Submitted by Raniere Barreto (raniere.barros@ufvjm.edu.br) on 2018-04-12T16:59:34Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lara_franca_colares.pdf: 2530593 bytes, checksum: f6b37bbb6a95d44536fc10c983a32afa (MD5) / Rejected by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br), reason: Verificar: "Ara?jo e Silva" e alterar, deixando Silva. Inserir as keywords adicionado campos devidos. UFVJM n?o ? ag?ncia financiadora. on 2018-04-20T14:32:10Z (GMT) / Submitted by Raniere Barreto (raniere.barros@ufvjm.edu.br) on 2018-05-15T17:49:49Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lara_franca_colares.pdf: 2530593 bytes, checksum: f6b37bbb6a95d44536fc10c983a32afa (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-05-16T11:27:14Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lara_franca_colares.pdf: 2530593 bytes, checksum: f6b37bbb6a95d44536fc10c983a32afa (MD5) / Made available in DSpace on 2018-05-16T11:27:14Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) lara_franca_colares.pdf: 2530593 bytes, checksum: f6b37bbb6a95d44536fc10c983a32afa (MD5) Previous issue date: 2016 / Plantas medicinais s?o comumente usadas por comunidades tradicionais, principalmente em regi?es com menor desenvolvimento humano. Algumas esp?cies vegetais possuem entre seus componentes subst?ncias denominadas inibidores de proteases. Os inibidores de proteases se destacam na prote??o de fluidos e tecidos contra sua degrada??o por prote?lise e poss?veis falhas na degrada??o de prote?nas de meia-vida que podem interferir de forma dr?stica nas fun??es celulares. Diante do exposto, esse estudo objetivou identificar e caracterizar inibidores de proteases em cinco esp?cies vegetais nativas do Cerrado e da Mata Atl?ntica. As esp?cies de Punica granatum L. (Rom?), Plantago major L. (Tansagem), Ocimum gratissimum L. (Alfavaca), Anadenanthera colubrina Vellozo (Angico) e Stryphnodendron adstringens Mart. Coville (Barbatim?o) foram selecionados nas cidades de Pot?, Ladainha, Atal?ia, Te?filo Otoni e Ara?ua?, devido ao seu uso tradicional como anti-inflamat?rio. A sequencia gen?mica de inibidores de proteases foi pesquisada para essas esp?cies vegetais no GenBank, mas nenhuma sequencia foi descrita para as esp?cies selecionadas. As amostras provenientes dos procedimentos de extra??o foram submetidas ?s quantifica??o de prote?nas e a presen?a de inibidores de proteases foi detectada por eletroforese em gel de poliacrilamida 12% SDS-PAGE. Somente os extratos das sementes de Punica granatum e das folhas do Anadenanthera colubrina tiveram detec??o satisfat?ria de inibidores de proteases e foram submetidos ? an?lise por cromatografia l?quida de alta efici?ncia em sistema de HPLC. Este trabalho demonstra pela primeira vez a detec??o e extra??o de inibidores de proteases em folhas de Anadenanthera colubrina e sementes de Punica granatum. / Disserta??o (Mestrado Profissional) ? Programa de P?s-Gradua??o em Tecnologia, Sa?de e Sociedade, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / Traditional communities, especially in regions with lower human development, commonly use medicinal plants. Some plant species have among their components substances called protease inhibitors. Protease inhibitors act protecting fluids and tissues from degradation by proteolysis and possible failures in the degradation of half-life proteins that can drastically interfere with cellular functions. This study aimed to identify and characterize protease inhibitors in five native plant species of Cerrado and Atlantic Forest. The species of Punica granatum L. (Rom?), Plantago major L. (Tansagem), Ocimum gratissimum L. (Alfavaca), Anadenanthera colubrina Vellozo (Angico) and Stryphnodendron adstringens Mart. Coville (Barbatim?o) were selected in the cities of Pot?, Ladainha, Atal?ia, Te?filo Otoni and Ara?ua? due to their traditional use. The genomic sequence of protease inhibitors was screened for these plant species in GenBank, but no sequence was described for the selected species. Samples from the extraction procedures were subjected to protein quantification and the presence of protease inhibitors was detected by 12% SDS-PAGE polyacrylamide gel electrophoresis. Only the extracts of the seeds of Punica granatum and of the leaves of Anadenanthera colubrina had satisfactory detection of proteases inhibitors and were submitted to the analysis by high performance liquid chromatography system. This work demonstrates for the first time the detection and extraction of protease inhibitors in leaves of Anadenanthera colubrina and seeds of Punica granatum.

