Dissertations / Theses on the topic 'Compost extracts'

Thesis (M.S.)--West Virginia University, 2005.
Title from document title page. Document formatted into pages; contains viii, 70 p. : ill. Includes abstract. Includes bibliographical references (p. 64-70).

One of the motivating reasons for the development of hydroponics was avoidance of root pathogens. Hydroponics involves growing crops in relatively sterile media, isolated from the underlying soil which may have disease pressure. However, even when hydroponics is coupled with controlled environments such as high tunnels and climate-controlled greenhouses, soil-borne pathogens can enter the growing area and proliferate due to optimal environmental conditions for pathogen growth. Control of root pathogens is difficult and usually achieved through synthetic fungicides since few biocontrol options are available. Compost water extracts (CWE) have recently been gaining the attention of greenhouse growers because they may be a low-cost, environmentally friendly approach to control root disease. CWE are mixtures of compost and water incubated for a defined period of time, either with or without aeration, and with or without additives intended to increase microbial populations, which in turn suppress disease. Much anecdotal, but very little scientific, evidence exists describing CWE effect on suppressing soil-borne pathogens. The present study 1) examined the effect of an aerated CWE on disease suppression at the laboratory scale and in container studies using different soilless substrates, 2) investigated a phenotypic change at the root level caused by CWE that may be associated with disease suppression, and 3) isolated some factors in the production of CWE that affect the ability of a CWE to suppress disease. The common model pathogen-host system of Fusarium solani f.sp. pisi and pea was used to examine CWE-induced disease suppression, with information then being translatable to similar patho-systems involved in greenhouse crop production. In the first study, laboratory-based root growth and infection assays resulted in 100% suppression of F. solani when roots were drenched in CWE. These protected seedlings were then taken to a greenhouse and transplanted into fine coconut coir, watered with hydroponic nutrient solution, and grown for five weeks. At the end of the experiment, 23% of the shoots of the pathogen-inoculated, CWE-drenched seedlings remained healthy while only 2% of the inoculated seedlings without CWE drench remained healthy. All of the roots of the inoculated seedlings developed lesions, even those drenched in CWE. However, 29% of the CWE drenched roots were able to recover from disease, growing white healthy roots past the lesion, while only 2% recovered naturally. A shorter-term container study was conducted in the laboratory to determine the effects of CWE-induced suppression when peas were grown in different substrates and to determine if the hydroponic nutrient solution had an effect on the suppression. Peas were grown in sterilized fine and coarse coconut coir fiber and sand irrigated with water, with a second set of fine coir irrigated with hydroponic nutrient solution. Pea seeds with 20-25mm radicles were inoculated with pathogen and sown directly into CWE-drenched substrate and grown for three weeks. At the end of the experiment, 80%, 60%, 90%, and 50% of the shoots of the inoculated, CWE-drenched seedlings remained healthy when grown in fine coir, coarse coir, sand, and fine coir irrigated with hydroponic nutrient solution, respectively. Nearly 100% of the roots grown in coconut coir substrates again developed necrotic lesions but 83%, 87%, 100%, and 87% grew healthy roots beyond the disease region. The hydroponic nutrient solution had a negative effect on suppression, with a reduction of at least 30 percentage points. Sand demonstrated a natural ability to suppress F. solani. Only 23% of inoculated seedlings had dead or dying shoots by the end of the experiment (compared to 77-80% in coir substrates) and although all but one of the roots developed lesions, all were able to recover on their own with CWE. CWE further increased shoot health and also prevented 57% of the roots from developing lesions. In a second study, two different CWE were used to examine the effect on root border cell dispersion and dynamics in pea, maize, cotton, and cucumber and its relation to disease suppression. Dispersal of border cells after immersion of roots into water or CWE was measured by direct observation over time using a compound microscope and stereoscope. Pictures were taken and the number of border cells released into suspension were enumerated by counting the total number of cells in aliquots taken from the suspension. Border cells formed a mass surrounding root tips within seconds after exposure to water, and most cells dispersed into suspension spontaneously. In CWE, >90% of the border cell population instead remained appressed to the root surface, even after vigorous agitation. This altered border cell phenomena was consistent for pea, maize, and cotton and for both CWE tested. For most cucumber roots (n=86/95), inhibition of border cell dispersal in both CWE was similar to that observed in pea, maize, and cotton. However, some individual cucumber roots (8±5%) exhibited a distinct phenotype. For example, border cells of one root immersed into CWE remained tightly adhered to the root tip even after 30 minutes while border cells of another root immersed at the same time in the same sample of CWE expanded significantly within 5 minutes and continued to expand over time. In a previous study, sheath development over time in growth pouches also was distinct in cucumber compared with pea, with detachment of the sheaths over time, and root infection was reduced by only 38% in cucumber compared with 100% protection in pea (Curlango-Rivera et al. 2013). Further research is needed to evaluate whether this difference in retention of border cell sheaths plays a role in the observed difference in inhibition of root infection. In the third study, a series of investigations were conducted to isolate different factors that contribute to the suppression ability of a CWE by changing incrementally changing some aspect of the CWE production process. The basic aerated CWE recipe (with molasses, kelp, humic acid, rock phosphate, and silica) provided 100% protection of pea from root disease while the non-aerated basic recipe CWE provided 72% protection. Aerated CWE made of only compost and water resulted in 58% protection. It was found that molasses did not contribute to the suppression ability of the ACWE, while kelp contributed strongly. When soluble kelp was added by itself to the compost and water, the CWE provided 80% suppression. However, when all additives were included except molasses and kelp, suppression remained high (93%) indicating that humic acids, rock phosphate, and/or silica were also major contributors toward the suppression effect. Optimal fermentation time for ACWE was 24 hr to achieve 100% suppression, with increased time resulting in inconsistent suppression results. Optimal fermentation time for NCWE was 3 days or 8 days. These studies are important contributions to understanding the differences that might be expected in CWE suppression when growing in different substrates, some of the factors in the production of CWE that affects the ability of a CWE to suppress disease, and the phenotypic effect CWE has on the root zone of plants and the possible relationship between that effect and disease suppression.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Aiming the discovering of new products with less impact to the environment, there were made experiments aiming to evaluate the efficiency on control, as well as the feeding inibit result on soya caterpillar (Anticarsia gemmatalis).with different vegetation stratum in different commercial soya tillings in Uruguaiana and Santa Maria (RS), from january to March, 2005 under low, medium and high population pressure. There were evaluate nim oil at 5%, watered stratum of cinamomum at 1/6 (p/v), distilled of Timbó 1/1 (v/v), distilled of araticum 1/1 (v/v) standart treatment (permetrina), dosage of 0,05 L ha-1 and testifying treatment (water). In all the treatments adherent spreading at 0,1% was used. The doses were applied at tilling, of soya plants and there was noticed that the concentrations experienced had some insecticide activity on Anticasia gemmatalis causing high mortality. The mortality changes with the use of the different stratum grub state of the insect, population pressure and weather conditions of the place. There was noticed that the leaf eat away by caterpillars under the effect of stratum tested was little when compared with, those of tistiled treatment, even when made on high population pressure.
Objetivando a descoberta de novos produtos que sejam menos impactantes ao meio ambiente, foram conduzidos experimentos com o objetivo de avaliar a eficiência de controle de Anticarsia gemmatalis e nível de dano, em diferentes lavouras comerciais de soja, nos municípios de Uruguaiana e Santa Maria (RS), no período de janeiro a março de 2005 sob pressão populacional baixa, média e alta. Foram avaliados o óleo de nim a 5%, extrato aquoso de cinamomo 1/6 (p/v), destilado de timbó 1/1 (v/v), destilado de araticum 1/1 (v/v), tratamento padrão (permetrina), na dose de 0,05L ha-1 e tratamento testemunha (água). Em todos os tratamentos utilizou-se espalhante adesivo 0,1%. As doses foram aplicadas a campo, sobre as plantas de soja, e verificou-se que as concentrações testadas exerceram acentuada atividade inseticida sobre A. gemmatalis, provocando elevada mortalidade, porém, esta variou de acordo com o extrato testado, estádio larval do inseto, pressão populacional e condições meteorológicas do local. Verificou-se que o consumo foliar por lagartas submetidas à ação dos extratos testados foi reduzido, quando comparados àquelas do tratamento testemunha, mesmo quando realizado sob pressão populacional alta.

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas
A mudança de hábitos e costumes da sociedade que tem vindo a acontecer, principalmente nos últimos anos, conduziu a uma alteração dos padrões de alimentação e a um maior sedentarismo, o que resultou numa preocupação generalizada, por parte dos cidadãos, em tentar minorar problemas de saúde decorrentes deste estilo de vida. Como resposta a esta situação, a indústria alimentar reforçou a sua política de investigação, surgindo um novo tipo de alimentos, denominados alimentos funcionais. O interesse por parte dos consumidores por estes alimentos tem vindo a crescer, pois para além da função nutricional básica que possuem, apresentam benefícios para a saúde humana, tornando-se portanto um mercado bastante promissor. No nosso país existe já uma variedade de alimentos funcionais, como ovos enriquecidos com ácidos gordos ómega-3, tomate rico em licopeno, leite ou iogurtes com cálcio adicionado, entre outros. Portugal é um importante produtor de vinho, com as exportações a aumentar, o que demonstra o dinamismo do setor. O vinho e a vinha têm, na sua constituição, compostos fortemente antioxidantes e com um amplo espetro de propriedades biológicas passíveis de serem extraídas. Os componentes fenólicos são um exemplo de compostos bioativos de grande relevância, apresentando uma variedade de propriedades fisiológicas, tais como antioxidante, cardioprotetora, anticarcinogénica, antiinflamatória, antienvelhecimento, antimicrobiana. Por conseguinte, o interesse por parte da indústria alimentar por estas propriedades tem vindo a aumentar, assim como o dos consumidores, que cada vez são mais exigentes. Desta forma, pretende-se, para além de realizar uma revisão bibliográfica sobre alimentos funcionais no geral, conhecer a composição dos constituintes extraídos do vinho de uva ou da videira, que possam ser incorporados em alimentos já existentes, de forma a trazer benefícios para a saúde e bem-estar do individuo. De acordo com isto e com o desafio final deste trabalho, propôs-se um alimento funcional, mais concretamente, uma massa enriquecida com antioxidantes extraídos do vinho de uva ou da videira, assim como toda a conjuntura associada à extração e à quantificação dos compostos fenólicos dos extratos, o processo de desenvolvimento da massa e a avaliação da sua composição. The changing habits and mores of society that has been happening, especially in recent years led to a change in the eating patterns and to a more sedentary way of life, which resulted in a widespread concern from the citizens who try to minimize the health problems caused by this lifestyle. In response to this situation, the food industry reinforced its research policy and a new type of food emerged, the so-called functional food. The consumers show an increasing interest for this kind of food because, apart from its basic nutritional function, it brings benefits to human health, thus becoming a very promising market. In our country there are already a variety of functional foods as eggs fortified with omega-3, tomato rich with lycopene, milk or yoghurt with calcium added, among others. Portugal is an important wine-producing country; the exports are increasing, hence demonstrating the sector’s dynamism. The wine and the vine have in their constitution strong antioxidant compounds and a broad spectrum of biological properties that can be extracted. The phenolic compounds are examples of highly relevant bioactive compounds, showing a variety of physiological properties, such as antioxidant, cardioprotective, anti-carcinogenic, anti-inflammatory, anti-aging, and antimicrobial properties. Therefore, the interest from both the food industry and the consumers - who are more and more demanding - is increasing. Overall, this dissertation aims not only at performing a bibliographic revision on functional food as a whole, but also at getting to know the composition of the compounds extracted from the grape wine or the vine which may be added into existing food, so to bring benefits for the individuals’ health and well-being. According to this purpose and to the ultimate challenge of this dissertation, a functional food is suggested, a pasta enriched with antioxidants extracted from grape wine or vine, as well as the whole circumstances associated with the extraction and the quantification of the phenolic compounds of the extracts, the process to develop the pasta and the evaluation of its composition.

Existe um número muito limitado de estudos referentes à produção de componentes antimicrobianos ou bacteriocinas produzidas por espécies de bifidobactérias. Nesse âmbito, o objetivo deste trabalho foi avaliar a produção de bifidobacteriocina em leite desnatado (LD). Para tanto, o estudo foi dividido em três etapas. A primeira etapa constituiu na preparação dos meios de cultura Man, Rogosa e Sharpe (MRS), Bifidus Selective Medium (BSM) e LD suplementado com 1% (p/v) de Tween 80 (T80), Inulina (I) ou Extrato de levedura (YE). Nesta etapa, os processos fermentativos foram conduzidos em shaker, nas condições: 50 rpm/37ºC/48h. Foram realizadas análises de pH, concentração de açúcares e ácidos, crescimento celular e determinação da atividade da bifidobacteriocina pelo método de difusão em ágar contra L. monocytogenes. Na segunda etapa, e baseado nos resultados obtidos, foi desenhado um delineamento composto central (CCD) construído a partir dos seguintes parâmetros: temperatura (34, 37, 40 °C) e concentração de YE (0,5; 1,0; 1,5 g/L). Na terceira etapa do trabalho, foram realizados os cultivos em biorreator de 2 L, contendo 10% de leite desnatado, nas seguintes condições: 200 rpm, 36°C, 2,0 g/L de YE, 48h de incubação em anaerobiose. Obteve-se em LD suplementado com YE (1%), combinado ao método de difusão em placa modificado (prévia refrigeração das placas por 12h), contra L. monocytogenes (2130 AU/mL), com uma fase exponencial de 24h, µm de 0,604/h. A otimização feita através do CCD permitiu atingir níveis de atividade de 3.000 AU/mL a 3.100 AU/mL (ensaios 7, 11 e 14, blocos 3 e 1) contra L. monocytogenes, em condições ótimas de crescimento de YE: 2,0 g/L1 e T°C: 36°C. A análise de regressão mostrou ser estatisticamente significativa a relação entre as variáveis: \"concentração de \"YE e temperatura\". Os resultados indicaram que o leite desnatado é um meio adequado para produção de bifidobacteriocina.
There are few publications that have been reported about bacteriocin production by Bifidobacterium species. Therefore, the aim of this work was measure bacteriocin production in skim milk by B. lactis. Consequently, this work was divided in three stages. First, MRS, BSM and LD medium were tested with additives (Tween 80 (T80), Inuline (I) or Yeast extract (YE)) for bacteriocin production and cellular growth. Fermentation processes were conducted in shaker under specific conditions: 50 rpm/37ºC/48h. pH; sugars; acids; biomass, and bacteriocin activity against L. monocytogenes, L. plantarum, E. coli, L. sakei e S. aureus strains were analyzed . In the second stage, based on the obtained results, a central composite design (CCD) was created using the parameters: temperature (34, 37, 40 ºC), and concentration of YE (0.5, 1.0, 1.5 g/L). After, the activity was measured by two methods of plates pre-diffusion (cooling and addition of Tween 20). Third step consisted of 2 L bioreactor cultivations containing 10% skim milk diluted in 1.5 L of water (6.5 pH), under 200 rpm, 36 ºC, 2.0 g/L of YE, 48h, under anaerobic condition. Finally, the cultures supplemented with LD and YE (1%) with a modified plate diffusion method (cooling plates for 12 h) showed bacteriocin activity against L. monocytogenes (2130 AU/mL) with an exponential phase of 24 h, µm of 0.604/h. The optimization performed using CCD resulted in a higher level of activity 3000 AU/mL to 3100 AU/mL mL (Run 7, 11 and 14, blocks 3 and 1) against L. monocytogenes, also with ideal growth conditions of YE: 2,0 g/L1 and T °C: 36 °C. The pH value varied between 6.4 and 4.0. Concentration of produced acid lactic varied from 3.03 to 4.72 g/L and biomass concentration from 3.4 to 11.1 Lg UFC/mL. Regression analysis was significant to the variables: YE concentration and temperature. Results indicated that skim milk is a proper medium for \"Bifidobacteriocin\" production.

As plantas medicinais correspondem às mais antigas estratégias empregadas pelo homem no tratamento de enfermidades de todos os tipos. Devido à grande diversidade vegetal com potencial terapêutico, ao histórico e à cultura do povo brasileiro, plantas com potencial antioxidante, antidiabético, antimicrobiano e leishmanicida têm sido alvo de estudo por uma gama de pesquisadores. Com base nessas informações, a espécie vegetal Eugenia florida DC. é uma candidata promissora para tais estudos. Desta forma, o presente trabalho teve como objetivos identificar e/ou determinar os compostos químicos presentes no extrato hidroetanólico 70% (v/v) e nos macerados em metanol, acetato de etila, acetona e hexano das folhas da espécie em questão, bem como a avaliação das atividades antirradicalar, antimicrobiana, leishmanicida e antidiabética destes extratos. Inicialmente, fez-se uma triagem fitoquímica preliminar, por meio de cromatografia em camada delgada nos extratos secos, que permitiram a detecção de compostos como flavonoides, taninos, ácidos fenólicos, saponinas e triterpenos. Posteriormente, os extratos foram analisados em espectrômetro de massas, no modo negativo, o que possibilitou a identificação de alguns constituintes químicos da classe dos fenóis, dos terpenoides e das saponinas, baseando-se na massa molecular e nos padrões de fragmentação de algumas das moléculas. As análises químicas quantitativas demonstraram que os extratos hidroetanólico (EB) e o macerado em acetato de etila (M-EtOAc) foram os que apresentaram os maiores teores de compostos fenólicos, enquanto que EB e o macerado em metanol (M-MeOH) foram os que demonstraram conter a maior quantidade de flavonoides e taninos. Na determinação da atividade antirradicalar, o valor de CE50 para EB apresentou-se bem próximo ao obtido com o padrão de quercetina, indicando uma excelente atividade. Os macerados apresentaram atividade antirradicalar bem menor. Nas análises de atividade antimicrobiana, o M-MeOH foi o que se mostrou mais ativo contra as leveduras testadas, mas todos eles foram pouco efetivos para as bactérias usadas. Nenhum dos extratos foi efetivo para inibir o crescimento das formas promastigotas de Leishmania amazonensis utilizadas no teste. Na avaliação dos parâmetros bioquímicos de ratos diabéticos tratados com EB, observaram-se diminuições significativas nos níveis séricos de glicose, frutosamina e triglicerídeos, bem como o aumento dos níveis de colesterol HDL. A administração do EB não apresentou efeitos sobre a função renal, na atividade de AST e ALT e nos demais parâmetros do perfil lipídico. Porém, ocorreu melhora nos danos hepáticos e renais nos animais diabéticos tratados, através da redução do malondialdeído (MDA). Dessa forma, concluiu-se que a espécie E. florida DC. apresenta um potencial biológico interessante, devido, principalmente, aos seus metabólitos secundários bioativos, que demonstraram promissora atividade sobre os parâmetros bioquímicos em modelos de ratos diabéticos.
The medicinal plants correspond to the earliest strategies employed by man in the treatment of diseases of all types. Due to the great plant diversity with therapeutic potential, the history and culture of the Brazilian people, plants with potential antioxidant, antidiabetic, antimicrobial and leishmanicidal have been the study by a range of researchers. Based on this information, the plant species Eugenia florida DC. is a promising candidate for such studies. Thus, this study aimed to identify and/or determine the chemical compounds present in hydroethanolic extract 70% (v/v) and macerated in methanol, ethyl acetate, acetone and hexane of species leaves in question, and evaluating the antiradical, antimicrobial, antidiabetic and leishmanicidal activities. Initially, there was a preliminary phytochemical screening by means of thin layer chromatography in dry extracts, which allowed the detection of compounds such as flavonoids, tannins, phenolic acids, terpenoids and saponins. Subsequently, the extracts were analyzed in a mass spectrometer in negative mode, which allowed the identification of some chemical constituents of phenolics, terpenoids and saponins, based on the molecular weight and fragmentation mechanisms of some of the molecules. Quantitative chemical analysis showed that hydroethanolic extracts (EB) and macerated in ethyl acetate (M-EtOAc) were those with the highest levels of phenolic compounds, while EB and macerated in methanol (M-MeOH) were the demonstrated that contain the greater amount of flavonoids and tannins. In determining the antiradical activity, the EC50 value for EB presented very close to that obtained with standard quercetin, indicating an excellent activity. The macerated showed much lower antiradical activity. In the analysis of antimicrobial activity, the M-MeOH was more active against the yeast tested, but they were ineffective for bacteria used. None of the extracts was effective to inhibit the growth of promastigotes of Leishmania amazonensis used in the test. In evaluating the biochemical parameters of diabetic rats treated with EB, there were significant decreases in serum glucose, fructosamine and triglyceride levels and increasing HDL cholesterol levels. EB's administration has had no effect on renal function, in AST and ALT activity and other parameters of lipid profile. However, there was an upgrade in liver and kidney damage in diabetic animals treated through reduction of malondialdehyde (MDA). Thus, it was concluded that the species E. florida DC. presents an interesting biological potential, mainly due to its bioactive secondary metabolites, which have demonstrated promising activity on biochemical parameters in models of diabetic rats.
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES

