Neste trabalho foram estudadas as potencialidades eletroanalíticas de um material híbrido condutor baseado em acetato de celulose e grafite com eletrodeposição de filme condutor de Azul da Prússia (CA/G/PB) como sensor para espécies com importância biológica. O material híbrido foi preparado pelo processo de inversão de fase e caracterizado pelas técnicas de microscopia eletrônica de varredura acoplada com espectroscopia de energia dispersiva (SEM-EDS) e voltametria cíclica. O composto Azul da Prússia (PB) foi imobilizado na superfície do material por eletropolimerização, aplicando-se um potencial fixo de 20 mV em uma janela de -0,3 V a 1,2 V. Os estudos eletroquímicos do eletrodo modificado CA/G/PB foram realizados em solução de KCl 0,1 mol.L-1, sendo obtidos dois pares redox para a espécie eletroativa imobilizada com potenciais médios (E0) em 0,204 V e 0,842 V, indicando um comportamento quase-reversível. O eletrodo demonstrou alta estabilidade após 500 ciclos redox, não sendo observada lixiviação da espécie eletroativa da superfície da matriz modificada. Os dois pares redox do material híbrido CA/G/PB permaneceram praticamente constantes entre os pH 5,0 e 8,0, indicando que as intensidades de pico não são significativamente afetadas nessa faixa de pH. A correlação linear entre as intensidades de pico e a raiz quadrada da velocidade de varredura, indicou que o sistema possui um comportamento similar aqueles em que o processo é controlado por difusão das espécies eletroativas à superfície do eletrodo. O azul da Prússia imobilizado foi aplicado na determinação de dopamina (DP), ácido úrico (AU), ácido ascórbico (AA) e Paracetamol (PCT) através da técnica de voltametria cíclica, voltametria de pulso diferencial e cronoamperometria. / In this work, we studied the electroanalytical potential of a conductive hybrid material based on cellulose acetate and graphite with electrodeposition Blue conductor film of Prussia (CA / G / PB) as a sensor for species with biological importance. The hybrid material was prepared by phase inversion process and characterized by the techniques of scanning electron microscopy coupled with energy dispersive spectroscopy (SEM-EDS) and cyclic voltammetry. The dye of Prussian blue (PB) was immobilized on the surface of the material by electropolymerization applying a fixed potential of 20 mV in a interval from -0.3 V to 1.2 V. The electrochemical behavior of the modified electrode CA / G / PB were performed in solution of 0.1 mol L-1 KCl being obtained two redox couples for the electroactive species immobilized with midpoint potentials (E0) in 0.204 V and 0.842 V, indicating a quasi-reversible behavior. The electrode showed high stability after 500 redox cycles with no observed leaching of electroactive species to the surface of the modified electrode. The two redox pair of the CA / G / PB electrode was kept practically constant within pH 5.0 and 8.0, indicating that the peak intensities are not significantly affected in this pH range. The linear correlation between peak intensities and the square root of scan rate, indicated that the system has a similar behavior those in which the process is controlled by diffusion of electroactive species to the electrode surface. The immobilized Prussian blue was applied to determine dopamine (DP), uric acid (UA), ascorbic acid (AA) and paracetamol (PCT) by analytical techniques of cyclic voltammetry, differential pulse voltammetry and chronoamperometry.

Neste trabalho foram estudadas as potencialidades eletroanalíticas de um material híbrido condutor baseado em acetato de celulose e grafite com eletrodeposição de filme condutor de Azul da Prússia (CA/G/PB) como sensor para espécies com importância biológica. O material híbrido foi preparado pelo processo de inversão de fase e caracterizado pelas técnicas de microscopia eletrônica de varredura acoplada com espectroscopia de energia dispersiva (SEM-EDS) e voltametria cíclica. O composto Azul da Prússia (PB) foi imobilizado na superfície do material por eletropolimerização, aplicando-se um potencial fixo de 20 mV em uma janela de -0,3 V a 1,2 V. Os estudos eletroquímicos do eletrodo modificado CA/G/PB foram realizados em solução de KCl 0,1 mol.L-1, sendo obtidos dois pares redox para a espécie eletroativa imobilizada com potenciais médios (E0) em 0,204 V e 0,842 V, indicando um comportamento quase-reversível. O eletrodo demonstrou alta estabilidade após 500 ciclos redox, não sendo observada lixiviação da espécie eletroativa da superfície da matriz modificada. Os dois pares redox do material híbrido CA/G/PB permaneceram praticamente constantes entre os pH 5,0 e 8,0, indicando que as intensidades de pico não são significativamente afetadas nessa faixa de pH. A correlação linear entre as intensidades de pico e a raiz quadrada da velocidade de varredura, indicou que o sistema possui um comportamento similar aqueles em que o processo é controlado por difusão das espécies eletroativas à superfície do eletrodo. O azul da Prússia imobilizado foi aplicado na determinação de dopamina (DP), ácido úrico (AU), ácido ascórbico (AA) e Paracetamol (PCT) através da técnica de voltametria cíclica, voltametria de pulso diferencial e cronoamperometria. / In this work, we studied the electroanalytical potential of a conductive hybrid material based on cellulose acetate and graphite with electrodeposition Blue conductor film of Prussia (CA / G / PB) as a sensor for species with biological importance. The hybrid material was prepared by phase inversion process and characterized by the techniques of scanning electron microscopy coupled with energy dispersive spectroscopy (SEM-EDS) and cyclic voltammetry. The dye of Prussian blue (PB) was immobilized on the surface of the material by electropolymerization applying a fixed potential of 20 mV in a interval from -0.3 V to 1.2 V. The electrochemical behavior of the modified electrode CA / G / PB were performed in solution of 0.1 mol L-1 KCl being obtained two redox couples for the electroactive species immobilized with midpoint potentials (E0) in 0.204 V and 0.842 V, indicating a quasi-reversible behavior. The electrode showed high stability after 500 redox cycles with no observed leaching of electroactive species to the surface of the modified electrode. The two redox pair of the CA / G / PB electrode was kept practically constant within pH 5.0 and 8.0, indicating that the peak intensities are not significantly affected in this pH range. The linear correlation between peak intensities and the square root of scan rate, indicated that the system has a similar behavior those in which the process is controlled by diffusion of electroactive species to the electrode surface. The immobilized Prussian blue was applied to determine dopamine (DP), uric acid (UA), ascorbic acid (AA) and paracetamol (PCT) by analytical techniques of cyclic voltammetry, differential pulse voltammetry and chronoamperometry.