Made available in DSpace on 2016-08-19T12:59:16Z (GMT). No. of bitstreams: 1 Dissertacao-DayanneSilvaFreitas.pdf: 3246572 bytes, checksum: 53f34629ab61070b06e48c3c8284366d (MD5) Previous issue date: 2016-03-14 / The Euterpe oleracea Mart seed. (Acai) has demonstrated different biological activities, such as antioxidant, anti-inflammatory, antihypertensive, antinociceptive and antineoplastic. In this new study, the aim was to analyze the antineoplastic activity of fractions derived from hydroalcoholic extract of Euterpe oleracea Mart. seed in cell line MCF-7, and to identify the compounds responsible for the antineoplastic action by mass spectrometry using electrospray ionization source, and a ciclotrônico analyzer coupled to a Fourier transform (ESI-FT-ICR MS). The MCF-7 cell line was treated with 10, 20, 40 and 60 μg L-1 with fractions hexane (FH), chloroform (FC) and ethyl acetate (FAE) of the hydroalcoholic extract acai seed for 24, 48 and 72 hours. After treatment the cell viability was measured using the assay with 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) and the cell type of death was assessed using annexin - Iodide propidium (PI) assay. Data were analyzed statistically by analysis of variance (ANOVA) or Student's t test, us appropriate. It was observed that all the fractions caused significant reduction of cell viability of MCF-7, but the FAE was the most cytotoxic (p <0.001). In the test Annexin-PI, there was no significant labeling annexin but increased PI staining was significant (p <0.001). This study showed that the FAE was more effective in reducing cell viability, following necroptose mechanism in MCF-7 cell. The FAE is composed of epicatechin, proanthocyanidin A2 and trimeric and tetrameric procyanidins. / A semente de Euterpe oleracea Mart. (açaí) tem demonstrado diferentes atividades biológicas, tais como: antioxidante, anti-inflamatória, anti-hipertensiva, antinociceptiva e antineoplásica. Neste estudo, o objetivo foi analisar a atividade antineoplásica de frações do extrato hidroalcoólico da semente de Euterpe oleracea Mart. na linhagem celular MCF-7, assim como identificar os compostos responsáveis pela ação antineoplásica por espectrometria de massas utilizando fonte de ionização por eletrospray e um analisador ciclotrônico acoplado a uma transformada de Fourier (ESI- FT-ICR MS). A linhagem MCF-7 foi tratada com 10, 20, 40 e 60 μg L-1 com as frações hexânica (FH), clorofórmica (FC) e de acetato de etila (FAE) do extrato hidroalcoólico da semente do açaí por 24, 48 E 72 horas. Após o tratamento a viabilidade celular foi mensurada usando o ensaio com 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) e o tipo de morte celular foi avaliado com o ensaio Anexina - Iodeto de propídeo (PI). Os dados foram analizados estatisticamente por análise de variância (ANOVA) ou por teste T student, quando apropriado. Foi observado que todas as frações causaram redução significativa da viabilidade celular da MCF-7, porém a FAE foi a mais citotóxica (p < 0,001). No ensaio de anexina-pi, não foi observado marcação significativa para anexina porém o aumento da marcação de PI foi significativo (p<0,001). O presente estudo demonstrou que a FAE foi a mais eficaz na redução da viabilidade celular, seguindo mecanismo de necroptose na célula MCF-7. A FAE é composta por epicatequina, proantocianidina A2 e procianidinas trimérica e tetramérica.

Made available in DSpace on 2014-08-20T14:37:52Z (GMT). No. of bitstreams: 1 dissertacao_ciciane_fernandes.pdf: 1291718 bytes, checksum: 7919ec746c53767068a313c9fd4b16f2 (MD5) Previous issue date: 2013-02-08 / The aim was to evaluate the healing of open cutaneous wounds of Wistar rats treated with Carapa guianensis (andiroba) and Caesalpinia ferrea Mart. (Jucá) and their repellent action against flies of the Calliphoridae family. It were studied 160 Wistar rats with open wounds in the back that were treated daily with Carapa guianensis at concentrations of 20% (A20) and 50% (A50), Caesalpinia ferrea Mart. concentrations at 20% (J20) and 50% (J50) and Vaseline (control group). Clinical, histological and morphometric studies were carried out after four, seven, 14 and 21 days of treatment, as well as a tensiometric study after 21 days. In order to study the repellency, W.O.T traps (Wind Oriented Trap) containing deteriorated bovine liver and cream with the herbal medicine Carapa guianensis and Caesalpinia ferrea Mart. at concentrations of 20% and 50% were used to catch flies. Clinically, after four days of treatment, the wounds of the control group presented exudate, differing statistically from the other groups (p=0.0065). In the morphometric study, J50 and J20 groups had higher average area (p=0.0001) after seven days of treatment, whereas after 14 days, the wounds of the control group had higher average area (p=0.0000) compared with the other groups. In the histopathological study, the differences between groups were seen after four days of treatment, with the A20 group showing a greater number of wounds in the proliferative phase (p=0.0000). In the tensiometric study, the control group showed better results (3.52MPa) in tension when compared to other groups (p=0.0055). In the repellency study, the traps containing J50 (p<= 0.010) and D20 (p<= 0.010) showed a higher repellency 97% and 100%, respectively, followed by the A50 group (p<= 0.010) with 93.4% and the A20 group with 56.5%. The results led to the following conclusions: Carapa guianensis at a concentration of 20% accelerates the phases of the healing process in the first days after tissue injury, whereas Carapa guianensis at 50% and Caesalpinia ferrea Mart. at concentrations of 