Les composés phénoliques sont des métabolites secondaires chez les végétaux, qui possèdent un large éventail de propriétés biologiques et fonctionnelles, par exemple antioxydantes, antimicrobiennes. Comme ils sont très répandus dans le règne végétal, les sous-produits issus de la transformation végétale représentent une source intéressante et peu coûteuse des polyphénols qui pourraient être utilisés comme additifs naturels, inoffensifs et multifonctionnels, avec une valeur de marché ajoutée. Toutefois, leur polarité limite leur utilisation dans des systèmes émulsionnés complexes, tels que ceux rencontrés en formulations cosmétiques, alimentaires, ou pharmaceutiques. Sur cette base, la lipophilisation s’est avérée être une stratégie efficace pour améliorer le caractère hydrophobe de ces composés bioactifs et, par conséquent, leurs propriétés fonctionnelles.Le principal objectif de ce travail a été d'ajouter de la valeur à des sources végétales riches en polyphénols par l'extraction, caractérisation et enfin, par lipophilisation des principaux polyphénols issus de ces sources. Dans ce contexte, Hibiscus sabdariffa L. ou roselle, cultivée dans le monde entier et considérée comme une importante source d’anthocyanes, avec des propriétés colorantes uniques, ainsi que des propriétés antioxydantes et antimicrobiennes, a été utilisée pour l'extraction assistée par ultrasons, suivi d’une seule étape de fractionnement par chromatographie en phase liquide à basse pression, pour isoler ces principaux anthocyanes. Les composés ainsi obtenus ont été utilisés pour évaluer leur possibilité à être lipophilisés en utilisant différentes longueurs de chaînes alkyles. Egalement, les propriétés colorantes des anthocyanes issues d’hibiscus ont été calculées par la méthode de pH jumps. Ainsi, les constantes de pseudo équilibre (pK'a 2,9), d’acidité (pKa 3,7) et d’hydratation (pK'h 3,0) de la delphinidine-3-O-sambubioside, ont été estimées.La delphinidine-3-O-sambubioside et la cyanidine-3-O-sambubioside ont été les principaux anthocyanes obtenus à partir de l’extrait d'hibiscus. Ces composés ont été lipophilisés avec succès pour la première fois, par voie chimique. Les produits ont été caractérisés par des techniques de chromatographie et spectrométrie. L'acide protocatéchique (PA), qui peut être extrait de la fleur d'hibiscus, a été utilisé comme molécule modèle, et il a été lipophilisé par voie chimique avec une série homologue d'alcools. Afin d'évaluer l'impact de la lipophilisation sur leurs propriétés fonctionnelles, leur activités antioxydantes (DPPH, ORAC, CAT), et antifongique (méthode de microdilution) ont été évaluées avant et après lipophilisation. De manière générale, les molécules estérifiées avec des alcools de chaîne courte et moyenne (de C1 à C6) ont montré des activités améliorées par rapport au composé non estérifié.En conclusion, cette étude a montré que la lipophilisation, en utilisant une longueur de chaîne optimale, pourrait avoir un impact positif sur les propriétés antioxydantes et antifongiques des composés phénoliques. Ces travaux de thèse ont également démontré que cette stratégie peut être appliquée à différentes familles de composés phénoliques, tels que les anthocyanes et les acides phénoliques, pour l’obtention des molécules pouvant être potentiellement utilisées comme additifs dans des produits de formulation. / Phenolic compounds are a group of secondary plant metabolites with a broad range of inherent biological and functional properties e.g. antioxidants and antimicrobial agents, all bundled within the phenol functional group. Since they are one of the most widely distributed molecules in the vegetal kingdom, by-products issued from the vegetal processing represent an interesting and inexpensive source of polyphenols that could be used as natural, innocuous and multifunctional ingredients, with an added market value. However, the polarity of these phenolic compounds limits their use in complex emulsified systems, such as food, pharmaceutical or cosmetic formulations. Based on this, lipophilization has showed to be an efficient strategy to enhance the hydrophobicity of these bioactive compounds and, consequently, their functional properties. The aim of this work was to add value to polyphenol-rich plant sources by the extraction, characterization and finally, lipophilization of their predominant polyphenols. In this context, Hibiscus sabdariffa L. or roselle flower, an inexpensive, worldwide produced source of anthocyanins, with unique color, and, antioxidant and antimicrobial properties, was used for ultrasound-assisted extraction and single-step fractionation by low-pressure liquid chromatography, to isolate its main anthocyanins. The obtained compounds were used to evaluate their suitability for lipophilization with different chain length alkyl donors. Moreover, the color properties of hibiscus anthocyanins were studied by the pH jumps method, which allowed the calculation of the pseudo equilibrium (pK'a 2.9), acidity (pKa 3.7) and hydration constants (pK'h 3.0).The obtained hibiscus anthocyanins (delphinidin-sambubioside and cyanidin-sambubioside) were successfully chemically lipophilized. The products were characterized by means of LC/ESI-MS. Moreover, protocatechuic acid (PA), which can be extracted from hibiscus flowers, was used as model molecule, and it was chemically lipophilized with a homologous series of alcohols. In order to evaluate the impact of the lipophilization on their functional properties, protocatechuates were assessed for their antioxidant (DPPH, ORAC, CAT), and antifungal (microdilution method) activity, before and after chemical lipophilization. In general, protocatechuates with short to medium alkyl chain lengths (from C1 to C6) showed improved activities compared to the parent compound. In conclusion, this study showed that, lipophilization using the optimal chain length could have a positive impact in the functional properties, e.g. antioxidant and antifungal activities, of phenolic compounds. Here was also demonstrated that this strategy can be applied to different families of phenolic compounds, such as anthocyanins and phenolic acids, representing an alternative to synthesize molecules that might have potential use as additives in formulated products.

