Investigou-se a influência de íons metálicos Al(III), Ca(II), Fe(III), Pb(II), Cu(II) e Cr(III) (10-6 a 10 -3 mol L-1 na intensidade relativa de fluorescência (IRF) do ácido húmico (AH, 10 mg L-1). os íons Cu(ii), Pb(II) e Cr (III) produziram um efeito de supressão, enquanto os íons Fe(III), Al(III) e Ca(II) não influenciaram significativamente nos valores de IRF. A partir dos valores de IRF foram calculadas a constante de estabilidade (K) dos complexos e a capacidade complexante do AH com Cu(II), Pb(II) e Cr(III). Os valores de K seguiram a ordem: Cr(III)> Pb(II) > Cu(II) e a capacidade complexante: Cr(III) ~ Pb(II)> Cu(II). Desenvolveu-se um novo método analítico baseado na reação quimiluminescente da oxidação do AH com peroxomonosulfato de potássio (PMS; 0,6 mol L-1</SUP) ) em meio básico (NaOH 1,0 mol L-1 ). A intensidade máxima de radiação emitida (Imáx ) e a área em função do tempo foram lineares com a quantidade de AH em solução na faixa de 0,5 a 20 mg L-1, com o limite de detecção de 0,24 mg L-1 , com limite de detecção de 0,24 mg L-1 . Um estudo comparativo foi realizado utilizando-se H2O2 (0,58 mol L-1 na presença de CH2O (0,44 mol L-1) em meio básico (NaOH 0,16 mol L-1) com limite de detecção de 0,4 mg L-1. As adições dos íons Cl-, NO3- , PO43- , CO32- , Fe(III), Cu(II), Cr(III) e Ca(II) na reação quimiluminescente de AH com PMS, não interferiram significativamente no sinal. Adições de 50 mg L-1 de Co(II) ou Mn(II) à solução de AH Aldrich, na forma de complexos de EDTA ou cloretos, aumentaram o valor Imáx devido à formação de espécies fortemente oxidantes como SO5°-, SO4°- HO° durante a reação de decomposição do PMS. O método foi aplicado para determinar a concentração de AH em amostra de água do Rio Miranda - MS, obtendo-se valores entre 2,9 a 12,3 mg L-1 . As reações de oxidação, com emissão de radiação, de alguns compostos orgânicos com PMS em meio básico foram estudadas. Com os polifenóis obtiveram- se intensidades de radiação mais significativas e os valores de área seguiram a seguinte ordem: floroglucinol >> ácido fúlvico > ácido húmico > resorcinol > ácido pirogálico > catecol > hidroquinona. Desta forma, como polifenóis existem na estrutura de substâncias húmicas, acredita-se que essas porções da molécula devem ser as responsáveis pela produção de quimiluminescência. / The influence of Al(III), Ca(II), Fe(III), Pb(II), Cu(II), and Cr(III) metal ions (10-6 to 10-3 mol L-1) on the relative fluorescence intensity (RFI) of humic acid (HA, 10 mg L-1) was investigated. Cu(II), Pb(II), and Cr(III) ions produced a quenching effect, while Fe(III), Al(III), and Ca(II) ions did not significantly interfere with RFI values. Stability constants (K) and complexing capacities of HA with Cu(II), Pb(II), and Cr(III) were obtained using RFI values. Stability values of complexes followed the order: Cr(III)> Pb(II) > Cu(II) and the complexing capacity values: Cr(III) ~ Pb(II) > Cu(II). A new analytical method was developed based on the chemiluminescent oxidation of HA by peroxymonosulfate (PMS; 0.6 mol L-1) in basic medium (NaOH 1.0 mol L-1). The intensity of radiation emission (Imax) and area vs. time were linear functions of HA concentration range of 0.5-20 mg L-1. The detection limit was 0.24 mg L-1. A comparative study was conducted using H2O2 (0.58 mol L-1) in the presence of CH2O (0.44 mol L-1) in basic medium (NaOH 0.16 mol L-1). The detection limit was 0.4 mg L-1 of HA. The addition of Cl-, NO3-, PO43-, CO32-, Fe(III), Cu(II), Cr(III), and Ca(II) to the chemiluminescent reaction of HA with PMS did not interfere with the signal. The addition of 50 mg L-1 Co(II) or Mn(II) (as EDTA complexes or chloride salts) to the HA sample enhanced radiation emission, due to the formation of strong oxidant species such as SO5o-, SO4o-, and HOo during PMS decomposition reaction. This method was applied to determine HA concentration in a sample of river water (Miranda River - MS), obtaining values between 2.9 and 12.3 mg L-1. Oxidation reactions of some organic compounds by PMS in basic medium were studied, which produced radiation emission. Polyphenols produced the highest emissions and the area values followed the order: phloroglucinol> fulvic acid> humic acid> resorcinol> pyrogalic acid> cathecol> hydroquinone. Since polyphenolic groups are supposed to exist within humic acid structure, those portions of the molecule are most likely to be responsible for that chemiluminescence.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-09-09T17:33:04Z No. of bitstreams: 1 lianamoraisviannateutschbein.pdf: 2292842 bytes, checksum: e71aad5773fd914f4bbeb7bc101caa7e (MD5) / Approved for entry into archive by Diamantino Mayra (mayra.diamantino@ufjf.edu.br) on 2016-09-13T13:50:13Z (GMT) No. of bitstreams: 1 lianamoraisviannateutschbein.pdf: 2292842 bytes, checksum: e71aad5773fd914f4bbeb7bc101caa7e (MD5) / Made available in DSpace on 2016-09-13T13:50:13Z (GMT). No. of bitstreams: 1 lianamoraisviannateutschbein.pdf: 2292842 bytes, checksum: e71aad5773fd914f4bbeb7bc101caa7e (MD5) Previous issue date: 2011-04-06 / O cerrado brasileiro é um dos maiores biomas de diversidade biológica do planeta, abrigando uma ampla variedade de espécies medicinais utilizadas como parte da tradição e costume das comunidades locais. Os objetivos deste estudo foram determinar os teores de polifenóis e flavonóides; avaliar a atividade antioxidante; correlacionar os teores de polifenóis e flavonóides entre si e com a atividade antioxidante e avaliar a atividade cicatrizantes de três espécies deste bioma: Lippia sidoides; Synadenium grantii e Stachytarpheta gesnerioides. Através dos extratos etanólicos (EE), obtidos das folhas, foram quantificados espectrofotometricamente os teores de polifenóis e de flavonóides e, ainda, avaliada a atividade antioxidante (DPPH) e as correlações. Os EE foram incorporados ao creme base (10% de L. sidoides; 