20% and 50% do not show satisfactory results as a cicatrizant. As repellent action, Caesalpinia ferrea Mart. at concentrations of 50% and 20% and Carapa guianensis at 50% have repellent effect against Chrysomya albiceps, Chrysomya megacephala, Lucilia cuprina, Lucilia eximia, Lucilia sericata and Sarconesia chlorogaster flies, species of the Calliphoridae family, while the Carapa guianensis at 20% has less repellent action. / Objetivou-se avaliar a cicatrização de feridas cutâneas abertas de ratos Wistar tratadas com Carapa guianensis (andiroba) e Caesalpinia ferrea Mart. (jucá) e, suas ações repelentes frente a moscas da família Calliphoridae. Foram estudados 160 ratos da linhagem Wistar, com realização de feridas cutâneas abertas no dorso, tratadas diariamente com Carapa guianensis nas concentrações de 20% (A20) e 50% (A50), Caesalpinia ferrea Mart. nas concentrações de 20% (J20) e 50% (J50) e vaselina (grupo controle), sendo realizadas avaliações clínicas, morfométricas e histológicas aos quatro, sete , 14 e 21 dias de tratamento e estudo tensiométrico aos 21 dias. Para estudo da repelência, foram utilizadas armadilhas W.O.T. (Wind Oriented Trap) para captura de moscas contendo fígado bovino deteriorado e creme contendo os fitoterápicos Carapa guianensis e Caesalpinia ferrea Mart. nas concentrações de 20% e 50%. Clinicamente, aos quatro dias de tratamento, as feridas tratadas apresentaram formação de crosta, diferindo estatisticamente do grupo controle que ainda apresentava exsudato nas feridas (p=0,0065). No estudo morfométrico, aos sete dias de tratamento, os grupos J50 e J20 apresentaram maior média de área (p=0,0001), enquanto que aos 14 dias, as feridas do grupo controle apresentaram maior média de área (p=0,0000) comparada com os demais grupos. Na histopatologia foram observadas diferenças entre os grupos aos quatro dias de tratamento, com o grupo A20 apresentando maior número de feridas na fase proliferativa (p=0,0000). No estudo tensiométrico, o grupo controle apresentou melhor resultado (3.52MPa) na tensão comparado aos demais grupos (p=0,0055). No estudo sobre repelência, as armadilhas contendo J50 (p<= 0,010) e J20 (p<= 0,010) apresentaram maior repelência respectivamente 97 e 100%, seguida do grupo A50 (p<= 0,010) com 93,4% e grupo A20 com 56,5%. Os resultados levaram as seguintes conclusões: Carapa guianensis na concentração de 20% acelera o processo cicatricial nos primeiros dias após injúria tecidual, enquanto que Carapa guianensis a 50% e Caesalpinia ferrea Mart. nas concentrações de 20% e 50% não apresentam resultados satisfatórios como cicatrizante. Como ação repelente, Caesalpinia ferrea Mart. nas concentrações de 50% e 20% e Carapa guianensis na concentração de 50% apresentam efeito repelente frente às moscas Chrysomya albiceps, Chrysomya megacephala, Lucilia cuprina, Lucilia eximia, Lucilia sericata e Sarconesia chlorogaster, espécies da família Calliphoridae, enquanto que Carapa guianensis a 20% apresenta menor ação repelente.

Submitted by biblioteca unifenas (biblioteca@unifenas.br) on 2018-03-02T17:52:47Z No. of bitstreams: 1 Maysa Eduarda de Castro Dissertação.pdf: 1155468 bytes, checksum: c65b96b4486f0cf6c4642c0a68aa6335 (MD5) / Made available in DSpace on 2018-03-02T17:52:47Z (GMT). No. of bitstreams: 1 Maysa Eduarda de Castro Dissertação.pdf: 1155468 bytes, checksum: c65b96b4486f0cf6c4642c0a68aa6335 (MD5) Previous issue date: 2018-02-21 / Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG / Handroanthus impetiginosus (Mart. ex DC.) Mattos (HI) it has been widely used for an extended period in traditional medicine, and several studies have shown the presence of chemical compounds and phytotherapeutic potentials of this plant. The objective was to evaluate the genotoxicity of the extract of H. impetiginosus and lapachol (LAP), one of the main compounds found in this plant, using the mouse bone marrow micronucleus assay. Experimental groups consisting of male and female Swiss albinus mice (Unib: SW) were evaluated after 24-48h (HI) e 24h (LAP) post treatment with Cyclophosphamide (CYCLO, 50 mg.kg-1), NaCl (150 mM), HI (0.5; 1.2 g.kg-1), LAP (0.075, 0.15, 0.30 g.kg-1). For HI, analysis of the MNPCEs showed significant differences between treatment doses (500–2,000 mg.kg-1) and NaCl. PCE/NCE showed significant differences between treatment doses (500–2,000 mg.kg-1) or NaCl compared to CP (50 mg.kg-1). This research suggests presence of genotoxicity in treatment doses 2,000 mg.kg-1, and mild genotoxicity in the others treatment doses (500–1,000 mg.kg-1) of HI, sex and time-independent and absence of toxicity doses-, time- and sex-independent. However, for lapacho, analysis of the MNPCEs showed significant differences between treatment dose (300 mg.kg-1) and NaCl. PCE/NCE showed significant differences between treatment doses (500–2,000 mg.kg-1) or NaCl compared to CP (50 mg.kg-1). This research suggests presence of genotoxicity of LAP, dose-dependent (300 mg.kg-1), but time- and sex-independent and absence of toxicity doses-, time- and sex-independent. / Handroanthus impetiginosus (Mart. ex DC.). Mattos (HI) tem sido muito utilizada por um extenso período na medicina tradicional e vários estudos têm mostrado a presença de compostos químicos e potenciais fitoterapêuticos dessa planta. O objetivo foi avaliar a genotoxicidade do extrato de H. impetiginosus e do lapachol (LAP), um dos principais compostos encontrados nessa planta, usando o ensaio do micronúcleo em medula óssea de camundongos. Grupos experimentais constituídos de camundongos machos e fêmeas Swiss albinus (Unib: SW) foram avaliados após 24-48h (HI) e 24h (LAP) de tratamento com Ciclofosfamida (CICLO; 