Orientador: Roberto Herminio Moretti
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Resumo: A elaboração de extrato por maceração de madeira de carvalho triturada com destilado alcoólico simples de cana-de-açúcar (DASCA) para maturação de aguardente de cana mostra vantagens de processo e econômicas por racionalizar a utilização e otimizar a extração de compostos da madeira, reduzir o tempo de maturação da bebida, prescindindo da utilização de tonéis de carvalho. A utilização de embalagens de PET mostra viabilidade para o acondicionamento e/ou maturação de destilados em substituição aos tonéis de madeira tradicionalmente utilizados, por suas características de integridade estrutural, permeabilidade seletiva e inocuidade com a bebida. Foram obtidos 13 extratos por macerações contínuas em batelada a partir de mesma quantidade de madeira de carvalho triturada, nas formas tostada e não tostada, com DASCA de teor alcoólico de 55% em volume, a 20°C. O extrato de ca da maceração foi estudado quanto ao extrato seco, compostos fenólicos totais, pH, acidez volátil, fixa e total e cor. A unificação dos extratos integrais das 5 primeiras macerações originaram um extrato resultante do qual foram obtidas as dosagens de 20, 40 e 60mL utilizadas na maturação de aguardente de cana em embalagens de PET de 0,250, 2 e 20L e vidro de 4,7L, durante 4, 8 e 12 meses. Aguardente de cana sem extrato foi maturada paralelamente em barris de carvalho de 200L, durante 4, 8, 12, 36 e 48 meses. A maturação da bebida foi acompanhada com o estudo de perda de massa, teor alcoólico real, pH, acidez volátil, fixa e total, compostos fenólicos totais, cor, álcoois superiores (isoamílico, isobutílico e n-propílico), aldeídos e ésteres totais em cada período de maturação correspondente. As aguardentes de cana maturadas em embalagens de PET, vidro e barris de carvalho, em cada período de maturação, incluindo o tempo zero, foram submetidas ao teste de aceitação com 81 consumidores, avaliando-se os atributos cor, aroma, sabor e impressão global, utilizando escala hedônica não estruturada de 9cm. Os resultados das análises físico-químicas e sensorial de aceitabilidade foram tratados com análise de variância e comparação de médias pelos testes de Duncan (p£0,05) e Tukey (p£0,10), respectivamente. O atributo impressão global da análise de aceitação foi complementarmente analisado pelo método Mapa de Preferência Interno (MDPREF). O extrato resultante originado da mistura dos extratos integrais das cinco primeiras macerações (acumulando 41 dias de maceração), no contexto de 13 macerações (acumulando 346 dias de maceração), resultou na extração de 74,5% de extrato seco, 67,5% de compostos fenólicos totais, 83,9% de acidez fixa e 67,7% de acidez total. A maturação da aguardente de cana pela incorporação de 20, 40 e 60mL deste extrato/L de aguardente de cana alterou significativamente (p£0,05) o perfil físico-químico e sensorial da bebida, agregando cor e compostos fenólicos totais, reduzindo o pH, incrementando a acidez total, volátil e fixa e a composição orgânica de aldeídos e ésteres totais e de álcoois superiores isoamílico, isobutílico e n-propílico. O período de maturação de 12 meses em embalagens de PET de 0,250, 2 e 20L alterou o perfil físico-químico da aguardente de cana, diminuindo a acidez volátil e total e de aldeídos totais, incrementando a acidez fixa e o teor de ésteres totais. O aumento de volume das embalagens de PET refletiu em menores perdas de massa e maiores incrementos de ésteres totais. A aguardente de cana maturada 12 meses em embalagens de PET e vidro com a incorporação de extrato de carvalho mostrou similar aceitabilidade pelos consumidores, em relação à bebida maturada em barris de carvalho, entre 12 até 48 meses. A incorporação na aguardente de cana de dosagens de extrato de 20, 40 e 60mL/L determinou incrementos significativos (p£0,10) de aceitabilidade da bebida maturada em embalagens de PET e vidro, independentemente do tempo de maturação. O atributo cor, originado pela incorporação de distintas dosagens de extrato à aguardente de cana, foi o de melhor aceitabilidade pelos consumidores, seguido pelo aroma, impressão global e sabor
Abstract: The elaboration of extract through maceration of brazilian sugar-cane spirit (BSCS) with oak wood grinded for brazilian sugar-cane spirit maturation shows advantages of process as well as economic ones, such as rationalization of the utilization, optimization of wood compounds extraction, reduction of the maturation time of the drink, renouncing the utilization of oak wood barrels. The utilization of PET packages shows its viability for storage and/or maturation of distilled beverages in substitution to the traditionally used wood structures for its structural integrity characteristics, selective permeability and innocuousness with the content. It was obtained 13 oak extracts by sequential units macerations using fixed quantity of toasted and not toasted grinded wood with BSCS 50°GL/20°C. Each extract was studied with the dry extract, total phenolic compounds, fixed, volatile and total acidity and colour. The unification of the integral units extracts from the five initial macerations produced the resultant extract for which were obtained the 20, 40, 60mL dosages for BSCS maturation in 0.250, 2, and 20L PET packages and in 4,7L glass packages during 4, 8 and 12 months. This study went along with the maturation of BSCS in 200L oak barrels during 4, 8, 12, 36 and 48 months. The maturation process of BSCS was physical-chemically accompanied by the study of mass losses, real alcoholic level, fixed, volatile and total acidity, total phenolic compounds, colour, superior alcohols (isoamilic, isobutilic, and n-propilic), total esters and aldehydes, in each specific times, included at zero time. An acceptance test also took place involving 81 BSCS consumers through their manifestation concerning the attributes colour, aroma, flavour and global impression using a nonstructured 9cm hedonic scale. The physical-chemical and acceptance tests results were statistically treated with variance analysis and Duncan (p=0.05) and Tukey (p=0.10) tests for average comparisons, respectively. The global impression attribute of the acceptance test was supplementary treated by Multidimensional Preference Analysis (MDPREF) method. The resultant extract obtained of the mixture of the five initial units macerations (totalizing 41 days of maceration) in context of thirteen units macerations (totalizing 346 days of maceration) resulted in 74.5% dry extract, 67.5% total phenolic compounds, 83.9% fixed acidity and 67.7% total acidity extraction. The incorporation of 20, 40, and 60mL of this extract per liter for BSCS maturation were significant effect (p£0.05) on physical-chemical profile of the beverage, associating to it colour and total phenolic compounds, reducing pH, adding volatile and fixed total acidity, the organic composition of superior alcohols, total esters and aldehydes. At 12 months BSCS maturation time in 0.250, 2 and 20L PET packages altered the physical-chemical profile of the beverage, decreasing the total and volatile acidity and total aldehydes and increasing the fixed acidity and total esters. The volume increase in PET packages reflected smaller indexes of mass losses and larger increments of total esters. The BSCS matured in PET and glass packages with the incorporation of oak extract during 12 months showed similar acceptability by consumers, in relation to the BSCS matured in oak barrels, during 12 to 48 months. The incorporation of the 20, 40, and 60mL oak extract dosage per liter for BSCS maturation determined significant increments (p=0.10) of acceptance of the BSCS matured in PET and glass packages independently of maturation time. The colour attribute originated by the distinct dosages of extract was the better acceptance by the consumers, followed by aroma, global impression and flavour
Doutorado
Tecnologia de Alimentos
Doutor em Tecnologia de Alimentos

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES
Although the Acacia mearnsii tree is widely cultivated and used in the industry, its flowers are seldom researched. Considering the extensive availability of A. mearnsii flowers and the growing interest in natural products, this work aimed at the obtainment of extracts from these flowers and investigation of their composition and biochemistry properties. In this work, essential oil and aqueous extract by steam distillation and extracts were obtained with supercritical fluid. The essential oil was characterized by CG-MS and olfactometry. The main compounds identified in the essential oil were 8-heptadecene, heptadecane, nonadecane e kaurene. The main aroma detected by olfactometric analysis was roast nutty, as well as coffee, sweet, floral, vegetal and spicy. Antioxidant activity was determined and the aqueous extract showed an activity superior to rutin standard. LC-qTOF-MS identified 42 likely phenolic compounds, such as quercitrin, naringenin, myricetin, rutin, and robinetin. The extracts obtained by supercritical extraction and isolated fractions presented activity against S. aureus. The yield of the supercritical extraction was optimized using factorial planning and mass transfer parameters were obtained through mathematical models that represent the extraction curve. Thus, it was concluded that the flowers of A. mearnsii can be exploited by extractive processes used in this study to generate a plant by-product. The results indicate the aromatic and antioxidant potential of the essential oil and aqueous extract, respectively, obtained with 2.0 bar steam distillation, while the supercritical extract has antibacterial action.
A Acacia mearnsii, apesar de ser amplamente cultivada e utilizada industrialmente, n?o tem o potencial de suas flores explorado. Considerando a disponibilidade de flores da A. mearnsii e o crescente interesse por produtos naturais, esse trabalho consistiu na obten??o de extratos destas flores e na investiga??o de suas composi??es e a??es bioqu?micas. A pesquisa compreendeu a extra??o do ?leo essencial e a obten??o do extrato aquoso de flores da planta via destila??o por arraste a vapor, al?m da extra??o com fluido supercr?tico. O ?leo essencial foi analisado por GC-MS e apresentou, entre os 31 compostos identificados, 8-heptadeceno, heptadecano, nonadecano e caureno como compostos majorit?rios. Na an?lise olfatom?trica (GC-O), os principais aromas detectados no ?leo essencial foram caf?, doce, floral, vegetal e refrescante, com destaque para ?castanha/defumado?. O extrato aquoso das flores da A. mearnsii apresentou atividade antioxidante superior ? da rutina. Utilizando cromatografia de alta resolu??o (LC-qTOF/MS), foram verificados 42 prov?veis compostos fen?licos, entre eles, quercitrina, naringenina, miricetina, rutina, robinetina. Os extratos obtidos via extra??o supercr?tica e fra??es isoladas apresentaram atividade frente a S. aureus. O rendimento da extra??o supercr?tica foi otimizado utilizando planejamento fatorial e par?metros da transfer?ncia de massa foram obtidos atrav?s de modelos matem?ticos que representam a curva de extra??o. Assim, concluiu-se que as flores de A. mearnsii podem ser exploradas pelos processos extrativos empregados nesse estudo a fim de gerar um co-produto da cadeia produtiva da planta, uma vez que o ?leo essencial e o extrato aquoso, obtidos pela extra??o por arraste a vapor a 2,0 bar, possuem, respectivamente, potencial arom?tico e poder antioxidante, enquanto o extrato obtido com fluido supercr?tico apresenta a??o antibacteriana.

Conselho Nacional de Desenvolvimento Científico e Tecnológico
Mortadella is one of the meat products with good nutritional value and excellent consumer acceptance. In addition to protein and lipid source, a plethora of non-meat ingredients have been used in the preparation of emulsified products. The use of natural compounds to increase the shelf life of meat products is a promising alternative, since many plant substances possess antioxidant and antimicrobial properties. The ora-pro-nobis is considered a nutritional supplement because of its protein content, fiber, iron, calcium, among others. This study aimed to obtain extracts from the leaves of ora-pro-nobis and evaluate the antioxidant and antimicrobial activity, in addition, apply the extract with antioxidant activity in bologna, aiming as natural antioxidant potential. First, it applied the chemical and mineral composition of the leaves of ora-pro-nobis. Extracts were obtained by conventional stirring, varying the solvent (water and ethanol), time (1 and 24h) and temperature (95 °C and 25 °C). In the extracts were carried out analysis of total phenolics, total flavonoids, antimicrobial and antioxidant activity in vitro through the DPPH methods, FRAP and ABTS radical ● +. The extract obtained by extraction by shaking using distilled water as a solvent and 95 °C showed the best antioxidant properties, so it has been applied in different concentrations in bologna. After the finished product were carried out physico-chemical, sensory and microbiological analysis. The bologna presented within the requirements of legislation for both the chemical composition and for microbiological stability. They showed no oxidative changes during storage. He observed a tendency of increasing the pH of all treatments during the storage time. The mortadella added 0.2% sodium erythorbate and 2.0% extract showed the lowest lipid oxidation after 70 days of storage. The sensory evaluation showed a good acceptance by consumers. It can be concluded that the extract from the leaves of ora-pro-nobis presented with antioxidant potential to be tapped by carnea industry.
A mortadela é um dos produtos cárneos com bom valor nutritivo e excelente aceitação pelo consumidor. Além da fonte proteica e lipídica, uma infinidade de ingredientes não cárneos tem sido utilizados na elaboração dos produtos emulsionados. O uso de compostos naturais para aumentar a vida útil dos produtos cárneos é uma alternativa promissora, uma vez que muitas substâncias vegetais possuem propriedades antioxidantes e antimicrobianas. A ora-pro-nóbis é considerada um complemento nutricional devido ao seu conteúdo proteico, fibras, ferro, cálcio, dentre outros. Este estudo teve como objetivo obter extratos das folhas de ora-pro-nóbis e avaliar a atividade antioxidante e antimicrobiana, além disso, aplicar o extrato com maior atividade antioxidante em mortadelas, visando como potencial antioxidante natural. Primeiramente, foi realizada a composição química e mineral das folhas de ora-pro-nóbis. Os extratos foram obtidos por agitação convencional, variando o solvente (água e etanol), tempo (1 e 24h) e temperatura (95 °C e 25 °C). Nos extratos obtidos foram realizadas análises de compostos fenólicos totais, flavonoides totais, atividade antimicrobiana e atividade antioxidante in vitro através dos métodos DPPH, FRAP e radical ABTS●+. O extrato obtido através de extração por agitação utilizando água destilada como solvente e temperatura de 95 °C foi o que apresentou as melhores características antioxidantes, então o mesmo foi aplicado em diferentes concentrações em mortadelas. Após o produto acabado foram realizadas as análises físico-químicas, sensoriais e microbiológicas. As mortadelas apresentaram dentro dos requisitos da legislação tanto para a composição centesimal quanto para estabilidade microbiológica. Não apresentaram alterações oxidativas durante o armazenamento. Observou uma tendência de aumento nos valores de pH de todos os tratamentos, durante o tempo de armazenamento. As mortadelas adicionadas de 0,2% de eritorbato de sódio e 2,0% de extrato foram as que apresentaram menor oxidação lipídica após os 70 dias de armazenamento. A avaliação sensorial demonstrou uma boa aceitabilidade pelos consumidores. Pode-se concluir que o extrato das folhas de ora-pro-nóbis apresentou com potencial antioxidante a ser aproveitado pela indústria cárnea.

Cette thèse porte sur la caractérisation structurale de nouveaux polyphénols présents dans le Sorghum. A partir d’un extrait, nous avons mis au point un protocole conduisant à leur fractionnement, à leur séparation et à leur détection. L’utilisation de techniques analytiques comme la chromatographie liquide haute performance (HPLC) couplée à la spectroscopie UV-visible et à la spectrométrie de masse (MS) nous a ainsi permis de détecter de nombreux flavonoïdes, dont certains sont connus mais dont la plupart ont des structures nouvelles. L’isolement par HPLC, puis l'identification par MS et RMN 1D et 2D ont permis de caractériser trois composés avec une structure rétrodihydrochalcone. De la même manière, une série de trois pigments phénoliques appartenant à la classe des pyrano-3-déoxyanthocyanidines a été structuralement caractérisée, ainsi qu’un hétéromère coloré à deux unités. Ces composés colorés ont comme unité de base l’apigéninidine, une 3-déoxyanthocyanidine spécifique du Sorghum. L’ensemble de ces polyphénols a été identifié pour la première fois non seulement dans le Sorghum, mais plus généralement dans le monde végétal. Sur la base des informations issues des fragments caractéristiques obtenus par MSn pour les flavonoïdes élucidés, et ensuite, à partir des fragments de composés inconnus obtenus par LC/MSn, il nous a été possible de proposer des structures précises pour neuf nouveaux flavonoïdes présents dans l’extrait de Sorghum. Par ailleurs, la détection d’une nouvelle série d’hétéromères de flavonoïdes de trois à six unités a pu être réalisée grâce à l’utilisation de la spectrométrie de masse par MALDI/TOF. La grande diversité des polyphénols dans le Sorghum et l’identification structurale de ces nouveaux flavonoïdes pourraient éventuellement servir de base pour une utilisation accrue du Sorghum dans les industries alimentaire, cosmétique et pharmaceutique
This thesis is centred on the structural characterization of new polyphenols in Sorghum plant. From an extract, we have developed a protocol leading to their fractionation, separation and detection. The use of analytical techniques such as high performance liquid chromatography (HPLC) coupled with UV-Visible spectroscopy and with mass spectrometry (MS), allowed us to discern numerous flavonoids, most of them unknown. The isolation by HPLC, the identification by MS and RMN 1D and 2D allowed the characterisation of a series of retrodihydrochalcones. In the same way, a series of three phenolic pigments, belonging to the pyrano-3-deoxyanthocyanidins class, have being structurally characterized along with a two units coloured heteromer. These coloured compounds have as basic unit apigeninidin, a 3-deoxyanthocyanidin specific of the Sorghum plants. All these polyphenols have been identified for the first time in Sorghum and in plant in general. The information obtained from MSn fragmentation of identified compound, along with the data obtained from analysis of the Sorghum extract by LC/MSn, have allowed a precise structural determination for nine novel flavonoids. Moreover, the analysis by MALDI/TOF mass spectrometry has led to the detection of a new series of flavonoids heteromers. The wide diversity of polyphenols in Sorghum and the structural identification of new products could eventually lead to an improve use of Sorghum in food, cosmetic and pharmaceutical industries

