.80 ± 0.42 mg/ml) show significantly (P < 0.0001) higher concentrations in the case of the stem samples.Results demonstrate that the production of secondary metabolites are influenced by environmental factors like seasonal harvesting, as indicated by the variation in the chemical constituent composition of Lessertia frutescens depending on the season collected in. Moreover, the storage of Lessertia frutescens for a period of one year resulted in an increase of two of the four constituents being monitored. There was slight variations in the chemical constituents, depending on whether the leaf or stem material of Lessertia frutescens was being used. Finally, the type of chemical constituent being monitored was also important in the consideration of this study. Therefore, this study can be seen as a starting point to further investigations of these aspects, which are of clinical, pharmacological and economic importance.

As biotécnicas da reprodução animal, amplamente difundidas no Brasil, tem possibilitado maior controle em relação à transmissão de agentes patogênicos, porém, a transmissão de doenças continua a ser uma preocupação justificável para a busca de melhores meios de controle. O objetivo do trabalho foi o de avaliar a diminuição da replicação viral (BoHV-1 Colorado) em embriões murinos após tratamentos com Produtos Naturais (PN). Trabalhamos com 3 grupos de PN, totalizando 18 tratamentos: Oléos Essenciais (OE) de Chamomilla recutita, Allium sativum, Zingiber officinale, Syzygium aromaticum, Illicium verum Hook f., Punica granatum e Melissa officinalis; Extratos Etanólicos (EE) das mesmas plantas anteriores com acréscimo de Pterodon emarginatus (casca e semente) e Extrato Aquoso (EA) de Própolis e Agaricus blazei. Inicialmente encontramos concentrações não tóxicas em células MDBK (Madin Darby Bovine Kidney) relacionada a cada PN (ampla sintonia) para posteriormente verificarmos as concentrações não tóxicas para os zigotos/embriões murinos (sintonia fina). Após encontrarmos as concentrações não tóxicas, os zigotos foram divididos em quatro grupos: G1 (Controle), G2 (expostos aos vírus BoHV-1 Colorado à 108 TCID50/mL), G3 (expostos aos PN) e G4 (expostos aos vírus e PN). Os grupos foram mantidos a 37,5 ºC em meio TCM199 (100 &mu;L) com 10% de soro fetal bovino em estufa a 5% de CO2 e 95% de umidade. Após 24h, analisamos a taxa de clivagem (Teste Exato de Fisher_p&lt;0,05), a morfologia (por microscopia óptica), a n-PCR e a titulação dos embriões em co-cultura com células MDBK após mais 72h do tratamento com os PN (Teste de Mann Whitney_ p&lt;0,05). Dentre as 18 análises, os embriões murinos tratados com EA de Própolis, EE de Punica granatum e de Pteron emarginatus (Casca) apresentaram resultados satisfatórios: sem alterações morfológicas, taxa de clivagem semelhante aos controles e apesar da constatação da partícula viral junto aos embriões pela n-PCR, houve diminuição da concentração viral após tratamentos com estes extratos, o que sugere interferência destes tratamentos no ciclo viral do BoHV-1 Colorado. / The biotech Animal reproduction widespread in Brazil, has allowed greater control over the transmission of pathogens, however, the disease transmission remains a justifiable concern for the pursuit of the best means of control. The aim of this study was to evaluate the reduction of viral replicarion (Colorado BoHV-1) in murine embryos after treatments with natural products (NP). We work with groups \'3 \'PN, totaling 18 treatments: essential oils (EO) of Chamomilla recutita, Allium sativum, Zingiber officinale, Syzygium aromaticum, Illicium verum Hook f., Melissa officinalis and Punica granatum; ethanol extracts (EE) of the same plants prior to adding Pterodon emarginatus (skin and seed) and Aqueous Extract (EA) Propolis and Agaricus blazei. Initially we find non-toxic concentrations in MDBK (Madin Darby Bovine Kidney) related to each PN (broad line) then we check for the non-toxic concentrations for zygotes / embryos murine (fine tuning). After finding the non-toxic concentrations, zygotes were divided into four groups: G1 (control), G2 (exposed to viruses BoHV-1 at 108 Colorado TCID50/mL), G3 (exposed to PN) and G4 (exposed to viruses and PN .) The groups were maintained at 37.5 °C in TCM199 (100 mL) with 10% fetal bovine serum in an incubator at 5% CO2 and 95% humidity. After 24h, we analyzed the cleavage rate (Exact Test Fisher_p &lt;0.05), morphology (light microscopy), the n-PCR and titration of embryos in co-culture with MDBK cells after 72 h of treatment with more NPs (Mann Whitney_ p &lt;0.05). Among the 18 analyzes, murine embryos treated with EA Propolis, EE Punica granatum and Pteron emarginatus (bark) showed satisfactory results: no morphological changes, cleavage rate similar to controls, and despite the finding of the viral particle with the embryos by n-PCR, decreased the viral concentration after treatment with these extracts, suggesting interference of these treatments in the viral cycle of BoHV-1 Colorado.

