Memoria para optar al Título Profesional de Ingeniero Agrónomo / Durante la temporada 2010/2011 se realizó un ensayo con granados ´Wonderful´ en Huechún conducido como multieje a 5 x 3 m, región Metropolitana; Cerrillos de Tamaya como copa a 6 x 4 m y Vicuña como copa y eje a 3 x 1,5 m en la región de Coquimbo. El objetivo fue evaluar las relaciones entre el peso promedio de frutos, productividad, intercambio gaseoso, incidencia de golpe de sol, partidura, color y ramaleo en diferentes sistemas de conducción y localidades. Para ello, se seleccionaron diez plantas para cada combinación sistema de conducción-localidad, cada uno sometido a diferentes niveles de carga frutal, regularmente distribuidos entre muy baja y muy alta, ajustad luego del cuaje de la segunda cohorte de floración. La carga frutal se expresó como frutos por unidad de superficie de suelo, área de sección transversal de tronco (ASTT) y fracción de radiación interceptada (FI). La FI se midió con un Ceptómetro lineal un mes después del ajuste de carga frutal. El intercambio gaseoso se registró mediante un analizador de gases infrarrojo portátil durante la mañana (AM), el mediodía (MD) y el atardecer (PM) durante el período de activo crecimiento del fruto y pre-cosecha. La fruta cosechada, se clasificó de acuerdo a su calibre. Los daños comerciales como golpe de sol, partidura, color y ramaleo, se expresaron como porcentaje. Se encontró una relación lineal positiva entre la fracción de radiación interceptada y el rendimiento. El peso promedio disminuyó, y la productividad aumentó al aumentar la carga frutal. El intercambio gaseoso aumentó al incrementar la carga frutal. El golpe de sol disminuyó al aumentar la carga mientras la partidura aumentó. El color y el daño por ramaleo no fueron afectados por la carga frutal. El huerto de Huechún tuvo un alto rendimiento promedio pero con una mayor proporción de frutos pequeños, mientras que en Cerrillos de Tamaya la productividad fue baja. En Vicuña, tanto el sistema de copa como eje, presentaron una alta precocidad, buen tamaño de fruto, baja partidura de frutos, por lo que esta zona y sistema de conducción, si se controla el golpe de sol, se proyectan como adecuadas para la producción de granados. Palabras clave: intercambio gaseoso, fracción de radiación interceptada, carga frutal, peso promedio, eficiencia productiva.

Tesis presentada para optar al Título Profesional de Ingeniero Agrónomo y al Grado de Magíster en Ciencias Agropecuarias, Mención en Producción Frutícola / Con el fin de conocer la acumulación y variación estacional de la biomasa de órganos y estructuras en plantas jóvenes y en producción de granado var. Wonderful, se utilizaron relaciones alómetricas que permitieron estimar los coeficientes de reparto (CR), el índice del área foliar (IAF) y la eficiencia del uso de la radiación (EUR) en ambos huertos. Para construir las relaciones alométricas se utilizó un método directo, en donde se extrajeron 4 árboles de tres años y 2 árboles de cinco años, en los periodos de brotación, cuaja, crecimiento intermedio del fruto, cosecha y receso. Los árboles fueron separados en raíz, estructura caulinar, hojas, frutos y caídas, obteniéndose el peso fresco y seco de cada compartimiento. Antes de la extracción de cada grupo de plantas, se midió su radiación interceptada (PARf) y su área de sección transversal del tronco (ASTT), variables independientes que se relacionaron con el peso seco de los órganos extraídos. Las alometrías (método indirecto) obtenidas, fueron