Le diabète est reconnu comme un problème majeur de santé publique causant des conséquences humaines et économiques redoutables. La phytothérapie s’offre comme une nouvelle avenue thérapeutique pour le contrôle de la glycémie. Le grenadier, Punica granatum, a servi de remède contre le diabète dans le système Unani de la médecine pratiquée en Inde et au Moyen Orient. Des études ont démontré un effet hypoglycémiant des extraits de grenadier via divers mécanismes notamment par une amélioration de la sensibilité à l’insuline et la régénération des cellules béta-pancréatiques. Cependant, aucune étude n’a démontré à ce jour, l’effet de grenadier sur le transport de glucose dans le muscle, étape cruciale dans la régulation de l’homéostasie glucidique postprandiale. De plus, l’effet de la maturation sur le potentiel antidiabétique du fruit de grenadier n’a pas été étudié. Ainsi, le but de ce projet est d’évaluer l’effet antidiabétique des extraits de grenadier sur le transport de glucose dans les cellules musculaires C2C12 en fonction de la variété et du stade de maturation du fruit et d’élucider les mécanismes d’action. Le choix des variétés du grenadier tunisien (Espagnoule [EP] et Gabsi [GB]) a été orienté pour leur pouvoir antioxydant et leur consommation locale. Deux parties de la plante ont été utilisées, les fleurs et les fruits à 3 stades de maturation soit 2, 4 et 6 mois. Les résultats ont montré que seule la variété du grenadier Gabsi stimule significativement le transport de glucose par rapport au contrôle (DMSO), et ceci sans être toxique. Cet effet est plus prononcé au stade de fruit mûr (à 6 mois) que celui de la fleur. De plus, l’extrait de fleurs stimule la voie insulino-indépendante de l’AMPK et augmente le niveau d’expression des transporteurs spécifiques de glucose (GLUT-4). Par contre, l’extrait de fruits mûrs, en plus de ces deux mécanismes, active fortement aussi la voie insulino-dépendante de l’AKT. En conclusion, cette étude présente un nouveau mécanisme d’action antidiabétique de grenadier (plus particulièrement du fruit mûr) qui est dépendant de la variété. / Diabetes is a major public health problem worldwide with astounding human and economic consequences. The seed and the flower of pomegranate (Punica granatum), a native plant of Central Asia and the Mediterranean regions, exhibited a hypoglycaemic effect in in vivo studies. However, the underlying mechanisms have not yet been elucidated. The aim of this project was to evaluate the effect of the flower and the fruit (at 3 maturation stages) of pomegranate on glucose transport in skeletal muscle cells and to determine the molecular mechanisms involved in this effect. To accomplish this, we chose two varieties of pomegranate cultivated in Tunisia (Gabsi [GB] and Espagnoule [EP]), which have been shown to be highly consumed in that area and to possess high antioxidant activity. Differentiated C2C12 cells were treated for 18 hours with 80% ethanolic extract of the flowers and fruits (at 2, 4, and 6 months) of each variety. Our results showed that the Gabsi variety of pomegranate significantly enhances glucose uptake, without any toxicity. This effect is more pronounced in the ripe fruit (6 months) than in the flower. In parallel, the ripe fruit stimulated both the insulin-dependent pathway (Akt) and the insulin-independent pathway (AMPK), while the flower stimulated the latter only. In addition, both flower and ripe fruit treatment resulted in enhanced expression level of GLUT-4 glucose transporter in the muscle. Hence, these results suggest that regulation of glucose transport in skeletal muscle is one of the components involved in the anti-diabetic effect of Tunisian pomegranate.