Submitted by Carvalho Dias João Paulo (joao.dias@famerp.br) on 2018-04-04T13:34:51Z No. of bitstreams: 1 lucianigaspardetoledo_dissert.pdf: 1518015 bytes, checksum: e7bd4f850932330d46b398509046745b (MD5) / Made available in DSpace on 2018-04-04T13:34:51Z (GMT). No. of bitstreams: 1 lucianigaspardetoledo_dissert.pdf: 1518015 bytes, checksum: e7bd4f850932330d46b398509046745b (MD5) Previous issue date: 2016-06-24 / Introduction: Candida spp are opportunistic pathogens isolated from human biota in the gastrointestinal tract, oral and vaginal mucosa, which can lead to the development of superficial lesions to disseminated infections, especially in immunosuppression. The high toxicity, the high cost of treatment and the emergence of resistant strains justify the search for new therapeutic agents. The plant biodiversity is rich in active ingredients that have contributed to the development of new and effective drugs, less expensive treatments and population access. Cymbopogon nardus (L.) Rendle is a plant popularly known as citronella and cultivated in subtropical and tropical areas of Asia, Africa and America, including Brazil. Essential oils present in the Cymbopogon genus plants have been widely studied, but there are few studies involving chemical analysis and microbiological ethanol extract of C. nardus. Objective: The objective of this study was to evaluate the antifungal potential, in vitro, of ethanol extract (EE) and essential oil (EO) from the leaves of Cymbopogon nardus (L.) Rendle (citronella) clinical isolates against of Candida spp. Material and Methods: In this study the species Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, Candida parapsilosis sensu stricto and C. orthopsilosis were selected. EE was obtained by extraction ultrasonic bath and analyzed by ultra-performance liquid chromatography (UPLC-ESI-QTOF-MS). The EO was extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometer (GC-MS). The antifungal activity of EE and EO was performed by determination of the minimum inhibitory concentration (MIC), time-kill assay inhibition of hyphal growth of C. albicans and inhibit mature biofilm. Additionally, the cytotoxic evaluation (determination of IC50) was assessed in HepG-2 cell lines (hepatic) and MRC-5 (fibroblast). Results: The results of the chemical analysis of EE showed presence of glycosylated flavones and glycosylated phenylpropanoids. According to the EO chemical analysis, the main compounds observed were monoterpenes containing-oxygen: citronellal, geranial, geraniol, citronellol and neral. Biological assays showed effective antifungal activity of EE (MIC 1000 to 125 μg / ml) and of EO (MIC 1000 the 250 μg/ml). In the time-kill assay was observed inhibition of growth of the species tested for EE and EO. The hyphal growth of C. albicans was inhibited by EE (1000 to 31 μg/ml) and the EO (1000 to 15.8 μg/ml) for 12 and 24 hours. The EE and EO inhibit mature biofilm species C. albicans, C. krusei, and C. parapsilosis at concentrations of 50xCIM and 10xCIM, respectively. EE exhibited the lower IC50 values for HepG-2 (322 μg/ml) and MRC-5 (181.1 μg/ml) than essential oil that showed IC50 values for HepG-2 (96.6 μg/ml) and MRC-5 (33.1 μg/ml). Conclusions: The EE and EO from C. nardus present as a promising