50 mg.Kg-1), NaCl (150 mM), HI (0,5; 1; 2 g.Kg-1), LAP (0,075; 0,15; 0,30 g.Kg-1). As análises de MNPCEs do grupo tratado com HI mostraram diferenças (p  0,05) entre todas as doses de tratamento (500–2.000 mg.Kg-1) e controle negativo (NaCl). As proporções PCE/NCEapresentaram diferenças significativas (p  0,05) entre as doses de HI (500–2.000 mg.Kg-1) ou controle negativo (NaCl), frente ao controle positivo ciclofosfamida (50 mg.Kg-1).Os resultados sugeremum efeito potencialmente genóxico dependente da dose (2.000 mg.Kg-1) e levemente genotóxico nas demais doses (500–1.000 mg.Kg-1) do extrato de HI, independentemente do tempo de tratamento (24 e 48 h) e do sexo (macho e fêmea), e ausência de toxicidade sistêmica do HI dose, sexo e tempo-independente. Contudo, as análises de MNPCEs do grupo tratado com LAP apresentaram diferenças significativas (p  0,05) entre a dose de tratamento (300 mg.Kg-1) e controle negativo (NaCl), jáas proporções PCE/NCEapresentaram diferenças significativas (p  0,05) entre as doses de LAP (75–300 mg.Kg-1) ou controle negativo (NaCl), frente ao controle positivo ciclofosfamida (50 mg.Kg-1), sugerindopresença de genotoxicidade do LAP, dependentemente da dose de administração fitoterapêutica (300 mg.Kg-1), mas independente do tempo de tratamento (24 e 48 h), e sexo (macho e fêmea), e ausência de toxicidade sistêmica do LAP nas condições do ensaio MN, dose, sexo e tempo-independente.

En dépit de progrès considérables, le cancer reste l'une des principales causes de morbidité et de mortalité dans le monde. Actuellement, 90% des décès liés au cancer sont causés par la propagation de cellules cancéreuses vers des organes distants. Une fois implantées et disséminées, les métastases sont beaucoup plus difficiles à détruire par les moyens de la chimiothérapie.A la suite d’un processus d’intravasation, certaines cellules tumorales s’échappent de la tumeur primaire et empruntent les systèmes circulatoires avant d’être ensuite extravasées, puis distribuées et finalement disséminées dans divers organes. Ainsi, dans l’environnement circulatoire, ces cellules tumorales circulantes (CTCs) se trouvent particulièrement accessibles aux agents thérapeutiques. Dans ce cadre, nous avons imaginé d’utiliser des nanoparticules à architecture contrôlée, afin d’intercepter de manière sélective ces cellules dans l’environnement sanguin.Dans cet objectif, nous avons synthétisé par ouverture de cycle de la lactone correspondante des copolymères amphiphiles di- et tri-blocs du poly(glutamate de benzyle). Leur auto-assemblage a permis d'obtenir des nanoparticules amphiphiles de taille inférieure à 100 nm et de potentiel ζ négatif, dont la géométrie contrôlable va de la forme sphérique (rapport d'aspect 1.3) à la forme ellipsoïdale (oblats) (rapport d'aspect 2,6) et qui présentant en surface des chaînes de PEG sous des conformations et des densités de surface contrôlées.En raison de leur capacité de circuler dans le compartiment sanguin, ces nanoparticules ont une probabilité d’interaction optimale avec les CTCs.L’impact de la modification de leur architecture a été établi en étudiant les capacités d’interactions des différentes nanoparticules préparées, d’une part avec les protéines plasmatiques et d’autre part, avec les différents types cellulaires rencontrés dans le compartiment sanguin.Les résultats les plus marquants montrent que l’élongation des nanoparticules (oblats) et l’anisotropie de leur surface, caractérisée par leur balance hydrophile/lipophile, gouvernent profondément leurs interactions. De manière fort intéressante, il apparaît que l’élongation des particules dont la surface est uniformément hydrophile diminue l’intensité de leur capture par les différents types cellulaires modèles étudiés (HUVECs modèle de cellules endothéliales), cellules RAW 276.7 (modèle de macrophages) et cellules PC3 (cancer de la prostate) et B16 (mélanome). En revanche, lorsque ces nanoparticules présentent une anisotropie de surface, leur capture par ces différents types cellulaires est augmentée avec l’élongation des particules (facteur d’élongation de 2,1).Dans un dernier volet expérimental, ces nanoparticules ont été modifiées par greffage de la protéine MART1 à leur surface. Ces immuno-nanoparticules ont montré une certaine capacité de reconnaissance des cellules B16 (modèle du mélanome). Leur efficacité après injection intraveineuse devra toutefois être précisée in vivo. / Despite the considerable progress, cancer remains one of the leading causes of morbidity and mortality worldwide. Currently, 90% of cancer deaths are caused by the spread of cancer cells to distant organs. Once implanted and disseminated, metastases are much more difficult to destroy by means of chemotherapy.Following a process of intravasation, some tumor cells escape from the primary tumor and migrate through the circulatory systems before being extravasated, then distributed and finally disseminated in various organs. Thus, in the circulatory environment, these circulating tumor cells (CTCs) are particularly accessible to therapeutic agents. In this context, we have imagined the use of nanoparticles with controlled architecture, in order to selectively intercept these cells in the blood environment.For this purpose, we have synthesized by ring opening of the corresponding lactone, amphiphilic di- and tri-block copolymers of poly (benzyl glutamate). Their self-assembly made it possible to obtain amphiphilic nanoparticles smaller than 100 nm in size and with a negative ζ potential, whose controllable geometry ranges from spherical (aspect ratio 1.3) to ellipsoidal (oblates) (aspect ratio 2, 6) and having PEG chains on the