A biodiversidade da flora brasileira abre a possibilidade de estudos com extratos naturais para formulações de soluções irrigantes bactericidas e biocompatíveis para uso na terapia endodontia. Este estudo teve como objetivo analisar o potencial antimicrobiano de extratos da Punica granatum L. (P. granatum) contra micro-organismos presentes nas infecções do sistema de canais radiculares. Foram preparados extratos hidroetanólicos (50% etanol e 50% água) na concentração de 100 mg/mL, a partir da liofilização da casca, folha e semente da P. granatum, contra os seguintes microrganismos: Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis e Candida albicans. Os grupos foram distribuídos de acordo com o extrato de P. granatum: G1 etanol 50% (controle positivo), G2 - hipoclorito de sódio 1% (NaOCl) (controle negativo), G3 Extrato da casca, G4 Extrato da folha, G5 Extrato da semente. A capacidade antimicrobiana dos extratos foi avaliada pelo teste de difusão em ágar, concentração inibitória mínima (CIM) e concentração bactericida/fungicida mínima (CBM/CFM). Os dados, em mm, foram analisados por Analise de Variância e teste de Tukey (p<0,05). Pelo teste de difusão em ágar, verificou-se que, independentemente do micro-organismo, o maior halo de inibição foi encontrado para o controle positivo (30,76 &plusmn; 4,73 a), seguido da casca e folha, que não apresentaram diferença significante entre si (23,32 &plusmn; 3,65 b e 21,08 &plusmn; 2,28 b, respectivamente). A semente apresentou o menor halo de inibição microbiana (17,83 &plusmn; 6,92 c) (p<0,05). Na comparação isolada para cada micro-organismo, os extratos da casca e folha apresentaram atividade antimicrobiana semelhante ao NaOCl para a bactéria S. mutans (p=0,0000), além de exercerem bom efeito antimicrobiano sobre os demais micro-organismos. Para as bactérias E. faecalis e E. coli, os extratos (casca, folha e semente) apresentaram halos de inibição semelhantes entre si (p>0,05), porém inferiores aos obtidos pelo controle positivo. Com relação a CIM e CBM/CFM, encontrou-se efeito bactericida e bacteriostático para todos os extratos. Verificou-se maior zona de inibição para o extrato da casca contra a C. albicans<i/> (CIM = 6,25 mg/mL e CFM = 12,5 mg/mL), seguido da S. aureus (CIM e CBM =12,5 mg/mL). Concluiu-se que, os extratos hidroalcoólicos de P. Granatum L. apresentaram atividade antimicrobiana contra os patógenos orais comuns nas infecções endodontias, com a maior atividade para os obtidos da casca e folha / Brazilian flora biodiversity opens possibility of studies with natural extracts for bactericidal and biocompatible solutions for use in endodontic therapy. The purpose of this study was to investigate the antimicrobial effect of Punica granatum L. (pomegranate) against pathogens of infections from root canal system. Ethanolic solutions (50% ethanol and 50% water) were prepared in 100 mg/mL of lyophilized extracts derived from the peel, seed and leaf of pomegranate, against the following oral microorganisms: Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Enterococcus faecalis and Candida albicans. The groups were distributed according to the extract tested for root canal irrigation: G1 - 50% ethanol (positive control), G2 - 1% sodium hypochlorite (NaOCl) (negative control), G3 - Solution of the peel, G4 Seed solution, G5 - Solution of the leaf. The antimicrobial capacity of the extracts was evaluated by the agar diffusion test, minimum inhibitory concentration (MIC) and minimum bactericidal / fungicidal concentration (MCB/MFC). The data were analyzed by ANOVA (extracts and microorganisms) and Tukey test (p<0.05). By the diffusion test in agar, it was verified that, independently of the microorganism, the largest inhibition zone was found for the positive control (30.76 &plusmn; 4.73 a), followed by peel and leaf, which did not presented significant difference between them (23.32 &plusmn; 3.65 and 21.08 &plusmn; 2.28 b, respectively). The seed had the lowest of microbial inhibition zone (17.83 &plusmn; 6.92 c) (p<0.05). In the isolated comparison for each microorganism, peel and leaf extracts presented antimicrobial activity similar to NaOCl for S. mutans (p = 0.0000), besides having a good antimicrobial effect on the other microorganism. For the E. faecalis and E. coli, the extracts (peel, leaf and seed) had similar inhibition zones (p>0.05), but lower than those obtained by the positive control. Regarding MIC and MBC/MFC, it was found a bactericidal and bacteriostatic effect for all extracts. The P. granatum peel extract had greater inhibition zone against C. albicans (MIC = 6.25 mg/mL and MFC = 12.5 mg/mL), followed by S. aureus (MIC and MBC = 12.5 mg/mL). It may be concluded that the hydroalcoholic extracts of P. granatum L. presented antimicrobial activity against oral pathogens common in endodontic infections, with the highest activity for those obtained from the peel and leaf