source of new molecules with antifungal activity especially to the inhibition of the main virulence factors such as formation of hyphae and biofilm. / Introdução: Leveduras do gênero Candida são patógenos oportunistas isolados da biota humana no trato gastrointestinal, mucosa oral e vaginal, que podem levar ao desenvolvimento de lesões superficiais até infecções disseminadas, especialmente em situações de imunossupressão. O alto custo para tratamento de infecções, a elevada toxicidade, e o surgimento de cepas resistentes justificam a busca de novos agentes terapêuticos. A biodiversidade vegetal é rica em princípios ativos que têm contribuído com o desenvolvimento de novos e efetivos medicamentos, mais econômicos e de fácil acesso populacional. Cymbopogon nardus (L.) Rendle, é uma planta popularmente conhecida como citronela e cultivada em áreas subtropicais e tropicais da Ásia, África e América, incluindo o Brasil. Os óleos essenciais presentes em plantas do gênero Cymbopogon têm sido amplamente estudados, porém, a analise química e microbiológica do extrato etanólico de C. Nardus,é pouco explorada Objetivo: O objetivo deste estudo foi avaliar o potencial antifúngico, in vitro, do extrato etanólico (EE) e do óleo essencial (OE) das folhas de Cymbopogon nardus (L.) Rendle (citronela) frente isolados clínicos de Candida spp. Material e Métodos: Foram consideradas as espécies: Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis, Candida parapsilosis sensu stricto e C. orthopsilosis. O EE foi obtido por extração em banho ultrassônico e analisado por cromatografia liquida de ultra performance (UPLC-ESI-QTOF-MS). O OE foi extraído por hidrodestilação e analisado por cromatografia gasosa acoplada a espectrômetro de massa (CG-EM). A atividade antifúngica do EE e do OE foi realizada através da determinação da concentração inibitória mínima (CIM), ensaio time-kill, inibição do crescimento hifal de C. albicans e inibição de biofilme maduro. Adicionalmente, a avaliação citotóxica (determinação de IC50) foi estabelecida em linhagens celulares HepG-2 (hepática) e MRC-5 (fibroblasto). Resultados: Os resultados da análise química do EE evidenciaram presença de flavonas e fenilpropanoides glicosilados. De acordo com a análise química do OE, os principais compostos observados foram monoterpenos contendo oxigênio: citronelal, geranial, geraniol, citronelol e neral. Os ensaios biológicos mostraram importante atividade antifúngica para o EE (CIM de 1000 a 125 μg/mL) e, para o OE (CIM de 1000 a 250 μg/mL). A inibição do crescimento ocorreu para todas as espécies avaliadas frente aos produtos, EE e OE. O EE (1000 até 31 μg/mL) e OE (1000 até 15,8 μg/mL) inibiram o desenvolvimento da hifa de C. albicans, durante 12 e 24 horas, além de inibir o biofilme maduro das espécies de C. albicans, C. krusei e C. parapsilosis, nas concentrações de 50xCIM e 10xCIM, respectivamente. O EE apresentou os menores valores de IC50 para HepG-2 (322 μg/mL) e MRC-5 (181,1 μg/mL) em comparação com o óleo essencial - IC50 para HepG-2 (96,6 μg/mL) e MRC-5 (33,1 μg/mL). Conclusões: O EE e OE de C. nardus apresentam-se como uma fonte promissora de novas moléculas com atividade antifúngica com destaque à inibição dos principais fatores de virulência: formação de hifa e biofilme.