surface under controlled surface conformations and densities.Due to their ability to circulate in the blood compartment, these nanoparticles have an optimal probability of interaction with CTCs.The modification impact of their architecture has been established by studying the interaction capacities of the different nanoparticles prepared. On the one hand with the plasma proteins and on the other hand, with the different cell types encountered in the blood compartment.The most striking results show that the elongation of the nanoparticles (oblates) and the anisotropy of their surface, characterized by their hydrophilic / lipophilic balance, strongly govern their interactions. Interestingly, it appears that the elongation of particles whose surface is uniformly hydrophilic decreases the intensity of their capture by the different types of cell models studied (HUVEC model endothelial cells), RAW 276.7 cells (macrophage model) and cells PC3 (prostate cancer) and B16 (melanoma). Although, when these nanoparticles exhibit surface anisotropy, their capture by these different cell types is increased with the elongation of the particles (elongation factor of 2.1).In a final experimental part, these nanoparticles were modified by grafting the MART1 protein on their surface. These immuno-nanoparticles showed a certain recognition capacity of B16 cells (melanoma model). However, their efficacy after intravenous injection should be specified in vivo.

Published Article / In response to the Human Immunodeficiency Virus (HIV) epidemic in the country, the South African Government started with the provisioning of Antiretroviral Therapy (ART) in the public health sector. Monitoring and evaluating the effectiveness of the ART programme is of the utmost importance. The current patient information system could not supply the required information to manage the rollout of the ART programme. A data warehouse, consisting of several data marts, was developed that integrated several disparate systems related to HIV/AIDS/ART into one system. It was, however, not possible to trace a patient across all the data marts in the data warehouse. No unique identifiers existed for the patient records in the different data marts and they also had different structures. Record linkage in conjunction with a mapping process was used to link all the data marts and in so doing identify the same patient in all the data marts. This resulted in a longitudinal patient record of an ART patient that displayed all the treatments received by the patient in all public health care facilities in the province.

知識經濟時代的來臨，科技與速度是成功的關鍵，營運效率成為優勝劣敗的決定因素。企業要經得起全球化的挑戰，必須以各類取得的資訊做客觀的分析、策略性的思考，來規劃及發展出具前瞻性、適切性的企業經營新模式，而其重點即為將企業內、外部資源作有效的整合，在技術、產品與策略上推陳出新，因時因地的調整經營策略，進而轉型、再造、創新，以提升企業之競爭力及傲人的營運績效，以促成傲人的資訊流、物流、金流與商流的整合。 資訊流應達到電腦化、金流應達到即時流通順暢化、商流結構亦應達到完整性大目標外，最其要者即屬物流應達成即時管理。而其顯著的特點是走向系統化及即時化、電腦化的資訊管理使供應鏈物流活動之間的生產資訊控制，訂貨、儲存、搬運、進出庫、發貨、運輸、結算等物流環節之間的自動化資訊控制更為合理及流暢。 因其中物流成本占全部營銷成本的比例極高，因此個人以為降低物流成本應可成為企業第三獲利來源。 數位經濟的時代，在網際網路與資訊科技一日千里進步中，物流資訊化、網路化、全球化、虛擬化、整合化的方向，努力建構具核心競爭力的資訊整合實為當務之急的核心，如何改變手工資料登錄，使輸入的品質和速度提高，條碼就在這樣的環境下應運而生，其中RFID(Radio Frequency Identification)射頻技術的條碼自動識別技術以電腦、光電技術和通訊為發展基礎的綜合性技術最具競爭優勢及潛力。 本研究希望能從企業導入 RFID 現況與發展趨勢、應用於供應鏈所展現之核心競爭力及Wal-Mart 個案來分析，並從以下三重點來探討： 1. 企業導入RFID 對核心競爭力的影響。 2. 供應鏈物流系統如何運用RFID化以提升競爭力。 3. 政府推動RFID計畫以加速臺灣運籌中心之附加價值的建議。 / The era of knowledge economy has emerged. The key success factors are new technology and speed, and the main decision element is operation excellence. The enterprise has to be continually challenged by globalization through integrated information, strategic thinking, future planning, and new business plans, to integrate all the above resources internally and externally. In addition, the enterprise needs to refresh workable resources, technology, product and strategy and continually make changes with re-modeling, re-engineering, and creativity to promote the competitive strength and operation excellence so as to synergize information flow, material flow, cash flow and business process flow. Information flow should be computerized, cash flow should be smooth, and business process flow should be to meet the most integrated goal: ON-LINE Management. It is important that these processes are systematically linked with on-line computerized information within the supply chain management model. For instance: ordering, stocking, transportation, warehousing, tracking and cashier etc. Nevertheless, logistics cost constitutes a high percentage of operation cost. As such, I view that logistics cost should be brought down so as to reap enormous benefits from a third resource. With the advent of the digital economy and the development of web-base technology, logistics information flow has to be systematic, web-based, virtual, and integrated. Striving to build up a competitive information integration is very important. In order to change the data-entering from a manual mode to a systematic mode the quality and speed are highly expected. RFID (Radio Frequency Identification) development depends largely on high technology of computers, optoelectronics, electronic, telecommunication. Thus, it has tremendous potential to become a synergistic competitive strength in the market This research paper will address RFID implementation, status and future development potential, the application in supply chain management and its core competencies. A case review on Wal-Mart’s success with reference to the following points will be discussed in this paper: 1. What are the core competencies required if one applies RFID? 2. How to apply RFID in the field of Supply Chain Management? 3. What is the add-value if Taiwan Government promotes RFID plans?