Le diabète est reconnu comme un problème majeur de santé publique causant des conséquences humaines et économiques redoutables. La phytothérapie s’offre comme une nouvelle avenue thérapeutique pour le contrôle de la glycémie. Le grenadier, Punica granatum, a servi de remède contre le diabète dans le système Unani de la médecine pratiquée en Inde et au Moyen Orient. Des études ont démontré un effet hypoglycémiant des extraits de grenadier via divers mécanismes notamment par une amélioration de la sensibilité à l’insuline et la régénération des cellules béta-pancréatiques. Cependant, aucune étude n’a démontré à ce jour, l’effet de grenadier sur le transport de glucose dans le muscle, étape cruciale dans la régulation de l’homéostasie glucidique postprandiale. De plus, l’effet de la maturation sur le potentiel antidiabétique du fruit de grenadier n’a pas été étudié. Ainsi, le but de ce projet est d’évaluer l’effet antidiabétique des extraits de grenadier sur le transport de glucose dans les cellules musculaires C2C12 en fonction de la variété et du stade de maturation du fruit et d’élucider les mécanismes d’action. Le choix des variétés du grenadier tunisien (Espagnoule [EP] et Gabsi [GB]) a été orienté pour leur pouvoir antioxydant et leur consommation locale. Deux parties de la plante ont été utilisées, les fleurs et les fruits à 3 stades de maturation soit 2, 4 et 6 mois. Les résultats ont montré que seule la variété du grenadier Gabsi stimule significativement le transport de glucose par rapport au contrôle (DMSO), et ceci sans être toxique. Cet effet est plus prononcé au stade de fruit mûr (à 6 mois) que celui de la fleur. De plus, l’extrait de fleurs stimule la voie insulino-indépendante de l’AMPK et augmente le niveau d’expression des transporteurs spécifiques de glucose (GLUT-4). Par contre, l’extrait de fruits mûrs, en plus de ces deux mécanismes, active fortement aussi la voie insulino-dépendante de l’AKT. En conclusion, cette étude présente un nouveau mécanisme d’action antidiabétique de grenadier (plus particulièrement du fruit mûr) qui est dépendant de la variété. / Diabetes is a major public health problem worldwide with astounding human and economic consequences. The seed and the flower of pomegranate (Punica granatum), a native plant of Central Asia and the Mediterranean regions, exhibited a hypoglycaemic effect in in vivo studies. However, the underlying mechanisms have not yet been elucidated. The aim of this project was to evaluate the effect of the flower and the fruit (at 3 maturation stages) of pomegranate on glucose transport in skeletal muscle cells and to determine the molecular mechanisms involved in this effect. To accomplish this, we chose two varieties of pomegranate cultivated in Tunisia (Gabsi [GB] and Espagnoule [EP]), which have been shown to be highly consumed in that area and to possess high antioxidant activity. Differentiated C2C12 cells were treated for 18 hours with 80% ethanolic extract of the flowers and fruits (at 2, 4, and 6 months) of each variety. Our results showed that the Gabsi variety of pomegranate significantly enhances glucose uptake, without any toxicity. This effect is more pronounced in the ripe fruit (6 months) than in the flower. In parallel, the ripe fruit stimulated both the insulin-dependent pathway (Akt) and the insulin-independent pathway (AMPK), while the flower stimulated the latter only. In addition, both flower and ripe fruit treatment resulted in enhanced expression level of GLUT-4 glucose transporter in the muscle. Hence, these results suggest that regulation of glucose transport in skeletal muscle is one of the components involved in the anti-diabetic effect of Tunisian pomegranate.