Tyrimo tikslas - parinkti neetanolinio propolio ekstrakto technologiją ir atlikti veikliųjų medžiagų analizę. Uždaviniai: nustatyti propolio koncentracijos, ekstrakcijos tirpiklių bei jų koncentracijų, ekstrakcijos laiko, temperatūros, ekstrakcijos metodo įtakas fenolinių junginių išsiskyrimui. Nustatyti propolio ekstrakto antiradikalinį aktyvumą ir flavonoidų kiekį bei efektyviosios skysčių chromatografijos metodu nustatyti propolio ekstrakte esančius cheminius junginius ir jų antiradikalinį aktyvumą. Metodika. Bendras fenolinių junginių kiekis, buvo nustatytas naudojant farmakopėjinį Folino Ciocalteu metodą pagal galo rūgšties ekvivalentą. Flavonoidų kiekis nustatytas naudojant spektrofotometrinį metodą pagal rutino ekvivalentą. Antioksidacinis aktyvumas spektrofotometriškai buvo nustatomas naudojant DPPH radikalą. Efektyviosios skysčių chromatografijos su elektrocheminiu detektoriumi būdu buvo identifikuoti fenoliniai junginiai ir nustatytas jų antiradikalinis aktyvumas. Rezultatai. Didinant propolio koncentraciją, naudojant tirpiklius didėja bendras fenolinių junginių kiekis (p<0,05). Technologiniai procesai t.y. ekstrakcijos trukmė, temperatūros padidinimas daro reikšmingą įtaką fenolinių junginių išsiskyrimui (p<0,05). Didžiausias bendras fenolinių junginių kiekis 35,60 mg/ml buvo gautas naudojant ultragarsu skatinamą ekstrakciją 10 min esant 70 ⁰C temperatūrai, ekstrakcijos tirpikliu naudojant 30 proc. makrogolio ir vandens mišinį. Šiame ekstrakte nustatytas 88,76±1... [toliau žr. visą tekstą] / The aim of survey - to select the technology of nonethanolic propolis extract and analyse the active substances. Tasks: To examine the influence of concentration, solvent and its concentration, extraction time, temperature, the extraction method on release of total phenolic compounds. Determine the radical scavenging activity and flavonoid content of propolis extract and using high performance liquid chromatography method identify active substances and its antiradical activity. Methods: Total phenolic compounds were determined by using pharmacopoeial Folin Ciocalteu method expressed in gallic acid equivalents. Flavonoids were determined using a spectrophotometric method in rutin equivalent. Radical scavenging activity was determined by spectrophotometry using DPPH radical. High performance liquid chromatography with electrochemical detection were used to identify phenolic compounds and its radical scavenging activity. Results: Increasing of propolis concentration made a significance impact on the amount of total phenolic content (p<0,05). Extraction time, increase of temperature made a significance impact on increase of the amount of total phenolic content (p<0,05). The highest amount of total phenolic compounds 35,60 mg/ml were obtained using ultrasonic induced extraction for 10 minutes at 70 ⁰ C temperature and using 30 % macrogol – water solution as solvent. The radical scavenging capacity of this extract was 88,76 ± 1,27% and amount of flavonoids were 3,89 ± 0,43 mg/ml... [to full text]

122 f. / Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-03-22T17:20:17Z No. of bitstreams: 1 Tese corrigida final.pdf: 1322795 bytes, checksum: a277616fb5634d49040e3b569eae8dd2 (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-06-05T17:38:45Z (GMT) No. of bitstreams: 1 Tese corrigida final.pdf: 1322795 bytes, checksum: a277616fb5634d49040e3b569eae8dd2 (MD5) / Made available in DSpace on 2013-06-05T17:38:45Z (GMT). No. of bitstreams: 1 Tese corrigida final.pdf: 1322795 bytes, checksum: a277616fb5634d49040e3b569eae8dd2 (MD5) Previous issue date: 2012 / CAPES / No presente trabalho, desenvolveu-se um método alternativo para determinação de cobre, ferro, níquel e zinco, usando a espectrometria de fluorescência de raios-X com energia dispersiva (EDXRF), em amostras de etanol combustível e cachaça, após procedimento de pré-concentração. O procedimento consistiu na retenção dos analitos em papel cromatográfico de troca iônica, suporte sólido que se mostrou apropriado para medições diretas por EDXRF. Algumas variáveis foram otimizadas e os melhores resultados obtidos foram: vazão de amostragem de 2,0 mL min-1, pH entre 5,0 e 7,5 e volume de amostragem de 10 mL. No procedimento, com auxílio de