Orientador: Adriana Zerlotti Mercadante
Texto em português e Inglês
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Resumo: Neste trabalho foi avaliada a capacidade antioxidante de carotenoides em sistema homogêneo e em lipossomas, de carotenoides microencapsulados e de extratos de maná-cubiu (Solanum sessiflorum) frente às espécies reativas de oxigênio (ROS) e espécies reativas de nitrogênio (RNS) de importância biológica. Devido à insolubilidade de carotenoides em solução tampão, utilizada como meio de reação no conhecido método oxygen radical absorbance capacity (ORAC), foi desenvolvido também um novo método para determinar a capacidade de carotenoides de desativarem o radical peroxila (ROO). Em geral, as microcápsulas de goma arábica e de maltodextrina (20 DE) contendo compostos antioxidantes (trolox, -tocoferol, ß-caroteno, apo-8¿-carotenal e apo-12¿-carotenal) foram capazes de desativar as ROS e RNS, sendo a eficiência de desativação influenciada pelo material de parede, pela espécie reativa (ROO, peróxido de hidrogênio (H2O2), radical hidroxila (HO), ácido hipocloroso (HOCl) e ânion peroxinitrito (ONOO-) e pela estrutura do composto antioxidante. Interessantemente, as microcápsulas vazias também foram capazes de desativar as ROS e RNS, sendo as de goma arábica mais eficazes que as de maltodextrina. A incorporação de apo-8¿-carotenal promoveu o maior aumento na capacidade das microcápsulas de desativarem ROS e RNS, variando de 50% a 132% e de 39% a 85% para goma arábica e maltodextrina, respectivamente. Este fato sugere que o apo-8¿-carotenal apresenta o melhor balanço entre a sua localização no interior das microcápsulas e a reatividade frente às espécies reativas. A localização também foi um fator importante na eficiência dos carotenoides desativarem ROS e RNS em lipossomas. Neste sistema, os carotenoides com grupos hidroxila foram geralmente mais potentes na desativação do ROO, HO e HOCl que carotenos, com destaque para a astaxantina, enquanto o ß-caroteno foi mais eficiente na desativação do ONOO-. Para o estudo dos carotenoides em sistema homogêneo foi desenvolvido e validado com sucesso um novo método para avaliação da capacidade de carotenoides de desativarem o ROO, o qual é baseado na perda de fluorescência da sonda ácido 4,4-difluoro-5-(4-fenil-1,3-butadienil)-4-bora-3a,4a-diazo-s-indaceno-3-undecanoico (C11-BODIPY581/591) devido à oxidação pelo ROO, que é gerado pela termodecomposição do azobisisobutironitrila (AIBN). A aplicação deste novo método permitiu o estudo da relação entre a estrutura química de diferentes carotenoides e a capacidade de desativar o ROO. Foi demonstrado que neste sistema os carotenoides são potentes desativadores desta espécie reativa, sendo a eficiência influenciada principalmente pela abertura do anel -ionona e pela extensão do cromóforo. O licopeno foi o mais potente desativador de ROO (8,67 0,74), porém não tão eficiente quanto os carotenoides do maná-cubiu (9,80 0,80). Os compostos bioativos de maná-cubiu foram determinados por cromatografia líquida de alta eficiência acoplada aos detectores de arranjo de diodos e espectrômetro de massas (HPLC-DAD-MS/MS). Esta fruta apresentou -caroteno (7,15 g/g) e luteína (2,41 g/g) como carotenoides majoritários. Por fim, o extrato fenólico de maná-cubiu, contendo o ácido 5-cafeoilquínico (1298 g/g) como composto fenólico principal, apresentou grande eficiência em desativar o H2O2 (IC50 = 305 17 g/mL) e o HOCl (IC50 = 13 0,8 g/mL). Os resultados do presente estudo mostram que os carotenoides foram capazes de desativar todas as ROS e RNS de relevância biológica avaliadas neste trabalho, sendo a eficiência de desativação influenciada pela concentração e estrutura química do carotenoide, pelo nível de organização do sistema (homogêneo ou multifásico) e pelo tipo de espécie reativa
Abstract: In the present study, the antioxidant capacity of carotenoids in homogeneous system and in liposomes, of microencapsulated carotenoids and of extracts from mana-cubiu (Solanum sessiflorum) were evaluated against reactive oxygen (ROS) and reactive nitrogen (RNS) species of biological relevance. Due to the insolubility of carotenoids in aqueous buffers, which is used as reaction medium in the well known oxygen radical absorbance capacity method (ORAC), a novel method was also developed to determine the peroxyl radicals (ROO) scavenging capacity of carotenoids. In general, the gum arabic and maltodextrin (20 DE) microcapsules containing antioxidant compounds (trolox, -tocopherol, ß-carotene, apo-8¿-carotenal and apo-12¿-carotenal) were capable to scavenge ROS and RNS. The scavenging efficiency of the microcapsules was influenced by the wall material, the reactive species (ROO, hydrogen peroxide (H2O2), hydroxyl radical (HO), hypochlorous acid (HOCl) and peroxynitrite anion (ONOO-) and the structure of the antioxidant compound. Interestingly, the empty microcapsules were also able to scavenge the ROS and RNS, being the gum arabic microcapsules more efficient than the maltodextrin ones. The incorporation of apo-8¿-carotenal resulted in the largest increase in the microcapsules scavenging capacity, varying from 50% to 132% and from 39% to 85% in the gum Arabic and maltodextrin microcapsules, respectively. These findings suggest that the apo-8¿-carotenal presented the best balance between its location in the microcapsules interior and the reactivity against the reactive species. The location was also an important factor influencing the efficiency of carotenoids to scavenge ROS and RNS in liposomes. In this system, the carotenoids with hydroxyl groups were usually more potent ROO, HO and HOCl scavengers than the carotenes, especially astaxanthin, whilst ß-carotene was the most efficient ONOO- scavenger. To study the carotenoids in homogeneous system, a new ROO scavenging assay, based on the loss of fluorescence of the probe 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid (C11-BODIPY581/591) due to its oxidation induced by ROO generated by the thermo decomposition of azobisisobutyronitrile (AIBN), was successfully developed and validated. This new method allowed the establishment of the relationship between the structure of different carotenoids and their ROO scavenging capacity. In this system, carotenoids showed to be potent scavengers of this reactive species, and their scavenging capacity was influenced mainly by the opening of the -ionone ring and by the chromophore extension. Lycopene was the most potent ROO scavenger (8.67 0.74); however, it was not so efficient as the carotenoids from mana-cubiu (9.80 0.80). The bioactive compounds of mana-cubiu were determined by high performance liquid chromatography coupled to diode array and mass spectrometry detectors (HPLC-DAD-MS/MS). The major carotenoids of this fruit were -carotene (7.15 g/g) and lutein (2.41 g/g). Finally, the main phenolic compound found in the phenolic extract from mana-cubiu was 5-caffeoylquinic acid (1298 g/g), which showed to be a good H2O2 (IC50 = 305 17 g/mL) and HOCl (IC50 = 13 0.8 g/mL) scavenger. The results of this thesis showed that the carotenoids are able to scavenge all the evaluated ROS and RNS of biological relevance and that their efficiency depends on the carotenoid structure and concentration, level of organization of the reaction medium (homogeneous or multiphase) and type of reactive species
Doutorado
Ciência de Alimentos
Doutor em Ciência de Alimentos

La contamination des aliments par les mycotoxines est une source très importante de gaspillage de nourriture. Depuis des années, la stratégie classique pour lutter contre ces contaminants est l’utilisation de pesticides. Cependant, il a été montré que ces produits pouvaient avoir des effets nocifs sur la biodiversité et la santé humaine et animale. Il est donc nécessaire de développer de nouvelles stratégies, plus écologiques, pour lutter contre les mycotoxines. L’utilisation de produits naturels pourrait représenter une alternative intéressante et plus respectueuse de l’environnement. En effet, certains produits naturels sont capables d’inhiber la production de mycotoxines. Cependant, le mécanisme d’action mis en jeu est souvent mal compris. Un des objectifs principaux de ce travail a consisté à identifier des produits naturels capables d’inhiber la production de mycotoxines et d’élucider leur mécanisme d’action moléculaire. Pour ce faire, un outil moléculaire a été conçu pour comprendre les mécanismes permettant à des produits naturels d’inhiber la production d’Aflatoxine B1 (AFB1) chez Aspergillus flavus. L’étude de cette mycotoxine est importante puisque c’est un cancérigène puissant chez l’homme et les animaux. Un outil moléculaire permettant l’analyse simultanée de l’expression de 60 des principaux gènes impliqués dans la synthèse de l’aflatoxine B1 par q-*-PCR a été développé. Il permet d’étudier l’ensemble des gènes du cluster d’AFB1 mais également 33 autres gènes codant pour des facteurs de régulation liés à l’environnement dans lequel se trouve la moisissure. Par cette approche, le mécanisme d'action moléculaire de l’eugénol et la pipérine, ainsi que celui de 3 extraits naturels de plantes a été identifié et l'impact de ces composés sur les gènes impliqués dans la production d'AFB1 a été élucidé. L'un des principaux résultats de cette étude a été de montrer que les différents produits naturels modulent systématiquement plusieurs gènes au cours de l'inhibition de la production d’AFB1. Notre étude montre que les produits naturels représentent une alternative possible pour limiter la contamination des aliments par l’AFB1. L'élucidation du mécanisme d'action des produits naturels ouvre de nouvelles perspectives sur les méthodes à employer pour inhiber la production de la toxine
Mycotoxin’s contamination represents an important source of food spoilage that has to be taken in consideration. For years pesticides, have been used as a common strategy to combat mycotoxin contamination. However, such products were also demonstrated to be harmful to humans and animals’ health. Therefore, it is necessary to find new strategies in order to avoid mycotoxin contamination and the use of natural products could be a promising alternative. Indeed, these compounds may be eco-friendly and several of them are demonstrated aseffective agents against toxin production. Nevertheless, their precise mechanism of action is poorly documented. One of the principal aim of this work consisted in the identification of natural sources capable to inhibit mycotoxin production and in the elucidation of their molecular mechanism of action. For that, we developed a molecular tool aiming the analysis of the impact of natural extracts on the expression of several genes related to Aflatoxin B1 synthesis in Aspergillus flavus. The study of this mycotoxin is an important issue since it is one of the most dangerous compounds inducing cancer in humans and animals. Taking advantage of the well-studied genome of Aspergillus flavus and considering that AFB1 production involves a great number of genetic elements, a q-PCR approach including 60 of the principal genes involved in toxin biosynthesis was developed. This tool simultaneously studies the entire AFB1 gene cluster but also 33 regulatory factors coding for external stimuli to which fungus is exposed. Using this molecular approach, the study of already known but also, new sources of anti-aflatoxigenic compounds was performed. The molecular mechanism of action of 2 isolated molecules and 3 whole plant extracts were determined and the impact of these compounds on the genes involved in AFB1 production was analysed. One of the innovative findings consisted in the demonstration that natural products systematically modulate several genes within AFB1’s inhibition. Taken together, our approach demonstrated that the use of natural products against mycotoxin production can represent an alternative strategy to inhibit food contamination. The elucidation of the mechanism of action of natural products allowed a better understanding of the fungal machinery through which toxin can be inhibited

En agriculture biologique, peu de produits sont autorisés pour améliorer la croissance des plantes. Cette étude qualifie les éléments solubles d’extraits de compost (EC) sur la croissance du soya. En présence ou non d’aération, six composts ont macéré dans l’eau pendant 14 jours et caractérisé. Quatre EC ont été sélectionnés, fortifiés et vaporisés sur le feuillage du soya cultivé dans du sable et traité avec de l’eau ou une solution nutritive. Les éléments solubles des EC évoluent en fonction du temps, dont les nitrates qui diminuent en fonction de l’aération. La teneur en éléments minéraux des EC est optimale après 2 jours et sans aération. L’EC «Fumier de vers» a démontré une augmentation du développement, des biomasses, et du prélèvement en éléments minéraux du soya traité avec une solution nutritive. L’effet bénéfique de cet EC semble lié à des substances de croissance solubles, autres que les éléments minéraux solubles.
In organic agriculture, there are few certified products that can be used to favor plant growth. This study was conducted to determine the soluble compounds from water extract of compost (WEC) that could benefit soybean growth. In the presence or the absence of aeration, six composts were macerated into water up to 14 days. Following the characterization of WEC, four were selected, enriched and applied on foliage of soybean grown in sand and watered with a nutrient solution or water. Over time, most soluble mineral salts went into solution, but N could be lost trough denitrification. To enhance mineral contents and minimize N loss, WEC should be produced within two days. When treated with the nutrient solution, the water extract of vermicompost enhanced soybean development, biomasses, and plant mineral contents. This WEC appears to favor plant growth through the provision of soluble growth substances, other than soluble mineral elements.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
As condições ambientais, assim como o contraste sazonal nas fitofisionomias do Cerrado entre a época seca e úmida colaboram para a diversificada sintetize e seleção de metabólitos secundários em plantas. Os metabólitos secundários atuam como uma interface química entre plantas e o meio, desta forma a sua síntese pode ser comumente alterada pelas condições ambientais. Fatores como temperatura, disponibilidade hídrica, luminosidade, características do solo, nutrientes e poluição atuam diretamente sobre a produção e liberação desses compostos. Os microrganismos do solo podem transformar os compostos tóxicos liberados, desativando-os ou dando origem a compostos ainda mais tóxicos, assim como esses compostos podem alterar as características e atributos do solo quando presentes em quantidades adequadas. Dentre as plantas nativas do Cerrado destaca-se a espécie Vernonia ferruginea, uma agressiva infestante de pastagem, que possui indícios de produção de compostos com potencial tóxico. Desta forma, esta pesquisa foi desenvolvida para verificar a atividade tóxica de V. ferruginea e se a mudança de algumas características como disponibilidade hídrica, tipo de solo, condições de luminosidade e concentração de alumínio no solo influenciam a produção desses compostos. Para tal, plantas de V. ferruginea foram desenvolvidas sobre diferentes condições para posterior extração e/ou fracionamento do material vegetal para verificação de atividade biológica por meio de bioensaios e identificação de grupos químicos. Considerando os resultados dos parâmetros de germinabilidade e de desenvolvimento das espécies testadas é possível indagar sobre a possibilidade de V. ferruginea ter a capacidade de liberar substâncias tóxicas, uma vez que, consequências diretas da presença desses compostos nos extratos foram verificadas no bioensaio, como: a inibição/redução da germinação de sementes, injúria ou necrose no sistema radicular e redução do desenvolvimento e do acúmulo de massa seca das plântulas. Os resultados também dão base para constatar que algumas interações da espécie com as alterações nas suas condições de desenvolvimento causam desvios de rotas biossintética de metabólitos modificando e/ou potencializando sua capacidade fitotóxico.
The environmental conditions, as well as the seasonal contrast in the Cerrado phytophysiognomies between the dry and wet seasons, contribute to the diversified synthesis and selection of secondary metabolites in plants. Secondary metabolites act as a chemical interface between plants and the environment, so their synthesis can be commonly altered by environmental conditions. Factors such as temperature, water availability, luminosity, soil characteristics, nutrients and pollution act directly on the production and release of these compounds. Soil microorganisms can transform the released toxic compounds, deactivating them or giving rise to even more toxic compounds, as these compounds can alter the characteristics and attributes of the soil when present in adequate amounts. Among the native plants of the Cerrado, the species Vernonia ferruginea, an aggressive pasture weed, stands out, with indications of production of compounds with toxic potential. Thus, this research was developed to verify the toxic activity of V. ferruginea and if the change of some characteristics such as water availability, soil type, light conditions and aluminum concentration in the soil influence the production of these compounds. For this, plants of V. ferruginea were developed on different conditions for later extraction and / or fractionation of the vegetal material for verification of biological activity by means of bioassays and identification of chemical groups. Considering the results of the parameters of germinability and development of the tested species it is possible to inquire about the possibility of V. ferruginea being able to release toxic substances, since direct consequences of the presence of these compounds in the extracts were verified in the bioassay, such as: The inhibition / reduction of seed germination, injury or necrosis in the root system and reduction of the development and accumulation of dry mass of the seedlings. The results also provide evidence that some interactions of the species with changes in their developmental conditions cause biosynthetic pathway deviations of metabolites modifying and / or potentiating their phytotoxic capacity.
FAPESP: 12/25298-0

SOUSA, Karolina Costa de. Atividades antioxidante, antimicrobiana, antibiofilme e citotóxica de extratos da macroalga marinha parda Lobophora variegata (J. V. Lamouroux). 2017. 99 f. Dissertação (Mestrado em Engenharia de Pesca)-Universidade Federal do Ceará, 2017.
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The diversity of marine macroalgae is directly associated to environmental conditions so that, in adverse circumstances, they can synthesize metabolites for selfprotection. Flavonoids constitute one of the main groups of phenolic compounds. Found in large quantities in plants, they perform defense functions and are potential sources to prevent damages associated with cellular oxidation and to avoid attacks of microorganism. The objective of this work was to investigate the biological activities of dichloromethane (DCM) and 70% ethanol (EtOH 70%) extracts of the brown marine alga Lobophora variegata, collected at Paracuru beach-Ceará. Additionally, the extracts were examined for total phenolic content (TPC), total flavonoid content (TFC) and in-vitro antioxidant activity, measured through DPPH radical scavenging activity (DPPH), ferrous ion chelating activity (FIC), ferric ion reduction power (FRAP), -carotene bleaching (BCB), and also for antibacterial, antifungal, antibiofilm and cytotoxic activities. Oven-dried alga was used for preparation of DCM and EtOH 70% extracts, which were re-suspended in 50% methanol (MeOH 50%) to achieve a concentration of 1000 µg mL-1, which after successive dilutions resulted in concentrations ranging from 500 to 7.8125 µg mL-1. Both antioxidant and cytotoxic activities were determined for the DCM and EtOH 70% extracts at concentrations ranging from 7.8125 to 1000 µg mL-1. Subfractions obtained from column chromatography were analyzed for antimicrobial and antibiofilm activities. The yield of DCM and EtOH 70% extracts were 3.1% and 4.8%, respectively. EtOH 70% extracted the highest quantities of TPC and TFC and showed the highest antioxidant activity. Two subfractions of DCM extract were effective against Escherichia coli (ATCC25922) at 100 and 1000 g mL-1, while three others were effective against Staphylococcus aureus (ATCC25923) at the highest concentration. No subfraction showed antifungal activity against Aspergillus niger nor Saccharomyces cerevisiae. Neither did they contribute to the inhibition of the formation of Pseudomonas spp. biofilm. DCM extract at concentrations of 125, 250 and 500 g mL-1 showed a dose-dependent cytotoxic activity with Artemia sp. nauplii, killing 100% of individuals after 48 h exposure. Cytotoxic activity results for the EtOH 70% extract were not shown because the percentage of dead nauplii in the control was higher than the acceptable maximum of 10%. Lobophora variegata presents different biological activities, and further investigation of its potential should be carried out on subfractions in order to verify their therapeutic efficacy
A diversidade das macroalgas marinhas está diretamente associada com as condições ambientais de modo que, em situações adversas, elas são capazes de sintetizar metabólitos para sua proteção. Dentre estes, os compostos fenólicos são os principais metabólitos secundários sintetizados em resposta a pressões do meio ambiente. Os flavonoides são considerados um dos principais grupos de compostos fenólicos encontrados em abundância nos vegetais, desempenhando funções de defesa, por isso são considerados fontes potenciais para a prevenção de danos associados á oxidação celular e ao ataque de micro-organismos. Uma variedade desses compostos apresenta diversas atividades biológicas, por exemplo, antimicrobiana, antialérgica, antidiabética e antioxidante. O objetivo deste trabalho consistiu em investigar a atividade biológica dos extratos diclorometano (DCM) e etanol a 70% (EtOH 70%) da macroalga marinha parda Lobophora variegata, coletada na Praia do ParacuruCeará, avaliando especificamente o conteúdo de compostos fenólicos totais (CFT), o teor de flavonoides totais (TFT), a atividade antioxidante in vitro, medida pela capacidade de sequestro do radical DPPH, habilidade de quelação de íons ferrosos (FIC), poder de redução de íons férricos (FRAP) e branqueamento do -caroteno (BCB) e as atividades antibacteriana, antifúngica, antibiofilme e citotóxica. A macroalga marinha desidratada foi usada para a preparação dos extratos com DCM e EtOH 70%, os quais foram ressuspendidos em metanol a 50% (MeOH 50%) obtendo-se a concentração de 1.000 g mL-1 que após sucessivas diluições resultaram nas concentrações de 500 a 7,8125 g mL-1. Nos extratos DCM e EtOH 70% com concentração de 7,8125 a 1.000 g mL-1 foram determinadas as atividades antioxidantes e citotóxicas. As subfrações obtidas das cromatografias em coluna foram utilizadas para as atividades antimicrobiana e antibiofilme. Os rendimentos dos extratos DCM e EtOH 70% foram 3,1% e 4,8%, respectivamente. CFT e TFT foram extraídos em maiores quantidades pelo EtOH 70%, que apresentou maior atividade antioxidante. Duas frações do extrato DCM foram efetivas nas concentrações de 100 e 1.000 g mL-1 contra Escherichia coli (ATCC25922), enquanto três foram efetivas contra Staphylococcus aureus (ATCC25923) na maior concentração. Nenhuma fração apresentou atividade antifúngica contra Aspergillus niger ou Saccharomyces cerevisiae, nem contribuiu para a inibição da formação de biofilme de Pseudomonas spp. O extrato DCM nas concentrações 125, 250 e 500 g mL-1 apresentou dose dependência no ensaio de citotoxicidade com náuplios da Artemia sp., matando 100% dos indivíduos após 48 h de exposição. Os resultados para o extrato EtOH 70% não foram apresentados porque o percentual de mortos no controle foi superior ao máximo admitido de 9 10%. A macroalga marinha parda Lobophora variegata apresenta diferentes atividades biológicas com potencial de investigação havendo a necessidade de estudos mais aprofundados das frações obtidas a fim de verificar a sua eficácia terapêutica.