Made available in DSpace on 2016-08-11T14:41:32Z (GMT). No. of bitstreams: 1 GeruziaMTQ_DISSERT.pdf: 1649490 bytes, checksum: ad9e3fac3a63c7899a5596551eb9d334 (MD5) Previous issue date: 2015-02-10 / Medicinal plants are a therapeutic resource and are being increasingly used at societies worldwide. It constitutes a preservation tool of the cultural environment, of the economic and social development, promoting improvements in the population's quality of life. The aim of this study was to evaluate the use of phytotherapy as a therapeutic alternative in the framework of the public health of Mossoró/RN, considering the popular and scientific knowledge. The data collection instrument regarding the informations about the phytotherapy was the structured questionnaire, administered to a population of 453 subjects, among which, users (269) and SUS s health professionals (184), enrolled in the Basic Health Units. For evaluation of the use of vegetable species, on specific illness, was made the determination of the antibacterial activity of the plants s extracts cited in ITU (Phyllanthus niruri L, Punica granatum L and Zea mays L), against clinical isolates of people from the community and ATCC s standard strains of Escherichia coli, Klebsiella pneumoniae and Staphylococcus aureus, utilizing the disk diffusion in solid medium method. The crude extracts were tested at concentrations of 100, 200 and 300 mg/mL having DMSO as solvent. To the analysis of the data obtained with the questionnaire was used Fisher's exact test for contingency tables, as for the comparison of the diameter of the extracts inhibition halos, was used the variance analysis in the design in randomized blocks, and for the comparison of the halos means the Student-Newman-Keuls test, with the R software with a 5% significance level. The results showed that 86.41% of health professionals and 89.96% of public health services users know, accept and utilize the phytotherapy as a therapeutic option, and the Punica granatum extract as the most effective and with the greater spectrum of action regarding the tested bacterias, confirming the popular knowledge about the antibacterial property of these plants, enabling the implantation of a municipal health public policy, focused on phytotherapy as a way to ensure the right to health of the population, without necessarily depend on allopathy, having the common sense as a search instrument of potentially therapeutic plants in the region / As plantas medicinais são um recurso terapêutico e vêm sendo cada vez mais utilizadas nas sociedades em todo o mundo. Constitui uma ferramenta de preservação do ambiente cultural, de desenvolvimento econômico e social, promovendo melhorias na qualidade de vida da população. O objetivo desse estudo foi avaliar o uso da fitoterapia como alternativa terapêutica no âmbito da saúde pública de Mossoró/RN, considerando o conhecimento popular e científico. O instrumento de coleta de dados quanto a informações sobre a fitoterapia foi um questionário estruturado, aplicado a uma população de 453 sujeitos, dentre os quais, usuários (269) e profissionais de saúde (184) do SUS, arrolados nas Unidades Básicas de Saúde. Para avaliação do uso de espécies vegetais, em enfermidades específicas, fez-se a determinação da atividade antibacteriana dos extratos de plantas citadas em ITU (Phyllanthus niruri L, Punica granatum L e Zea mays L), sobre isolados clínicos de pessoas da comunidade e cepas padrão ATCC de Escherichia coli, Klebsiella pneumoniae e Staphylococcus aureus, utilizando-se o método de Disco Difusão em meio sólido. Os extratos brutos foram testados nas concentrações de 100, 200 e 300 mg/mL tendo como solvente o DMSO. Para a análise dos dados obtidos com os questionários foi utilizado o teste exato de Fisher para tabelas de contingência, já para a comparação do diâmetro dos halos de inibição dos extratos, utilizou-se a análise de variância no delineamento em blocos aleatorizados, e para a comparação das médias dos halos o teste de Student-Newman-Keuls, por meio do software R, com um nível de significância de 5%. Os resultados mostraram que 86,41% dos profissionais da saúde e 89,96% dos usuários dos serviços públicos de saúde conhecem, aceitam e utilizam a fitoterapia como opção terapêutica, e o extrato de Punica granatum como o mais eficaz e com maior espectro de ação quanto às bactérias testadas, confirmando o conhecimento popular quanto à propriedade antibacteriana destas plantas, possibilitando a implantação de uma política pública de saúde municipal, voltada para a fitoterapia como forma de assegurar o direito à saúde da população, sem necessariamente depender da alopatia, tendo o senso comum como instrumento de busca de plantas potencialmente terapêuticas na região