10% de Synadenium grantii e 10% de Stachytarpheta gesnerioides) e avaliados quanto à atividade cicatrizante, empregando o modelo de indução de úlceras dérmicas em coelhos albinos da Nova Zelândia (machos 1.5-2.0 kg; n=6). As úlceras foram analisadas macroscopicamente quanto ao aspecto da lesão e a contração de sua área durante 10 dias. Após este período, foi feito estudo histológico em lâminas coradas com hematoxilina-eosina para comparação do número de células inflamatórias; fibroblastos; vasos sanguíneos; área de colágeno e área de matriz extracelular entre os grupos e imunoistoquímica com a contagem dos miofibroblastos. Foram realizadas análises estatísticas descritivas para avaliar a significância das diferenças entre as médias dos resultados dos grupos, foi aplicada a análise de variância (ANOVA) e teste post hoc de Tukey, utilizando o SPSS versão 14.0 e p < 0,05. Foi avaliada a correlação de Pearson entre os teores de polifenóis e flavonóides e entre esses teores e a atividade antioxidante. Todos os EE das espécies utilizadas apresentaram presença de polifenóis, flavonóides e atividade antioxidante. Foi demonstrada uma correlação positiva entre os teores de polifenóis e flavonóides e atividade antioxidante. Não foi observada diferença na contração da lesão quando comparados os grupos controle e tratados e em relação ao número de miofibroblastos os grupos testados foram estatisticamente similares ao controle positivo. O tratamento com L. sidoides apresentou número de vasos sanguíneos, área de colágeno e área de matriz extracelular similar ao controle positivo; o número de fibroblastos e células inflamatórias foi 24% (p=0,002) e 54% (p < 0,001) menor, respectivamente, em relação ao controle positivo. O tratamento com Synadenium grantii apresentou número de fibroblastos, área de colágeno e área de matriz extracelular semelhante ao controle positivo e reduziu o número de células inflamatórias (61%) e vasos sanguíneos (54%) (p < 0,001 para ambos), em relação ao controle positivo. O tratamento com Stachytarpheta gesnerioides apresentou número de fibroblastos; área de colágeno e área de matriz extracelular semelhante ao controle positivo e reduziu o número de células inflamatórias (65%, p < 0,001) e vasos sanguíneos (42%, p=0,005) em relação ao controle positivo. Em relação ao número de miofibroblastos, todos os tratamentos foram estatisticamente similares ao controle positivo. A análise conjunta dos resultados inéditos sugere a realização de novos estudos científicos relacionados à atividade cicatrizante de L. sidoides. / The Brazilian cerrado is one of the planet’s major biological diversity biomes, hosting a wide variety of medicinal plants used as part of the tradition and customs of local communities. The objectives of this study were to determine the levels of polyphenols and flavonoids; evaluate the antioxidant activity; correlate the levels of polyphenols and flavonoids among themselves and with the antioxidant activity and to evaluate the healing activity of the ethanol extracts (EE) of three species of the cerrado: Lippia sidoides, Synadenium grantii and Stachytarpheta gesnerioides. With the ethanol extracts (EE), obtained from leaves, it were spectrophotometrically quantified the concentrations of polyphenols and flavonoids, and it was also evaluated the antioxidant activity (DPPH) and their correlations. The EE obtained from the leaves were incorporated into the cream base (10% of L. sidoides, 10% of Synadenium grantii and 10% of Stachytarpheta gesnerioides) and their healing activity was evalluated. It was used as experimental model the induction of dermal ulcers in New Zealand albino rabbits (males 1.5-2.0 kg; n = 6). The ulcers were analyzed for the appearance of the lesion and the contraction of the wound area during 10 days. After this period, the histology was conducted using histological sections stained with hematoxylin-eosin to compare the number of inflammatory cells, fibroblasts and blood vessels and immunohistochemistry to compare the number of myofibroblasts among the experimental groups. Descriptive statistics analysis were conducted and, to assess the significance of differences between the mean results of groups, It was also applied the analysis of variance (ANOVA) and Tukey's post hoc test, using SPSS version 14.0 and p <0.05. It was evaluated the Pearson correlation between the levels of polyphenols and flavonoids and between these levels and the antioxidant activity. All the EE of the used species showed the presence of polyphenols, flavonoids and antioxidant activity. It was demonstrated a positive correlation among these constituents and antioxidant activity. No difference was observed in the contraction of the lesion when compared to the control and treated groups. At microscopic analysis, treatment with L. sidoides showed the number of blood vessels, collagen area and area of extracellular matrix similar to the positive control; the number of fibroblasts and inflammatory cells was 24% (p = 0.002) and 54% (p <0.001) lower, respectively, compared to positive control. Treatment with Synadenium grantii presented number of fibroblasts, collagen area and area of extracellular matrix similar to the positive control and reduced the number of inflammatory cells (61%) and blood vessels (54%) (p <0.001 for both) compared the positive control. Treatment with Stachytarpheta gesnerioides presented number of fibroblasts, collagen area and area of extracellular matrix similar to the positive control and reduced the number of inflammatory cells (65%, p <0.001) and blood vessels (42%, p = 0.005) compared the positive control. In relation to the myofibroblasts, all treatments presented number of myofibroblasts similar to the positive control A combined analysis of these results suggest to perform further scientific studies related to healing activity of L. sidoides.