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / In aquatic organisms, changes in salinity in water provoke a variety of physiological responses, so this study aimed to evaluate the effect of water salinity on weight gain, survival and expression of the genes Catalase (CAT), Glutathione Reductase (GSR), Glutathione Synthase (GSS), Glutaione peroxidase (GPX), and heat shock protein (HSP70) in the liver of Nile Tilapia (Oreochromis niloticus). For the conduction of the experiment, 160 tilapias with a mean weight of 20.8 g ( ± 4.02) were used. The experiment was conducted in a completely randomized design (DIC) composed of four treatments and four replicates, being: T1 = Treatment 1 (salinity 0 + basal diet); T2 = Treatment 2 (salinity 7 + basal diet); T3 = Treatment 3 (salinity 21 + basal diet); T4 = treatment 4 (salinity 21 + diet with 1500 mg of vitamin C / kg of feed). The parameters of water quality were monitored daily and were kept within the range of comfort for the species. For the analysis of gene expression samples of liver of tilapia were collected in two distinct periods, 24 hours and 14 days after the beginning of the experiment. Gene expression quantification was performed by qRT-PCR. There was a significant effect of the treatments on weight gain, treatment 1 (4.10 g) and treatment 2 (6.42 g) had the highest weight improve. Animals from T4 had greater weight gain than animals from T3. Higher survival was observed in treatment 2 followed by control (T1). HSP 70 and GSS were more expressed in the 24 h period and GSRR was more expressed in the 14 day period, whereas CAT and GPX did not differ from one period to the other (P <0.05). The salinity variation had an effect under the expression of the genes evaluated in the liver of O. niloticus with the exception of catalase for the 24 hour period. After 14 days of evaluation, animals raised in salinity 21 and fed with vitamin C supplementation presented greater expression of GPX, GSR and GSS, suggesting that animals raised at this level of salinity had greater need of action of glutathione system, and that the supplementation of vitamin C under these conditions allowed greater expression of these genes. It was concluded that salinity had an effect on the expression of the antioxidant defense system genes evaluated, which varied considerably over the 24-hour period and presented a more defined pattern in the 14-day period. Supplementation of vitamin C contributed, in parts, for better development of tilapia created in salinity 21, which can be observed, through the results of weight gain and gene expression. / Em organismos aquáticos, mudanças de salinidade na água provocam uma variedade de respostas fisiológicas, assim, este estudo objetivou avaliar o efeito da salinidade da água sobre o ganho de peso, a sobrevivência e a expressão dos genes Catalase (CAT), Glutatona Redutase (GSR), Glutationa Sintetase (GSS), Glutationa peroxidase (GPX) e o da proteína de choque térmico (HSP70) no fígado de Tilápias do Nilo (Oreocrhomis niloticus). Para a condução do experimento foram utilizadas 160 tilápias com peso médio 20,8 g (± 4,02). O experimento foi conduzido em delineamento inteiramente casualizado (DIC) compostos por quatro tratamentos e quatro repetições, sendo: T1 = Tratamento 1 (salinidade 0 + dieta basal); T2= Tratamento 2 (salinidade 7 + dieta basal); T3= Tratamento 3 (salinidade 21 + dieta basal ); T4 = tratamento 4 (salinidade 21 + dieta com 1500 mg de vitamina C/ kg de ração). Os parâmetros de qualidade de água foram monitorados diariamente e se mantiveram dentro da faixa de conforto para a espécie. Para as análises de expressão gênica foram coletadas amostras do fígado de tilápias em dois períodos distintos, 24 horas e 14 dias após o início do experimento. A quantificação da expressão gênica foi realizada por qRT-PCR. Houve efeito significativo dos tratamentos sobre o ganho de peso, tilápias do tratamento 1 (4,10g) e tratamento 2 (6,42 g) obtiveram os maiores ganhos de peso. Animais do tratamento 4 (salinidade 21 + acréscimo de vitamina C na ração) tiveram maior ganho de peso que os animais do tratamento 3 (salinidade 21 + dieta basal). Maior sobrevivência foi observada no tratamento 2 seguido pelo controle. HSP 70 e GSS foram mais expressos no período de 24 h e GSR sendo mais expressa no período de 14 dias, enquanto que CAT e GPX não diferiram de um período para o outro. A variação de salinidade teve efeito sob a expressão dos genes avaliados no fígado de O.niloticus com exceção da catalase para o período de 24 horas. Após 14 dias de avaliação animais criados em salinidade 21 e alimentados com suplementação de vitamina C apresentaram maior expressão de GPX, GSR e GSS, sugerindo que animais criados nesse nível de salinidade, tiveram maior necessidade de ação do sistema glutationa, e que a suplementação de vitamina C nessas condições permitiu maior expressão desses genes. Conclui-se que a salinidade apresentou efeito sobre a expressão dos genes do sistema de defesa antioxidantes avaliados, os quais variaram bastante no período de 24 horas e apresentaram um padrão mais definido no período de 14 dias. A suplementação de vitamina C contribuiu em partes, para melhor desenvolvimento das tilápias criadas em salinidade 21, o que pode ser observado, pelos resultados de ganho de peso e expressão gênica.