Précédemment, il a été démontré au laboratoire qu’une vaccination des souris avec des lipopolyplexes (LPR) contenant l’ARNm de l’antigène de mélanome MART1 permet d’induire la formation de lymphocytes T cytotoxiques spécifiques et de retarder le développement de mélanomes B16F10 et de métastases pulmonaires. Les LPR sont des complexes ternaires constitués d’ARNm, d’un polymère cationique histidylé et de liposomes cationiques histidylés. L’objectif de ma thèse était d’améliorer cette vaccination antitumorale en développant de nouveaux liposomes capables de cibler les cellules dendritiques (DC). Le ciblage a été réalisé en incorporant un glycolipide mannosylé aux liposomes afin de favoriser leur reconnaissance par le récepteur mannose. A partir de ces liposomes, des formulations de complexes ternaires à base d’ADN (LPD mannosylés ou Man11-LPD100) ou à base d’ARN (LPR mannosylés ou Man11- LPR100) ont été mis au point. Les résultats montrent que : in vitro les formulations Man11-LPD100 sont mieux internalisés et transfectent plus efficacement les DC que les LPD100 non mannosylés. Les formulations Man11-LPR100 transfectent avec une plus grande efficacité les DC par rapport aux Man11- LPD100. Par ailleurs, une forte réduction de la toxicité des formulations a été obtenue en dialysant les liposomes. Il est également possible de conserver les formulations sous forme déshydratée. Une imagerie par scintigraphie effectuée chez la souris a permis de constater que 9% des LPD sont captés dans la rate après une injection IV. Nous avons mis en évidence après un isolement de DC spléniques que les formulations Man11-LPR100 transfectent 4 fois plus de DC que les LPR non manosylés. Enfin, l’immunisation des souris avec Man11-LPR100 contenant l’ARNm MART1 permet une vaccination plus efficace contre la tumeur B16F10 et une meilleure survie. En conclusion, les LPR Man11-LPD100 sont de bons vecteurs pour cibler et transfecter les DC spléniques avec l’ARNm d’un antigène tumoral et pour induire la réponse immune contre les cellules tumorales. / Previously, it has been demonstrated that mice vaccination with lipopolyplexes (LPR) containing melanoma antigen MART1 mRNA can induce the generation of specific cytotoxic T cells and delay B16F10 melanoma growth and lung metastases. LPR are ternary complexes consisting of mRNA, a histidylated cationic polymer and histidylated cationic liposomes. The objective of my thesis was to enhance this antitumor vaccination through the development of new liposomes that can target specifically dendritic cells (DC). The targeting of DC was achieved by incorporating a mannosylated glycolipid within liposomes to enhance their recognition by the mannose receptor. From these liposomes, formulations based ternary complexes of DNA (mannosylated-LPD or Man11-LPD100) or formulations based on mRNA (mannosylated LPR or Man11 LPR100) were developed. The results show that formulations made with Man11-LPD100 are better internalized and transfect efficiently DC than LPD100. Man11 LPR100 transfect with greater efficiency DC compared to DNA based formulation (Man11-LPD100). Furthermore, a strong reduction of the toxicity of LPD was obtained by liposomes dialysis. It is also possible to preserve their activity by freeze-drying. Mice scintigraphy revealed that 9% of LPD are captured in the spleen following IV injection. We demonstrated after isolation of splenic DC that Man11-LPR100 transfect DC 4 times more than LPR100. Finally, immunization of mice with Man11-LPR100 containing mRNA MART1 allows a more effective vaccination against B16F10 tumor and a better mice survival than non-mannosylated ones. In conclusion, Man11-LPR100 are promising vectors to target and transfect splenic DC with a tumor antigen mRNA aiming to an induction of an immune response against tumor cells.