aplicadas en otros 6 árboles de ambos huertos en base a la medición de dichas variables alométricas desde brotación a receso. Además, se marcaron 16 frutos recién cuajados por árbol para realizar el seguimiento de diámetro ecuatorial hasta cosecha. Los resultados indican que las alometrías obtenidas determinaron con un R2 =0,92 la biomasa de granados. El crecimiento anual de biomasa de árboles en producción fue de 14 kg en peso seco. Los frutos representaron el 55% de la biomasa total, alcanzando un CR máximo de 0,8, previo a la cosecha y su crecimiento siguió un patrón de doble sigmoidea. El IAF máximo fue de 2,2 con un PARf de 45% y la EUR se estimó en 1,22 g·MJ-1. Los árboles jóvenes acumularon 2,5 kg en peso seco y los coeficientes de reparto al desarrollo de raíces, caulinar y hojas fueron similares. Su IAF máximo fue 0,6 con un PARf de 22% y una EUR de 0,8 g·MJ-1 / The accumulation and seasonal variation of the biomass of organs and structures, in young and fully-producing pomegranate plants var. Wonderful, were estimated using allometric relationships which allowed to estimate the partition coefficients (CR), the leaf area index (LAI) and the radiation use efficiency (RUE) in both orchards. In order to build the allometric relationships a direct method was used, which included four three year old and two five year old trees which were uprooted during the periods of bud break, fruit set, intermediate fruit growth, harvest and winter dormancy. The fresh and dry weight of the organs of the tree, which were pooled into five compartments (roots, caulinar structures, leaves, fruits and litter) was then measured. Before uprooting the plants, their radiation interception (PARf) and trunk cross-sectional area (TCSA) were measured, which were related to the dry weight of the aboveground organs. The allometric (indirect method) hereby obtained were applied in 6 trees in both orchards using the same biometric measurements from sprouting to winter dormancy. In addition, the equatorial diameter of 16 fruits was tracked from fruit set to harvest. The results gave a determination coefficient of 0.92 for the estimation of the whole plant biomass of pomegranates from the allometric functions. The annual growth of the fruit-bearing trees was estimated at 14 kg dry weight. The fruits accounted for 55% of the total biomass, reaching a maximum CR of 0.8 before harvest and their growth curve followed a double sigmoid pattern. The maximum LAI was 2.2 with a PARf of 45% and the RUE was estimated at 1.22 g·MJ-1. Young trees accumulated 2.5 kg dry weight over the growing season and equally devoted the partition coefficients of root development, caulinar structures and leaves. The maximum LAI of these trees was of 0.6 with a PARf of 22% and a RUE 0.8 g·MJ-1.

Memoria para optar al título profesional de: Ingeniero Agrónomo Mención Manejo de suelos y aguas / Debido a que el granado (Punica granatum L.) var. Wonderful se cultiva principalmente en zonas de suelos marginales y con problemas de escasez de agua, y que la producción de fruta está relacionada con el agua que se suministre, se hace necesario conocer los efectos del déficit hídrico sobre variables fisiológicas, de crecimiento y productivas de la especie. Con dicho fin se aplicaron cuatro tratamientos con distintas tasas de riego en granados, en un predio ubicado en la comuna de Til-til, Región Metropolitana. En un suelo delgado con textura superficial franco arenosa y presencia de sustrato aluvial con matriz arenosa en profundidad.