Acredita-se que o uso de agentes antioxidantes poderia auxiliar na restituição da resistência adesiva diminuída após o clareamento dental. O objetivo deste estudo in vitro foi avaliar o efeito do hidrogel de ascorbato de sódio a 10% e 20% e hidrogel de ácido ascórbico a 10%, aplicados por 15 minutos imediatamente após o clareamento com peróxido de hidrogênio a 35% (Lase Peroxide Sensy-DMC) (PH) na resistência adesiva imediata (24 horas) e tardia (7 dias) de uma resina composta (RC) ao esmalte bovino. Foram utilizadas 45 superfícies de esmalte de incisivos bovinos e distribuídas em 9 grupos com 5 em cada uma, sobre as quais foram confeccionados 4 corpos de prova (n= 20), de acordo com o tratamento: G1: sem clareamento (controle) + RC; G2: PH + 24h + RC; G3: PH + 7 d + RC; G4: PH + hidrogel de ascorbato de sódio 10% + 24h + RC; G5: PH + hidrogel de ascorbato de sódio a 10% + 7 d + RC; G6: PH + hidrogel de ascorbato de sódio a 20% + 24h + RC; G7: PH + hidrogel de ascorbato de sódio a 20% + 7 d + RC; G8: PH + hidrogel de ácido ascórbico a 10% + 24h + RC; G9: PH + hidrogel de ácido ascórbico a 10% + 7 d + RC. Foram confeccionados cilindros de (0.8x1mm), utilizando o sistema adesivo Single Bond 2 e a resina composta Z100 (3M ESPE) e submetidos ao teste de resistência adesiva ao microcisalhamento na máquina de ensaios universal (EMIC) com célula de carga de 50N a uma velocidade de (0,5mm/min). Posteriormente foram observados os tipos de fraturas em um estereomicroscópio com aumento de 40x. A seguir, os valores de resistência adesiva ao microcisalhamento foram avaliados estatisticamente através da Análise de Variância a um critério (ANOVA) e do teste de Tukey para comparações múltiplas, com nível de significância de 5%. Os resultados foram: G1: 24,34ab ± 5,182; G2: 15,27c ± 7,170; G3: 16,65c ± 7,614; G4: 20,30bc ± 7,071; G5: 17,77bc ± 8,135; G6: 18,03bc ± 4,016; G7: 19,60bc ± 6,396; G8: 20,03bc ± 3,941; G9: 17,63bc ± 3,390. De acordo com os resultados obtidos na presente pesquisa pode-se concluir que: a técnica de clareamento dental com peróxido de hidrogênio a 35% promoveu uma diminuição estatisticamente significante na resistência adesiva da resina composta ao esmalte bovino nos procedimentos realizados 24 horas e 7 dias após o clareamento dental. Tanto o tratamento com hidrogel de ascorbato de sódio como com o hidrogel de ácido ascórbico, independente da concentração e tempo de espera, foram capazes de melhorar a resistência adesiva da resina composta ao esmalte bovino clareado. Entretanto, ambos os tratamentos com agentes antioxidantes não foram capazes de recuperá-la. / It is referred that the use of antioxidants may improve the decreased bond strength after bleaching treatment. The aim of this in vitro study was to evaluate the effect of 10% and 20% sodium ascorbate hydrogel and 10% ascorbic acid hydrogel applied for 15 minutes immediately after 35% hydrogen peroxide bleaching (HP) (Lase Peroxide Sensy-DMC). 45 bovine incisors enamel surfaces divided into 9 groups were used. Four specimens were made on each surface (n = 20) according to the different treatments: G1: without bleaching (control group) + RC; G2 : HP + 24 h + RC; G3: HP + 7 d+ RC; G4: HP + 10% sodium ascorbate hydrogel + 24 h + RC; G5: HP + 10% sodium ascorbate hydrogel + 7 d + RC; G6: HP + 20% sodium ascorbate hydrogel + 24h + RC; G7: HP + 20% sodium ascorbate hydrogel + 7 d + RC; G8: HP + 10% ascorbic acid hydrogel + 24 h + RC; G9: HP + 10% ascorbic acid hydrogel + 7 d + RC. Cylinder restorations (0.8x1mm) were made using Single Bond 2 and Z100 (3M ESPE). Microshear bond strength test was performed using a universal testing machine (EMIC) with a 50N load at a crosshead speed of 0.5mm/min. Failure modes were assesed using a stereomicroscope (40x) showing mainly adhesive failures. Data were analyzed by ANOVA and Tukey analysis for multiple comparisons at the significance level of 5%. The results were: G1: 24.34 ± 5.182 ab G2: 15.27 ± 7.170 c; G3: 16.65 ± 7.614 c, G4: 20.30 ± 7.071 bc; G5: 17.77 ± 8.135 bc G6: ± 18.03 bc 4.016; G7: 19.60 bc ± 6.396; G8: 20.03 bc ± 3.941; G9: 17.63 ± 3.390 bc. According to the results obtained it can be concluded that: the 35% hydrogen peroxide tooth whitening technique promoted a statistically significant decrease in bond strength of resin composite to enamel performed 24 hours and 7 days after bleaching. Both treatment with sodium ascorbate hydrogel and ascorbic acid hydrogel, independent of concentration and time, were able to improve the bond strength of composite resin to bleached bovine enamel, however, both antioxidants were not able to retrieve it.

FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / A cicatrizaÃÃo de feridas à uma cascata de eventos celulares e moleculares que interagem para que ocorra a reconstituiÃÃo do tecido. Neste estudo avaliou-se o efeito de um composto de triglicerÃdeos de cadeia mÃdia, Ãcido linolÃico, lecitina de soja e vitaminas A e E (DersaniÂ), e tambÃm da Ãgua de coco liofilizada, nas formas aquosa e gel, no processo de cicatrizaÃÃo de Ãlceras cutÃneas experimentais. Utilizaram-se 105 ratos, machos, da linhagem Wistar, nos quais foi removido um segmento cutÃneo de espessura total com 4 cm2 de Ãrea. Os animais foram distribuÃdos aleatoriamente em grupos constituÃdos de 15 ratos compreendendo trÃs experimentos constituÃdos dos grupos Controle e ReferÃncia e dos grupos Amorfa-aquosa e Cristalina-aquosa (Experimento 1), Amorfa-gel e Cristalina-gel (Experimento 2) e TriglicerÃdeo (Experimento 3). Tais grupos foram tratados por via tÃpica, respectivamente, com soluÃÃo salina 0,9%, clostebol + sulfato de neomicina, Ãgua de coco amorfa e cristalina-aquosa, Ãgua de coco amorfa e cristalina-gel e DersaniÂ. As Ãreas das feridas foram mensuradas por planimetria digital nos dias zero, 3, 7 e 12 de pÃs-operatÃrio, a partir das quais foram calculados o grau e a taxa mÃdia de reparaÃÃo. AlÃm disso, o processo de cicatrizaÃÃo foi avaliado atravÃs dos critÃrios de Myers e da quantificaÃÃo de colÃgeno e angiogÃnese utilizando mÃtodo histoquÃmico. No 3o dia observou-se uma expansÃo da Ãrea da ferida no grupo referÃncia e uma leve contraÃÃo nos grupos controle e testes. Nos dias subseqÃentes o processo de reparaÃÃo, medido pela variÃvel grau de reparaÃÃo, evoluiu de forma linear, de modo que, no 12o dia, a Ãrea reparada alcanÃou 77,95% da regiÃo ulcerada inicial no grupo Controle, 78,40% no ReferÃncia, 82,24% no Amorfa-aquosa, 84,12% no Cristalina-aquosa 84,12% (experimento 1), 85,09% no grupo Amorfa-gel, 84,20% no Cristalina-gel (experimento 2) e 83,49% no TriglicerÃdeo (experimento 3), nÃo sendo constatadas diferenÃas estatisticamente significantes em nenhum dos experimentos. Igualmente semelhantes foram os valores da taxa mÃdia de reparaÃÃo referente aos 12 dias de tratamento: 25,79 mm2/dia no grupo Controle, 25,42 mm2/dia no ReferÃncia, 26,48 mm2/dia no Amorfa-aquosa, 27,89 mm2/dia no Cristalinaaquosa, 26,47 mm2/dia no Amorfa-gel, 27,04 mm2/dia no Cristalina-gel e 27,38 mm2/dia no TriglicerÃdeo. Entretanto, nos grupos Amorfa-aquosa (P<0,01), Amorfa-gel (P<0,05), Cristalina-gel (P<0,05) e TriglicerÃdeo (P<0,01) a densidade de colÃgeno foi significatemente maior do que a verificada no grupo Controle. Apenas no grupo TriglicerÃdeo a densidade vascular, a deposiÃÃo de colÃgeno e de tecido de granulaÃÃo foram significantemente maior que nos grupos Controle e ReferÃncia (P<0,01). Estes achados permitem concluir que o tratamento com o Dersani à um potencializador do processo de reparo tecidual / Healing of wounds is a cascade of cellular and molecular events which interact with one another ensuing tissue reconstruction. This study assessed the effect of a compound medium-chain triglycerides, linoleic acid, soy lecithins and vitamins A and E (Dersan) and also lyophilized coconut water in the aqueous and gel forms in the process of healing of experimental cutaneous ulcers. 105 male Wistar rats were employed in which a 4cm2 full thickness skin segment was removed. The animals were randomly distributed into groups constituted of 15 rats comprising three experiments consisting of Control and Reference groups, as well as Amorphous-Aqueous and Crystalline-Aqueous groups (Experiment 1), Amorphous-gel and Crystalline-gel groups (Experiment 2) and Triglycerides (Experiment 3). Such groups were treated via topic, respectively with 0.9% saline, clostebol+neomycin sulphate, amorphous and crystalline coconut water, amorphous and crystalline-gel coconut water and Dersani. The wound areas were measured by digital planimetry at post-operative days zero, 3, 7 and 12; from them the degree and average rate of repair were calculated. Moreover, the healing process was assessed through the criteria of Meyers and the amount of collagen and angiogenesis by means of histochemical methods. At day 3, an expansion of the wound area in the Reference Group and a slight contraction of Control Groups and tests were noted. On the subsequent days the process of repair, measured through the variable degree of healing evolved in a linear form, so that on the twelfth day, the repaired area achieved 77.95% of the initial ulcerated region in the Control Group, 78.40% in the Reference Group, 82.24%, in the Amorphous-Aqueous one, 84.12% in the Crystalline- Aqueous (Experiment 1), 85.09% in the Amorphus-gel Group, 84.20% in the Crystalline-gel Group (Experiment 2) and 83.49% in that of Triglyceride (Experiment 3); no statistically significant differences were found in the experiments. Equally similar were the values for the average rate of repair in regard to the 12 days of treatment: 25,79mm2/day in the Control Group, 25,42mm2/day in the Reference Group, 26.48mm2/day in the Amorphous-Aqueous one, 27.89mm2/day in the Crystalline-Aqueous Group, 26,47mm2/day in the Amorphous-gel Group, 27.04mm2/day in the Crystalline-gel and 27.38mm2/day in the Triglyceride Group. However, as to the Groups Amorphous-aqueous (P<0,01), Amorphous-gel (P<0.05), Crystalline Gel (P<0,05) and Triglycerides (P<0,01), the density of collagen was significantly higher than that observed in the Control Group. Only the Group of Triglycerides showed significantly higher vascular density, deposition of collagen and the granulation tissue than Control and Reference groups (P<0, 01). These findings allow the conclusion that Dersani have potential to the process of tissue repair

