Submitted by Natalie Mendes (nataliermendes@gmail.com) on 2015-07-28T23:56:29Z No. of bitstreams: 1 Disserta??o_?ngara Keisle S?o Paulo Barretto Miranda.pdf: 984548 bytes, checksum: 4e0290fe54977157fb35593ccdb7ef99 (MD5) / Made available in DSpace on 2015-07-28T23:56:29Z (GMT). No. of bitstreams: 1 Disserta??o_?ngara Keisle S?o Paulo Barretto Miranda.pdf: 984548 bytes, checksum: 4e0290fe54977157fb35593ccdb7ef99 (MD5) Previous issue date: 2014-03-25 / Funda??o de Amparo ? Pesquisa do Estado da Bahia - FAPEB / Bromelain is an enzyme complex with great therapeutic importance attributed both to it?s proteolytic activity, as the non-proteolytic components in the extract. Thus, the enzyme was extracted from different parts of Ananas comosus var comosus, from two pineapple with different codes: BGA 771 and BGA 772. The proteolytic and specific activities of bromelain from each part (stem, crown, peel, pulp and leaves) were investigated as well as their kinetic characteristics by measuring the optimum temperature and the optimum pH, and finally was studied it?s effect in vitro on murine melanoma B16F10 cells. In BGA 771, the highest proteolytic activity was found in the stem (81.10 mg/mL/min), while the highest specific activity was found in the pulp and peel (1983.44 and 1822.33 UA/mg of bromelain, respectively). In general, the optimum temperature varied between of 50 to 80 ?C and pH between 6 and 8. Bromelain all parts were effective more than 50% in inhibiting the melanoma B16F10 cells proliferation after 48 hours. The most significant inhibition was find on crown bromelain with 72.86%, and a time-dependent effect was observed on crown bromelain. At BGA 772 the higher activity was find in the pulp (35.58 mg/mL/min and 1446.60 UA/mg bromelain). The optimum temperature in the extracts ranged between 40 and 90 ?C, while the pH 6 was the best for all parties, while the leaf bromelain had two great points: 6 and 8. Only the bromelain peel was able to inhibit the melanoma B16F10 proliferation of, being 66.37% after 48 hours of contact, and a dose-dependent activity was observed. / A bromelina ? um complexo enzim?tico de grande import?ncia terap?utica, atribu?da tanto ? sua atividade proteol?tica, como aos componentes n?o proteol?ticos contidos no extrato. Dessa forma, a enzima foi extra?da de diferentes partes de Ananas comosus var comosus, a partir de dois abacaxizeiros com identifica??es distintas: BGA 771 e BGA 772. Foram investigadas as atividades proteol?tica e espec?fica da bromelina de cada parte (caule, coroa, casca, polpa e folha), bem como suas caracter?sticas cin?ticas atrav?s da medida da temperatura e do pH ?timos, investigando-se, por fim, o seu efeito in vitro sobre c?lulas tumorais de melanoma murino B16F10. No BGA 771, a maior atividade proteol?tica foi encontrada no caule (81,10 ?g/mL/min), enquanto a maior atividade espec?fica foi encontrada na polpa e na casca (1983,44 e 1822,33 UA/mg de bromelina, respectivamente). De um modo geral, a temperatura ?tima variou entre as partes de 50 a 80 ?C e o pH ?timo entre 6 e 8. A bromelina de todas as partes foi eficaz em inibir a prolifera??o das c?lulas de melanoma B16F10 ap?s 48 horas em mais de 50%, sendo a da coroa a mais significativa, com 72,86% de inibi??o, tendo sido observada uma atividade tempo-dependente. No BGA 772, as maiores atividade foram encontradas na polpa (35,58 ?g/mL/min e 1446,60 UA/mg de bromelina). A temperatura ?tima nos extratos variou entre 40 e 90?C, enquanto o pH 6 foi o melhor para todas as partes, exceto para a folha que teve dois pontos ?timos: 6 e 8. Somente a bromelina da casca foi capaz de inibir a prolifera??o do melanoma B16F10, sendo de 66,37% ap?s 48 horas de contato, tendo sido observada uma atividade dose-dependente.

The omega-3 polyunsaturated fatty acid eicosapentaenoic acid (EPA), in the free fatty acid (FFA) form, has been demonstrated to reduce adenoma number and size in patients with familial adenomatous polyposis. However, the mechanistic basis of the antineoplastic activity of EPA in the colorectum remains unclear. We tested the hypothesis that EPA-FFA negatively modulates synthesis of and signaling by prostaglandin (PG) E(2) in human colorectal cancer (CRC) cells. EPA-FFA induced apoptosis of cyclooxygenase (COX)-2-positive human HCA-7 CRC cells in vitro. EPA-FFA in cell culture medium was incorporated rapidly into phospholipid membranes of HCA-7 human CRC cells and acted as a substrate for COX-2, leading to reduced synthesis of PGE(2) and generation of PGE(3). Alone, PGE(3) bound and activated the PGE(2) EP4 receptor but with reduced affinity and efficacy compared with its "natural" ligand PGE(2). However, in the presence of PGE(2), PGE(3) acted as an antagonist of EP4 receptor-dependent 3',5' cyclic adenosine monophosphate induction in naturally EP4 receptor-positive LoVo human CRC cells and of resistance to apoptosis in HT-29-EP4 human CRC cells overexpressing the EP4 receptor. We conclude that EPA-FFA drives a COX-2-dependent "PGE(2)-to-PGE(3) switch" in human CRC cells and that PGE(3) acts as a partial agonist at the PGE(2) EP4 receptor.

Submitted by Rosivalda Pereira (mrs.pereira@ufma.br) on 2017-05-17T20:53:38Z No. of bitstreams: 1 DayaneFreitas.pdf: 3248256 bytes, checksum: ea021e1cfc7eeadb7ac1ec7bc8cfd5a3 (MD5) / Made available in DSpace on 2017-05-17T20:53:38Z (GMT). No. of bitstreams: 1 DayaneFreitas.pdf: 3248256 bytes, checksum: ea021e1cfc7eeadb7ac1ec7bc8cfd5a3 (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 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.