Orientador: Maria Angela de Almeida Meireles / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-21T20:58:56Z (GMT). No. of bitstreams: 1 Veggi_PriscillaCarvalho_D.pdf: 5176961 bytes, checksum: 389c029fe6c05f6c9fcc19436840506e (MD5) Previous issue date: 2013 / Resumo: Este estudo teve como finalidade utilizar a extração supercrítica (SFE = supercritical fluid extraction) para obtenção de compostos fenólicos presentes em plantas nativas brasileiras de elevado potencial econômico. As plantas estudadas foram: Cipó-de-são-joão (Pyrostegia venusta), Nó-de-cachorro (Heteropteris aphrodisiaca O. Mach), Ingá Cipó (Inga edulis), Jatobá (Hymenaea courbaril var. stilbocarpa) e Feijão (Phaseolus vulgaris L.). Experimentos de rendimento global (X0,S/F=50) utilizando CO2 puro, CO2 + etanol (10 %, massa) e CO2 + água (10 %, massa) como solventes foram realizados a 35 MPa e 323 K; a razão da massa de CO2 para a massa de alimentação foi mantida constante (50 kg/kg). Posteriormente os extratos foram analisados por cromatografia em camada delgada (CCD); foram determinadas também a atividade antioxidante, o teor de polifenóis e o de flavonoides totais. Após a avaliação do comportamento dos extratos quanto ao tipo de solvente utilizado e análises realizadas, observou-se que o solvente CO2 + H2O resultou em maior rendimento em extrato, especialmente para os de jatobá; adicionalmente, o extrato desta planta teve excelentes características antioxidantes. Desta forma, os estudos posteriores foram realizados com o jatobá. Para selecionar a temperatura e a pressão ótimas de operação, isotermas de rendimento global (IRG's) foram determinadas nas temperaturas de 323 e 333 K e pressões de 15, 25 e 35 MPa. Em função do maior teor de polifenóis e taninos totais obtidos, foi selecionada a pressão de 15 MPa e temperatura de 323 K para estudos da cinética de extração. Curvas cinéticas (OEC = overall extraction curve) foram construídas visando à determinação dos parâmetros de processo através do ajuste das OECs a um spline linear com 3 retas. Em seguida, realizou-se um estudo do aumento de escala, mantendo constantes a razão entre massa de solvente e massa de matéria-prima (S/F) e tempo de processo. As análises químicas dos extratos obtidos por SFE foram realizadas por Cromatografia Líquida de Ultra Eficiência acoplada ao Espectrômetro de Massas em série (UPLC-MS/MS) e Espectrometria de Massas com Ionização por Eletrospray (IES-MS/MS). De acordo com os resultados, os extratos apresentaram perfil químico de procianidinas. Por último, foi determinado o custo de manufatura (COM) dos extratos das cinco plantas obtidos nos ensaios de rendimento global (X0, S/F=50) para os três solventes: CO2 puro, CO2 + etanol (10 %, massa) e CO2 + água (10 %, massa), e do extrato de jatobá no processo otimizado (OEC's) para o solvente CO2 + H2O. O COM estimado demonstrou a viabilidade do processo SFE para unidades industriais com extratores de 50 e 500 L, dependendo da matéria-prima a ser processada. Segundo o COM estimado, a obtenção de extratos ricos em taninos das cascas de jatobá foi economicamente viável para o solvente CO2 + H2O. Adicionalmente, extratos da casca de jatobá foram obtidos utilizando a extração assistida por ultrassom (UAE). As extrações foram realizadas com potência de ultrassom de 20 e 60 W durante 40 min, utilizando a água em uma razão S/F igual a 20 (otimizada em um ensaio de extração por maceração). Maior quantidade de polifenóis foi obtida com a potência de 60 W. A eficiência da UAE na extração de polifenóis foi avaliada através de extrações em corrente cruzada em três estágios; foi estudado o aumento de escala em extrator de 3 litros. Os resultados foram comparados com os obtidos com extração por agitação (AGE) e sonda de ultrassom (UP) utilizando as mesmas condições da UAE. Foi realizado ainda um estudo cinético avaliando a obtenção de polifenóis da casca de jatobá para os processos UAE e AGE. Os extratos foram analisados por CCD; a atividade antioxidante e teor de polifenóis dos extratos foram determinados. Os resultados provaram que a UAE é um método eficiente para obter extratos ricos em compostos fenólicos e com elevada atividade antioxidante. Através da análise química com Cromatografia de Alta Eficiência com detecção por arranjo de diodos (CLAE/DAD), o perfil de compostos fenólicos da casca de jatobá obtidos por UAE constituiu de procianidinas, a (+)-catequina e a (-)-epicatequina / Abstract: The objective of this study was to use supercritical fluid extraction (SFE) to extract phenolic compounds from indigenous Brazilian plants with high economic potential. The plants studied were: Flame wine (Pyrostegia venusta), Dog's knot (Heteropteris aphrodisiaca O. Mach), Ice-cream bean tree (Inga edulis), Jatoba (Hymenaea courbaril var. stilbocarpa) and beans (Phaseolus vulgaris L.). Global yields experiments (X0,S/F=50) using pure CO2, CO2 + ethanol (10 %, mass) and CO2 + water (10 %, mass) as solvents were performed 35 MPa and 323 K; the ratio of the CO2 mass to feed mass was maintained constant (50 kg/kg). The extracts were analyzed by thin layer chromatography (TLC). Their antioxidant activity, total polyphenolic and flavonoids content were also determined. After evaluating the behavior of the yield and chemical composition of the extracts, it was observed that the solvent mixture CO2 + H2O provided higher yield, especially