uma bomba peristáltica, alíquotas da solução padrão ou amostra foram passadas em papel cromatográfico de troca catiônica, que foi cortado em círculos com 13 mm de diâmetro e adaptados em suporte para filtração. Após a secagem dos papéis cromatográficos, foram realizadas medições diretas no papel empregando espectrômetro de fluorescência de raios-x de energia dispersiva. As curvas de calibração apresentaram resposta linear no intervalo de 0,050 a 1,50 μg mL-1 dos analitos. As repetibilidades, expressas em termos de RSD das soluções-padrão contendo 0,25 μg mL-1 de Cu, Fe, Ni e Zn, calculadas a partir de 15 determinações consecutivas, foram de 2,5, 2,8, 3,0 e 2,7%; e 3,1, 2,9, 2,8 e 2,7%, para matrizes com 96% e 50% de etanol, respectivamente. Obteve-se limites de quantificação (LQ) iguais a 43, 50, 50 e 40; e 47, 54, 59 e 45 μg mL-1, para matrizes com 96% e 50% de etanol, respectivamente. O procedimento foi verificado através de testes de recuperação, adicionando-se 100 μg L-1 de cada metal nas diferentes amostras, com resultados entre 92 e 99%. Os resultados obtidos pelo método proposto foram comparados com os resultados obtidos por um método comparativo usando ICP OES, após digestão das amostras, não apresentando diferenças significativas, com um nível de confiança de 95%. O método proposto é simples e não exigiu que as amostras fossem submetidas a qualquer pré-tratamento drástico ou demorado. / Salvador

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Spiranthera odoratissima A. ST. HIL. (RUTACEAE), conhecida como manacá, é uma planta medicinal localizada na região do Cerrado Brasileiro. É utilizada na medicina tradicional como anti-inflamatório, ansiolítico e para o tratamento de doenças renais, hepáticas, reumatismo, dores de cabeça e de estômago. A Rutaceae apresenta-se amplamente distribuída pelo mundo, com cerca de 150 gêneros e 1700 espécies, no Brasil há cerca de 30 gêneros nativos e aproximadamente 192 espécies, sendo rica em metabólitos secundários e suas respectivas atividades farmacológicas e biológicas. O objetivo deste trabalho foi avaliar o potencial gastroprotetor do extrato etanòlico das folhas da S. odoratissima (SOL) através de modelos de úlcera gástrica em ratos bem como sua atividade antimicrobiana e toxicidade aguda. A análise da ressonância magnética nuclear mostrou sinais característicos de hidrogênio para clases de terpenos, limonoides, esteroides, cumarinas, alcaloides. Foram realizados modelos experimentais de úlcera gástrica que, com base em suas respectivas especificações, foram incluídos dois grupos controles, sendo um positivo (Carbenoxolona 100 mg.kg-1 ou Cimetidina 100 mg.kg-1) e um negativo (Veículo – Tween 80® 12%, 10 mL.kg-1). Após cada metodologia, foi realizado a eutanásia por overdose de CO2, os estômagos removidos, abertos na região da maior curvatura e fotografados para quantificação da área de lesão ulcerativa por meio do programa AVSoft®. O SOL, na dose de 100 mg.kg-1 (a menor dose mais efetiva), apresentou atividade antiulcerogênica contra lesões gástricas induzidas por etanol absoluto (93% de inibição; p<0,05) e pelas drogas anti-inflamatórias não esteroidais (DAINEs) (73% de inibição;p<0,05). Não apresentou atividade antimicrobiana nas cepas padrão utilizada neste estudo. Em adição, nenhum sinal de toxicidade foi observado, considerando os parâmetros analisados. / Spiranthera odoratissima A. ST. HIL. (RUTACEAE), knowns as manacá, is a medicinal plant present in Brazilian Cerrado region. It is used in traditional medicine as antiinflammatory, anxiolytic and for the treatment of renal diseases, hepatic diseases, rheumatism, headache and stomachache. The Rutaceae presents widely distributed around the world, with about 150 genera and 1700 species, in Brazil there are about 30 native genera and approximately 192 species, is rich in secondary metabolites with diverse pharmacological and biological activities. The objective of this study was to evaluate the gastroprotective potential of ethanolic extract of the leaves of S. odoratissima(SOL) through gastric ulcer models in rats and its antimicrobial activity and acute toxicity. Analysis of the nuclear magnetic resonance showed characteristic signs of hydrogen to terpenes, limonoids, steroids, coumarins