Submitted by Jean Medeiros (jeanletras@uepb.edu.br) on 2016-05-16T15:16:09Z No. of bitstreams: 1 PDF - Érika Ponchet Alves.pdf: 1299396 bytes, checksum: 3c0c4042f58e9e0ee9edd8547eef555f (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-21T20:42:38Z (GMT) No. of bitstreams: 1 PDF - Érika Ponchet Alves.pdf: 1299396 bytes, checksum: 3c0c4042f58e9e0ee9edd8547eef555f (MD5) / Approved for entry into archive by Secta BC (secta.csu.bc@uepb.edu.br) on 2016-07-21T20:42:48Z (GMT) No. of bitstreams: 1 PDF - Érika Ponchet Alves.pdf: 1299396 bytes, checksum: 3c0c4042f58e9e0ee9edd8547eef555f (MD5) / Made available in DSpace on 2016-07-21T20:42:48Z (GMT). No. of bitstreams: 1 PDF - Érika Ponchet Alves.pdf: 1299396 bytes, checksum: 3c0c4042f58e9e0ee9edd8547eef555f (MD5) Previous issue date: 2015-07-31 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Medicinal plants represent a promising source of active compounds which can be used in the development of new drugs. There is a diversity of plants that are used by the population for therapeutic purposes, among which are the Guapira graciliflora Mart., popularly known as “joão-mole”, and Schinus terebinthifolius Raddi, known as “aroeira-da-praia”. These plants stand out in folk medicine due to its therapeutic properties in inflammatory and infectious processes. This study evaluated in vitro antimicrobial and antiproliferative activities besides, toxicity and chemical composition of the hydroalcoholic extract of the leaves of G. graciliflora. and the essential oil from unripe fruits of S. terebinthifolius. Antimicrobial activity was evaluated by microdilution technique in broth, with determination of minimum inhibitory concentration (MIC) and bactericidal/ Fungicide minimum concentration (MBC / MFC) using to bacterial strains: Staphylococcus aureus ATCC 6538, Escherichia coli ATCC 11775 and Salmonella enteritidis ATCC 13076, and the yeast Candida albicans ATCC 10231. The extracts were analyzed againstC. albicans biofilm after 48 hours of expositions, by means of the CFU / ml count and examination of cell morphology byfluorescence microscope (FM) using Calcofluor White. The antiproliferative activity was evaluated against nine tumor cell lines (U251, MCF7, NCI / ADR-RES, 786-0, NCI-460, HT29, K562, and PC-03 OVCAR-3) and a non-tumor cell line of keratinocytes (HaCat ). Preliminar toxicity was evaluated through the hemolysis assay. Phytochemical analysis of the extract G. graciliflora was performed by thin-layer chromatography, and the essential oil of S. terebinthifoliuswas analysed by Gas Chromatography hifenized with Mass Spectrometry (GC-MS). The G. graciliflora andS. terebinthifoliussamplesshowed potential antifungal against C. albicans with the MIC values of 0.25mg/mL and 1mg/mL, respectively, and no antibacterial activity (MIC>2mg/mL). Both samples (1mg/mL) reduced the number of CFU/mL of C. albicans biofilm. In fluorescence microscopy it was, observed the maintenance of cellular architecture of C.albicansafter the treatments. The G. gracilifloraextract presented cytocidal effect for all cells of the tumor lines except for the K562 (leukemia) while the essential oil of S. terebinthifolius proved cytocidal for all strains. Both samples did not produce hemolysis over 50% up to a concentration of 16mg/mL. Phytochemical analysis of the extract G. graciliflora revealed the predominance of flavonoid compounds, and the major compound identifiede onS. terebinthifolius essential oil were to, terpene, alpha- and beta- phellandrene. The extract from the leaves of G. graciliflora and the essential oil of the fruit of S. terebinthifolius showed potential antifungal against C. albicans in their planktonic form and organized in biofilms; besides antiproliferative activity across the human tumor cell lines and low toxicity on red blood cells. They have been considered promising sources of active compounds, for product development and medicine, anticandida and antiproliferative action. This way, other studies may extend these results, aiming the development to the studies of herbal medicines to clinical use. / As plantas medicinais representamuma fonte promissora de compostos ativos, que podem ser utilizados no desenvolvimento de novos fármacos. Existe uma diversidade de plantas que são usadas pela população para fins terapêuticos, dentre elas, encontram-se a Guapira graciliflora Mart., popularmente conhecida como joão-mole, e a Schinus terebinthifolius Raddi, conhecida como aroeira- da-praia. Essas plantas se destacam na medicina popular em função das suas propriedades terapêuticas em processos inflamatórios e infecciosos. Esteestudo avaliou in vitro as atividadesantimicrobiana eantiproliferativa, além da toxicidade, e da composição química do extrato hidroalcoólico das folhas da G. graciliflorae do óleo essencial dos frutos verdes da S. terebinthifolius. A atividade antimicrobiana foi avaliada por meio da técnica de microdiluição em caldo, com determinação da Concentração Inibitória Mínima (CIM) e Concentração Bactericida/Fungicida Mínima (CBM/CFM), frente às cepas bacterianasStaphylococcus aureus ATCC 6538, Escherichia coli ATCC 11775 e Salmonella enteritidis ATCC 13076, e a levedura Candida albicans ATCC 10231. Foi analisada a ação das duas amostras vegetais sobre o biofilme monoespécie de C. albicans, de 48 horas, por meio da contagem de UFC/mL e análise da morfologia celular em microscópio de fluorescência (MF), usando o Calcofluor White. A atividade antiproliferativa foi realizada contra nove linhagens de células tumorais humanas (U251, MCF7, NCI/ADR-RES, 786-0, NCI-460, HT29, k562, PC-03 e OVCAR-3) e uma linhagem não tumoral de queratinócitos (HaCat). Para o teste de toxicidade preliminar,realizou-se o ensaio de hemólise.A análise fitoquímica do extrato da G. graciliflora foi realizada através da Cromatografia de Camada Delgada, e a do óleo essencial da S. terebinthifoliuspor meio da Cromatografia Gasosa com Espectrometria de Massa (CG-EM). A G. graciliflorae a S. terebinthifoliusapresentaram potencial antifúngico frente a C. albicans, com valores da CIM0,25 mg/mL e 1mg/mL, respectivamente, e ausência de atividade antibacteriana (CIM>2 mg/mL). As duas substâncias vegetais (1 mg/mL) reduziram o número de UFC/mL no biofilme de C. albicans. Na microscopia de fluorescência, observou manutenção da arquitetura celular da C.albicans. A G. graciliflora apresentou efeito citocida para todas as células das linhagens tumorais, exceto para a K562 (leucemia); e o óleo essencial da S. terebinthifoliusmostrou-se citocida para todas as linhagens. As substâncias vegetais não produziram hemólise acima de 50% até a concentração de 16 mg/mL. A análise fitoquímica do extrato da G. graciliflorarevelou a predominância de compostos flavonoides, e no óleo essencial da S. terebinthifolius, de terpenos alfa e beta-felandreno. O extrato das folhas da G. graciliflora e o óleo essencial dos frutos da S. terebinthifolius apresentaram potencial antifúngico frente a C. albicans, em sua forma planctônica e organizada em biofilme; atividade antiproliferativa frente às células de linhagens tumorais humanas e baixa toxicidade frente às hemácias. Podem representar promissoras fontes de compostos ativos, para o desenvolvimento de produtos ou medicamentos, com ação anticandida e/ou antiproliferativa. Desta forma, são necessários outros estudos,para ampliar os conhecimentos a respeito dessas espécies, seguindo para os estudos de riscos e eficiência do uso clínico dessas substâncias.