O objetivo deste trabalho foi otimizar a extração dos compostos fenólicos presentes na casca do fruto de Punica granatum L. (romã) com diferentes locais de coletas e adquiridas comercialmente, bem como desenvolver e validar método analítico para separação e quantificação de punicalaginas totais por cromatografia líquida de alta eficiência (CLAE) e cromatografia líquida de ultra eficiência (CLUE), ensaios de pureza como determinação de água e cinzas totais e testar a atividade anti-inflamatória pela avaliação antiquimiotáxica in vitro do extrato e fração enriquecida de punicalagina. Para a otimização da extração com amostra coletada em Alegrete/15, diferentes técnicas extrativas foram testadas, entre elas: maceração, ultrassom, ultraturrax e refluxo, sendo esta última à escolhida. Análises de fatores críticos que influenciam na resposta, sendo eles temperatura, granulometria, porcentagem do solvente extrator, relação massa vegetal/solvente e tempo, foram realizadas, utilizando software estatístico MiniTab®. O método por CLAE foi desenvolvido, permitindo a separação e quantificação dos anômeros punicalagina α e β, com o emprego de coluna cromatográfica Kromasil® C18 – 5μm x 4.6mm x 150mm e fase móvel constituída por água acidificada com ácido trifluoroacético : acetonitrila acidificada com TFA.. O método pôde ser considerado eficiente e confiável, sendo linear, preciso, robusto, exato e específico, podendo ser utilizado para análise do extrato de P. granatum. O extrato bruto, bem como a fração de punicalaginas foram testados e reduziram significativamente a migração leucocitária nas concentrações mais elevadas (10 e 1 μg/mL) em relação ao controle positivo, indicando potencial atividade inibitória da migração leucocitária. / The objective of this work was to optimize the extraction of the phenolic compounds present in the fruit bark of Punica granatum L. (pomegranate) with different collection sites and commercially acquired, as well as to develop and validate an analytical method for the separation and quantification of total punicalagins by liquid chromatography (HPLC) and ultra-high performance liquid chromatography (HPLC), purity tests such as water determination and total ashes, and to test the anti-inflammatory activity by the in vitro antichemistoxic evaluation of the extract and enriched fraction of punicalagin. For the optimization of the extraction with sample collected in Alegrete / 15, different extractive techniques were tested, among them: maceration, ultrasound, ultraturrax and reflux, the latter being chosen. Analysis of critical factors influencing the response, being temperature, particle size, solvent extractor percentage, vegetable mass / solvent ratio and time, were performed using MiniTab® statistical software. The HPLC method was developed allowing separation and quantification of the punicalagin α and β anomers using a Kromasil® C18 - 5μm x 4.6mm x 150mm chromatographic column and mobile phase consisting of water acidified with trifluoroacetic acid: acetonitrile acidified with TFA The method could be considered efficient and reliable, being linear, precise, robust, exact and specific, and it can be used for the analysis of the extract of P. granatum. The crude extract as well as the fraction of punicalaginas were tested and significantly reduced the leukocyte migration at the highest concentrations (10 and 1 μg / mL) in relation to the positive control, indicating potential inhibitory activity of leukocyte migration.

Pomegranate extract (PE) is a standardized whole fruit extract of pomegranate (Punica granatum L.), a fruit with known anti inflammatory and anti cancer properties. In the present study, the effects of PE on two different cell lines of human pancreatic cancer cells, AsPC-1 and PANC-1 were examined. Both cell lines are epithelial cancers but differ morphologically and in their response to treatment with PE. PE inhibited proliferation of both cancer cell lines in a dose dependent and time dependent manner. The reason for this was the potency of PE to cause significant cell cycle arrest in the pancreatic cancer cells, which was noted to be cell specific. AsPC-1 was arrested in G₂ phase while PANC-1 was arrested in G₀/G₁phase of the cell cycle. PE also checked the viability of confluent cancer cells in a concentration dependent manner. In case of AsPC-1 this was due to the action of PE to induce apoptosis, as demonstrated by the Annexin-V assay. In case of PANC-1, a highly invading cancer greatly affected by the presence of surface stem cell markers CD44⁺/CD24⁺, PE was capable of modifying the stem cell markers and induce an increase in the non tumorigenic sub population of cells. A study was also conducted to look at the action of PE in causing inhibition of pancreatic cancer cell proliferation, in comparison to a clinically used chemotherapeutic agent, Paclitaxel. The results indicated that the effect of PE, at clinically feasible doses was superior to that of clinically used doses of Paclitaxel. This data suggests that PE, which is proven to be a safe dietary supplement, looks promising in the prevention and treatment of human pancreatic cancer cells without the side effects of standard drug therapy and could possibly have a role in controlling stem cancer cell population / text