and alkaloids. Foram performed experimental models of gastric ulcer, based on their respective specifications, two control groups were included, one positive (carbenoxolone 100 mg.kg-1 or Cimetidine 100 mg.kg-1) and negative (vehicle - Tween 80® 12%, 10 mL.kg-1). After each methodology was performed euthanasia by CO2 overdose, the stomachs removed, opened in the region of greater curvature and photographed for quantification of ulcerative lesion area through AVSoft® program. SOL at a dose of 100 mg.kg-1 (the lower most effective dose) showed antiulcer activity against gastric lesions induced by absolute ethanol (93% inhibition;p<0,05) and the indomethacin, an anti-inflammatory drug nonsteroidal, (73% inhibition;p<0,05). Howerver, the treatment with SOL does not showed antimicrobial activity in the bacterial strains tested. Additionally, no signs of toxicity were observed, considering the parameters analyzed. / Dissertação (Mestrado)

Orientadores: Silmara Marques Allegretti, Vera Lúcia Garcia Rehder / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T02:12:21Z (GMT). No. of bitstreams: 1 Oliveira_ClaudineideNascimentoFernandesde_D.pdf: 6645819 bytes, checksum: c09a778f460110b77585bd22b8da5344 (MD5) Previous issue date: 2012 / Resumo: A propagação da esquistossomose e a ameaça de tolerância e resistência ao fármaco de escolha, o praziquantel, têm intensificado as pesquisas utilizando plantas medicinais com o intuito de promover o desenvolvimento de novos fármacos esquistossomicidas. A planta Phyllanthus amarus (quebra-pedra) possui atividades antiinflamatória e hepatoprotetora já comprovadas cientificamente, o que fez com que a mesma fosse selecionada para este estudo, uma vez que a principal patologia da esquistossomose é a formação de granulomas (processo inflamatório) no fígado. Esse trabalho teve como objetivo fazer um fracionamento biomonitorado do extrato etanólico de P. amarus por meio de ensaios in vitro e in vivo com o intuito de verificar a ação esquistossomicida dessa planta contra o S. mansoni linhagem BH. Para a realização dos testes in vitro, os vermes adultos coletados foram incubados em placas contendo meio de cultura RPMI 1640, um casal de verme e amostras de extrato etanólico bruto, frações de diferentes polaridades ou lignanas isoladas nas concentrações 200, 100, 50 e 25 ?g/mL. Os vermes foram observados por um período de 72 horas, sendo avaliados a taxa de mortalidade, a oviposição, o acasalamento e as alterações tegumentares. A melhor atividade in vitro foi observada com a fração 2 butanólica, pois foi letal para 100% dos vermes em 48 h de observação, sendo assim selecionada para os testes in vivo. A fração 2 butanólica é composta majoritariamente por lignanas, por isso algumas delas (nirantina, filantina + nirantina e filantina + filtetralina + nirtetralina), mesmo não sendo efetivas nos testes in vitro, foram selecionadas para avaliação in vivo. Nos testes in vivo camundongos Balb/c foram tratados oralmente, 45 ou 60 dias após a infecção. No 45° dia de infecção os animais foram tratados com 100 e 200 mg/kg da fração 2 butanólica, 100 mg/kg das lignanas nirantina, filantina + nirantina e filantina + filtetralina + nirtetralina em dose única e 100 mg/kg da fração 2 butanólica distribuídos em 3 dias consecutivos. Já no 60° dia de infecção o tratamento foi feito em dose única com a fração 2 butanólica, nirantina, filantina + nirantina e filantina + filtetralina + nirtetralina (100mg/kg). A atividade in vivo foi avaliada com base nos seguintes parâmetros: ação sobre os vermes adultos, ovos eliminados nas fezes, oograma, formação das reações granulomatosas, e ação sobre o tegumento (feita por microscopia eletrônica de varredura - MEV). Os tratamentos mais efetivos no 45° dia de infecção foram apresentados pelos grupos tratados com filantina + nirantina e nirantina (100mg/kg). A lignana nirantina apresentou taxa de redução do número de ovos de 90,3% e a associação da filantina + nirantina, 63,8%. Os resultados referentes aos demais parâmetros (redução do total de vermes, de vermes fêmeas e do número de granulomas) foram semelhantes, apresentando taxas de redução em torno de 58%. Além dessas alterações, as imagens obtidas por MEV mostraram extensas