Thesis (PhD(Agric))--Stellenbosch University, 2009.
ENGLISH ABSTRACT: The global movement in agriculture is towards more environmentally friendly, sustainable production practices, since the role of soil microbial functions in ensuring crop production and soil fertility has become more evident in agricultural systems. Furthermore, with the impeding phase-out of methyl bromide, apple replant disease (ARD) is becoming an increasingly important problem and biological management practises are needed. Since microbial activity is generally carbon-limited in agricultural soil, it is widely accepted that management practices providing a range of organic compounds on a regular basis will tend to maintain an active and diverse microbial population. It was hypothesised that the application of various biological amendments can affect soil microbial numbers and function, thereby having a positive effect on fruit tree growth and yield. The effect of continued applications of organic material, various microbial inoculants and biostimulants on tree performance were evaluated in conventional management systems. Field trials were established in a conventional pear orchard, potential apple replant disease sites, as well as an optimally managed, high density apple orchard under controlled fertigation. The use of compost, compost extracts, a Bacillus inoculant and humates were investigated intensively. Furthermore, to improve our understanding of soil biological systems a combination of simple, practical methods were used to evaluate the effect of biological amendments on soil microbial properties and effects were related to tree performance. Regular application of compost extract in combination with compost showed the most significant effect in improving tree performance in commercial pome fruit orchards under various conditions. In the pear orchard, cumulative yield over the first two seasons was improved by more than 50% compared to controls, while in the fertigated orchard yield was improved by 22%. Biological amendments also showed improved growth in orchards suffering from stunted growth symptoms typical of ARD. However, in severe ARD cases methyl bromide fumigation showed the most consistent effects. Other biological amendments which showed positive effects on yield were application of Bacillus inoculants (Biostart®) in combination with a labile C source and a low dosage humate product, as well as a combination of compost and humates. It was clear that a combination of labile organic matter and a diverse group of microorganisms showed most promise. Although for some specific treatments increased microbial numbers and activity may have resulted in improved tree performance, in general, changes in culture-based plate counts, soil enzyme activity and carbon utilisation profiles could not be used as an indicator of yield. It was suggested that improved synchronisation of nutrient release and plant uptake, as well as microbial phytohormone production, may play an important role in improving tree performance with application of biological amendments. More research is needed on the exact mechanisms through which compost extracts improve yield and studies on root growth proliferation, as well as effects in the rhizosphere are recommended.
AFRIKAANSE OPSOMMING: Binne lanbouverband is daar tans wêreldwyd die neiging om die uitwerking van produksie-praktykte op die omgewing in ag te neem en sodoende meer verantwoordelik op te tree. Omdat die belangrike rol wat grondmikro-organisme funksionering in volhoubare verbouingspraktyke speel nou deeglik besef word, word meer volhoubare bestuurspraktyke bepleit. Hiermee saam, noodsaak aspekte soos die uitfasering van metielbromied vir die beheer van appelhervestigingsiekte, dat biologiese bestuurspraktyke meer aandag geniet. Daar word geredelik aanvaar dat gereelde toediening en aanvulling van organiese materiaal ‘n aktiewe, diverse mikrobe populasie in die grond tot gevolg sal hê. Die hipotese is gestel dat die toediening van ‘n verskeidenheid biologiese produkte grondmikrobe getalle en werking gunstig kan beïnvloed. Dit kan moontlik weer aanleiding gee tot positiewe reaskies wat die groei en drag van vrugtebome betref. In hierdie studie is die uitwerking van voortgesette toedienings van organiese materiaal, mikrobiese inokulante, asook biostimulante, op die prestasievermoë van vrugtebome ondersoek. Veldproewe is uitgelê in ‘n konvensionele peerboord, verskeie boorde met moontlike appelhervestigingsiekte probleme, asook ‘n hoëdigtheidsaanplanting appelboord onder optimale bestuur. ‘n Deeglike ondersoek is gedoen met betrekking tot die gebruik van kompos, komposekstrak, Bacillus-inokulante en humate. Eenvoudige, praktiese metodes is aangewend om vas te stel hoe biologiese toevoegings grondmikrobe eienskappe beïnvloed en of dit verband hou met veranderinge in boomprestasie. Die studie het aangetoon dat die gereelde toediening van komposekstrak saammet kompos, betekenisvolle verbetering in boomprestasie van kernvrugboorde teweeg bring onder verskeie omstandighede. Die kumulatiewe opbrengs van ‘n peerboord is oor twee seisoene met meer as 50% verhoog teenoor die kontrole. In ‘n optimaal bestuurde appelboord onder sproeibemesting, is opbrengs met 22% verhoog in vergelyking met die kontrole. Biologiese toevoegings het ook groei verbeter in boorde waar appelhervestigingsiekte bome se groei vertraag het. In die geval van ernstige appelhervestigingsimptome het metielbromied egter steeds die mees konstante positiewe uitwerking gehad. Ander biologiese toevoegings wat ‘n gunstige uitwerking op opbrengs getoon het, was ‘n kombinasie van Bacillus inokulante, ‘n lae dosis humaat en ‘n aktiewe koolstofbron, asook kompos in kombinasie met humate. Dit is duidelik dat ‘n kombinasie van ‘n maklik afbreekbare koolstofbron (soos kompos) tesame met ‘n diverse groep mikroorganismes mees belowend is vir gebruik in biologiese verbouingssisteme. Resultate toon dat veranderings in aantal organismes gemeet deur plaattellings, die aktiwiteit van grondensieme, en verbruikspotensiaal van verskillende koolstofbronne, nie as ‘n aanduiding van boomprestasie gebruik kan word nie. Daar is voorgestel dat verbeterde sinkronisasie van voedingselementvrystelling en plantopname, sowel as produksie van plantgroeihormone deur mikrobe, moontlik ‘n rol speel by boomreaksies op biologiese toevoegings. Meer navorsing wat verband hou met die meganisme waardeur komposekstrak opbrengs verbeter, is nodig. Verder word studies op fynwortelontwikkeling sowel as aspekte van die wortelrisosfeer aanbeveel.

Mestrado em Biologia Molecular e Celular
Pterospartum tridentatum (L.) WillK., leguminosa que cresce espontaneamente em Portugal, é usada na medicina popular, no tratamento de irritações de garganta e em misturas herbais de várias plantas para o tratamento da diabetes. A formação de radicais livres que causam stress oxidativo pode constituir a base de muitas doenças. Por esta razão, o uso de ervas ou de compostos naturais delas isoladas é cada vez mais comum, com vista aos seus efeitos benéficos. O objectivo do presente trabalho consistiu em estudar os efeitos histopatológicos de extractos de folhas e flores de P. tridentatum no fígado, baço e rim de ratinhos e a sua acção protectora após a administração de um tóxico, o CCl4. No estudo de toxicidade, utilizaram-se 27 ratinhos ICR-CD1 machos, divididos em 7 grupos: o grupo controlo negativo (NaCl 0,9%), 3 grupos aos quais foram injectados extractos de folhas nas concentrações 10 mg/kg, 100 mg/kg e 1000 mg/kg e outros 3 grupos aos quais foram administrados extractos de flores com as mesmas concentrações. Estes 7 grupos receberam uma injecção subcutânea diária durante 6 dias. No estudo de protecção, 25 ratinhos ICR-CD1 foram divididos em 5 grupos: o grupo controlo positivo (CCl4 2ml/kg) recebeu duas injecções subcutâneas, uma no primeiro dia de ensaio e a segunda após 72 horas; 4 grupos foram tratados com as duas injecções de CCl4 e injecções diárias de extractos de folhas nas concentrações de 5 mg/kg e 10 mg/kg e extractos de flores nas concentrações de 5 mg/kg e 10 mg/kg, durante 10 dias. Após sacrifício dos animais, o fígado, o baço e o rim foram removidos, fixados em solução de Bouin e preparados para estudos histológicos. Os cortes histológicos dos órgãos dos animais expostos aos extractos de folhas e flores revelaram lesões hemorrágicas mais frequentes nas concentrações de 100 mg/kg e 1000 mg/kg. O estudo de protecção revelou uma recuperação significativa nos vários órgãos quando comparado com o controlo positivo (CCl4). Em conclusão, o presente estudo revelou que os extractos de folhas e flores de P. tridentatum na concentração de 10 mg/kg não induziram alterações histopatológicas significativas ao nível do fígado, baço e rim e que após a administração de CCl4, os mesmos extractos nas concentrações de 5 mg/kg e 10 mg/kg demonstraram ter um efeito protector nestes órgãos. Sugere-se que a actividade protectora se deve à presença de compostos bioactivos, como os compostos fenólicos, cuja actividade antioxidante é bem conhecida. ABSTRACT: Pterospartum tridentatum (L.) Willk., a Leguminosae that grows spontaneously in Portugal, is used in folk medicine for throat irritation treatment, and herb mixtures for diabetes. The formation of free radicals that cause oxidative stress is on the base of some pathologies and the use of herbal or natural compounds isolated from plants to reduce them is increasingly common. The aim of the present work was to study the histopathological effects of P. tridentatum extracts of leaves and flowers in the liver, spleen, and kidney of mice and their protective action after administration of the toxic CCl4. In the toxicity study, 27 ICR-CD1 mice were used and divided into 7 groups: the negative control group (NaCl 0.9%), 3 groups were injected with leaves extracts in concentrations of 10 mg/kg, 100 mg/kg and 1000 mg/kg, and 3 other groups were administered with flowers extracts with the same concentrations. These 7 groups received a subcutaneous daily injection for 6 days. In the study of protection, 25 ICR-CD1 mice were divided into 5 groups: the positive control group (CCl4 2 ml/kg) received two subcutaneous injections, one on the first day of testing and the second one 72 hours later; 4 groups were treated with two CCl4 injections and daily injections of leaves extracts at a concentration of 5 mg/ kg and 10 mg/kg and flowers extracts at a concentration of 5 mg/kg and 10 mg/kg for 10 days. After sacrifice, the liver, spleen and kidney were removed, fixed in Bouin's solution and prepared for histological studies. Sections of organs exposed to leaves and flowers extracts showed more frequent hemorrhagic lesions in concentrations of 100 mg/kg and 1000 mg/kg. The protection study revealed a significant recovery in the various organs when compared with the positive control (CCl4). In conclusion, this study showed that extracts of P. tridentatum leaves and flowers at a concentration of 10 mg/kg did not induce significant histopathological changes in the liver, spleen and kidney and, after administration of CCl4, the same extracts in concentrations of 5 mg/kg and 10 mg/kg supported the organs protection. We suggest that the protective activity is due to some bioactive elements, such as phenolic compounds, whose antioxidant activity is well known.

Thesis (MTech (Horticulture))--Cape Peninsula University of Technology, 2017.
The exact responses to the concentration of compost tea extract and methods of irrigation application were not previously measured on Hypoxis hemerocallidea and Siphonochilus aethiopicus. Commercial exploitation, habitat loss and degradation, overharvesting, and enhancement of their medicinal properties, have led to this investigation and the need to replenish both these valuable plant species. This is crucial for plant survival, especially in the wild and for use of the traditional medicinal plants. Hypoxis hemerocallidea and Siphonochilus aethiopicus, known as star flower and wild ginger respectively continue to decline, due to overharvesting from their natural habitat. Both these species have tremendous traditional medicinal value among localized African people. To enhance their commercial cultivation, compost tea extracts, in the following ratios (no catalyst added (control1); T 1000:1, T750:1, T500:1, and T250:1L) were applied in equal dosages to determine an optimal compost tea extract ratio. The experiment was conducted in a temperature controlled greenhouse. Mushroom compost (500 g, per brew) was used for all extracts. Brewing was done with no catalyst added (Control 1), and 24 hours later another brew was done with catalyst added, weekly for 20 weeks. The Control treatment received water only. Both species were slow growing and comparatively, the Hypoxis plants responded faster than the Siphonochilus plants. In this investigation, plant growth parameters such as plant height, number of leaves, leaf width, leaf length and leaf colour, were measured and evaluated. Despite the plants positive response to the mushroom compost tea extracts, across all the above plant growth parameters, no significant differences were noted between the treatments during the twenty-week application period. Leaf chlorophyll content peaked in week 11 of the hypoxis plants and was the highest in week 14 of the Siphonochilus plants with no significant interaction between weeks vs. treatment over the twenty-week experiment. The chlorophyll readings indicate that both species increased their chlorophyll production over time. Although the total wet leaf length, root length, corm diameter, leaf weight, corm weight and root weight of Hypoxis were non-significant between treatments, T500:1 total wet weight was significantly higher when compared to the rest of the treatments. The total dry weight analysis of hypoxis was non-significant. Control Calcium level was significantly lower between the control and the treatments of the Hypoxis total dry leaf nutrient analysis. The Hypoxis dried roots nutrient analysis was non-significant across treatments.

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O pinhão manso (Jatropha curcas L.) é uma planta versátil que ultimamente tem sido utilizada como opção em diversas áreas e na medicina popular. Estudos demonstram seu potencial para o tratamento de constipação, doenças parasitárias, como antimicrobiano, antifúngico, antiinflamatório, analgésico, antipirético, antiviral, antioxidante e atividade contra algumas linhagens tumorais. Dentre os diversos fatores extrínsecos e intrínsecos apontados como responsáveis para o surgimento do câncer, um dos mais discutidos atualmente têm sido os radicais livres ou espécies reativas de oxigênio (ERO), moléculas instáveis, quimicamente reativos produzidos ao longo do metabolismo celular normal, no entanto, podem causar estresse oxidativo que tem sido descrito como um dos principais precursores de doenças como aterosclerose, catarata, doenças neurodegenerativas em especial o câncer. Antioxidantes são moléculas que têm como propriedade o bloqueio, a inibição ou o retardo da deterioração oxidativa, reduzindo ação de radicais livres. O arsenal terapêutico antineoplásico atualmente disponível não são específicos levando à morte de células cancerígenas, como de células normais desencadeando o aparecimento dos efeitos colaterais. Assim, novos compostos que apresentem seletividade são requeridos e recentemente foram introduzidos quimioterápicos de origem natural no tratamento do câncer, o que valida à busca de novos alvos farmacológicos a partir de produtos naturais, principalmente, direcionado às plantas usadas na medicina popular. Nesse contexto, este estudo objetivou avaliar o perfil fitoquímico, a atividade antioxidante e a ação antitumoral de extratos etanólicos de Jatropha curcas L., sob linhagem tumoral in vitro. Para tanto, foram preparados extratos frescos e secos de raiz, caule, folha e sementes por dois métodos de extração (maceração e uso de Soxhlet), foi então realizada a identificação dos grupos fitoquímicos e a triagem destes extratos através do efeito antiproliferativo sob a linhagem HepG2. Para avaliação da atividade antioxidante foram realizadas análises através dos métodos de sequestro do radical livre 2,2-difenil-1- picrilhidrazil (DPPH) e sequestro do radical livre 2,2′-Azino-bis (3-etilbenzotiazolina-6-ácido sulfônico) (ABTS). Os extratos de forma geral apresentaram a presença de flavonoides, taninos catéquicos e fenóis simples. Os rendimentos dos extratos obtidos por ambas as metodologias de extração mostraram diferenças significativas em termos de percentual. O rendimento dos extratos obtidos pelo método de soxhlet apresentou de forma geral menor percentual em relação ao rendimento dos extratos obtidos por maceração, variando entre 1,24 a 24, 96 %, e não tendo relação direta com a atividade antioxidante do extrato. Verificou-se que o extrato preparado com soxhlet de amostras secas de folha de J. curcas apresentou melhor atividade antioxidante atingindo o percentual superior a 80%, quando avaliado pelo método do DPPH enquanto que os extratos obtidos por maceração de sementes frescas (98,54%) e por soxhlet de sementes secas (97,69%) apresentaram melhor capacidade antioxidante usando o método do ABTS. O extrato de folhas secas obtido por soxhlet apresentou atividade antioxidante pelo método do DPPH de 83,1% / EC50 - 47,46 μg/mL,enquanto que o extrato obtido por maceração de folhas frescas foi de 68,1% / EC50 – 52,88 μg/mL, correspondendo aos menores valores de EC50 dos extratos brutos avaliados pelo método do DPPH. Os extratos etanólicos secos de raiz, caule e folhas obtidos por soxhlet, apresentaram melhor efeito antitumoral sobre as células tumorais da linhagem HepG2 e não foram citotóxicos quando avaliados em macrófagos peritoneias. Conclui-se que extratos etanólicos brutos de folhas de Jatropha curcas L. possui atividade antioxidantee extratos etanólicos de raiz e caule promovem efeito antiproliferativo em células tumorais da linhagem HepG2. Conclui-se que a atividade antintumoral sob linhagem HepG2 e antioxidante dos extratos etanólicos de Jatropha curcas L. avaliada através do método de sequestro do radical DPPH e do ensaio ABTS varia em função do método de preparo do extrato, das amostras utilizadas (peso úmido e peso seco) e das partes botânicas analisadas. Este trabalho confirma o potencial antioxidante e antitumoral de extratos etanólicos de Jatropha curcas L.

Notre travail a porte sur l'etude de plantes medicinales repertoriees dans la pharmacopee traditionnelle chinoise pour leur action cicatrisante. Il s'est deroule en trois parties : tout d'abord l'extraction des differentes plantes selectionnees dans la medecine traditionnelle, suivi par la mise en evidence d'activites immunostimulantes dans plusieurs extraits aqueux, et ensuite une importante partie consacree a la purification de ces fractions actives, puis a leur analyse structurale. Nous avons mis au point les methodologies de purification et d'analyse sur l'extrait aqueux d'imperata cylindrica, puis nous les avons employees pour les autres extraits actifs. La purification a ete suivie par la mise en oeuvre d'un pharmacoguidage a l'aide d'un test de proliferation de splenocytes murins. Elle a permis d'isoler apres plusieurs chromatographies, des fractions actives de nature polysaccharidique. L'analyse des monosaccharides par plusieurs techniques (cpg, hpaec-pad) a ensuite ete realisee. Nous avons pu mettre en evidence six monomeres dans chaque fraction analysee : glucose, galactose, mannose, rhamnose, arabinose, xylose, seules les proportions entre eux varient d'une fraction a l'autre. En conclusion, cette these porte sur l'isolement et la caracterisation de polysaccharides de plantes ayant des activites immunostimulantes. Ces macromolecules biologiques pourraient effectivement jouer un role dans la cicatrisation, ce qui expliquerait leur utilisation en medecine traditionnelle.

O nitrogênio é um componente universal de combustíveis fósseis, geralmente associados à matriz orgânica. A grande maioria destes compostos nitrogenados presentes no petróleo e resíduos tem efeito muito importante na estabilidade de processos catalíticos. Por exemplo, eles podem causar o envenenamento de catalisadores, além da formação de gomas na gasolina. Um grande número de compostos básicos apresenta propriedades tóxicas, especialmente os aza heterocíclicos e aminas aromáticas primárias, que têm sido citadas como carcinogênicas. Os compostos nitrogenados neutros são em geral menos tóxicos do que os básicos, porém vários dibenzocarbazóis têm sido citados como substâncias carcinogênicas. Assim o conhecimento mais detalhado dos tipos e das concentrações dos compostos nitrogenados em produtos de petróleo e derivados se faz necessário para removê-los de maneira segura. O petróleo é constituído por mais de uma centena de compostos, constituindo assim uma matriz bastante complexa o que impossibilita a caracterização seus derivados sem uma etapa de pré-fracionamento tal como extração líquido/líquido, extração com solvente ou cromatografia líquida preparativa. O objetivo deste trabalho foi desenvolver procedimentos analíticos para a separação e identificação de compostos nitrogenados em gasóleo pesado (GOP) e resíduo atmosférico (RAT) de petróleo brasileiro. Para a extração dos compostos nitrogenados do GOP e RAT foi utilizado o pré-fracionamento com alumina neutra, resultando em quatro frações. A terceira fração, isolada com diclorometano, que potencialmente contém os compostos nitrogenados, foi re-fracionada utilizando: resinas Amberlyst A 15 e A27; sílica modificada com KOH e HCI e Chromosorb W/FeCb. Foi avaliada a dessorção dos compostos nitrogenados da alumina, das resinas, da sílica modificada e do Chromosorb W/FeCb por eluição em coluna e no ultra-som. A sílica modificada com dessorção dos compostos nitrogenados em batelada no ultra-som mostrou ser uma boa técnica para separar compostos nitrogenados básicos, enquanto o Chromosorb W/FeCb em coluna mostrou-se um bom adsorvente para os compostos nitrogenados neutros. A cromatografia gasosa com detector de nitrogênio e fósforo (GC/NPO) apresentou-se mais sensível para a detecção de compostos em baixa concentração. A cromatografia gasosa com detector seletivo de massas (GC/MSD) permitiu a determinação dos compostos nitrogenados do GOP e RA T e a identificação de alquil quinolinas, alquil benzoquinolinas, alquil tetrahidrodibenzoquinolinas, alquil carbazóis e alquil benzocarbazóis nestas amostras.
Nitrogen is an universal component of fossil fuels, generally associated with the organic portion of crude materiais. With the possible exception of some petroleum products of commercial value, nitrogen compounds in petroleum and residue adversely affect many important catalyst processes and product stability. For example, they cause catalyst poisoning and are involved with gum and color formation in products. Moreover, a number of basic compounds are toxic; several of the aza heterocycles and aromatic primary amines are known, or suspected, to be carcinogens. Neutral nitrogen compounds appear, in general, to be less toxic than basic compounds, but several dibenzocarbazoles have been reported as carcinogenic substances. A detailed knowledge of the types and concentration of nitrogen compounds present in petroleum products is clearly desirable to optimize methods for their removal and specify methods for the safe handling of such materiais Petroleum is constituted of more than a hundred of compounds, being a quite complex matrix. Which sends impossible the characterization of crude oil products without a good pre-fractionation step, such as liquid-liquid extractiàn, solvent extraction or preparative liquid chromatography. The goal of this work was to develop different procedures for the reliable separation and identification of nitrogen compounds in heavy gas oil (GOP) and atmospheric residue (RAT) from Brazilian petroleum A pre-fractionation scheme was used with neutral alumina for the extraction of the nitrogen compounds from GOP and RA T, resulting in four fractions. The third fraction, isolated with dichloromethane, which potentially contains the nitrogen compounds, was fractionated by using ion exchange resins (Amberlyst A 15 and A2.7), modified silica (Si02/KOH and Si02/HCI) and Chromosorb W modified with FeCh. The desorption of the nitrogen compounds from the stationary phases (neutral alumina, resins, silica and Chromosorb W/FeCh) was performed by elution in column and by sonication. Modified silica desorbed by sonication was shown to be a good technique to separate basic nitrogen compounds, while Chromosorb W/FeCh in column was better for the separation of neutral nitrogen compounds. Gas Chromatography with Nitrogen and Phosphorus Detector (GC/NPD) showed to be more sensitive for the detection of compounds at low concentration. Gas Chromatography with Mass Spectrometry Detector (GC/MSD) allowed the analysis of the nitrogen compounds of GOP and RA T and the identification of alkyl quinolines, alkyl benzoquinolines, alkyl tetrahydrodibenzoquinolines, alkyl carbazoles and alkyl benzocarbazoles in these samples.