Memoria para optar al título profesional de: Ingeniero Agrónomo Mención: Fruticultura / La prolongada floración del granado tiene como consecuencia una marcada heterogeneidad en el estado de desarrollo de los frutos al momento de la cosecha, lo cual implica que un alto porcentaje de ellos no llega a madurez. Este hecho evidencia la necesidad de raleo de frutos en los huertos de granado, práctica que, en la actualidad, se realiza de forma manual. El objetivo de este estudio fue evaluar diferentes reguladores de crecimiento como posibles raleadores químicos de flores y/o frutos de granado. Con este fin se realizaron cinco ensayos en un huerto comercial de granados de la variedad Wonderful ubicado en la Región Metropolitana, que consistió en la evaluación de cinco productos químicos, en cinco concentraciones diferentes cada uno: ácido naftalenacético (ANA), en un rango de concentraciones entre 0 y 40 mg L-1; ethephon (ET), en un rango de concentraciones entre 0 y 400 mg L-1; benciladenina (BA), en concentraciones entre 0 y 400 mg L-1; ácido giberélico (AG), en concentraciones entre 0 y 100 mg L-1; y aminoetoxivinilglicina (AVG), en un rango de concentraciones entre 0 y 200 mg L-1. Se realizaron dos aplicaciones del producto químico con un lapso de dos semanas entre ellas, comenzando la tercera semana de enero. Se evaluó la dinámica de caída de flores y frutos durante las seis semanas siguientes a la primera aplicación. En la cosecha se recolectó toda la fruta presente en el árbol, clasificando cada fruto según cobertura externa de color, cohorte de floración a partir de la cual se desarrolló y peso fresco. Los ensayos se analizaron mediante correlaciones y regresiones. El ET fue el único producto evaluado que presentó efecto raleador, disminuyendo la proporción de frutos de la segunda y tercera cohorte de floración. Adicionalmente, el ET incrementó el color de los frutos y redujo el peso fresco de los frutos de la primera cohorte de floración.

O objetivo deste trabalho foi otimizar a extração dos compostos fenólicos presentes na casca do fruto de Punica granatum L. (romã) com diferentes locais de coletas e adquiridas comercialmente, bem como desenvolver e validar método analítico para separação e quantificação de punicalaginas totais por cromatografia líquida de alta eficiência (CLAE) e cromatografia líquida de ultra eficiência (CLUE), ensaios de pureza como determinação de água e cinzas totais e testar a atividade anti-inflamatória pela avaliação antiquimiotáxica in vitro do extrato e fração enriquecida de punicalagina. Para a otimização da extração com amostra coletada em Alegrete/15, diferentes técnicas extrativas foram testadas, entre elas: maceração, ultrassom, ultraturrax e refluxo, sendo esta última à escolhida. Análises de fatores críticos que influenciam na resposta, sendo eles temperatura, granulometria, porcentagem do solvente extrator, relação massa vegetal/solvente e tempo, foram realizadas, utilizando software estatístico MiniTab®. O método por CLAE foi desenvolvido, permitindo a separação e quantificação dos anômeros punicalagina α e β, com o emprego de coluna cromatográfica Kromasil® C18 – 5μm x 4.6mm x 150mm e fase móvel constituída por água acidificada com ácido trifluoroacético : acetonitrila acidificada com TFA.. O método pôde ser considerado eficiente e confiável, sendo linear, preciso, robusto, exato e específico, podendo ser utilizado para análise do extrato de P. granatum. O extrato bruto, bem como a fração de punicalaginas foram testados e reduziram significativamente a migração leucocitária nas concentrações mais elevadas (10 e 1 μg/mL) em relação ao controle positivo, indicando potencial atividade inibitória da migração leucocitária. / The objective of this work was to optimize the extraction of the phenolic compounds present in the fruit bark of Punica granatum L. (pomegranate) with different collection sites and commercially acquired, as well as to develop and validate an analytical method for the separation and quantification of total punicalagins by liquid chromatography (HPLC) and ultra-high performance liquid chromatography (HPLC), purity tests such as water determination and total ashes, and to test the anti-inflammatory activity by the in vitro antichemistoxic evaluation of the extract and enriched fraction of punicalagin. For the optimization of the extraction with sample collected in Alegrete / 15, different extractive techniques were tested, among them: maceration, ultrasound, ultraturrax and reflux, the latter being chosen. Analysis of critical factors influencing the response, being temperature, particle size, solvent extractor percentage, vegetable mass / solvent ratio and time, were performed using MiniTab® statistical software. The HPLC method was developed allowing separation and quantification of the punicalagin α and β anomers using a Kromasil® C18 - 5μm x 4.6mm x 150mm chromatographic column and mobile phase consisting of water acidified with trifluoroacetic acid: acetonitrile acidified with TFA The method could be considered efficient and reliable, being linear, precise, robust, exact and specific, and it can be used for the analysis of the extract of P. granatum. The crude extract as well as the fraction of punicalaginas were tested and significantly reduced the leukocyte migration at the highest concentrations (10 and 1 μg / mL) in relation to the positive control, indicating potential inhibitory activity of leukocyte migration.