Made available in DSpace on 2014-08-20T13:42:05Z (GMT). No. of bitstreams: 1 Tese_Cristina_Simoes_da_Costa.pdf: 676580 bytes, checksum: 3d29b81ccb9b1d19211d238ccd303fe3 (MD5) Previous issue date: 2009-07-30 / Strawberry is a highly perishable fruit, presenting a post-harvest shelf life relatively short due to physiologic changes and incidence of fungal decay, which limitate its fresh commercialization. Chitosan based edible coating are a good alternative on controlling post harvest changes associated do strawberry decay. This work was divided in two experiments. In first article, the application of chitosan edible coatings added of calcium and fat acid to promote preservation of the quality of strawberry cv. Aromas during refrigerated storage. Three coatings were studied: chitosan + calcium chloridre (QC), . chitosan + calcium chloride + oleic acid (QCAO), and chitosan + calcium chloride + stearic acid (QCAE). After coatings were applied, the fruits were stored at + 2°C e 75 ± 5%HR for 10 days. Firmness, pH, titratable acidity, content of soluble solids and, color weren t affected significantly at the end of storage, and no differences between treatment were observed. Coating affected significantly fungal decay on QC coated fruits when compared to non coated fruits, promoting 83%. Fruits that received QC or QCAE coating obtained same acceptability of control fruits. In the second article, the ability of chitosan edible coating, added of calcium and/or ascorbic acid to maintain strawberry post-harvest quality under refrigerated (0°C) and atmospheric (25ºC) storage was evaluated. Four coatings were studied: chitosan (Q); chitosan + calcium chloridre (QC), chitosan + ascorbic acid (QA), and chitosan + calcium chloride + ascorbic acid (QCA). After coatings were applied, the fruits were stored for 15 days at 0 + 2°C e 75 ± 5%HR (refrigerated) and for 7 days at 25ºC + 2ºC (atmospheric). Firmness, pH, titratable acidity, content of soluble solids and, color weren t affected significantly at the end of storage, and no differences between treatment were observed. On refrigerated condition, chitosan coatings promoted the maintenance of titrable acidity, reduction of loss of firmness and of fungal decay, which was completely inhibited on the fruits covered with QA and QC until the twelve day. Soluble solids content, pH, color (L*, C*, h*), anthocyanins content and phenolic compounds content weren´t affected by coating. On the strawberry stored under atmospheric temperature, coatings promoted retention of firmness and controlled fungal decay, showing no effects on the other attributes evaluated. In general, post-harvest application of chitosan based edible coatings preserve strawberry quality during storage. Incorporation of ascorbic acid or calcium chloride on coating promotes additional gain on the control of fungal decay, being observed a higher acceptability of coatings containing ascorbic acid. The employ of this coating can extend the shelf life of refrigerated or atmospheric stored strawberry until 12 and 3 days, respectively. / O morango é um fruto altamente perecível, apresentando período de conservação pós-colheita relativamente curto em virtude de alterações fisiológicas e incidência de podridão fúngica, o que limita sua comercialização in natura. As coberturas comestíves à base de quitosana tem-se mostrado como alternativas no controle das alterações pós-colheita responsáveis pela deterioração do morango. Este trabalho foi dividido em dois ensaios experimentais. No primeiro artigo, avaliou-se a aplicação de coberturas comestíveis à base de quitosana contendo cálcio e ácidos graxos para promover a manutenção da qualidade pós-colheita de morangos cv. Aromas durante o armazenamento refrigerado. Três coberturas foram estudadas: quitosana + cloreto de cálcio (QC); quitosana + cloreto de cálcio + ácido oléico (QCAO); quitosana + cloreto de cálcio + ácido esteárico (QCAE). Após aplicação das coberturas, os frutos foram mantidos por 10 dias sob condições de 0+2°C e 75+5%UR. A firmeza, o pH, a acidez titulável, o conteúdo de sólidos solúveis e a cor não apresentaram variação significativa ao final do armazenamento, não sendo verificada diferença entre os tratamentos quando comparados ao controle. A cobertura apresentou efeito significativo na redução da podridão fúngica dos frutos cobertos com QC, verificando-se uma redução de 83%. Os frutos que receberam as coberturas QC ou QCAE apresentaram mesma aceitação que os frutos controle. No segundo artigo, avaliou-se o efeito de coberturas comestíveis à base de quitosana, combinada ou não com cálcio e ácido ascórbico, na manutenção da qualidade pós-colheita de morangos durante o armazenamento refrigerado (0°C) e sob temperatura ambiente (25°C). Quatro coberturas foram estudadas: quitosana (Q); quitosana + cloreto de cálcio (QC); quitosana + ácido ascórbico (QA) e quitosana + cloreto de cálcio + ácido ascórbico (QCA). Após aplicação das coberturas, os frutos foram mantidos por 15 dias, a 0+2°C e 75+5%UR (refrigerado) e à 25°C + 2ºC (ambiente) durante 7 dias. Nas amostras refrigeradas, a utilização de coberturas promoveu manutenção da Lacidez titulável, redução de perda de firmeza e do desenvolvimento fúngico, sendo este completamente inibido nos frutos cobertos com QA e QC até o décimo segundo dia de armazenamento. O pH, o conteúdo de sólidos solúveis, a cor (L*, C* e h*), o conteúdo de antocianinas e o de compostos fenólicos não sofreram influência da aplicação da cobertura. Nos frutos armazenados à temperatura ambiente, foi verificado o efeito da cobertura na manutenção da firmeza, e no controle do desenvolvimento fúngico, não sendo observado efeito sobre a perda de massa e demais parâmetros estudados. De forma geral, pode-se concluir que a aplicação pós-colheita de coberturas à base de quitosana em morangos preserva sua qualidade durante o armazenamento. A incorporação de ácido ascórbico ou cloreto de cálcio na cobertura possibilita ganho adicional no controle do desenvolvimento fúngico, obtendo-se maior aceitação para as coberturas contendo ácido ascórbico. O uso dessa cobertura permite estender a vida útil de morangos armazenados sob refrigeração e à temperatura ambiente, por até doze e três dias, respectivamente.