for jatobá. Additionally, this extract presented high antioxidant activity. Based on these results, global yields isotherms (GYI) of jatoba were determined at temperature of 323 and 333 K and pressures of 15, 25 and 35 MPa in order to select the optimum operational conditions. Due to the higher polyphenols and tannins content, the optimum conditions selected were 15 MPa and 323 K. Overall extraction curves (OEC¿s) were obtained under these conditions in order to determine the kinetic parameters by fitting the OECs to a linear spline with three straight lines. For the scale-up study, the criterion adopted was keeping constant the solvent to feed ratio (S/F) and process time. The chemical analyses of SFE extracts were performed by Ultra performance liquid chromatography - diode array detection (UPLC-MS/MS) and electrospray ionization mass spectrometry (ESI-MS/MS). According to the results, the extracts showed chemical profile of procyanidins. Finally, it was evaluated the cost of manufacturing (COM) of the extracts for all five plants obtained in the GYI experiment (X0, S/F=50) for the three solvent: CO2 pure, CO2 + ethanol (10 %, mass) e CO2 + water (10 %, mass); and for the jatobá extract obtained at optimized process conditions using CO2 + water (10 %, mass). The estimated COM demonstrated the feasibility of the SFE process for industrial scale extractors of 50 and 500 L, depending on the raw material to be processed. Moreover, according to the estimated COM, obtaining jatobá bark extracts rich in tannins was economically feasible with CO2 + H2O. Jatoba extracts were also obtained using ultrasound assisted extraction (UAE). The extractions were performed at the potency of 20 and 60 W during 40 min, using water at S/F ratio of 20 (optimized in a maceration extraction assay). High content of polyphenols were obtained using the potency of 60 W. The efficiency of UAE in extracting polyphenols was evaluated using crosscurrent extraction in three stages. The scale-up of the UAE process was studied in an extractor of three liters. It was also performed a kinetic study to evaluate the extraction of polyphenols from jatobá via UAE and AGE. The results were compared to those obtained by extraction under agitation (AGE) and ultrasound probe (UP), in the same conditions used for the UAE experiment. The extracts were analyzed by TLC. The antioxidant activity and polyphenols content of the extracts were also determined. The results proved that UAE is an efficient method to obtain extracts rich in phenolic compounds and with high antioxidant activity. The chemical analysis of the extracts by high performance liquid chromatography - diode array detection (HPLC-DAD), showed that the phytochemical profile of jatobá bark extracts presented procyanidins, (+)-catechin and (-)-epicatechin / Doutorado / Engenharia de Alimentos / Doutora em Engenharia de Alimentos

Submitted by Biblioteca da Faculdade de Farmácia (bff@ndc.uff.br) on 2017-03-28T17:01:20Z No. of bitstreams: 1 Bedê, Teresa Palmisciano [Dissertação, 2015].pdf: 1365884 bytes, checksum: 2fb9e05630c2ebc5ce56e4099bbd7366 (MD5) / Made available in DSpace on 2017-03-28T17:01:20Z (GMT). No. of bitstreams: 1 Bedê, Teresa Palmisciano [Dissertação, 2015].pdf: 1365884 bytes, checksum: 2fb9e05630c2ebc5ce56e4099bbd7366 (MD5) / Introdução: A busca da população por melhor qualidade de vida contribui para os estudos acerca de alimentos com propriedades funcionais e de prevenção de doenças. Destacam-se o suco de uva tinto integral (SUTI) e o vinho tinto (VT) que, ricos em polifenóis, vem demonstrando potente ação antioxidante e antinflamatória. Objetivo: Avaliar os efeitos do consumo de SUTI, de VT e de solução de resveratrol (SR) sobre a função e integridade hepática de ratos alimentados com dieta hiperlipídica. Materiais e Métodos: Foi realizado ensaio biológico com 50 Rattus Novergicus Wistar Albino, fêmeas adultas, divididas em 5 grupos: controle (GC)- ração caseína + água; hiperlipídico (GH)- ração hiperlipídica + água; vinho tinto (GV)- ração hiperlipídica + 10mL/dia de VT; suco de uva (GS)- ração hiperlipídica + 15mL/dia de SUTI e, resveratrol (GR)- ração hiperlipidica + 15mL/dia de SR. Durante 60 dias, ração e água foram ofertados em livre demanda e o peso corporal, o consumo alimentar e a pressão arterial foram verificados e registrados. Ao final do estudo, os animais foram sacrificados, amostras de sangue foram coletadas para análises bioquímicas e o fígado foi retirado para avaliações histológicas. Para comparação das médias entre grupos foi utilizado Anova one-way e Tukey como pós-teste, considerando um nível de significância de 5%. Resultados: A concentração de polifenóis totais (mg EAG -1) e de trans-resveratrol (mg/L) foi menor (p<0,05) no SUTI do que nas demais bebidas, mas a capacidade antioxidante não diferiu entre elas. Não houve diferença no peso corporal e no consumo alimentar dos grupos. A pressão arterial sistólica (mmHg) foi menor (p<0,05) no GS e GC. O GH apresentou maior (p<0,05) variação de glicemia (mg/dL) e o GR foi o único que apresentou diminuição de glicemia ao longo do estudo. A concentração (mg/dL) de colesterol total foi menor (p<0,05) no GS do que no GC, mas não houve diferença na concentração (mg/dL) de lipoproteína de alta densidade – HDL e, a concentração (mg/dL) de triglicerídeos foi maior (p<0,05) no GC. Não houve diferença