Orientador: Florencia Cecilia Menegalli
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Resumo: O objetivo do presente trabalho foi desenvolver revestimentos ativos de pectina incorporados com extratos de frutas a fim de entender e avaliar a estabilidade e a difusão dos nutrientes incorporados aos mesmos, assim como o seu efeito antioxidante quando aplicados em alimentos. Na primeira etapa foram desenvolvidos filmes ativos de pectina. Extratos alcoólicos e aquosos de acerola, caju, mamão, morango e pequi foram obtidos. Estes foram caracterizados e selecionados de acordo com teores de vitamina C, carotenoides, antocianinas, compostos fenólicos e atividades antioxidantes (ABTS, DPPH e FRAP). Como os extratos alcoólicos de acerola, caju e morango foram os com maior recuperação de nutrientes e, consequentemente, maiores atividades antioxidantes (969,0 ± 0,7, 163,6 ± 0,2 e 138,1 ± 0,5 mg de Trolox/g de extrato seco, respectivamente), eles foram incorporados aos filmes. Também foi elaborada uma formulação com a mistura dos extratos. Os filmes foram avaliados quanto às suas atividades antioxidantes, propriedades ópticas e pela capacidade de reter nutrientes através de um estudo de estabilidade. Dentre os filmes estudados, o de acerola possuiu a maior atividade antioxidante (36±2 ?g de Trolox/g de filme seco) com um tempo de meia vida de 99 dias. A segunda parte do trabalho avaliou a migração dos compostos fenólicos (nutrientes majoritários) para meios simulantes de alimentos a fim de entender o comportamento difusional desses compostos. Foram utilizados como simulantes líquidos, o metanol e a água, e como sólidos, géis de gelatina (com e sem adição de celulose). Os filmes foram aditivados com os extratos alcoólicos de fruta. Para os simulantes líquidos, o metanol foi o mais efetivo na extração de compostos fenólicos, enquanto que para o simulante sólido foram observadas maiores retenções para os filmes em contato com o gel contendo celulose (retenções entre 41 % e 45%) devido à maior resistência mecânica causada pela adição da fibra de celulose. Na terceira etapa do trabalho, foram preparadas coberturas de pectina, de acordo com as formulações testadas para os filmes na primeira parte do trabalho. Estas coberturas foram aplicadas a fatias de kiwi previamente ao processo de secagem. O estudo avaliou a influência das coberturas aditivadas sobre as características químicas e físicas do kiwi, após o processo de secagem e ao longo do armazenamento. As influências das coberturas nas isotermas de sorção e sobre as cinéticas de secagem também foram avaliadas. A aplicação das coberturas contendo antioxidantes contribuiu para minimizar os efeitos indesejáveis da oxidação de nutrientes sem influenciar no processo difusional da água, além de promover uma melhoria da manutenção das características de qualidade dos kiwis ao longo da estocagem por 31 dias. No geral, os resultados revelaram que a incorporação de extratos de frutas nos revestimentos de pectina, potencialmente, produz filmes e coberturas ativos, podendo ser utilizados para diferentes aplicações em produtos alimentares. Além de atuar como barreira anti-oxidante, podem conferir nutrientes aos produtos em questão
Abstract: This study aimed to develop pectin films and coatings incorporated with fruit extracts, evaluate the stability and diffusion of nutrients in these films and coatings, and understand the antioxidant effect of fruit extracts on food. The first stage of the study consisted in developing active pectin films. Evaluation of the alcoholic and aqueous extracts of five fruits (acerola, cashew, papaya, strawberry, and pequi) in terms of nutrient content (vitamin C, carotenoids, anthocyanins, and phenolic compounds) and antioxidant capacity (ABTS, DPPH, and FRAP) showed that the alcoholic extracts of acerola, cashew, and strawberry promoted the highest recovery of nutrients and, consequently, higher antioxidant activity (969.0 ± 0.7, 163.6 ± 0.2 and 138.1 ± 0.5 mg Trolox /g of dry extract, respectively) which justified their incorporation in the films. Next, a formulation with a mixture of these alcoholic extracts was prepared. The resulting films were assessed in terms of their antioxidant activity, optical properties, and ability to retain nutrients along a stability study. The film incorporated with acerola extract had the highest antioxidant capacity (36 ± 2 ?g of Trolox/g of dry film); its half-life was 99 days. In an attempt to understand the diffusion behavior of nutrients, the second part of this study investigated how phenolic compounds (major nutrients) migrated from films incorporated with alcoholic fruit extracts to different food simulants. Methanol and water served as liquid simulants; gelatin gels with and without cellulose were the solid simulants. In the case of liquid simulant systems, methanol extracted phenolic compounds more effectively than water. As for solid simulant systems, films in contact with the gel containing cellulose, which had increased mechanical strength, retained the nutrients more satisfactorily (retentions between 41% and 45%). The third stage of this work dealt with the preparation of pectin coatings using the formulation previously tested for the films. These coatings were then applied to kiwi by immersing kiwi slices in the filmogenic solution prior to the drying process. The influence of these coatings on the chemical and physical characteristics, sorption isotherms, and drying kinetics of kiwi was evaluated after drying and during storage. Coatings containing antioxidants significantly minimized the oxidation of nutrients without affecting water diffusion during the drying process; these coatings maintained the characteristics of kiwi for 31 days. Overall, incorporation of fruit extracts into pectin films and coatings could add active nutrients and become an alternative antioxidant barrier in food products
Doutorado
Engenharia de Alimentos
Doutora em Engenharia de Alimentos

L’induction de réponses de défense via la Pathogen-Associated Molecular Pattern (PAMP)-Triggered Immunity fournit à toutes les plantes un arsenal de composés antimicrobiens contre les agents pathogènes. La stimulation de ces défenses par des éliciteurs exogènes serait une stratégie pour réduire les pesticides mais nécessite une meilleure compréhension des mécanismes de l’induction qui pourrait dépendre de l'éliciteur et du génotype. Deux génotypes de pomme de terre partiellement résistants à Phytophthora infestans ont été traités avec un PAMPs de P. infestans, un filtrat de culture concentré (CCF), une molécule synthétique, l’acide ß-aminobutyrique (BABA) et un extrait d’Ulva spp. Les métabolites induits dans les folioles, 48h plus tard, ont été comparés par une approche de métabolomique non ciblée (UPLC-qTOF-MSe) puis par des approches ciblées demétabolomique et transcriptomique. Les résultats ont montré que (i) chaque éliciteur induit une reprogrammation spécifique du métabolome confirmée partiellement par les transcrits : BABA et CCF ont induit les benzylisoquinoléine alcaloïdes et l'extrait d'Ulva, les stérols alcaloïdes et les phénylpropanoïdes; ii) la signalisation dépend de la voie du JA pour l’extrait d'Ulva, et de celle du SA pour BABA et CCF ; (iii) l’abondance de certains métabolites est génotype-dépendant. Nos résultats suggèrent que l’induction des composés antimicrobiens dépend de la perception spécifique de l’éliciteur et de l'interaction éliciteur/génotype. Afin d'améliorer leur efficacité sur le terrain, une attention particulière doit être portée à la si
The induction of response defences via the Pathogen-Associated Molecular Pattern (PAMP)-Triggered Immunity provides all plants an arsenal of antimicrobial compounds against pathogens. Stimulation of these defences by exogenous elicitors could be a strategy to reduce the pesticides but this requires a better understanding of the induction mechanisms. This induction could depend on both the origin of the elicitor and the genotype. Two potato genotypes partially resistant to Phytophthora infestans were treated with a PAMPs from P. infestans, the Concentrated Culture Filtrate (CCF), a synthetic molecule, the ß-aminobutyric acid (BABA); and an Ulva spp extract. 48h after treatment, the induced metabolites in the leaflets were first compared by a non target metabolomic using a UPLC-qTOF-MSe, then through targeted metabolomic and transcriptomic approaches.The results showed that (i) each elicitor induced a specific metabolic profile reprogramming, partially confirmed by transcript analysis: the BABA and the CCF induced the benzylisoquinoline alkaloids and the Ulva extract, the alkaloid sterols and phenylpropanoids; (ii) the signalling depended on the JA pathway for the Ulva extract while it is rather SA pathway-dependent for the BABA and CCF; (iii) the abundance of several antimicrobial metabolites was genotype-dependent. Our results suggest that the induction of the antimicrobial compounds depends on the elicitor/genotype interaction and on the specific perception of the elicitor. In order to improve their efficiency in the field, special attention should be paid on the defence

La résistance aux antimicrobiens est l’un des défis majeurs du XX1eme siècle. Pseudomonas aeruginosa est inscrit sur la liste des organismes pathogènes qui deviennent résistants aux antibiotiques conventionnels. De nouvelles stratégies visant à atténuer la virulence sans perturber la croissance et la viabilité bactériennes, également connues sous le nom de stratégie anti-virulence, sont développées. Les plantes sont connues pour produire de nombreux métabolites secondaires. Des extraits de fruits de Pistachia Lentiscus originaires d'Algérie et de 40 extraits de plantes originaires du Nord-Chili ont été criblés pour leur capacité à atténuer la production de la pyocyanine, un facteur de virulence majeur de P. aeruginosa, dans le but d’évaluer leur potentiel effet antivirulence. Les extraits sélectionnés (Pistacia lentiscus, Azorella atacamensis, Baccharis grisebachii, Haplopappus rigidus et Parastrephia terestiucula), ont été fractionnés et l’ensemble de ces extraits et fractions a montré une atténuation de la production d’autres facteurs de virulence (élastase, rhamnolipides), qui a pu être attribuée, au moins partiellement à une diminution de la communication bactérienne via le mécanisme du quorum sensing. Ces extraits et fractions altèrent également la fluidité membranaire de P. aeruginosa. Cet effet anti-virulence a été validé dans un modèle d'infection cellulaire, et sur le nématode Caenorhabditis elegans. Dans toutes ces conditions, la croissance de P. aeruginosa n'a pas été affectée. Un profilage chimique des extraits et fractions de P. lentiscus et d'A atacamensis a révélé la présence d'acide gingkolique et de diterpenoides de type azorellane/mulinane comme potentiels composés bioactifs. De futures études visent à identifier les composés bioactifs sur P. aeruginosa H103, ainsi que sur un panel de souches cliniques, et à évaluer un potentiel effet potentialisateur de l'activité des antibiotiques. Ces travaux visent in fine à proposer ces composés d’origine végétale comme adjuvants dans le traitement des infections à P. aeruginosa
Antimicrobial resistance has become a great challenge in therapeutic medicine so much so that the World health organization forecasts the possibility of a post-antibiotic era where minor injuries may lead to mortality. Pseudomonas aeruginosa is among the list of organisms that are highly resistant to conventional antibiotics, partly due to its broad genome, which facilitates the elaboration of virulence determinants and rapid adaptation to various environments, in addition to its inherent resistance mechanisms. In view of this, alternative measures of controlling microbial virulence activities using novel approaches that do not disturb its growth and viability, also known as anti-virulence strategy, are gaining wider attention. Since plants are repositories of several metabolites with chemical defense system against environmental pathogens, through ethnobotanical led studies, the effect of Pistacia lentiscus fruit extracts originating from Algeria and forty plant extracts originating from North-Chile were biologically and chemically evaluated with the aim of deciphering their anti-virulence effects against P. aeruginosa. Furthermore, this study tried to gain more insight into the bioactive compounds and possible mechanism of action. From the results obtained, selected plant extracts attenuated P. aeruginosa mainly pyocyanin activity and /or elastase and rhamnolipids virulence production which appears to be associated with the inhibition of quorum sensing activities and the alteration in membrane activities. The anti-virulence effect of the selected extracts (P. lentiscus, Azorella atacamensis, Baccharis grisebachii, Haplopappus rigidus and Parastrephia terestiucula) were also validated in biological models of infections where they mediated the toxicity of P. aeruginosa towards A549 human monolayer cells and/or Caenorhabditis elegans nematode. Interestingly, growth of the pathogen was not affected. Further chemical profiling of P. Lentiscus, and A atacamensis extracts revealed the presence of gingkolic acid and azorellane/mulinane diterpenoids as the putative bioactive compounds. Future studies intend to explore these extracts and their derived compounds on the potentiation of antibiotic activity in a panel of clinical strains. In general, this study sets the pace for the possible use of these plant extracts as adjuvants in treatment of P. aeruginosa infections

ABSTRACT Effects of Compost, Legume Cover Cropping and Vermicompost Extract Foliar Applications on Nutrition and Yield of Washington Navel Oranges William Payton Carling An experiment was conducted to test the effects of four treatments on Washington navel orange (Citrus sinensis) trees in regards to nutrient content of the leaves and fruit, soil nutrient content and properties, and fruit yield. The four treatments included: compost (C) and vermicompost extract foliar (VEF) applications, legume cover cropping (LCC) and VEF applications, VEF applications, and a control. The treatments were implemented from February 2010 to April 2011 and sampling occurred in May 2011. Fruit Growers Laboratory, Inc. (FGL) conducted the soil, leaf, and fruit analyses. The test site was located in San Isidro, Baja California Sur, Mexico. The compost used was made onsite with livestock manure, carbon-based farm waste, and water, and applied around the drip-lines of 8 WNO trees once every two months. The vermicompost and vermicompost extract was made onsite by introducing red wiggler worms (Eisenia fetida) into horse manure, allowing 1 month for casting content to build up, and collecting the extract as water was filtered through the material. Vermicompost extract was applied using a backpack foliar sprayer on 8 WNO trees twice a month. The legume cover crop treatment consisted of two plantings at a rate of approximately 72 seeds per square foot around the drip-lines of 8 WNO trees. Black-eyed peas (Vigna unguiclata) were planted in April 2010 and minimally incorporated using a hoe and shovel in August 2010. Fababeans (Vicia faba) and dry peas (Pisum sativum) were planted in January 2011 and minimally incorporated into the soil in March 2011. WNO leaf and soil analyses were compared to FGL optimum ranges. The C+VEF treatment showed trends of increased soil and leaf nitrogen, phosphorus, potassium, calcium, zinc, and copper levels and improved soil properties by raising soil organic matter percentage, saturation percentage, and moisture percentage. The control treatment had low or deficient values in these nutrients and soil properties. The LCC+VEF treatment increased nitrogen in the soil and leaf content but decreased organic matter percentage. The VEF treatment increased nitrogen and potassium content in the leaf but increased sodium beyond FGL optimum range. The C+VEF treatment had the highest yield and the greatest amount of nutrients removed as a result of yield. The C+VEF treatment had more available nutrients in the soil for WNO tree uptake and future crops.

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Universidade Federal de Minas Gerais
About 70% of the area cultivated with sugarcane in Brazil are on acid soils, characterized by having severe limitations in terms of fertility. Organic acids from residues decomposition of sugarcane, due to mechanized harvest without the use of fire, are able to interact with elements of the soil solution. This research project was aimed at investigating the influence of the addition of plant extracts residue of sugarcane in the complexation of Al phytotoxic and expansion of the root system of the crop . The experiment was conducted in laboratory and greenhouse, with the following specific objectives: (a) characterize chemically and determine qualitatively and quantitatively the organic acids present in plant extracts of the residue of sugarcane harvested mechanically to select the plant extract with greater neutralizing capacity of Al; (b) test doses of plant extracts of of sugarcane for complexation of toxic Al; (c) determine the depth of root development of varieties of sugarcane under conditions of high Al saturation; (d) verify that the plant extracts applied on columns filled with soil naturally acid, cultivated with sugar cane were able to promote the elevation of the pH of soil. The results of laboratory analyzes of plant extracts showed greater power to carry exchangeable cations along the soil profile and neutralizing Al phytotoxic to the plant extract of the residue variety RB867515. The results of the experiment at home a greenhouse indicated that the varieties of sugarcane differed in Al tolerance of the soil, on the accumulation of biomass and aluminum aerial part and root system, as well as deepening the roots. The plant extract residue of variety RB867515 had a positive effect on root development of varieties, in raise soil pH, alleviating Al toxicity and the availability of basic cations to the sublayers of the soil columns. The dosage of the plant extract, which promoted effective improvement in soil properties, was the equivalent of 21 tha- 1. The varieties were classified as tolerance to Al, as follows : RB966928 > RB855453 > RB867515.
Cerca de 70% das áreas cultivadas com cana-de-açúcar no Brasil estão sobre solos ácidos, caracterizados por possuírem severas limitações em termos de fertilidade. Os ácidos orgânicos provenientes da decomposição da palhada de cana-de-açúcar, decorrente da colheita mecanizada sem o uso de fogo, são capazes de interagir com elementos da solução do solo. Este projeto de pesquisa teve por objetivo principal a investigação da influência da adição de extratos vegetais de palhada de cana-de-açúcar na complexação do Al fitotóxico e na expansão do sistema radicular da cultura. O experimento foi conduzido em condições de laboratório e casa-de-vegetação, com os seguintes objetivos específicos: (a) caracterizar quimicamente e determinar quantitativa e qualitativamente os ácidos orgânicos presentes nos extratos vegetais da palhada de cana-de-açúcar colhida mecanicamente para selecionar o extrato vegetal com maior capacidade de neutralização do Al; (b) testar dosagens de extratos vegetais da palhada para complexação de Al tóxico; (c) determinar a profundidade de desenvolvimento radicular de variedades de cana-de-açúcar submetidas ao estresse pelo Al; (d) verificar se os extratos vegetais aplicados em colunas preenchidas com solo naturalmente ácido, cultivadas com cana-de-açúcar foram capazes de promover a elevação do pH deste solo. Os resultados das análises laboratoriais indicaram maior poder de transporte de cátions trocáveis e de neutralização do Al fitotóxico para o extrato vegetal da palhada da variedade RB867515. Os resultados do experimento em casa-de-vegetação indicaram, que as variedades de cana-de-açúcar diferiram quanto à tolerância ao Al do solo, quanto ao acúmulo de biomassa e de alumínio da parte aérea e do sistema radicular, assim como no aprofundamento das raízes. A aplicação do extrato vegetal da palhada da veriedade RB867515 teve efeito positivo no desenvolvimento radicular das variedades, promoveu a elevação do pH do solo, amenização da toxicidade do Al e, a disponibilidade de cátions básicos até às subcamadas das colunas de solo. A dosagem do extrato vegetal, que proporcionou melhora efetiva nos atributos do solo, foi o equivalente a 21 tha-1. As variedades foram classificadas, quanto a tolerância ao Al, da seguinte forma: RB966928> RB855453> RB867515.

Orientador: Pedro Luiz Rosalen
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba
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Resumo: Este estudo avaliou a atividade antimicrobiana de extratos da Rheedia brasiliensis contra Streptococcus mutans e o potencial anticárie do seu composto ativo isolado. Os extratos hexânicos (EH), acetato de etila e etanólico (concentrações entre 6,25-800 µg/ml) do fruto (¿bacupari¿) foram testados contra S. mutans UA159 por meio de determinação das concentrações inibitória mínima (CIM) e bactericida mínima (CBM). Biofilmes de 5 dias de formação foram tratados com os extratos ativos (100 x CIM), por 0, 1, 2, 3 e 4 h, e plaqueados para contagem das UFC/ml (time kill). Os extratos ativos foram submetidos à análise química por métodos espectroscópicos, e o composto ativo isolado (0,625-80 µg/ml) foi testado contra células de S. mutans por meio de CIM/CBM. A influência do composto isolado (12,5-100 µg/ml) sobre a síntese de glucanos foi avaliada através da atividade da enzima glucosiltranferase (GTF) B em superfície de hidroxiapatita. A ação do composto sobre biofilmes (125 e 250 µg/ml) foi testada com os ensaios de queda de pH e inibição de formação de biofilme. O potencial anticárie do composto (250 µg/ml) foi avaliado em ratas Wistar sob alto desafio cariogênico. Os EH (casca e semente) inibiram S. mutans em baixas concentrações (CIM:12,5-25 µg/ml) e reduziram células viáveis dos biofilmes após 4 h de exposição. O composto ativo presente foi identificado como a 7- epiclusianona (7-epi), a qual inibiu o crescimento do S. mutans (CIM:1,25-2,5 µg/ml), a atividade da GTF B em superfície (48% de inibição) e a produção de ácidos do S. mutans em biofilme, porém não reduziu a formação e o acúmulo dos biofilmes. No estudo animal, a 7-epi reduziu a incidência e a severidade de cárie e a microbiota total, mas não diminuiu a porcentagem de S. mutans. A 7-epi diminuiu a virulência do S. mutans e inibiu o desenvolvimento de cárie in vivo, sugerindo ser um agente terapêutico promissor para o controle do biofilme dental cariogênico. Palavras-chave: Rheedia brasiliensis, 7-epiclusianona, Streptococccus mutans, fatores de virulência, cárie animal
Abstract: The present study evaluated the antimicrobial activity of Rheedia brasiliensis extracts against Streptococcus mutans and the anticaries properties of its isolated bioactive compound. Hexane (HE), ethyl-acetate and ethanolic extracts (concentrations ranging from 6.25 to 800 µg/ml) of R. brasiliensis fruits (¿bacupari¿) were tested against S. mutans UA159 by determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). S. mutans 5-days-old biofilms were treated with active extracts (100 x MIC), for 0, 1, 2, 3 e 4 h (time-kill), and plated for colony counting (UFC/ml). Such extracts were submitted to exploratory chemical analyses to isolate and identify the bioactive compound using spectroscopic methods. The compound (0.625-80 µg/ml) was then tested against S. mutans cells using MIC/MBC assays. The influence of the bioactive compound (12.5-100 µg/ml) on glucans synthesis was evaluated by testing the activity of glucosyltranferase (GTF) B on hydroxyapatite surface. The effects of the compound (125 e 250 µg/ml) on biofilms were analyzed using glycolytic pH-drop and inhibition of formation assays. The anticaries activity of such compound (250 µg/ml) was evaluated in Wistar rats submitted to a high cariogenic challenge. HE (peel and seeds) reduced S. mutans cells at low concentrations (MIC:12.5-25 µg/ml) and also biofilm viability after four hours, confirming the presence of the bioactive compound. This compound was identified as 7-epiclusianone (7-epi), which inhibited the S. mutans growth (MIC:1.25-2.5 µg/ml), the activity of GTF B (48% of inhibition) and the acid production, not interfering with the formation and accumulation of biofilms. In the animal study, 7-epi additionally reduced the incidence and severity of caries and also the total microbiota, however no reduction on the percentage of S. mutans was observed. In conclusion, 7-epi may be considered a promising agent to control cariogenic dental biofilm. Keywords: Rheedia brasiliensis, 7-epiclusianone, Streptococccus mutans, virulence factors, animal caries
Mestrado
Farmacologia, Anestesiologia e Terapeutica
Mestre em Odontologia

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Organismos marinhos são importantes fontes de novos produtos naturais e compostos bioativos com funções variadas. Entre as principais fontes de novas moléculas esta o filo dos Cnidários, com uma importante contribuição no numero de compostos descritos nos últimos anos. Bunodossoma caissarum é uma espécie de anêmona endêmica do Brasil, cujo perfil químico foi analisado. Para essa análise foram realizados uma extração hidroalcoólica e o isolamento de determinadas moléculas com técnicas cromatográficas. Os compostos isolados foram aqueles que apresentaram absorbância no comprimento de onda no UV em 310 nm e foram identificados a partir de técnicas de espectrometria de massas e ressonância magnética. Entre as substâncias encontradas estão sais piridinicos, núcleos imidazóis, caissarona e a etilglicose, moléculas com importantes papeis fisiológicos.
Marine organisms are importante sources of new natural products and bioactive compounds with various functions. The phylum Cnidaria is among the main sources of new molecules, with na importante contribution to the number of compounds described in the last years. The anemone species Bunodossoma caissarum is endemic to Brazil and had it's chemical profile analyzed. For the purpose of this analysis were done a hydroalcoolic extraction and the isolation of certain molecules by chomatographic techniques. The isolated compounds were those that displayed absorbance in the 310nm wavelength of the UV spectrum and were identified on the basis of mass spectrometry and magnetic ressonance. Among the substances found are pyridinic salts, imidazole nucleus, caissarone and ethyl glucose, which are molecules with important physiological roles.