lesões no tegumento dos vermes machos. No 60° dia de infecção, a associação de lignanas filantina + nirantina e a fração 2 butanólica (100mg/kg) apresentaram as taxas de redução mais significativas: 50,8% e 46,7% para o número total de vermes, 47,2% e 42,7% para o número de vermes fêmeas, 87,5% e 98,3% para o número de ovos e 44% e 18% para o número de granulomas, respectivamente. Assim sendo, de acordo com os parâmetros biológicos avaliados neste trabalho, a associação das lignanas filantina: nirantina na concentração de 100 mg/kg revelou um efeito esquistossomicida promissor, uma vez que foi efetiva nos dois períodos estudados / Abstract: The dissemination of schistosomiasis and the threat of its causing agents becoming resistant to the drug of choice, i.e., praziquantel, have intensified the research with medicinal plants to promote the development of new schistosomicidal drugs. Phyllanthus amarus (stone-breaker) is a plant whose anti-inflammatory and hepatoprotective activities have already been attested, which is the reason why it was chosen for this study, as the main pathology of schistosomiasis is the formation of granulomas (inflammatory process) in the liver. The aim of this work was to carry out a bioguided fractionation of the ethanol extract of P. amarus by means of in vitro and in vivo assays in order to verify the schistosomicidal potential of that plant against S. mansoni, BH strain. To carry out the in vitro assays, the collected adult worms were incubated in plates containing RPMI 1640 medium, a pair of mating worms, and samples of crude ethanol extract, fractions of different polarities or isolated lignans at the concentrations of 200, 100, 50 and 25 ?g/mL. The worms were observed over a period of 72 hours, in which mortality rate, egg laying, mating, and tegumentary changes were evaluated. The best in vitro activity was provided by butanol fraction 2, as it proved lethal for 100% of the worms over 48 hours of observation, and so it was used in the in vivo assays. Butanol fraction 2 is mainly composed of lignans, and some of them (nirantin, filantin: nirantin, and filantin + filtetralin + nirtetralin), albeit not effective in the in vitro assays, were selected for in vivo evaluation. In the in vivo assays, Balb/c mice were treated orally 45 or 60 days following infection. On the 45th day following infection, the animals were treated with 100 and 200 mg/kg of butanol fraction 2, 100 mg/kg of filantina:nirantin and filantin + filtetralin + nirtetralin in a single dose, and 100 mg/kg of butanol fraction 2 distributed over 3 consecutive days. On the 60th day following infection, the treatment was carried out in a single dose with 100 mg/kg of butanol fraction 2, nirantin, filantina:nirantin, and filantin + filtetralin + nirtetralin. The in vivo activity was evaluated based on the following parameters: action on adult worms, eggs eliminated in the stool, egg counting, granulomatous reactions, and action on the tegument of the worms (using scanning electron microscopy (SEM). The most effective treatments on the 45th day were those carried out with 100 mg/kg of filantina:nirantin, and nirantin. The association of filantina:nirantin provided a reduction of 63.8% in the number of eggs, whereas nirantin achieved a reduction rate of 90.3%. The results for the other parameters (reduction in the total number of worms, number of females, and number of granulomas) were similar with reduction rates around 58%. In addition to such changes, images obtained by SEM showed extensive lesions on the tegument of male worms. On the 60th day following infection, the filantin + nirantin association and butanol fraction 2 at 100 mg/kg achieved the most significant xxiii reduction rates: 50.8% and 46.7% in the total number of worms, respectively; 47.2% and 42.7% in the number of females, respectively; 87.5% and 98.3% in the number of eggs, respectively; and 44% and 18% in the number of granulomas, respectively. Therefore, according to the biological parameters evaluated in this work, the association of the lignans filantina:nirantin at 100 mg/kg has a promising schistosomicidal activity, as it was effective over two periods of treatment / Doutorado / Parasitologia / Doutora em Parasitologia