na concentração (U/L) de aspartatotransaminase, mas o GS foi o único que apresentou-se dentro do intervalo de normalidade. Todos tiveram concentração (U/L) de alaninatransaminase acima do recomendado, mas o GS apresentou concentração numericamente menor e, a concentração de fosfatase alcalina (U/L) do GR foi menor (p<0,05) do que o GH e GV. A concentração de interleucina 6 (pg/mL) foi menor (p<0,05) no GS do que no GV. O peso do fígado (g/100gPC) foi semelhante entre os grupos, mas a concentração de gordura hepática (g%) foi menor (p<0,05) no GS e GC, e o teor de proteínas no fígado (g%) do GS foi menor (p<0,05) do que no GV e GR. A área dos núcleos dos hepatócitos (μm²) foi menor (p<0,05) no GC do que no GH, GV e GS, mas o IOD mostrou-se menor (p<0,05) no grupo GC, seguido pelo GS e maior no GH e GV. Conclusões: O consumo de SUTI parece ser capaz de minimizar os efeitos da dieta hiperlipídica sobre o tecido hepático. Mesmo diante do menor teor de polifenóis, o SUTI pode ter conferido proteção sobre certos parâmetros bioquímicos avaliados, assim como sobre o desenvolvimento de hipertensão arterial e da doença hepática gordurosa não alcoólica, o que não foi observado com o VT e a SR / Introduction: The search for people for better quality of life contributes to the studies of food with properties functional and disease prevention. Stand out from the full red grape juice (JG) and red wine (RW) that are rich in polyphenols, has demonstrated potent antioxidant action and antinflammatory. Objective: To evaluate the effects of consumption of JG, RW and resveratrol solution (SR) on liver function and integrity of mice fed high-fat diet. Materials and Methods: This was a biological assay with 50 Rattus Novergicus Wistar Albino adult females divided into 5 groups: control (GC) - casein diet + water; hyperlipidic (GH) - fat diet + water; red wine (GW) - fat diet + 10 mL of RW / day; juice (GJ) - fat diet + 15mL of JG / day and resveratrol (GR) - fat diet + 15 mL of SR / day. During 60 days, feed and water were offered on free demand and, body weight, food intake and blood pressure were checked and recorded. At the end of the study, the animals were sacrificed, blood samples were collected for biochemical analysis and the liver was removed for histological evaluation. To compare the means between groups we used one-way ANOVA and Tukey as post-test, considering a 5% significance level. Results: The concentration of total polyphenols (mg EAG -1) and trans-resveratrol (mg/ L) was lower (p<0.05) in JG than the other drinks, but the antioxidant capacity did not differ between them. There was no difference in body weight and food consumption groups. Systolic blood pressure (mmHg) was lower (p<0.05) in the GJ and GC. GH had higher (p<0.05) blood glucose change (mg/ dL), and GR is the only one that decreases in blood glucose throughout the study. Concentration (mg / dL) of total cholesterol was lower (p<0.05) in GJ than in the GC, but there was no difference in the concentration (mg/ dL) of high-density lipoprotein - HDL and the concentration (mg/ dL) triglyceride was higher (p<0.05) in GC. There was no difference in the concentration (U/L) aspartatotransaminase, but the GJ was the one who introduced himself within normal range. All had concentration (U/L) alaninatransaminase above recommended, but the GJ presented numerically lowest concentration, and the concentration of alkaline phosphatase (U/L) of GR was smaller (p<0.05) than GH and GW. The concentration of interleukin-6 (pg/mL) was lower (p<0.05) in GJ than in the GW. Liver weight (g /100g body weight) was similar between the groups, but the concentration of liver fat (g%) was lower (p<0.05) in the GJ and GC, and the protein content in the liver (g%) GJ was lower (p <0.05) than the GW and GR. The area of the nuclei of hepatocytes (μm²) was lower (p<0.05) in the GC than in the GH, GW and GJ, but the IOD was lower (p <0.05) in the GC, followed by GJ and higher in GH and GW. Conclusions: Consumption of JG seems to be able to minimize the effects of high fat diet on liver tissue. Even with the lower polyphenol content, the JG may have conferred protection on certain biochemical parameters, as well as on the development of hypertension and non-alcoholic fatty liver disease, which was not observed with RW and SR

Le syndrome métabolique (MS), une association des plus dangereux facteurs de risque pour les maladies cardiovasculaires et le diabète de type 2, est devenu l'un des principaux défis cliniques et de santé publique dans le monde. Un nombre croissant d’évidences s’est accumulé au cours de la dernière décennie, démontrant l'efficacité de certains composés bioactifs alimentaires pour le traitement et la prévention du MS. Néanmoins, la plupart des études d'intervention administre les composés bioactifs sous forme de composés purs, sans considérer que l'interaction entre les bioactifs ajoutés et l'ensemble de la matrice alimentaire peut impacter la bioaccessibilité, la biodisponibilité et, par conséquent, l’efficacité de ces molécules bioactives.L'objectif principal de cette thèse, intégrée dans le projet européen PATHWAY-27, était de formuler des aliments enrichis en composés bioactifs potentiellement efficaces contre le MS, et d'étudier l'effet de la matrice alimentaire sur la bioaccessibilité et la biodisponibilité de ces bioactifs. Cette étude a mis l'accent