Au cours des procédés d’appertisation, qui impliquent des traitements thermiques généralement pratiqués à hautes températures, la qualité nutritionnelle des produits alimentaires est souvent altérée. En effet, certaines vitamines, telles que la vitamine C, sont très thermolabiles.L’objectif du projet PREDINUT était de proposer un outil d’aide à la décision afin de prévoir la quantité de vitamine C (marqueur de qualité nutritionnelle) détruite dans des conditions données de durée/température, afin de pouvoir optimiser les procédés.Un thermorésistomètre Mastia®, a été utilisé pour établir des conditions de température et pression représentatives des procédés d‘appertisation. L’impact de la variation de paramètres physico-chimiques tels que le pH, la concentration initiale en acide ascorbique ou encore la pression partielle en oxygène dans l’espace de tête, sur la dégradation de l’acide ascorbique a été étudié en solution modèle. Des études ont également été menées dans des produits alimentaires (purée de pomme, purée de carotte et jus de carotte) afin d’établir un profil de destruction de la vitamine C dans des aliments concrets. Dans ce projet, la plage de température étudiée est de 95 à 125 °C, le pH fixé en solution modèle est de 2,5 à 5,5 en conditions d’anaérobiose stricte, et de 3,5 en conditions d’aérobiose (3,5 étant la valeur de pH de la majorité des fruits), les concentrations initiales en acide ascorbique varient de 15 à 90 mg/100mL, et la pression partielle en oxygène de 30000 à 100000 Pa. Dans les produits alimentaires, les expériences ont été réalisées par enrichissement en acide ascorbique à hauteur de 450 mg/L, la valeur du pH ne subissant pas d’ajustement.Des modèles ont été établis pour décrire le profil cinétique de la dégradation de la vitamine C en aérobiose et en anaérobiose. En l’absence d’oxygène la dégradation de la vitamine C suit une cinétique d’ordre 1 (sauf à pH = 4,5). En présence d’oxygène (conditions aérobies) la vitesse de dégradation croit avec la pression partielle d’oxygène, mais très peu avec la température. Des ordres apparents variant de 0,5 à 0,75 en fonction de la pression partielle en oxygène dans l’espace de tête, s’avèrent mieux ajuster les données que l’ordre 1.Dans les aliments, le mécanisme de dégradation de la vitamine C apparaît beaucoup plus complexe, et la vitesse de réaction n’évolue pas de façon continue avec le temps, avec des phases de plateau suivies de réaccélérations. Une connaissance fine des mécanismes et des espèces chimiques impliquées sera nécessaire pour comprendre cette évolution. / Canning involves thermal treatments performed at high temperatures, which may alter nutrional quality of food products. Indeed, some vitamins, like vitamin C, are very heat sensitive.The aim of the project PREDINUT was to propose a decision support tool that predicts vitamin C (as indicator of nutritional quality) degradation in given conditions of time/temperature, in order to optimise processes.A thermoresistometer Mastia® was used to reach the temperature and pressure conditions encountered during canning. The impact of the variation pH, initial concentration of ascorbic acid or again partial pressure of oxygen in headspace, on ascorbic acid degradation was studied in model solution. Studies were also performed in apple puree, carrot puree and carrot juice in order to obtain a destruction profile of vitamin C in real foods. . The temperature range studied was from 95 to 125 °C, the fixed pH in model solution was from 2.5 to 5.5 in strict anaerobic conditions, and 3.5 in aerobic conditions (3.5 being the pH value of common fruits), initial concentrations of ascorbic acid varied from 150 to 900 mg/100mL, and the partial pressure of oxygen from 30000 to 100000 Pa. Foods were enriched in ascorbic acid (up to 450 mg/mL), and the pH value was not adjusted.Models were established to describe the kinetics of vitamin C degradation in aerobiosis and anaerobiosis. En absence of oxygen ascorbic acid degradation was well described by a first order kinetics (except at pH = 4.5). In presence of oxygen the degradation speed increased with head-space oxygen’s partial pressure but very little with temperature. Apparent reaction orders varying from 0.5 to 0.75, depending on the partial pressure of oxygen in the headspace, fitted the data better than first-order.In foods, degradation kinetics of vitamin C appeared more complex, with eventual presence of plateaus followed by acceleration of the reaction. Definitely more knowledge on mechanisms is needed to predict ascorbic acid degradation in real foods.