Myrianthus arboreus est une essence de la forêt tropicale d’Afrique dont les tissus contiennent des molécules bioactives intéressantes. Les données ethnopharmacologiques africaines attribuent à l’extrait aqueux d’écorces de racines de M. arboreus des effets thérapeutiques notamment antidiabétique. Le but de cette thèse était de valider ce potentiel à travers trois séries des bioessais cellulaires in vitro, d’identifier les principes actifs et d’élucider les mécanismes d’actions par lesquels la plante exerce son activité sur le diabète. Une étude préliminaire sur les différents extraits et fractions obtenus à partir de solvants de force de polarité croissante (dichlorométhane, éthanol, méthanol, eau) a été menée et a démontré un excellent potentiel antioxydant de l’extrait éthanolique et de sa fraction acétate d’éthyle comparable à celui de l’Oligopin®, un extrait commercial riche en proanthocyanidines. Ces résultats ont corrélé avec les teneurs en acides hydroxicimiques et en proanthocyanidines, deux classes de polyphénols présentes dans l’extrait et la fraction acétate d’éthyle. Ensuite, les résultats des bioessais cellulaires indiquent qu’aucun extrait ou fraction n’a été capable de stimuler le transport de glucose dans les cellules musculaires C2C12. A l’inverse, l’extrait éthanolique, sa fraction hexanique et sa fraction enrichie en alcaloïdes se sont démarqués par leur effet sur le contrôle de l’homéostasie hépatique du glucose. Parallèlement, la fraction acétate d’éthyle a uniquement démontré un profil plus prometteur en diminuant la production de glucose dans le foie. Plus précisément, ces extraits et fractions ont significativement inhibé l’activité du glucose-6-phostaptase (G6Pase) via la stimulation des voies insulino-dépendante (Akt) et – indépendante (AMPK), à l’exception de la fraction EAc qui n’active que l’AMPK. Ce dernier mécanisme est semblable à celui de la metformine, l’hypoglycémiant oral le plus utilisé dans le monde. D’autre part, les extraits et fractions bioactifs ont également augmenté l’activité de la glycogène synthétase (GS) suite à la phosphorylation de la glycogène synthase kinase-3. Le sous-fractionnement de la fraction hexanique guidé par les deux bioessais a conduit à l’isolation de sept composés majeurs, dont quatre nouveaux triterpènes pentacycliques ayant un fragment férulate. L’acide 3β-O-trans-féruloyl-2α, 19α- dihydroxyurs-12-en-28-oïque (H1), l’acide 2α-acetoxy-3-β-0-trans-féruloryl-19α-hydroxyl- 12-en-28-oïque (H3) et l’acide ursolique (H5) sont responsables, en grande partie, de l’activité antidiabétique de la fraction hexanique. Ils ont démontré le même mécanisme d’action que l’extrait éthanolique en diminuant l’activité de la G6Pase et en augmentant celle de la GS ainsi qu’en activant les voies de signalisation impliquées. Le H3 s’est démarqué par son effet le plus puissant et comparable à celui de l’extrait éthanolique et sa fraction hexanique dans toutes les expériences. D’autre part, le sous-fractionnement de la fraction acétate d’éthyle guidé par les même bioessais a permis d’isoler cinq composés phénoliques, dont deux regioisomères Cglycosyl flavone à savoir isoorientine et orientine, ainsi que trois composés phénoliques très connus tels que l’acide chlorogénique, l’acide protocatéchique et l’aldéhyde protocatéchique. Dans tous les bioessais, isoorientine s’est démarqué avec ses effets remarquables en diminuant l’activité de la G6Pase, et augmentant celle de la GS, comparable à la fraction acétate d’éthyle. En conclusion, les résultats de cette thèse soutiennent le potentiel antidiabétique de M. arboreus rapporté dans la médecine traditionnelle africaine. Cette activité implique essentiellement le contrôle de l’homéostasie de glucose au niveau du foie à travers les voies de signalisation médiées par l’Akt, l’AMPK et la GSK-3. Les composés actifs tels que les dérivés féruliques de triterpène (H1 et H3) ainsi que le C-glycoside du flavones l’isoorientine (E1), avec un potentiel anti-diabétique prometteur pourront servir comme nouveaux candidats thérapeutiques des préparations traditionnelles ou pour des fins thérapeutiques ultérieures. Ces molécules bioactives vont représenter les marqueurs importants pour toute homologation future de cet extrait naturel de l’ethnopharmacologie congolaise.
Myrianthus arboreus est une essence de la forêt tropicale d’Afrique dont les tissus contiennent des molécules bioactives intéressantes. Les données ethnopharmacologiques africaines attribuent à l’extrait aqueux d’écorces de racines de M. arboreus des effets thérapeutiques notamment antidiabétique. Le but de cette thèse était de valider ce potentiel à travers trois séries des bioessais cellulaires in vitro, d’identifier les principes actifs et d’élucider les mécanismes d’actions par lesquels la plante exerce son activité sur le diabète. Une étude préliminaire sur les différents extraits et fractions obtenus à partir de solvants de force de polarité croissante (dichlorométhane, éthanol, méthanol, eau) a été menée et a démontré un excellent potentiel antioxydant de l’extrait éthanolique et de sa fraction acétate d’éthyle comparable à celui de l’Oligopin®, un extrait commercial riche en proanthocyanidines. Ces résultats ont corrélé avec les teneurs en acides hydroxicimiques et en proanthocyanidines, deux classes de polyphénols présentes dans l’extrait et la fraction acétate d’éthyle. Ensuite, les résultats des bioessais cellulaires indiquent qu’aucun extrait ou fraction n’a été capable de stimuler le transport de glucose dans les cellules musculaires C2C12. A l’inverse, l’extrait éthanolique, sa fraction hexanique et sa fraction enrichie en alcaloïdes se sont démarqués par leur effet sur le contrôle de l’homéostasie hépatique du glucose. Parallèlement, la fraction acétate d’éthyle a uniquement démontré un profil plus prometteur en diminuant la production de glucose dans le foie. Plus précisément, ces extraits et fractions ont significativement inhibé l’activité du glucose-6-phostaptase (G6Pase) via la stimulation des voies insulino-dépendante (Akt) et – indépendante (AMPK), à l’exception de la fraction EAc qui n’active que l’AMPK. Ce dernier mécanisme est semblable à celui de la metformine, l’hypoglycémiant oral le plus utilisé dans le monde. D’autre part, les extraits et fractions bioactifs ont également augmenté l’activité de la glycogène synthétase (GS) suite à la phosphorylation de la glycogène synthase kinase-3. Le sous-fractionnement de la fraction hexanique guidé par les deux bioessais a conduit à l’isolation de sept composés majeurs, dont quatre nouveaux triterpènes pentacycliques ayant un fragment férulate. L’acide 3β-O-trans-féruloyl-2α, 19α- dihydroxyurs-12-en-28-oïque (H1), l’acide 2α-acetoxy-3-β-0-trans-féruloryl-19α-hydroxyl- 12-en-28-oïque (H3) et l’acide ursolique (H5) sont responsables, en grande partie, de l’activité antidiabétique de la fraction hexanique. Ils ont démontré le même mécanisme d’action que l’extrait éthanolique en diminuant l’activité de la G6Pase et en augmentant celle de la GS ainsi qu’en activant les voies de signalisation impliquées. Le H3 s’est démarqué par son effet le plus puissant et comparable à celui de l’extrait éthanolique et sa fraction hexanique dans toutes les expériences. D’autre part, le sous-fractionnement de la fraction acétate d’éthyle guidé par les même bioessais a permis d’isoler cinq composés phénoliques, dont deux regioisomères Cglycosyl flavone à savoir isoorientine et orientine, ainsi que trois composés phénoliques très connus tels que l’acide chlorogénique, l’acide protocatéchique et l’aldéhyde protocatéchique. Dans tous les bioessais, isoorientine s’est démarqué avec ses effets remarquables en diminuant l’activité de la G6Pase, et augmentant celle de la GS, comparable à la fraction acétate d’éthyle. En conclusion, les résultats de cette thèse soutiennent le potentiel antidiabétique de M. arboreus rapporté dans la médecine traditionnelle africaine. Cette activité implique essentiellement le contrôle de l’homéostasie de glucose au niveau du foie à travers les voies de signalisation médiées par l’Akt, l’AMPK et la GSK-3. Les composés actifs tels que les dérivés féruliques de triterpène (H1 et H3) ainsi que le C-glycoside du flavones l’isoorientine (E1), avec un potentiel anti-diabétique prometteur pourront servir comme nouveaux candidats thérapeutiques des préparations traditionnelles ou pour des fins thérapeutiques ultérieures. Ces molécules bioactives vont représenter les marqueurs importants pour toute homologation future de cet extrait naturel de l’ethnopharmacologie congolaise.
Myrianthus arboreus is a tree of the tropical rainforest of Africa the tissues of which contain promising bioactive compounds. African ethnopharmacological data attribute to the aqueous extract of M. arboreus root bark therapeutic effects including antidiabetic. The aim of this thesis was to validate this potential through three series of cell-bioassays, to identify the principle responsible for this activity and to elucidate the mechanisms of actions by which this plant extract exerts its antidiabetic activity. A preliminary study on different extracts and fractions performed with solvent of increasing polarity (dichloromethane, ethanol, methanol and water) has revealed an excellent antioxidant potential of ethanol extract and of its ethyl acetate fraction comparable to that of Oligopin®, a commercial extract rich in proanthocyanidins. This result was well correlated with the levels of hydroxycinnamic acids and proanthocyanins, two classes of polyphenols present in the extract and its antioxidant fraction. In addition, the results of cell bioassays indicate that no extract or fraction was able to stimulate glucose transport in C2C12 muscle cells. In contrast, the ethanolic extract, its hexane and the alkaloid-enriched fractions stood out by their effect on the control of hepatic glucose homeostasis. Similarly, the ethyl acetate fraction only demonstrated a more promising profile by decreasing glucose production in the liver. Specifically, these extracts and fractions significantly inhibited glucose-6-phostaptase (G6Pase) activity via insulindependent (Akt) and -independent (AMPK) pathway stimulation, except for the ethyl acetate ethanolic fraction which only activated AMPK. The latter mechanism is similar to that of metformin, the most widely used oral hypoglycemic drug in the world. On the other hand, bioactive extracts and fractions also increased the activity of glycogen synthase (GS) through the phosphorylation of glycogen synthase kinase-3. Sub-fractionation of the hexane fraction guided by the two bioassays resulted in the isolation of seven major compounds, including new pentacyclic triterpenes having a ferulate fragment. 3β-O-trans-feruloyl-2α, 19α-dihydroxyurs-12-en-28-oic acid (H1), 2α-acetoxy-3- β-O-trans-feruloryl-19α-hydroxyl-acid 12-en-28- oic acid (H3) and ursolic acid (H5) which are responsible, to a large extent, for the antidiabetic activity of the hexane fraction. The active constituents exhibit the same mechanism of action as the EtOH extract by decreasing the activity of G6Pase, increasing that of GS and activating signaling pathways involved. The H3 stood out for its most powerful effect which was determined to be comparable to that of the ethanolic extract and to its Hexane fraction in all experiments. On the other hand, bioassay-guided fractionation of ethyl acetate fraction led to the isolation of five phenolic compounds, including two regioisomers C-glycosyl flavons namely isoorientin and orientin, along with three well-known phenolic compounds, chlorogenic acid, protocatechuic acid and protoctehuic aldehyde. In all experiments, isoorientin (E1) stood out with its most powerful effects, by decreasing the activity of G6Pase and increasing the GS. Altogether, the results of the present thesis support the antidiabetic potential of M. arboreus reported by African folk medicine, which involves control of glucose homeostasis in the liver via Akt, AMPK and GSK-3 phosphorylation. The bioactive molecules ferulic derivatives of triterpenes (H1 and H3), and C-glycosyl flavone E1 determined in studied extracts and fractions were determined to have a promising anti-diabetic activity, can be used as new therapeutic candidates for traditional preparations or for future therapeutic purposes. These molecules can also be used as markers for future authentifications of the natural products based on Myrianthus arboreus root bark extracts.
Myrianthus arboreus is a tree of the tropical rainforest of Africa the tissues of which contain promising bioactive compounds. African ethnopharmacological data attribute to the aqueous extract of M. arboreus root bark therapeutic effects including antidiabetic. The aim of this thesis was to validate this potential through three series of cell-bioassays, to identify the principle responsible for this activity and to elucidate the mechanisms of actions by which this plant extract exerts its antidiabetic activity. A preliminary study on different extracts and fractions performed with solvent of increasing polarity (dichloromethane, ethanol, methanol and water) has revealed an excellent antioxidant potential of ethanol extract and of its ethyl acetate fraction comparable to that of Oligopin®, a commercial extract rich in proanthocyanidins. This result was well correlated with the levels of hydroxycinnamic acids and proanthocyanins, two classes of polyphenols present in the extract and its antioxidant fraction. In addition, the results of cell bioassays indicate that no extract or fraction was able to stimulate glucose transport in C2C12 muscle cells. In contrast, the ethanolic extract, its hexane and the alkaloid-enriched fractions stood out by their effect on the control of hepatic glucose homeostasis. Similarly, the ethyl acetate fraction only demonstrated a more promising profile by decreasing glucose production in the liver. Specifically, these extracts and fractions significantly inhibited glucose-6-phostaptase (G6Pase) activity via insulindependent (Akt) and -independent (AMPK) pathway stimulation, except for the ethyl acetate ethanolic fraction which only activated AMPK. The latter mechanism is similar to that of metformin, the most widely used oral hypoglycemic drug in the world. On the other hand, bioactive extracts and fractions also increased the activity of glycogen synthase (GS) through the phosphorylation of glycogen synthase kinase-3. Sub-fractionation of the hexane fraction guided by the two bioassays resulted in the isolation of seven major compounds, including new pentacyclic triterpenes having a ferulate fragment. 3β-O-trans-feruloyl-2α, 19α-dihydroxyurs-12-en-28-oic acid (H1), 2α-acetoxy-3- β-O-trans-feruloryl-19α-hydroxyl-acid 12-en-28- oic acid (H3) and ursolic acid (H5) which are responsible, to a large extent, for the antidiabetic activity of the hexane fraction. The active constituents exhibit the same mechanism of action as the EtOH extract by decreasing the activity of G6Pase, increasing that of GS and activating signaling pathways involved. The H3 stood out for its most powerful effect which was determined to be comparable to that of the ethanolic extract and to its Hexane fraction in all experiments. On the other hand, bioassay-guided fractionation of ethyl acetate fraction led to the isolation of five phenolic compounds, including two regioisomers C-glycosyl flavons namely isoorientin and orientin, along with three well-known phenolic compounds, chlorogenic acid, protocatechuic acid and protoctehuic aldehyde. In all experiments, isoorientin (E1) stood out with its most powerful effects, by decreasing the activity of G6Pase and increasing the GS. Altogether, the results of the present thesis support the antidiabetic potential of M. arboreus reported by African folk medicine, which involves control of glucose homeostasis in the liver via Akt, AMPK and GSK-3 phosphorylation. The bioactive molecules ferulic derivatives of triterpenes (H1 and H3), and C-glycosyl flavone E1 determined in studied extracts and fractions were determined to have a promising anti-diabetic activity, can be used as new therapeutic candidates for traditional preparations or for future therapeutic purposes. These molecules can also be used as markers for future authentifications of the natural products based on Myrianthus arboreus root bark extracts.