sur l'utilisation des anthocyanes, de l’acide docosahexaénoïque et, dans une moindre mesure, des bêta-glucanes comme ingrédients bioactifs pour enrichir des produits laitiers et des produits à base d'œuf. Une combinaison des modèles de digestion in vitro et in vivo (chez le porc) a été utilisée.La composition et la structure des matrices alimentaires ont impacté la libération et la solubilisation de substances bioactives au cours de la digestion (bioaccessibilité), tel que démontré in vitro et in vivo. La structure de / Metabolic Syndrome (MS), a constellation of the most dangerous cardiovascular disease and type 2 diabetes mellitus risk factors, has become one of the major clinical and public health challenges worldwide. During the last decade, many bioactives have been proposed as effective for the treatment and prevention of MS. However, most intervention studies administer bioactives as pure compounds, without consider that bioactive-food matrix interactions could deeply impact on the bioaccessibility, bioavailability and hence on the effectiveness of bioactives.The main objective of this thesis, included within the European project PATHWAY-27, was to formulate potential effective bioactive-enriched foods against MS and to investigate the effect of the food matrix on the bioaccessibility and bioavailability of dietary bioactives.In particular, this study focused in the use of dairy and egg-based products as matrices and in the addition of anthocyanins, docosahexaenoic acid and, to a lesser extent, beta glucan as bioactives. A combination of in vitro and in vivo (pig) digestion models was used.Both the structure and the composition of the food matrices impacted the release and the solubilisation of bioactives during digestion (bioaccessibility), as demonstrated in vitro and in vivo. In addition, the structure of the food matrix modulated the final amount of DHA into the systemic circulation of pigs (bioavailability). This study proves that understanding how dietary bioactives interact with the matrix in which they are included in is the basis for the production of effective bioact

Oenococcus oeni est la principale de bactérie lactique du vin. L’espèce comprend de nombreuses souches plus ou moins bien adaptées au vin et qui forment différents groupes génétiques. Récemment, deux groupes de souches ont été identifiés dans les vins rouges (VR) et vins blancs (VB) de Bourgogne. Il est probable que les bactéries se sont adaptées à l’un ou l’autre de ces vins en raison de leurs différences physico-chimiques et de composition. Néanmoins, d’autres souches peuvent s’y développer. Ce sont par exemple des levains commerciaux, qui ont une bonne tolérance aux stress du vin. L’objectif de cette étude était d’analyser l’adaptation des souches d’O. oeni au vin et en particulier aux polyphénols. Dans un premier chapitre, des souches des groupes VR et VB ont été étudiées. Leurs phénotypes ont été comparés dans des moûts et des vins à différents pH et en présence de polyphénols. Les résultats ont permis de mieux comprendre leur adaptation au vin blanc ou rouge. Dans les deuxième et troisième chapitres, la tolérance à l’acidité et aux polyphénols de souches commerciales a été étudiée. Leur résistance au vin et leurs capacités fermentaires ont été analysées, les cellules ont été observées en microscopie électronique et les mécanismes de résistance à l’acidité et aux polyphénols ont été recherché par protéomique quantitative. Un protocole de purification des protéines spécifique pour les essais en présence de polyphénols a été mis au point. Les résultats montrent des voies métaboliques et enzymes qui contribuent à l’adaptation des souches au vin. Ils permettent aussi l’exploration de nouvelles pistes pour la sélection des souches industrielles. / Oenococcus oeni is the main lactic acid bacteria species in wine. The species comprises many strains, which are more or less well adapted to wine and form different genetic groups. Recently, two groups of strains have been identified from Burgundy’s red (VR) and white (VB) wines. It is likely that the bacteria have adapted to one or the other of these wines because of their different physico-chemical properties and compositions. Nevertheless, other strains can also develop in these wines. They are for example commercial starters, which are well resistant to wine stressors. The objective of this study was to analyze the adaptation of O . oeni strains to wine and particularly to phenolic compounds. In the first chapter, strains of groups VR and VB were studied. Their phenotypes were compared in grape musts and in wines at different pHs and in the presence of phenolic compounds. The results provide insights on their adaptation to red or white wine. In the second and third chapters, the tolerance to acidity and phenolic compounds of commercial strains has been studied. Their survival in wine and their fermentative capacities were analyzed, the cells were observed by electron microscopy and the mechanisms of resistance to acidity and phenolic compounds were investigated by quantitative proteomics. A specific protein purification protocol has been developed for trials performed in the presence of polyphenols. The results show metabolic pathways and enzymes that contribute to the adaptation of strains to wine. They also open new tracks for the selection of industrial strains.