Made available in DSpace on 2015-09-25T12:23:25Z (GMT). No. of bitstreams: 1 PDF - Thiara Karine de Araujo.pdf: 1613163 bytes, checksum: a957f23a80b49c364a5d41f8caa2a13e (MD5) Previous issue date: 2014-07-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The plants have been a rich source for therapeutic agents, serving as basis for the drug's synthesis, making an important source of new medicines for diseases that are yet without cure like infections and malignancies, in this context the Brazilian flora, due it's diversity, plays an important role in the discovery of potential antimicrobial drugs and antitumor. This work proposes to evaluate the antimicrobial potential of the hydroalcoholic extract of the Guapira Graciliflora Mart. (EOHG) thought the method of the microdilution in broth against strains of the Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis, Streptococcus parasanguinis, Streptococcus mitis and Candida albicans, it's antiproliferative capacity in cell lines of squamous cell carcinome SCC 9, SCC 25 e HSC 3 by the essay MTT, additional to it's inducer potential of apoptosis observed by the test of annexin staining V/PI. The EOHG has presented antimicrobial activity over tested microorganisms, and the better results was observed in the bacteria S. mutans (12,50 µl/ µl), S. mitis (12,50 µl/ µl) and S. salivarius (6,25 µl/ µl), and in the clinical strains of Candida albicans (11) and Candida albicans (410), with CIMs of 0,5mg/ml and 2mg/ml, respectively. The EOHG, in the concentration of 200 µg/ml, have presented inhibition rates of 49% for the cells SCC9 and 63% for the cells HSC3 in the interval of 48h. The Rate of cellular apoptosis for HSC 3 was 19,29% on the time of 48h, additionally the tests of the annexin V/PI has revealed the induction of the cellular apoptosis, suggesting that the cellular membrane and nuclear have been damaged. Based in the results that have been presented, the Guapira Graciliflora Mart. is elected as a potential source of bioactive compound to be used in the treatment of oral infections and the CCEO. / As plantas têm sido uma rica fonte de agentes terapêuticos, servindo de base para síntese de fármacos, constituindo importante fonte de novos medicamentos para doenças que continuam sem cura como infecções e neoplasias malignas, nesse contexto a flora brasileira, por sua diversidade, desempenha papel importante na descoberta de potenciais drogas antimicrobianas e antitumorais. O presente trabalho se propôs a avaliar o potencial antimicrobiano do extrato hidroalcóolico da Guapira Graciliflora Mart. (EOHG) através do método da microdiluição em caldo frente a cepas de Streptococcus mutans, Streptococcus salivarius, Streptococcus oralis, Streptococcus parasanguinis, Streptococcus mitis e Candida albicans, sua capacidade antiproliferativa em linhagens celulares de carcinoma de células escamosas SCC 9, SCC 25 e HSC 3 pelo ensaio MTT, além de seu potencial indutor de apoptose observado pelo teste da coloração anexina V/PI. O EOHG apresentou atividade antimicrobiana sobre os microrganismos testados, s os melhores resultados foram observados nas bactérias S. mutans (12,50 µl/ µl), S. mitis (12,50 µl/ µl) e S. salivarius (6,25 µl/ µl), e nas cepas clínicas de Candida albicans (11) e Candida albicans (410), com CIMs de 0,5mg/ml e 2mg/ml, respectivamente. O EOHG, na concentração de 200 µg/ml, apresentou taxas de inibição de 49% para as células SCC9 e 63% para as células HSC3 no intervalo de 48h. A taxa de apoptose celular para HSC 3 foi de 19,29% no período de 48h, adicionalmente o teste da anexina V/PI revelou indução de apoptose celular, sugerindo que houve danos a membrana celular e nuclear. Com base nos resultados apresentados, a Guapira Graciliflora Mart. representa uma potencial fonte de compostos bioativos a serem utilizados no tratamento das infecções orais e do CCEO.