Estudos relataram a atividade antimicrobina de Equisetum giganteum e Punica granatum, reduzindo o risco do desenvolvimento da estomatite protética, doença relacionada principalmente à colonização das próteses pelo fungo Candida albicans. Diante disso, é necessário que essas plantas sejam bem toleradas pelos tecidos bucais e, se possível, auxiliem no processo de reparo. Objetivo: Realizar um estudo fitoquímico de substâncias potencialmente ativas de Equisetum giganteum e Punica granatum e avaliar a viabilidade de fibroblastos gengivais humanos frente a diferentes concentrações destas plantas, após diferentes períodos in vitro. Material e Métodos: Após a identificação dos compostos das plantas por HPLC-PAD, foram selecionadas as frações com melhor atividade antifúngica perante Candida albicans. Os extratos brutos e frações selecionadas de P. granatum e E. giganteum foram testados em diferentes concentrações (0.23, 0.47, 0.94, 1.88, 3.75, 7.5, 15 e 30 mg) e períodos (12, 24, 36 e 60 h) sobre fibroblastos gengivais humanos (FGH), para a avaliação de suas citotoxicidades através da análise da viabilidade dos FGH, pelo ensaio LIVE/DEAD. Por meio desse ensaio, analisamos quantitativamente a viabilidade celular através das leituras das unidades relativas de fluorescência (URF) e qualitativamente por meio da analise da morfologia e da densidade observadas pela microscopia invertida de fluorescência, após três experimentos independentes para cada período de avaliação. Os resultados quantitativos paramétricos foram representados como média e desvio padrão (SD) com análise de Variância ANOVAOne- Way, seguida da análise comparativa pelo teste de Tukey HSD. Os resultados quantitativos não paramétricos foram apresentados como mediana e quartis e foram submetidos ao teste de Kruskal-Wallis, seguido de análise comparativa pelo teste de Dunn, de acordo com as análises de normalidade (Teste de Kolmogorov-Smirnov). Valores de p< 0.05 foram indicativos de significância estatística. Conclusões: Foram identificados compostos fenólicos, como flavonoides e taninos em E.giganteum e P. granantum, respectivamente. Identificamos uma citotoxicidade concentração-dependente para as duas plantas testadas, independente do tempo. Sob as menores concentrações das plantas a viabilidade e a morfologia dos FGH foram preservadas. Dentro deste contexto, acreditamos na possibilidade de se utilizar essas plantas como terapia alternativa em diversas áreas de saúde, porém em concentrações biocompatíveis. / Studies have reported the antimicrobial activity of Equisetum giganteum and Punica granatum, reducing the risk of development of denture stomatitis, a disease mainly related to the colonization of prostheses by the fungus Candida albicans. In view of this, it is necessary that these plants be well tolerated by the oral tissues and, if possible, assist in the repair process. Objective: To carry out a phytochemical study of potentially active substances of E. giganteum and P. granatum and to evaluate the viability of human gingival fibroblasts against different concentrations of these two plants, after different periods in vitro. Material and Methods: After the identification of the plant compounds by HPLC-PAD, the fractions with the best antifungal activity against Candida albicans were selected. The crude extracts and selected fractions of P. granatum and E. giganteum were tested at different concentrations (0.23, 0.47, 0.94, 1.88, 3.75, 7.5, 15 and 30 mg) and periods (12, 24, 36 and 60 h) against human gingival fibroblasts (FGH), for the evaluation of their possible cytotoxicity through the analysis of the viability of FGH by the LIVE/DEAD assay. By means of this assay we quantitatively analyzed the cell viability through the readings of the relative units of fluorescence (URF) and qualitatively by analysis of the cellular morphology and density using inverted fluorescence microscopy, after three independent experiments. The parametric quantitative results were represented as mean and standard deviation (SD) with ANOVA-One-Way Variance analysis, followed by comparative analysis by the Tukey HSD test. The non-parametric quantitative results were presented as medians and quartiles and were submitted to the Kruskal-Wallis test, followed by comparative analysis by the Dunn test, according to normality tests (Kolmogorov-Smirnov test). Values of p <0.05 were indicative of statistical significance. Conclusions: Phenolic compounds, such as flavonoids and tannins, were identified in E. giganteum and P. granantum, respectively. We identified a concentration-dependent cytotoxicity for the two plants tested, regardless of the time. Under the lowest concentrations of plants the viability and morphology of FGH were preserved when compared to the control. Within this context, we believe in the possibility to use these plants as alternative therapy in several health areas, but in biocompatible concentrations.