Dans un contexte économique tendu pour les chocolatiers « premium », la maîtrise de la qualité organoleptique de la matière première est un impératif et un avantage compétitif. La qualité aromatique d'un chocolat est entre autre liée aux traitements post-récolte et particulièrement à l'étape de fermentation. Cette étape est essentielle au développement des caractéristiques sensorielles. Or, les outils de contrôle qualité du cacao sont basés principalement sur l'évaluation sensorielle d'échantillons. Cette méthode est onéreuse et chronophage. Le développement d'outils rapides et robustes d'analyse de la qualité devient nécessaire. Cette étude a pour objectif de caractériser le déroulement de la fermentation afin de sélectionner des marqueurs spécifiques de la qualité du cacao. L'étude s'est intéressée aux marqueurs mesurables chez le planteur (à travers l'évolution de la température et du pH), et à ceux issus des transformations biochimiques des fèves (de type azoté et polyphénolique). Ces marqueurs ont ensuite été confrontés aux données traditionnelles de qualité (cut-test et dosage de l'azote ammoniacal), et aux données sensorielles, afin de sélectionner les plus pertinents pour le pilotage de fermentation. L'analyse globale de l'ensemble des résultats a permis de faire ressortir huit bio-marqueurs analytiques et trois bio-marqueurs sensoriels, représentatifs de la qualité du cacao et en lien avec le développement de la fermentation. Par ailleurs, des recommandations de méthodes de mesure de ces bio-marqueurs à mettre en place et permettant de contrôler la qualité ont été proposés. / In a difficult economic context for "premium" chocolate makers, the control of the organoleptic quality of the raw material is an imperative and a competitive advantage.The aromatic quality of a chocolate is highly related to post-harvest treatment and particularly to the fermentation. This step is essential to the development of sensory characteristics.However, cocoa quality control tools are mainly based on sensory evaluation of samples. This method is expensive and time consuming.Fast and robust tools for quality analysis are needed. This study aims at characterizing the course of the fermentation in order to select specific markers of the quality of cocoa. The study focused on markers which can be measured on plantation (through the evolution of temperature and pH), and those from biochemical transformations of the beans (nitrogenous and polyphenol type). These markers have been then compared to data from traditional quality methods (cut-test and ammonia nitrogen quantification) and to sensory data in order to select the most relevant ones for the control of fermentation. The overall analysis of the database enabled to identify eight analytical bio-markers and three sensory bio-markers, representative of the quality of cocoa and in connection with the development of fermentation. In addition, recommendations for methods of measuring these bio-markers to implement and to monitor the quality have been presented.

L'obésité se définit comme une accumulation anormale ou excessive de masse grasse corporelle pouvant nuire à la santé. En effet, surpoids et obésité concourent au développement de pathologies graves comme le diabète de type 2 et les maladies cardiovasculaires qui représentent un véritable problème de santé publique dans de nombreux pays et à La Réunion. Aussi, la recherche des mécanismes physiopathologiques associés à l'obésité constitue un enjeu scientifique et médical majeur. Dans l'organisme, c'est au niveau du tissu adipeux que le stockage de gras se localise, plus précisément dans les cellules adipeuses (adipocytes) provenant elles-mêmes de la maturation de cellules précurseurs appelées préadipocytes. Chez le sujet obèse, le tissu adipeux stockant un excès de gras, est le siège d'un stress oxydatif et d'une inflammation chronique qui concourent à la résistance à l'insuline, engendrant un état diabétique. Plusieurs données bibliographiques montrent les effets antioxydants et anti-inflammatoires des produits végétaux riches en polyphénols et soulignent leur intérêt dans le cadre de thérapies innovantes vis-à-vis de l'obésité. La Réunion dispose d'une grande variété de ressources végétales dont des plantes médicinales qui présenteraient des propriétés anti-inflammatoires, antidiabétiques et « anti-obésité ». Cependant, il manque des données scientifiques pour valider leur éventuel bénéfice santé. L'objectif du travail de thèse était d'évaluer l'impact des extraits riches en polyphénols antioxydants de plantes médicinales de La Réunion sur la réponse métabolique et inflammatoire des cellules adipeuses dans le contexte de la pathologie de l'obésité. Pour ce faire, trois plantes médicinales ont été sélectionnées pour leur utilisation en pharmacopée traditionnelle, à savoir Antirhea borbonica (Bois d’Osto), Doratoxylon apetalum (Bois de gaulette) et Gouania mauritiana (Liane Montbrun). Nos résultats ont mis en évidence les propriétés antioxydantes et anti-inflammatoires des extraits végétaux sur des préadipocytes 3T3-L1 soumis à des médiateurs majeurs liés à l'obésité tels que l'agent pro-oxydant H2O2, la cytokine pro-inflammatoire TNF-α et l'agent bactérien LPS. Par ailleurs, nos données ont montré la capacité des extraits végétaux à potentialiser l'accumulation de gras induite par l'insuline, et à réduire la sécrétion de molécules pro-inflammatoires des adipocytes 3T3-L1soumis à un stress oxydatif. Une étude menée sur des souris rendues obèses par un régime riche en graisses contenant ou non l'extrait de la plante D. apetalum a confirmé la capacité de cet extrait végétal à améliorer le stockage de gras, le statut antioxydant et le profil inflammatoire du tissu adipeux des animaux obèses. Plusieurs cibles moléculaires ont été identifiées dont les facteurs de transcription PPARγ et NF-κB, l'enzyme antioxydante SOD ainsi que les adipokines de type