SILVA, Aline Holanda. Desenvolvimento e caracterização do extrato de Erythrina velutina para o tratamento de doença neurodegenerativa. 2012. 113 f. Dissertação (Mestrado em Ciências Farmacêuticas) - Universidade Federal do Ceará, Faculdade de Farmácia, Odontologia e Enfermagem, Fortaleza, 2012.
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Dentre as espécies do gênero Erythrina (Fabaceae), duas conhecidas popularmente como mulungu, E. velutina e E. mulungu, tem comprovado interesse social e econômico para o Brasil. E. mulungu ocorre no Sudeste, pertence à Relação Nacional de Plantas de Interesse para o SUS (RENISUS). Já E. velutina é uma árvore amplamente utilizada no Nordeste, cuja casca do caule é utilizada tradicionalmente no tratamento da ansiedade, agitação e insônia. Contudo, não existe um produto farmacêutico com qualidade agregada e estudos farmacológicos dessa espécie para justificar seu uso medicinal. Diante do exposto, o objetivo do presente trabalho foi realizar o desenvolvimento e o controle de qualidade do extrato de Erytrhina velutina e investigar as atividades antioxidante e neuroprotetora, visando seu emprego no tratamento de doença neurodegenerativa, como a Doença de Parkinson (DP). Inicialmente, foi validado método espectrofotométrico para dosagem de fenóis totais (FT) em produtos derivados de E. velutina, sendo este específico, linear, preciso, exato e robusto. Foi estabelecido o método de preparação (estufa com circulação e renovação de ar – 80°C; 24h) e especificações para o controle da droga vegetal. O extrato etanólico de E. velutina (EEEV) produzido por percolação foi caracterizado quando ao teor de FT (155,14 ± 3,31 µ EAG/mg de extrato) e perfil cromatográfico por CLAE-DAD (Fenóis: hesperidina - Tr: 18,8 min; abssinina - Tr: 22,9 min; homoesperidina Tr: 31,2 min; ácido rizônico (AR) - Tr: 32,1 min e sigmoidina C - Tr: 38,6 min). O EEEV não mostrou citotoxicidade no teste do MTT, mas aumentou (EEEV: 100 e 200 µg/mL) a atividade da LDH em neutrófilo humano. Em células 9L/lacZ, o EEEV (100, 400, 1000 mg/mL) reduziu significativamente a viabilidade celular, teste MTT. EEEV (0,0025 – 1 µg/mL) e AR (0,0025 – 1 µg/mL) não foram citotóxicos e inibiram parcialmente a neurotoxicidade induzida por 6-OHDA em células SH-SY5Y, modelo experimental de DP, observada pela redução significativa dos níveis de nitrito/nitrato. Na avaliação do potencial antioxidante, testes DPPH e NBT, o EEEV (10 – 200 µg/mL) apresentou atividade sequestradora de radicais livres no teste DPPH. Os resultados obtidos no presente estudo permitiram definir as condições ideais de preparação da droga vegetal e do extrato, os quais foram caracterizados, incluindo validação de método analítico e avaliação farmacológica, comprovando o potencial antioxidante e neuroprotetor do EEEV e AR.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
Rosmarinus officinalis L., popularly known as rosemary, is a medicinal and aromatic species used in the cosmetics and food processing industries, with interest for the herbal industry. This study aimed to determine the biomass production and volatile oil during the growth of rosemary plants cultivated under different salinity (nutritious solution concentrations) and different nitrogen (N) concentrations, and to evaluate the effect of the extracts and volatile oil of the species on the Allium cepa L. cell cycle and genetic material, as well as knowing the phenolic compounds present in the extracts and the chemical composition of volatile oil. The work was divided into two stages: the first aimed evaluating the production of biomass and volatile oil during the rosemary plants growth cultivated under different nutritious solution concentrations, as well as the effect of oil and extracts on the A. cepa cell cycle and genetic material, besides determining the chemical composition of the oil and the phenolic compounds present in the extracts; the second stage aimed evaluating biomass and volatile oil production during the rosemary plants growth cultivated under different N concentrations, as well as the effect of oil and the extracts on the A. cepa cell cycle and genetic material, and determining the chemical composition of the oil and the phenolic compounds present in the extracts as well. The rosemary leaves were collected and analyzed for dry matter, volatile oil extraction and preparation of aqueous extracts by infusion at 5, 20, and 50 g.L-1 concentrations for plants cultivated under different salinities, and 5 and 50 g.L-1 concentrations for plants cultivated at different N concentrations. The volatile oil was analyzed by Gas Chromatography to knowledge of its constitution, and the aqueous extracts were analyzed by High Performance Liquid Chromatography for phenolic compounds determination. The oil and extracts effect was evaluated on the A. cepa cell cycle and genetic material. The completely randomized design was used, and the biomass production and volatile oil yield data were subjected to analysis of variance with polynomial regression and the remaining data were compared by the Scott-Knott test at the 5% level of probability. The nutritious solution concentration, the N concentration and the plant age interfere into the biomass production, and consequently into the rosemary volatile oil yield. However, do not alter the oil and extracts effect on the A. cepa cell cycle and genetic material. The volatile oil and extracts inhibited cell division in all concentrations, and this effect is dose-dependent. In addition, extracts did not show genotoxicity, in spite of oil concentration of 10% being genotoxic. The majority compounds camphor, 1.8 cineole, α-pinene, β-myrcene and verbenone, beyond of β-pinene compounds only controlling the oil crop plants in the concentrations of 8:05, 13:05 and 15:55 mmol.L-1. The phenolic compounds present in the extracts were rosmarinic acid, chlorogenic acid, caffeic acid, carnosic acid, kaempferol, quercetin and rutin.
Rosmarinus officinalis L., popularmente conhecida como alecrim, é uma espécie medicinal e aromática utilizada na indústria de cosméticos e agroalimentícia, com interesse para a indústria de fitoterápicos. Objetivou-se determinar a produção de fitomassa, óleo volátil, composição química do óleo e compostos fenólicos presentes nos extratos, durante o crescimento de plantas de alecrim cultivadas com diferentes salinidades (concentrações de solução nutritiva) e diferentes doses de nitrogênio (N), além de avaliar o efeito dos extratos e óleo volátil da espécie sobre o ciclo celular e material genético de Allium cepa L.. Foram coletadas as folhas de alecrim para determinação da fitomassa, extração de óleo volátil e preparo de extratos aquosos por infusão. Avaliou-se a produção de fitomassa e óleo volátil durante o crescimento de plantas de alecrim cultivadas com diferentes salinidades (1.0, 2.0, 3.0, 4.0 e 5.0 dS.m-1) e em diferentes doses de N (5.55, 8.05, 10.55, 13.05 e 15.55 mmol.L-1), além de avaliar o efeito do óleo (3 e 10%) e dos extratos (nas concentrações de 5, 20, e 50 g L-1 para as plantas cultivadas em diferentes salinidades e nas concentrações de 5 e 50 g L-1 a partir das plantas cultivadas em diferentes doses de N), sobre o ciclo celular e material genético de A. cepa. O óleo volátil foi submetido à análise por Cromatografia Gasosa para conhecimento de sua constituição, e os extratos aquosos à análise por Cromatografia Líquida de Alta Eficiência para a determinação dos compostos fenólicos. As plantas cultivadas em diferentes salinidades foram coletadas aos 100 e aos 160 dias após o plantio (DAP) e as cultivadas em diferentes doses de N aos 90, 180, 270 e 365 DAP. Foi utilizado o delineamento experimental inteiramente casualizado, os dados de produção de fitomassa e rendimento de óleo volátil foram submetidos à análise de variância com regressão polinomial e os demais dados foram comparados pelo teste Scott-Knott (p<0,05). A salinidade, a dose de N e a idade fisiológica da planta interferem na produção de fitomassa e consequentemente no rendimento de óleo volátil de alecrim e não alteram o efeito do óleo e dos extratos sobre o ciclo celular e material genético de A. cepa. O óleo volátil e os extratos inibiram a divisão celular em todas as concentrações estudadas e esse efeito é dose dependente, mas sem apresentar genotoxicidade, exceto na concentração de 10% quando o óleo foi genotóxico. Os compostos majoritários do óleo foram cânfora, 1.8 cienol, α-pineno, β-myrceno e verbenona, além dos compostos β-pineno majoritários apenas no óleo de plantas cultivadas nas concentrações de 8.05, 13.05 e 15.55 mmol.L-1. Os compostos fenólicos presentes nos extratos foram ácido rosmarínico, ácido clorogênico, ácido cafeico, ácido carnósico, canferol, quercetina e rutina.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico
As macroalgas sÃo fontes de molÃculas bioativas com propriedades antioxidantes, muitas das quais apresentam variaÃÃes ao longo do ano. Cinco algas verdes do gÃnero Caulerpa (C. cupressoides, C. mexicana, C. prolifera, C. racemosa e C. sertularioides) foram coletadas mensalmente na Praia do Pacheco, de janeiro a dezembro de 2006. ApÃs as coletas, elas foram transportadas para o laboratÃrio, lavadas com Ãgua corrente para retirada de impurezas e epÃfitas e liofilizadas. Os extratos algÃceos foram preparados em metanol (MeOH) na proporÃÃo 1:20 (p:v) sob agitaÃÃo por 1 h a 20ÂC em um agitador orbital, em seguida, filtrados, e os resÃduos submetidos a mais duas extraÃÃes sucessivas. O material das trÃs extraÃÃes metanÃlicas foi reunido e concentrado em um evaporador rotativo. PorÃÃes de 10 mg do extrato concentrado foram suspensas em 10 mL de MeOH (50%). Esse extrato (1 mg/mL) foi utilizado para a determinaÃÃo do conteÃdo fenÃlico total (CFT). A atividade antioxidante in vitro foi medida atravÃs da capacidade de sequestrar o radical DPPH, do poder de reduÃÃo de Ãons fÃrricos (FRAP), do branqueamento do β-caroteno (BCB) e da habilidade de quelaÃÃo de Ãons ferrosos (FIC), nas concentraÃÃes 5, 50, 500 e 1.000 Âg/mL, preparadas a partir do extrato. O quelante de metais EDTA foi utilizado como controle positivo no FIC, e o antioxidante sintÃtico BHA, nas demais determinaÃÃes. Os valores de CFT foram maiores no segundo semestre do ano e variaram de 4 a 32 mg AGE/g extrato. Os resultados do sequestro do radical DPPH, do BCB e do FIC nÃo exibiram padrÃo de variaÃÃo e apresentaram atividades superiores a 25%, 70% e 10%, respectivamente. O FRAP nos extratos de C. cupressoides, C. racemosa e C. serlularioides foi maior no segundo semestre do ano, no de C. mexicana o maior valor ocorreu no primeiro semestre e no de C. prolifera, as variaÃÃes ao longo do ano nÃo obedeceram a um padrÃo como nos demais. Em todas as metodologias as atividades dos extratos algÃceos foram inferiores Ãquelas dos controles positivos. As cinco espÃcies de Caulerpa podem ser consideradas fonte de potenciais antioxidantes, com possÃveis usos, como na indÃstria alimentÃcia, nÃo requisitando de um perÃodo do ano especÃfico para a extraÃÃo desses compostos antioxidantes, jà que embora haja variaÃÃo ao longo do ano, ela nÃo à muito pronunciada.
Macroalgae are sources of bioactive molecules with antioxidant properties, many of which present seasonal variation. Five green algae of the genus Caulerpa (C. cupressoides, C. mexicana, C. prolifera, C. racemosa and C. sertularioides) were collected monthly at Pacheco Beach, from January to December 2006. After collection, they were transported to the laboratory, washed under running water to remove impurities and epiphytes, and freeze-dried. The algae extracts were prepared in methanol (MeOH) in the proportion 1:20 (w:v) under stirring for 1 h at 20ÂC in an orbital shaker followed by filtration, and the residues were submitted to two further successive extractions. The filtrate from the three methanolic extractions was gathered and concentrated in a rotary evaporator. Portions of the concentrated extract (10 mg) were suspended in 10 mL MeOH (50%). This extract (1 mg/mL) was used in the determination of the total phenolic content (TPC). In vitro antioxidant activity was measured by DPPH radical scavenging, ferric-reducing antioxidant power (FRAP), β-carotene bleaching (BCB), and ferrous ion chelating (FIC), at the concentrations 5, 50, 500 and 1000 Âg/mL, prepared from the extract. The metal chelator EDTA was used as a positive control in FIC, while the remaining essays used the synthetic antioxidant BHA. The TPC values were higher in the second semester of the year and varied from 4 to 32 mg GAE/g extract. The DPPH radical scavenging, BCB and FIC results did not exhibit variation patterns and presented activities greater than 25%, 70% and 10%, respectively. FRAP in the extracts of C. cupressoides, C. racemosa and C. sertularioides was higher in the second half of the year, while for C. mexicana the largest value was found in the first half of the year, and for C. prolifera variations during the year did not obey a pattern similar to the others. In all methodologies the activities of the alga extracts were inferior to the positive controls. The five species of Caulerpa can be considered potential sources of antioxidants with possible uses, such as in the food industry, not requiring a specific annual period for the extraction of these antioxidant compounds, as though there is variation across the year, it is not highly pronounced.

Microbial and oxidative changes occurring in meat and meat products cause loss of quality and consequent commercial devaluation. The consumption of meat products has caused concern among consumers by the presence of chemical preservatives, which are often questioned because of the risk of harmful side effects to health. In an attempt to control microbial and oxidative changes, researches have been performed with the purpose of isolating natural phytochemicals obtained from various plant substrates, which besides having the preference of the consumers, also have the ability to act helping in the prevention of diseases. Antioxidant and antimicrobial activities have been attributed to the phenolic compounds; persimmon (Diospyros kaky L.) was used in this study since it is rich in polyphenols. Initially, we compared the antioxidant activity of hydroethanolic extracts of Diospyros kaki, L. of the cultivars Quioto and Rama Forte in chicken meat. The effect of these cultivars on the color and sensory characteristics of the samples was also verified. The crude hydroethanolic extract of persimmon cv. Rama Forte was used in subsequent tests since it showed a higher content of phenolic compounds, higher antioxidant activity, and it did not affect the sensory characteristics of chicken meat. The hydroethanolic extract of persimmon cv. Rama Forte was fractionated using solvents with different polarities (n-hexane, chloroform, and ethyl acetate) and subjected along with the hexane, chloroform, and ethyl acetate fractions and the residual fraction to the determination of total phenolic compounds as well as antimicrobial and antioxidant activity in vitro. The antioxidant activity in beef was verified and compared with the activity of the oily extract of rosemary (Chr. Hansen ®). The extract and/or the most efficient fractions were evaluated for antioxidant and antimicrobial activity as well as for the influence on the sensory characteristics and color of meat products (beef burgers and salami). Based on the results obtained, the crude hydroethanolic extract of persimmon cv. Rama Forte and the residual and ethyl acetate fractions showed antioxidant activity in beef during the storage period, however, no antimicrobial activity was observed. In the salami samples, the crude hydroethanolic extract of persimmon cv. Rama Forte also showed antioxidant activity and did not alter the sensory characteristics or microbial counts performed after the manufacturing period of the salami. In the frozen beef burgers, crude hydroethanolic extract of persimmon cv. Rama Forte and residual and ethyl acetate fractions, when used in the concentrations of 0.5 and 0.7%, did not show antioxidant potential or antimicrobial characteristics. Moreover, they did not interfere in the sensory samples. However, the ethyl acetate fraction (0.5 and 0.7%) contributed to the retention and stability of the red color of the samples of beef burgers during the storage of the frozen product. Thus, based on the results obtained, it seems appropriate to assume that the crude hydroethanolic extract of persimmon cv. Rama Forte as well as residual and ethyl acetate fractions have antioxidant potential. Further studies should be performed mainly in relation to the antioxidant activity in frozen meat products.
As alterações microbianas e oxidativas indesejáveis que podem ocorrer na carne ou nos produtos cárneos causam perda de qualidade e conseqüente desvalorização comercial. O consumo de produtos cárneos tem causado preocupação nos consumidores pela presença de conservantes sintéticos, os quais são questionados freqüentemente devido aos riscos de apresentarem efeitos colaterais maléficos à saúde. Com o intuito de controlar as referidas alterações, pesquisas têm sido desenvolvidas, com o propósito de estudar fitoquímicos naturais, obtidos de diversos substratos vegetais, os quais além de terem a preferência dos consumidores, também possuem a capacidade de atuarem como auxiliares na prevenção de doenças. Aos compostos fenólicos tem sido atribuída atividade antimicrobiana e antioxidante, e considerando tal capacidade foi utilizado neste estudo o caqui (Diospyros kaky L.) que é rico em polifenóis. Inicialmente foi comparada a atividade antioxidante dos extratos hidroetanólicos de Diospyros kaki L. das cultivares Quioto e Rama Forte em carne de frango e verificado o efeito dos mesmos sobre a cor e as características sensoriais das amostras. O extrato hidroetanólico bruto de caqui da cultivar Rama Forte, que apresentou maior teor de compostos fenólicos, maior atividade antioxidante e que não interferiu nas características sensoriais da carne de frango foi empregado nos testes posteriores. O extrato hidroetanólico de caqui da cultivar Rama Forte foi fracionado usando solventes com diferentes polaridades (n-hexano, clorofórmio e acetato de etila) e submetido juntamente com as frações hexânica, clorofórmica, de acetato de etila e a fração residual a determinação de compostos fenólicos totais, atividade antioxidante e antimicrobiana in vitro. Também foi verificada a atividade antioxidante dos mesmos em carne bovina e comparada com a atividade do extrato oleoso de alecrim (Chr. Hansen®). O extrato e/ou frações mais eficientes foram avaliados quanto a atividade antioxidante e antimicrobiana, bem como quanto a influencia sobre as características sensoriais e a cor de produtos cárneos (hambúrguer e salame). Com base nos resultados obtidos pode-se verificar que o extrato hidroetanólico bruto de caqui da cultivar Rama Forte, bem como as frações residual e a de acetato de etila apresentaram atividade antioxidante na carne bovina durante o período de armazenamento, mas não apresentaram atividade antimicrobiana. Nas amostras de salame, o extrato hidroetanólico bruto de caqui da cultivar Rama Forte também apresentou atividade antioxidante e não alterou as características sensoriais nem as contagens microbianas realizadas após o período de processamento do salame. Já no hambúrguer congelado o extrato hidroetanólico bruto de caqui da cultivar Rama Forte e as frações residual e de acetato de etila, quando utilizados nas concentrações de 0,5 e 0,7%, não demonstraram potencial antioxidante e antimicrobiano, e não interferiram nas características sensoriais das amostras, enquanto que a fração acetato de etila (0,5 e 0,7%) contribuiu na retenção e estabilidade da cor vermelha das amostras de hambúrguer de carne bovina durante o período de armazenamento do produto congelado. Desta forma, com base nos resultados obtidos parece adequado supor que o extrato hidroetanólico bruto de caqui da cultivar Rama Forte e as frações residual e de acetato de etila apresentam potencial antioxidante, mas estudos mais aprofundados devem ser realizados principalmente no que se refere a atividade antioxidante em produtos cárneos congelados.

Orientador: Wagner Vilegas
Resumo: Organismos marinhos são importantes fontes de novos produtos naturais e compostos bioativos com funções variadas. Entre as principais fontes de novas moléculas esta o filo dos Cnidários, com uma importante contribuição no numero de compostos descritos nos últimos anos. Bunodossoma caissarum é uma espécie de anêmona endêmica do Brasil, cujo perfil químico foi analisado. Para essa análise foram realizados uma extração hidroalcoólica e o isolamento de determinadas moléculas com técnicas cromatográficas. Os compostos isolados foram aqueles que apresentaram absorbância no comprimento de onda no UV em 310 nm e foram identificados a partir de técnicas de espectrometria de massas e ressonância magnética. Entre as substâncias encontradas estão sais piridinicos, núcleos imidazóis, caissarona e a etilglicose, moléculas com importantes papeis fisiológicos
Abstract: Marine organisms are importante sources of new natural products and bioactive compounds with various functions. The phylum Cnidaria is among the main sources of new molecules, with na importante contribution to the number of compounds described in the last years. The anemone species Bunodossoma caissarum is endemic to Brazil and had it's chemical profile analyzed. For the purpose of this analysis were done a hydroalcoolic extraction and the isolation of certain molecules by chomatographic techniques. The isolated compounds were those that displayed absorbance in the 310nm wavelength of the UV spectrum and were identified on the basis of mass spectrometry and magnetic ressonance. Among the substances found are pyridinic salts, imidazole nucleus, caissarone and ethyl glucose, which are molecules with important physiological roles
Mestre

Die pilzlichen Erreger der Tomate Alternaria solani und des Manioks Colletotrichum gloeosporioides f.sp. manihotis verursachen große Ertragsausfälle in Westafrika. In der vorliegenden Arbeit wurden die Blattkrankheiten nach Applikationen mit wässerigen Extrakten aus Westafrika (Ausgangsmaterial: organischer Haushaltsmüll) unter kontrollierten Bedingungen unterdrückt. Im Pathosystem Tomate - A. solani erzielten die Extrakte eine Befallsverminderung um bis zu 94 % im Vergleich zur Kontrollvariante und im Maniok - C. gloeosporioides wurde die Anzahl der gebildeten Acervuli/cm² Blattoberfläche bis um 87 % reduziert. Der primäre Wirkmechanismus der Extrakte gegen die Pathogene ist Antagonismus, hervorgerufen von den Mikroorganismen in den Extrakten. Isolierte Bakterien aus den Extrakten hemmten das Wachstum der Pathogene in Plattentests, was auf die Bildung von fungistatischen Substanzen zurückzuführen war. Die Bakterien, die aus den Extrakten isoliert und identifiziert wurden, gehörten überwiegend zu den Bakteriengattungen Pseudomonas und Bacillus. Die Extraktionsbedingungen nahmen in Hinblick auf die Wirksamkeit der Extrakte gegen die Pathogene eine wichtige Stellung ein. Die Untersuchungen zeigten einen Zusammenhang zwischen gestiegener Mikroorganismenanzahl und Befallsreduktion. Durch die Auswahl optimaler Extraktionsbedingungen erhöhte sich die Anzahl der Koloniedichte in den Extrakten; dies führte zur einer stärkeren Befallsreduzierung an der Tomate und am Maniok. Hierbei waren die Umgebungstemperatur und die Häufigkeit des Durchmischens des Kompost-Wasser-Gemisches von besonderer Bedeutung. Extrakte, die während der Extraktionszeit einer Temperatur von 20°C ausgesetzt waren, erzielten eine stärkere Befallsreduzierung von A. solani an der Tomate und wiesen eine höhere Mikroorganismendichte auf als Extrakte, die einer Temperatur von 30°C und 35°C ausgesetzt waren. Tägliches Durchmischen des Wasser-Kompost-Gemisches während der Extraktionszeit erhöhte die Mikroorganismendichte um das Siebenfache in den Extrakten und die Wirksamkeit gegen A. solani und C. gloeosporioides im Vergleich zu den Extrakten, die nur zu Beginn durchmischt wurden. Die Lagerungszeit der fertigen Komposte hatte einen Einfluß auf die Mikroorganismendichte der wässerigen Extrakte sowie auf deren Wirksamkeit. Extrakte, die aus frischen Komposten hergestellt wurden, zeigten einen besseren suppressiven Effekt gegen die Pathogene und wiesen eine höhere Mikroorganismenanzahl auf als Extrakte, deren Komposte schon länger gelagert worden waren. Die Anreicherung der Extrakte mit dem Formulierungshilfsstoff Xanthan steigerte die Wirksamkeit der Extrakte gegen A. solani an Tomatenpflanzen und gegen C. gloeosporioides f.sp. manihotis an abgetrennten Maniokblättern. Angereicherte Extrakte reduzierten den Befall um bis zu 46 % mehr als der Extrakt ohne Formulierungshilfsstoffe, während die Steigerung der Wirksamkeit nach Anreicherung mit Alginat oder Zellulose eine Abhängigkeit von der Herkunft der Komposte zeigte. Eine erhöhte ß-1,3-Glucanase-Aktivität wurde am vierten Tag nach der Applikation von Extrakten an der Tomate und an der Batate festgestellt. Diese Ergebnisse belegen die Bedeutung von Spritzbehandlungen mit wässerigen Kompostextrakten gegen die Pathogene A. solani und C. gloeosporioides f.sp. manihotis und stellen eine erfolgversprechende Alternative zur Bekämpfung von Pilzkrankheiten in Westafrika dar.
Great yield losses of cassava in West Africa are caused by fungal pathogens that also affect tomato, e.g. Alternaria solani and Colletotrichum gloeosporioides f sp. manihotis. In the present study these foliar diseases were effectively suppressed under controlled conditions after application of watery compost extracts to tomato and cassava The disease incidence of A. solani on tomato was reduced up to 94 % and the number of the acervuli produced on the surface of cassava leaves was reduced up to 87% compared to the control treatment. Reduction is due to antagonistic effects of the microorganisms in the extracts towards the pathogens. Bacterial strains isolated from the extracts suppressed the growth of the pathogens in dual-culture-tests due to the production of antibiotics. Bacteria isolated and identified from the extracts were mostly pseudomonas and bacillus strains. The conditions used for extraction of compost played a significant role for their effectiveness against different pathogens. The investigations showed that optimisation of extract conditions increased the density of microorganisms in the extracts and led to a higher disease reduction on tomato and cassava plants. Temperature and stirring frequency of the compost - water mixture during the extraction period were very important factors for increased effectiveness. Extracts incubated at 20 °C during the extraction period caused a higher disease reduction on tomato and showed a higher density of microorganisms, than extracts that were incubated at 30 or 35 °C. Daily stirring of the compost - water mixture during the extraction period increased the density of microorganisms in the extracts sevenfold and increased the effectiveness against A. solani and C. gloeosporioides compared to the extract that was stirred only once at the beginning. The Effectiveness as well as the density of the microorganisms were depended on the incubation time of the compost extracts. Extracts produced from fresh composts showed greater suppressive effects against pathogens and higher densities of microorganisms than extracts of composts that were stored for a longer time. Enrichment of the extracts with the formulate substance xanthan increased the effectiveness against A. solani on tomato plants and against C. gloeosporioides on detached leaves of cassava and reduced the disease incidence up to 46% compared to the extract without formulate substances. The increase of the effectiveness after enriching with alginate was depended on the origin of the compost. An increased ß-1,3- glucanase activity were determined on tomato and sweet potato leaves 4 days after the application of the extracts onto the plants. The results of this study show that watery compost extracts can be successfully applied against the cassava and tomato pathogens A. solani and C. gloeosporioides f. sp. manihotis and offers easily applicable methods for the reduction of fungal diseases in West Africa.