Made available in DSpace on 2017-07-24T19:22:03Z (GMT). No. of bitstreams: 1 FABIANE SCHREINER.pdf: 1371612 bytes, checksum: 4c92bfc09a7b2ad93f1107f9b7633f68 (MD5) Previous issue date: 2016-08-12 / Due to endodontic treatment failure, virulence and increase of antibiotic resistant microorganisms bacterial infections had a greater incidence. Plant extracts are promising resources in the control of biofilms and can be considered a new way to prevent and / or treat infections. Among these natural products, there is the Punica granatum L. (pomegranate),which features bioactive compounds such as flavonoids and phenols. Objective: To analyze, in vitro and ex vivo, the antibacterial effect of auxiliary substances based on Punica granatum L. (pomegranate) against Enterococcus faecalis. Method: After developed the hydroglycolic peel extract of Punica granatum L. , agar diffusion tests, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) and analysis of toxicity (lethality in microcrustacean - Artemia salina were performed. With this data gel with hydroglicolic extract of P. granatum pericarp (0.5 g/mL) were developed and tested in agar diffusion, as auxiliary substance of endodontic instrumentation and as intracanal medication in infected dentin with E. faecalis by counting colony forming units per milliliter (cfu/mL) in three experimental times. Results: The MIC was 0.5 g/ml and P. granatum gel at this concentration showed 8 mm inhibition zones against E. faecalis. The P. granatum gel reduced 96% in cfu/mL after instrumentation (p = 0.005), with evidence of effectiveness for use as an intracanal medication (p = 0.047), reducing contamination by 65%. The brine shrimp mortality rate was 42%, with an LC50 of 62.7 mg/ml, being a sligthy toxic extract. Conclusion: The pomegranate gel 0.5 g/ml was effective against E. faecalis and represents promising proposal in Endodontics. / Devido ao insucesso do tratamento endodôntico, da virulência e do aumento dos microorganismos resistentes a antibióticos houve uma maior incidência de infecções bacterianas. Os extratos de plantas são recursos promissores no controle de biofilmes, podendo ser considerados uma nova forma de prevenir e/ou tratar infecções. Dentre esses produtos de origem natural, destaca-se a Punica granatum L. (romã), que apresenta compostos fitoativos como os flavonóides e os fenóis. Objetivo: Analisar, in vitro e ex vivo, a ação antibacteriana de substância auxiliar e medicação intracanal à base do extrato hidroglicólico do pericarpo de Punica granatum L. (romã) veiculado em gel, contra Enterococcus faecalis. Método: Após desenvolvido o extrato hidroglicólico do pericarpo de P. granatum, foram realizados testes de difusão em ágar, determinação de concentração inibitória mínima (CIM) e concentração bactericida mínima (CBM) e análise da toxicidade (letalidade em microcrustáceo - Artemia salina). Com estes dados foi desenvolvido gel com extrato hidroglicólico do pericarpo de P. granatum (0,5 g/mL), que foi testado por difusão em ágar e como substância auxiliar da instrumentação endodôntica e medicação intracanal, para a verificação da ação antimicrobiana em dentina infectada com E. faecalis, pela contagem de unidades formadoras de colônia por mililitro (ufc/mL) em três tempos experimentais. Resultados: A CIM foi de 0,5 g/mL e o gel de P. granatum nesta concentração apresentou halos de inibição de 8 mm, sobre E. faecalis.O gel de P. granatum reduziu em 96% a contagem de ufc/mL após instrumentação (p=0,005),com indício de efetividade para uso como medicação intracanal (p=0,047), reduzindo a contaminação em 65%. A taxa de mortalidade em microcrustáceos foi de 42%, com LC50 de 62,7 mg/mL, sendo o extrato levemente tóxico. Conclusão: O gel de P. granatum a 0,5 g/mL foi efetivo sobre E. faecalis, representando proposta promissora em Endodontia.