leptine, adiponectine, TNF-α, IL-6 et MCP-1. En conclusion, ce travail de thèse a permis d'évaluer, pour la première fois, l'impact d'extraits riches en polyphénols antioxydants de trois plantes médicinales locales sur la réponse métabolique et inflammatoire des cellules adipeuses, dans le contexte de l'obésité. L'utilisation des modèles cellulaires et animaux développés au cours de ce travail permettra de poursuivre l'exploration des voies moléculaires impliquées et d'identifier de possibles nouvelles cibles thérapeutiques. / During obesity, excess fat mass is accumulated in adipose tissue, more precisely in adipocytes, which the precursor cells are preadipocytes. Adipose tissue is the place of oxidative stress and chronic inflammation involved in insulin resistance and type 2 diabetes. Plants rich in polyphenol, known for their antioxidant and anti-inflammatory effects, are high of interest to fight against obesity. In La Réunion, medicinal plants are commonly used for these properties but few literature data exist. Our objective was to evaluate the impact of antioxidant polyphenol-rich extracts from medicinal plants of La Réunion, such as Antirhea borbonica (Bois d’Osto), Doratoxylon apetalum (Bois de gaulette), Gouania mauritiana (Liane Montbrun), on the metabolic and inflammatory response in adipose cells in the context of obesity. Our results have demonstrated antioxidant and anti-inflammatory properties of plant extracts on 3T3-L1 preadipocytes in presence of three mediators of inflammation. Plant extracts are able to potentiate fat accumulation induced by insulin reduce the secretion of pro-inflammatory molecules in 3T3-L1 adipocytes during oxidative stress. These results were confirmed by a study on animal model. The plant extract from D. apetalum improved storage fat, antioxidant status and inflammatory profile of adipose tissue of obese mice. This work lead to evaluated the impact of antioxidant polyphenol-rich extract from three medicinal plants on metabolic and inflammatory response in adipose cells in the context of obesity.

Perturbations to extravillous trophoblast (EVT) cell migration and invasion are associated with the development of placenta-mediated diseases. Dietary polyphenols have been shown to influence cell migration and invasion in models of tumorigenesis and non-cancerous, healthy cells; however, never shown in EVT cells. We hypothesize that polyphenols present in V. angustifolium leaves will promote trophoblast migration and invasion through ERK and AKT activation. Using the HTR-8/SVneo cell line as a model for EVT cells, the leaf extract increased trophoblast migration and invasion, in an ERK- and AKT-independent manner, and had no effect on cell proliferation or viability. One major polyphenol of the leaf extract was identified and may be an active compound. We have demonstrated for the first time that V. angustifolium leaf extract increases EVT migration and invasion in vitro, thus further investigations examining potential therapeutic applications of this extract in the context of placenta-mediated diseases are warranted.

Les maladies du bois de la vigne, par exemple le syndrome de l’esca et l’eutypiose, sont dues à des champignons qui se développent dans le xylème et dégradent les tissus ligneux. Elles sont en recrudescence depuis une vingtaine d’année dans tous les vignobles du monde. A ce jour, aucun traitement curatif n’est disponible alors que ces maladies engendrent généralement la mort de la plante. L’étude des relations hôte-parasite et notamment celle des réponses de défense de plante, est nécessaire pour la compréhension et le développement de nouvelles stratégies de lutte. Parmi les molécules de défense de la vigne, les stilbènes sont connus pour leur forte activité antifongique.Notre travail, basé sur des approches biochimiques et moléculaires apporte de nouvelles données sur le rôle potentiel des stilbènes en tant que molécules de défense contre les maladies du bois. Nous avons identifié et quantifié les stilbènes majoritaires des tissus ligneux de V. vinifera, évalué la sensibilité des agents de maladie du bois à l’effet antifongique de ces stilbènes et suivi leur éventuelle induction suite à l’inoculation d’agents de maladie du bois. Nous avons souligné l’importance des oligomères de stilbènes dont l’activité antifongique est élevée et montré qu’ils sont fortement induits dans le bois inoculé par un champignon pathogène de la famille des Botryosphaeriaceae. La comparaison des réponses de défenses entre des cépages tolérants et sensible, a montré que les cépages tolérants répondent globalement plus fortement et plus rapidement que le cépage sensible à une élicitation par un filtrat de milieu de culture d’un agent de l’esca. / Grapevine wood diseases, as esca syndrome and eutypiosis, are worldwide afflictions. They are due to xylem inhabiting fungi that cause the decay of woody tissues and finally the death of the plant. No curative treatment has been successfully developed until now. Studying grapevine-wood disease pathogen interaction is a prerequisite to the development of protection strategies. Phenolics, particularly stilbenoids, are antimicrobial molecules involved in the resistance of grapevine to fungal pathogens. Our work, based on biochemical and molecular approaches, reveals new data on the possible role of stilbenoids in grapevine defence against trunk diseases. We identified and quantified major stilbenoids of woody canes from V. vinifera. These compounds, especially stilbenoid oligomers, displayed in vitro antifungal activity against trunk disease agents and were induced in cuttings inoculated with an agent of the Botryosphaeriaceae family. We also compared the defence response of susceptible or tolerant cultivars to wood decay based on an elicitation assay with a culture filtrate of an esca pathogen. Defence responses of tolerant cultivars were more effective than those of the susceptible cultivar.