Iron is an essential nutrient for cellular activities including oxygen transport, electron transfer, and gene regulation. However, iron is potentially toxic via its redox reactions which generate reactive oxygen species (ROS). Oxidative damage to biomolecules can be modulated by antioxidants such as ascorbic acid (AA). However, it is well known that in the presence of redox-active iron, AA can act as a pro-oxidant in vitro and contribute to the formation of hydroxyl radicals. Based on the possible pro-oxidant interaction of iron and AA, we evaluated the manifestations of supplementation of iron associated with the ascorbic acid. The study was delineated by nine non-smoking male healthy volunteers, aged between 20 and 31 years. The volunteers were supplemented with a single dose containing 2g of AA (first group), 150mg of iron (second group) and 2g of AA plus 150mg of iron (third group). The 9 volunteers were submitted the all the treatments, which were alternate every 15 days. The volunteers were submitted to blood collections before the supplementation and 2, 5 and 24 hours after the supplementation. They were evaluated the levels of iron and ferritin, the activity of the antioxidants enzymes catalase (CAT), gluthatione peroxidase (GPx), superoxide dismutase (SOD), the level non-enzymatic antioxidants: AA, non-protein-SH, as well as markers of the oxidative stress Thiobarbituric acid reactive substances (TBARS), diclorofluorescein oxidation and delta-amino levulinate dehydratase (ALA-D) activity. The results showed that plasma AA levels were increased at 2, 5 and 24 hours after AA or AA plus iron ingestion. Plasmatic iron level was increased at 2 hours after iron ingestion and 2, 5 hours in the group AA plus iron. The erythrocytes TBARS levels decreased at 5 hours after AA and 5, 24 hours after AA plus iron ingestion. The erythrocytes CAT levels caused a significant increase 5 hours after supplementation with AA plus iron. The other results showed no significant different in the determinations. Thus, the present study does not support the hypothesis that the combination of high plasma oncentrations of AA and iron, or iron alone, causes oxidative damage in vivo. However, further studies are required to determine if iron and AA interactions could have a pro-oxidant effect in vivo. / O ferro é um nutriente essencial para atividades das células incluindo transporte de oxigênio, transferência de elétrons e regulação genética. Entretanto, esse mineral é potencialmente tóxico por participar de reações de óxido-redução, que favorecem a formação de espécies reativas ao oxigênio (ERO). O dano oxidativo nas biomoléculas pode ser modulado por antioxidantes como o ácido ascórbico (AA). Entretanto, sabe-se que na presença de ferro, o ácido ascórbico pode atuar como um pró-oxidante in vitro e contribuir para formação de radicais hidroxila. Baseado na possibilidade pró-oxidante da interação entre o ferro e o ácido ascórbico, for avaliados as manifestações da suplementação do ferro associado com o ácido ascórbico. O estudo foi delineado por 9 voluntários saudáveis, não tabagistas, entre 20 e 31 anos. Os voluntários foram suplementados com uma dose única contendo 2g de ácido ascórbico (primeiro grupo), 150mg de ferro (segundo grupo) e 2g de ácido ascórbico mais 150mg de ferro (terceiro grupo). Os 9 indivíduos foram submetidos a todos os tratamentos, os quais foram alternados a cada 15 dias. Os voluntários foram submetidos a coletas sanguíneas antes da suplementação e 2, 5 e 24 horas após a suplementação. Foram avaliados os níveis de ferro e ferritina, a atividade das enzimas antioxidantes catalase (CAT), glutationa peroxidase (GPx), superóxido dismutase (SOD), os níveis dos antioxidantes não-enzimáticos: ácido ascórbico, tiois não proteicos (NPSH), bem como os marcadores do estresse oxidativo: espécies reativas ao ácido tiobarbitúrico (TBARS), oxidação da diclorifluoresceína e a atividade da delta aminolevulinato desidratase (ALA-D) . Os resultados encontrados mostraram que os níveis plasmáticos de ácido ascórbico aumentaram significativamente em 2, 5 e 24 horas após a ingestão de ácido ascórbico mais ferro ou somente ácido ascórbico. Os níveis plasmáticos de ferro aumentaram significativamente 2 horas após a ingestão de ferro e 2 e 5 horas no grupo ferro mais ácido ascórbico. Os níveis de TBARS eritrocitário diminuíram significativamente em 5 e 24 horas após a ingestão de ferro, bem como, em 5 horas após a ingestão de ferro mais ácido ascórbico. A atividade da CAT eritrocitária aumentou significativamente em 5 horas após a ingestão de ácido ascórbico mais ferro. Os demais parâmetros avaliados não mostraram diferenças significativas. Com isso, o presente estudo não confirma a hipótese que a combinação de altas doses de ácido ascórbico e ferro, ou apenas ferro causam dano oxidativo in vivo. Entretanto, mais estudos são necessários para determinar se a interação entre o ferro e o ácido ascórbico pode causar efeito pró-oxidante in vivo.

L’acide ascorbique (AsA) est un antioxydant essentiel à la fois pour l’homme et les végétaux. L’AsA provenant des plantes représente la source principale de vitamine C dans l’alimentation quotidienne. Au-delà de son impact nutritionnel, augmenter la teneur en AsA dans le fruit de tomate serait susceptible d’influencer la qualité des fruits après la récolte, en termes de conservation mais également de résistance à des pathogènes. Bien que le métabolisme de l’AsA soit bien caractérisé, les mécanismes impliqués dans sa régulation restent jusqu'à présent peu compris. Des études récentes menées sur des feuilles d’Arabisdopsis thaliana montrent que certaines protéines seraient capables de réguler la teneur en AsA, en agissant au niveau transcriptionnel ou post-transcriptionnel. A ce jour, ce type de régulation n’a pas été encore décrit chez les fruits. Dans ce but, une approche de génétique directe a été développée afin d’étudier les mécanismes impliqués dans la régulation de la teneur en AsA et ceci dans le fruit de tomate (Solanum lycopersicum). L’analyse d’une population de mutants EMS de tomate Micro-Tom a permis l’identification de lignées de mutants présentant des teneurs en AsA de 2,5 à 4 fois plus importantes que celles observées dans les fruits de tomate sauvage. La caractérisation de ces lignées a conduit à des résultats prometteurs pour l’étude de la qualité des fruits après la récolte. Une stratégie de NGS-mapping a permis l’identification des mutations causales responsables du phénotype AsA observé. Ainsi, le criblage de mutants EMS a permis la découverte de nouvelles protéines inattendues, permettant de confirmer au niveau moléculaire l’existence d’une interaction directe en la signalisation lumineuse et la régulation de la voie de biosynthèse de l’AsA. / The ascorbic acid (AsA) is an essential antioxidant in both plants and humans. Plant-derived AsA is the major source of vitamin C in the human diet. In addition to its effect on tomato nutritional value, increasing tomato AsA content would likely affect postharvest storage and resistance to pathogens of the fruit. While AsA metabolism is well characterized, the mechanisms involved in its regulation remain poorly understood. Recent studies in Arabidopsis leaves indicate that few regulatory proteins can regulate this pathway at transcriptional and post-transcriptional levels. Still nothing equivalent has been described in fruits. In that aim, a forward genetic approach has been carried out to investigate the regulation of AsA in tomato (Solanum lycopersicum) fruit. The screening of an EMS tomato mutant population in the miniature cultivar Micro-Tom for identifying mutant lines with AsA-enriched fruits was done. Among the 500 M2 mutant families screened, four mutant lines with higher AsA content ranging from 2.5 to 4 fold were selected. These mutant lines have been characterized for postharvest traits quality and showed promising results. A method based on NGS-mapping allowed the identification of the putative AsA-enriched related gene. Thus, the screening of EMS mutants led to original findings such as the discovery of new unexpected proteins regulating AsA in plants, and particularly in fruits. Our work confirms at the molecular level the direct interaction between light signaling component and the regulation of the AsA biosynthesis pathway.