Estudos relataram a atividade antimicrobina de Equisetum giganteum e Punica granatum, reduzindo o risco do desenvolvimento da estomatite protética, doença relacionada principalmente à colonização das próteses pelo fungo Candida albicans. Diante disso, é necessário que essas plantas sejam bem toleradas pelos tecidos bucais e, se possível, auxiliem no processo de reparo. Objetivo: Realizar um estudo fitoquímico de substâncias potencialmente ativas de Equisetum giganteum e Punica granatum e avaliar a viabilidade de fibroblastos gengivais humanos frente a diferentes concentrações destas plantas, após diferentes períodos in vitro. Material e Métodos: Após a identificação dos compostos das plantas por HPLC-PAD, foram selecionadas as frações com melhor atividade antifúngica perante Candida albicans. Os extratos brutos e frações selecionadas de P. granatum e E. giganteum foram testados em diferentes concentrações (0.23, 0.47, 0.94, 1.88, 3.75, 7.5, 15 e 30 mg) e períodos (12, 24, 36 e 60 h) sobre fibroblastos gengivais humanos (FGH), para a avaliação de suas citotoxicidades através da análise da viabilidade dos FGH, pelo ensaio LIVE/DEAD. Por meio desse ensaio, analisamos quantitativamente a viabilidade celular através das leituras das unidades relativas de fluorescência (URF) e qualitativamente por meio da analise da morfologia e da densidade observadas pela microscopia invertida de fluorescência, após três experimentos independentes para cada período de avaliação. Os resultados quantitativos paramétricos foram representados como média e desvio padrão (SD) com análise de Variância ANOVAOne- Way, seguida da análise comparativa pelo teste de Tukey HSD. Os resultados quantitativos não paramétricos foram apresentados como mediana e quartis e foram submetidos ao teste de Kruskal-Wallis, seguido de análise comparativa pelo teste de Dunn, de acordo com as análises de normalidade (Teste de Kolmogorov-Smirnov). Valores de p< 0.05 foram indicativos de significância estatística. Conclusões: Foram identificados compostos fenólicos, como flavonoides e taninos em E.giganteum e P. granantum, respectivamente. Identificamos uma citotoxicidade concentração-dependente para as duas plantas testadas, independente do tempo. Sob as menores concentrações das plantas a viabilidade e a morfologia dos FGH foram preservadas. Dentro deste contexto, acreditamos na possibilidade de se utilizar essas plantas como terapia alternativa em diversas áreas de saúde, porém em concentrações biocompatíveis. / Studies have reported the antimicrobial activity of Equisetum giganteum and Punica granatum, reducing the risk of development of denture stomatitis, a disease mainly related to the colonization of prostheses by the fungus Candida albicans. In view of this, it is necessary that these plants be well tolerated by the oral tissues and, if possible, assist in the repair process. Objective: To carry out a phytochemical study of potentially active substances of E. giganteum and P. granatum and to evaluate the viability of human gingival fibroblasts against different concentrations of these two plants, after different periods in vitro. Material and Methods: After the identification of the plant compounds by HPLC-PAD, the fractions with the best antifungal activity against Candida albicans were selected. The crude extracts and selected fractions of P. granatum and E. giganteum were tested at different concentrations (0.23, 0.47, 0.94, 1.88, 3.75, 7.5, 15 and 30 mg) and periods (12, 24, 36 and 60 h) against human gingival fibroblasts (FGH), for the evaluation of their possible cytotoxicity through the analysis of the viability of FGH by the LIVE/DEAD assay. By means of this assay we quantitatively analyzed the cell viability through the readings of the relative units of fluorescence (URF) and qualitatively by analysis of the cellular morphology and density using inverted fluorescence microscopy, after three independent experiments. The parametric quantitative results were represented as mean and standard deviation (SD) with ANOVA-One-Way Variance analysis, followed by comparative analysis by the Tukey HSD test. The non-parametric quantitative results were presented as medians and quartiles and were submitted to the Kruskal-Wallis test, followed by comparative analysis by the Dunn test, according to normality tests (Kolmogorov-Smirnov test). Values of p <0.05 were indicative of statistical significance. Conclusions: Phenolic compounds, such as flavonoids and tannins, were identified in E. giganteum and P. granantum, respectively. We identified a concentration-dependent cytotoxicity for the two plants tested, regardless of the time. Under the lowest concentrations of plants the viability and morphology of FGH were preserved when compared to the control. Within this context, we believe in the possibility to use these plants as alternative therapy in several health areas, but in biocompatible concentrations.

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