Submitted by Geyciane Santos (geyciane_thamires@hotmail.com) on 2015-06-09T14:25:49Z No. of bitstreams: 1 Dissertacao - Nivea Maria Simões Falcão.pdf: 1851149 bytes, checksum: 9f64cfee0db539642d59a412a4bf9eea (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-09T19:30:11Z (GMT) No. of bitstreams: 1 Dissertacao - Nivea Maria Simões Falcão.pdf: 1851149 bytes, checksum: 9f64cfee0db539642d59a412a4bf9eea (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2015-06-09T19:32:50Z (GMT) No. of bitstreams: 1 Dissertacao - Nivea Maria Simões Falcão.pdf: 1851149 bytes, checksum: 9f64cfee0db539642d59a412a4bf9eea (MD5) / Made available in DSpace on 2015-06-09T19:32:50Z (GMT). No. of bitstreams: 1 Dissertacao - Nivea Maria Simões Falcão.pdf: 1851149 bytes, checksum: 9f64cfee0db539642d59a412a4bf9eea (MD5) Previous issue date: 2010-03-30 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Leishmaniasis is a world public health problem wide considered by the World Health Organization as one of five infectious endemic diseases of major relevancy. The search for leishmanicids agents with fewer side effects is one of the greatest challenges in modern research, and is increasingly related to the uses of medicinal plants and other natural products. The species Caesalpinia ferrea Martius widely distributed in Brazil it is popularly used for many therapeutic purposes, including the treatment of various infections like those caused by leishmaniasis, making it a promising tests of biological activity against leishmania. The objective of this study was to evaluate by bioassays the activity of hexane and methanol extracts against protozoa, analyzing semi-purified fractions obtained from the most active. The fruits of C. ferrea were collected at the National Institute of Amazon Research, V8’s Campus in the City of Manaus - AM and processed in the departments of Natural Products and leishmaniasis and Chagas Diseases at INPA. The studies were performed with the whole fruit and its separate parts, coconut shell and seed. Hexane e methanol crude extracts were prepared from the fruits, coconut shell and seeds, which were tested to evaluate the biological activity in vitro against promastigotes of the stationary phase of the protozoan Leishmania (Viannia) guyanensis. The extracts were analyzed by thin layer chromatography Comparison (CCDC) and Nuclear Magnetic Resonance (NMR) to determine the chromatographic profile. All biological assays were carried out experimentally in vitro in microplates containing 106/mL promastigotes to axenic cultures of L. (V.) guyanensis evaluating the inhibitory activity and stimulating parasite growth. The most active crude extract was submitted to CCDC and fractionation by partition líquid-líquid resulting in semi-purified fractions that were evaluated for their chemical composition. The results showed that the epicarp methanolic extract of C. ferrea, showed anti-leishmanial activity for L. (V.) guyanensis in the concentration of 20 mg/mL/24h. However, contrary to expectations, there was a stimulatory effect on L. (V.) guyanensis by the hexanic extract obtained from the fruit seed of C. ferrea. Chemically analyzing the coconut shell and fruits methanolics extracts of, which showed anti-leishmanial activity, we verified the presence of substances such as chlorophyll, flavonoids, xanthones, tannins, triterpenes and saponins as major constituents, and shows antioxidant activity. The results showed that the methanolics extracts of coconut shell and fruitsof C. ferrea can be considered as an promising alternative in the treatment of cutaneous leishmaniasis and should be undertaken pharmacological and toxicological tests in vivo of extract, making other fractioning in order to isolate the substances responsible for the anti-leishmanial activity. / A leishmaniose é um problema de saúde pública mundial considerada pela Organização Mundial da Saúde como uma das cinco doenças infecto-parasitárias endêmicas de maior relevância. A busca por agentes leishmanicidas com poucos efeitos colaterais é um dos maiores desafios na pesquisa moderna, e cada vez mais está relacionada aos usos de plantas medicinais e outros produtos naturais. A espécie Caesalpinia ferrea Martius largamente distribuída no Brasil, é usada popularmente para muitos fins terapêuticos, inclusive no tratamento de diversas infecções como as causadas pela Leishmaniose, tornando-a promissora para a realização de ensaios de atividade biológica contra a leishmania. O objetivo deste trabalho foi o de avaliar, através de bioensaios, a atividade de extratos hexânicos e metanólicos contra protozoários, analisando também frações semi-purificadas obtidas do extrato de melhor atividade leishmanicida. Os frutos de C. ferrea foram coletados no Instituto Nacional de Pesquisas da Amazônia, Campus do V8 na Cidade de Manaus-AM, e processados nos departamentos de Produtos Naturais e de Leishmaniose e Doença de Chagas no INPA. Os estudos foram realizados com os frutos inteiros e suas partes separadas, epicarpos e sementes. Foram preparados extratos brutos hexânicos, e metanólicos a partir dos frutos inteiros e também das partes separadas, epicarpos e sementes, os quais foram submetidos aos ensaios para avaliação da atividade biológica in vitro contra formas promastigotas da fase estacionária de protozoários da espécie Leishmania (Viannia) guyanensis. Os extratos brutos foram analisados através de Cromatografia em Camada Delgada Comparativa (CCDC) e Ressonância Magnética Nuclear (RMN) para determinação do perfil cromatográfico. Todos os ensaios biológicos foram realizados experimentalmente in vitro, em microplacas contendo 106/mL formas promastigotas de culturas axênicas de L. (V.) guyanensis (MHOM/BR/75/IM 4147) avaliando-se a atividade inibitória ou ativadora de crescimento parasitário. O extrato bruto de melhor atividade biológica foi submetido à CCDC e fracionamento por partição líquido-líquido obtendo-se frações semi-purificadas que foram avaliadas quanto a sua composição química. Os resultados mostraram que o extrato metanólico do epicarpo de C. ferrea, apresentou atividade leishmanicida para L. (V.) guyanensis na concentração de 20 mg/mL/24h. No entanto, ao contrário das expectativas, houve um efeito estimulante em L. (V.) guyanensis pelo extrato hexânico obtido das sementes de frutos de C. ferrea. Analisando quimicamente o extrato metanólico de frutos e epicarpos, os quais demonstraram atividade leishmanicida, verificou-se a presença de substâncias como a clorofila, flavonóides, xantonas, taninos, triterpenos e saponinas como constituintes majoritários, além de apresentar atividade antioxidante. Os resultados obtidos demonstraram que os extratos metanólicos de frutos e epicarpos de C. ferrea podem ser considerados como importantes alternativas no tratamento das leishmanioses, devendo ser realizado testes farmacológicos e toxicológicos in vivo desses extratos, realizando outros fracionamentos a fim de se confirmar alguma substância de melhor atividade leishmanicida.