Parmi les maladies affectant les pommes, la moisissure bleue causée par Penicillium expansum est une préoccupation majeure. Elle cause des pertes de rendement et de qualité dues également à la production de mycotoxines telles que la patuline (PAT) et la citrinine (CIT). La PAT est la plus alarmante en raison de ses propriétés cytotoxiques, génotoxiques et immunosuppressives.L'Union européenne (UE) a établi des réglementations spécifiques pour protéger la santé des consommateurs et des niveaux maximaux de 50 g / kg sont fixés pour les jus de fruits et les produits dérivés, 25 g / kg pour les purées de pommes et les compotes et 10 g / kg pour les aliments destinés aux bébés et aux jeunes enfants. En dépit de ces mesures, la PAT continue à être présente dans les aliments et / ou les boissons commerciaux, dépassant parfois les limites maximales. Des recherches supplémentaires sont par conséquent nécessaires pour minimiser la contamination des produits alimentaires par cette mycotoxine et son champignon producteur. Bien que la plupart des études sur P. expansum soient essentiellement centrées sur la PAT, le génomede ce champignon présente d'autres clusters de métabolites secondaires (SM) prédits dont certains peuvent être associés à des métabolites potentiellement toxiques. Afin de contrôler la synthèse de SM, l'étude des facteurs de transcription globaux régulant leur production est essentielle. Dans une première partie, le gène veA, appartenant à la famille des protéines du complexe velvet, a été caractérisé et son impact sur le développement du champignon, sa virulence et son métabolisme secondaire a été élucidé. La délétion de ce gène a conduit à une réduction de la production de PAT et de CIT et à une diminution de l'expression de leurs clusters de gènes. VeA a également un impact global sur le métabolisme secondaire, puisque 15 des 35gènes de structure présentent une régulation différentielle sur les milieux testés. Dans une deuxième partie, l’influence de l’acide ascorbique (AA) sur la concentration de PAT dans le jus de pomme trouble a été étudiée à la fois en laboratoire et en milieu semi-industriel. Une méthodologie analytique séparant la PAT et d'autres composés générés au cours de la réaction aété optimisée. Les conditions optimales d'action de l’AA sur la PAT ont été analysées. De plus,nous avons identifié des produits de dégradation moins toxiques que la PAT et résultant du traitement par l’AA. Pour conclure, cette thèse se rattache à la gestion des risques de la PAT dans le secteur des fruits ; elle apporte des connaissances et des améliorations significatives tant sur le plan fondamental que sur le plan pratique. Ces avancées résident principalement dans la description d'une souche mutée de P. expansum moins toxique que celle naturellement retrouvée dans la nature, et décrivant un additif alimentaire améliorant les qualités de nombreux produits transformés et diminuant la concentration de PAT en générant des composés moins toxiques. / Among diseases affecting apples, blue mould caused by Penicillium expansum is a major concern. It causes yield and quality losses, as well as food safety issues due to the production of mycotoxins such as patulin (PAT) and citrinin (CIT). PAT is the most worrying one and has cytotoxic, genotoxic and immunosuppressive properties. The European Union (EU) has established specific regulations to protect the consumer’s health and maximum levels of PAT of 5 g/kg is set for fruit juices and derived products, 25 g/kg for apple purees and compotes and 10g/kg for food intended for babies and young children. However, PAT is still found in commercial food and/or beverage products, sometimes exceeding the maximum limits and more research is needed to minimize contamination of food products by this mycotoxin and its fungus. Even though most studies on P. expansum have focused on PAT itself, the genome of this fungus exhibits other predicted secondary metabolite (SM) clusters, some of which may be associated with potentially toxic metabolites. In order to control the synthesis of SMs, the study of global transcription factors regulating their production is essential. In a first part, the veA gene, belonging to the velvet family, was characterised and its impact on the development of the fungus, its virulence and its secondary metabolism was elucidated. The disruption of this gene led to the failure in PAT and CIT production and a decrease in the expression of their gene cluster. It also revealed a global impact on the secondary metabolism, as 15 of 35 backbone genes showed differential regulation on the media tested. In a second part, the influence of ascorbic acid (AA) on the concentration of PAT in cloudy apple juice was studied on both lab and semi-industrial scale. An analytical methodology separating PAT and other compounds generated during the reaction was optimized. Optimal conditions of action of AA on PAT were studied. In addition, degradation products less toxic than PAT and resulting from AA treatment were identified. To conclude, this thesis is part of the risk management of PAT in the fruit sector; it provides significant improvements at both fundamental and practical levels. These advances are mainly characterized by the description of a mutated strain of P. expansum that is less toxic than that naturally occurring in nature, and the description of a food additive that improves numerous products qualities and affects PAT concentration, thusgenerating less toxic compounds