Le règne végétal est une ressource renouvelable d'une large gamme de métabolites secondaires biologiquement actifs. Ces travaux de thèse proposent une stratégie multidisciplinaire d'évaluation du potentiel de composés phénoliques antimicrobiens d'origine végétale pour la conservation des aliments. Un criblage de l'activité antimicrobienne in vitro vis-à-vis de 8 souches de la flore pathogène et d'altération des aliments d'une centaine de molécules pures et une soixantaine d'extraits végétaux a d'abord permis de sélectionner les plus actifs. Différents mécanismes d'action vis-à-vis de S. aureus ont pu être mis en évidence par cytométrie de flux couplée à l'utilisation de marqueurs de l'état physiologique des bactéries pour quelques uns des composés actifs sélectionnés. En vue d'une application à de la viande bovine, l'activité antibactérienne des composés phénoliques ou extraits végétaux les plus actifs a été réévaluée dans des milieux de culture plus complexes mimant leur teneur en protéines et en matières grasses. Les résultats de ce criblage et un suivi microbiologique de viande hachée de bœuf avec 1% (m/m) d'extrait ajouté ont permis d'observer que les pertes d'activité antibactérienne observées étaient notamment corrélées aux interactions des composés phénoliques avec les protéines ou les matières grasses. L'incorporation des composés phénoliques ou extraits végétaux dans des matériaux d'emballage en contact alimentaire a constitué la seconde voie de mise en œuvre envisagée. Des films plastiques conservant une activité antibactérienne ont ainsi pu être élaborés par voie fondue
The plant kingdom is a renewable resource of a wide range of biologically active secondary metabolites. This thesis proposes a multidisciplinary strategy for evaluating the potential of plant-derived antimicrobial phenolic compounds for food preservation. A screening of the antimicrobial activity in vitro against 8 strains of foodborne pathogenic and spoilage microorganisms of a hundred pure molecules and about sixty plant extracts allowed to select the most active. Different mechanisms of action with respect to S. aureus could be demonstrated by flow cytometry coupled with the use of probes of the physiological state of the bacteria for some of the selected active compounds. For application to beef, the antibacterial activity of the most active phenolic compounds or plant extracts has been re-evaluated in more complex culture media mimicking their protein and fat content. The results of this screening and a microbiological monitoring of minced beef with 1% (m / m) of added extract made it possible to observe that the observed losses of antibacterial activity were in particular correlated with the interactions of the phenolic compounds with the proteins or fat. Incorporation of phenolic compounds or plant extracts into packaging materials in contact with food constituted was the second proposed route of implementation. Plastic films that retain antibacterial activity have thus been able to be prepared by melting

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior
Dentre as espÃcies do gÃnero Erythrina (Fabaceae), duas conhecidas popularmente como mulungu, E. velutina e E. mulungu, tem comprovado interesse social e econÃmico para o Brasil. E. mulungu ocorre no Sudeste, pertence à RelaÃÃo Nacional de Plantas de Interesse para o SUS (RENISUS). Jà E. velutina à uma Ãrvore amplamente utilizada no Nordeste, cuja casca do caule à utilizada tradicionalmente no tratamento da ansiedade, agitaÃÃo e insÃnia. Contudo, nÃo existe um produto farmacÃutico com qualidade agregada e estudos farmacolÃgicos dessa espÃcie para justificar seu uso medicinal. Diante do exposto, o objetivo do presente trabalho foi realizar o desenvolvimento e o controle de qualidade do extrato de Erytrhina velutina e investigar as atividades antioxidante e neuroprotetora, visando seu emprego no tratamento de doenÃa neurodegenerativa, como a DoenÃa de Parkinson (DP). Inicialmente, foi validado mÃtodo espectrofotomÃtrico para dosagem de fenÃis totais (FT) em produtos derivados de E. velutina, sendo este especÃfico, linear, preciso, exato e robusto. Foi estabelecido o mÃtodo de preparaÃÃo (estufa com circulaÃÃo e renovaÃÃo de ar â 80ÂC; 24h) e especificaÃÃes para o controle da droga vegetal. O extrato etanÃlico de E. velutina (EEEV) produzido por percolaÃÃo foi caracterizado quando ao teor de FT (155,14  3,31  EAG/mg de extrato) e perfil cromatogrÃfico por CLAE-DAD (FenÃis: hesperidina - Tr: 18,8 min; abssinina - Tr: 22,9 min; homoesperidina Tr: 31,2 min; Ãcido rizÃnico (AR) - Tr: 32,1 min e sigmoidina C - Tr: 38,6 min). O EEEV nÃo mostrou citotoxicidade no teste do MTT, mas aumentou (EEEV: 100 e 200 Âg/mL) a atividade da LDH em neutrÃfilo humano. Em cÃlulas 9L/lacZ, o EEEV (100, 400, 1000 mg/mL) reduziu significativamente a viabilidade celular, teste MTT. EEEV (0,0025 â 1 Âg/mL) e AR (0,0025 â 1 Âg/mL) nÃo foram citotÃxicos e inibiram parcialmente a neurotoxicidade induzida por 6-OHDA em cÃlulas SH-SY5Y, modelo experimental de DP, observada pela reduÃÃo significativa dos nÃveis de nitrito/nitrato. Na avaliaÃÃo do potencial antioxidante, testes DPPH e NBT, o EEEV (10 â 200 Âg/mL) apresentou atividade sequestradora de radicais livres no teste DPPH. Os resultados obtidos no presente estudo permitiram definir as condiÃÃes ideais de preparaÃÃo da droga vegetal e do extrato, os quais foram caracterizados, incluindo validaÃÃo de mÃtodo analÃtico e avaliaÃÃo farmacolÃgica, comprovando o potencial antioxidante e neuroprotetor do EEEV e AR.

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CAPES
Spondias mombin L., Anacardiaceae, é cultivada em várias partes do mundo e tradicionalmente é utilizada para o tratamento de várias doenças, por apresentar propriedades medicinais. Os extratos de suas folhas apresentam atividades farmacológicas reconhecidas tais como, antimicrobiana, antiviral, leishmanicida, antioxidante, anticonvulsivante, antipsicótica, antihelmíntica e anti-inflamatória. Na medicina popular, as folhas de Spondias mombin também têm sido utilizadas para indução de aborto. Em nossa revisão bibliográfica, constatamos carência de informações toxicológicas detalhadas sobre a espécie. Nesse sentido, o objetivo do presente estudo foi avaliar a segurança do uso do extrato hidroalcoólico de Spondias mombin (EHSm) em ratos Wistar de ambos os sexos, visando dar suporte a futuros testes clínicos. Inicialmente foram caracterizados os constituintes químicos do EHSm por CLAE-DAD e foram realizados testes de toxicidade aguda (por via oral na dose de 5 g/kg), testes de doses repetidas sobre os parâmetros bioquímico, hematológico e morfológico nas doses de 200, 500 e 2000 mg/kg consecutivas durante 30 dias, além de ensaios de toxicidade reprodutiva, nas mesmas doses administradas durante o período de pré-implantação (1º ao 6º dia) da gestação. A análise por CLAE-DAD do EHSm demonstrou a ocorrência dos flavonoides: quercetina, canferol e isoquercetina, além do polifenol resveratrol. Os resultados obtidos mostraram que por via oral o EHSm não provocou mortes na dose de 5 g/kg nos animais. O tratamento de doses repetidas com o EHSm não produziu sinais significativos de toxicidade ou mortes, o consumo de água não foi alterado em ambos os sexos e o consumo de ração foi diminuído apenas na primeira semana para os machos tratados na dose de 500 mg/kg, entretanto, o ganho de massa corporal não foi afetado em ambos os sexos durante todo o tratamento. Os perfis bioquímico e hematológico não foram clinicamente modificados e estão dentro dos limites de referência para a espécie. Não foram observadas alterações nas massas absolutas e relativas dos órgãos analisados, bem como na morfologia macroscópica externa dos mesmos. A análise microscópica dos órgãos apontou discreto infiltrado linfocitário no fígado (EHSm 500 e 2000 mg/kg) de animais de ambos os sexos e discreto aumento do espaço glomerular nos rins das fêmeas (EHSm 500 e 2000 mg/kg) e machos (todos os grupos tratados) em relação ao grupo controle, contudo tais alterações não apresentam relevância clínica, visto que ocorreram de modo não-generalizado. Ratas prenhes tratadas no período da pré-implantação com EHSm não apresentaram alterações nos parâmetros reprodutivos, entretanto, o tratamento provocou redução no ganho de massa corporal no período do tratamento. Nenhuma morte e/ou malformações dos fetos foi observada com a dose mais alta sugerindo que o extrato não possui efeito abortivo. Os resultados encontrados nos levam a concluir que o EHSm não apresentou toxicidade por via oral, entretanto, há indícios de possível toxicidade materna durante o período de pré-implantação, que deve ser analisada posteriormente em maior detalhe.
Spondias mombin L., Anacardiaceae, is cultivated in several parts of the world and traditionally is used for treatment of several diseases, for presenting medicinal properties. The extracts of the leaves have recognised pharmacological activities such as antimicrobial, antiviral, leishmanicidal, antioxidant, anticonvulsant, antipsychotics, anthelmintic and anti-inflammatory. In folk medicine, the leaves of Spondias mombin have also been used to induce abortion. In our bibliographical review, we verified a lack of detailed toxicological information about the species. In this sense, the aim of the present study was evaluate the safety of use of hydroalcoholic extract of Spondias mombin (HESm) in Wistar rats of both sexes, in order to support future clinical trials. Initially were characterized the chemical constituents of the HESm by HPLC-DAD and it were accomplished the acute toxicity test (by oral route in dose of 5 g/kg), tests of repeated doses on biochemical, hematogical and morphologic parameters in doses of 200, 500 and 2000 mg/kg consecutives during 30 days, as well as reproductive toxicity tests in the same doses administered during the pre-implantation period (1.... to 6.... day) of gestation. The analysis by HPLC-DAD of the HESm demonstrated the occurrence of flavonoids: quercetin, kaempferol and isoquercetin, besides the polyphenol resveratrol. The results obtained showed that orally the HESm not caused deaths at a dose of 5 g/kg in animals. Treatment of repeated doses with the HESm did not produce significant signs of toxicity or death, the water intake has not changed in both sexes and food intake was reduced only in the first week for males treated in dose of 500 mg/kg, however, body weight gain was not affected in both sexes throughout the treatment. Biochemical and Hematological profiles were not clinically modified and are within the limits of reference to the species. Alterations were not observed on absolute and relative weight of organs analyzed, as well as in external macroscopic morphology of the same. Microscopic analysis of the organs showed discrete lymphocytic infiltrate in the liver (HESm 500 and 2000 mgkg) of both sexes and discrete increased glomerular space in the kidney of females (HESm 500 e 2000 mg/kg) and males (all treated groups), compared to the control group, however these changes do not present clinical relevance, since it occurred in a non-generalised. Pregnant rats treated in the pre-implantation period with HESm not showed alterations in reproductive parameters, however the treatment caused reduction in body weight gain during the period of treatment. No death and/or malformations of fetuses was observed with the highest dose suggesting that the extract does not have abortifacient effect. Based on our results, we conclude that HESm did not present oral toxicity, however there are indications of possible maternal toxicity during the pre-implantation period, which should be examined in greater detail later.

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FAPEAM - Fundação de Amparo à Pesquisa do Estado do Amazonas
The search for compounds with aromatic characteristics desirable and which have antioxidant activity, is widely exploited by industries, pharmaceutical and food flavoring. In order to find compounds that exhibit these characteristics, the fruits of the species Psidium acutangulum, Duroia macrophylla, Matisia cordata and Borojoa sorbilis were evaluated in this work. For the study of the volatile compounds was used the technique of solid-phase microextraction (SPME), using the gaseous chromatograph coupled to mass spectrometer for identification of constituents, and for assessment of antioxidant capacity were used the methods of capture of DPPH free radicals and ABTS. With the results obtained, it was possible to verify that the profile of volatile compounds identified in the peel and pulp of the fruit of Psidium acutangulum is highly complex, in the compound of the terpene class predominating, including β-elemene, α-copaene and α-selinene, the fruit pulp in the majority was the class of the esters, such as butyl octanoate, butyl n-hexanoate and ethyl octanoate. For the fruit of Duroia macrophylla, on peel and pulp, the main classes identified were acid, alcohol and ester, fatty acid, octanoic acid was the most abundant substance in both peel and pulp. In Matisia cordata the predominant class in the shell was of terpenes, highlighting the compound eucalyptol and pulp the main class was that of esters, ethyl benzoate was the main compound. For the Borojoa sorbilis fruit, the most abundant class on peel and pulp was the esters, methyl hexanoate, was the major compound. As the evaluation of antioxidant activity of fruits extracts Psidium acutangulum and Matisia cordata presented the best results compared to the standards used by the DPPH, ABTS and total phenols, highlighting the major activity of the extracts from the shells.
A busca por compostos com características aromáticas desejáveis e que possuam atividade antioxidante, é largamente explorada pelas indústrias de aroma, farmacêutica e alimentícia. Afim de encontrar compostos que apresentem tais características, os frutos das espécies Psidium acutangulum, Duroia macrophylla, Matisia cordata e Borojoa sorbilis foram avaliados neste trabalho. Para o estudo dos compostos voláteis foi utilizada a técnica de microextração em fase sólida (SPME), utilizando o cromatográfo gasoso acoplado ao espectrômetro de massa para identificação dos constituintes, e para avaliação da capacidade antioxidante foram utilizados os métodos de captura de radicais livres DPPH e ABTS. Com os resultados obtidos, foi possível verificar que o perfil de compostos voláteis identificados na casca e polpa do fruto de Psidium acutangulum é altamente complexo, na casca houve a predominância de compostos da classe terpeno, destacando β-elemeno, α-copaeno e α-selineno, já na polpa do fruto à classe majoritária foi a dos ésteres, como, butil octanoato, butil n-hexanoato e octanoato de etila. Para o fruto de Duroia macrophylla, na casca e polpa, as principais classes identificadas foram ácido, álcool e éster, o ácido graxo, ácido octanóico foi a substância mais abundante tanto na casca quanto na polpa. Em Matisia cordata a classe predominante na casca foi a dos terpenos, destacando o composto eucaliptol e na polpa a principal classe foi a dos ésteres, benzoato de etila foi o principal composto. Para o fruto de Borojoa sorbilis, a classe mais abundante na casca e polpa foi a dos ésteres, hexanoato de metila, foi o composto majoritário. Quanto a avaliação da atividade antioxidante os extratos dos frutos Psidium acutangulum e Matisia cordata apresentaram os melhores resultados em comparação com os padrões utilizados pelos métodos de DPPH, ABTS e fenóis totais, destacando a maior atividade advinda dos extratos das cascas.

Abstract Bone morphogenetic proteins (BMPs) constitute a large family of osteoinductive proteins. Different BMPs are widely used in animal experiments and increasingly in the field of bone surgery. However, the sterilization of BMPs and the choice of a suitable mode of delivery, which binds and slowly releases BMP molecules, are still under intensive investigation. The aims of this study were to evaluate the effects of ethylene oxide and gamma sterilizations and different delivery systems on the osteoinductivity of reindeer BMP extract by using heterotopic and orthopic animal models. Ethylene oxide gas (Steri-Vac 4XL, temperature 29 °C, exposure time 4 h, concentration 860 mg/l) and gamma (doses of 3.15 or 4.15 Mrad) sterilizations were used. The tested delivery systems for reindeer BMP were collagen (Lyostypt®), gelatine capsule (no.1) and composites containing collagen combined with tricalcium phosphate (TCP) or hydroxyapatite (HA) or biphasic tricalcium phosphate-hydroxyapatite (TCP/HA) or biocoral (NC) frames. The injectability of reindeer BMP was tested by using injections containing a saline or gelatine vehicle. Osteoinductivity was evaluated as ectopic bone formation in the thigh muscle pouches of mouse hind legs. The induced new bone was evaluated based on the incorporation of 45Ca or calcium yield, radiographs and histological examination three weeks after the operations. The effect of gamma sterilization on the bone healing capacity of reindeer BMP extract was evaluated in a rabbit radial bone defect model in comparison with non-sterilized reindeer BMP extract and recombinant BMP-2. Bone healing was evaluated after eight weeks based on radiographs, mechanical tests and peripheral computerized tomography (pQCT). All BMP implants induced new bone in vivo visible in radiographs, but no bone formation was seen in the control implants without reindeer BMP. Gamma sterilization did not decrease significantly the osteoinductivity of reindeer BMP extract, except when administered as an injection containing gelatine vehicle. Ethylene oxide sterilization decreased significantly the osteoinductivity of reindeer BMP extract and was significantly inferior compared to gamma sterilization. Reindeer BMP combined with collagen or composite containing collagen and TCP/HA frame induced new bone significantly better than reindeer BMP combined with composite containing collagen and TCP frame. BMP injections with gelatine or saline vehicles induced new bone effectively. Injections were easy to handle and well tolerated by the mice. Reindeer BMP extract administered with collagen carrier healed the bone defect of the rabbit radius significantly better than control implants without reindeer BMP or no treatment and its effect was comparable with rhBMP-2 treatment.

Les régions de Midi-Pyrénées (France) et de Chongqing (Chine) sont riches en plantes aromatiques et médicinales dites oubliées (ou médiévales). Afin de valoriser pleinement les différentes bio-molécules extractibles de ces plantes, le concept de MAP-raffinerie a été créé et appliqué à une sélection de plantes issues de ces deux régions. Plusieurs technologies d’extraction utilisant l’eau comme solvant vert (hydrodistillation, distillation à la vapeur et extraction par eau sub-critique) ont ainsi été employées et leur impact tant sur la composition des huiles essentielles que sur la récupération des molécules anti-oxydantes a été évalué. Dans un premier temps, une liste de plantes aromatiques et médicinales oubliées, voire sous-utilisées dans les deux régions a été établie selon des règles de sélection prédéfinies. Six plantes modèles de la région de Midi-Pyrénées (Tussilago farfara L., Calendula arvensis L., Robinia pseudoacacia L., Geranium robertianum L., Cytisus scoparius L. et Spartium junceum L.) et trois plantes de la région de Chongqing (Tussilago farfara L., Citrus aurantium L. et Saussurea costus) ont finalement été retenues. Puis, le concept de MAP-raffinerie a été appliqué à ces plantes afin d’étudier leur possible valorisation globale. L’étude des compositions chimiques des extraits volatils des racines de Tussilago farfara L. et de Calendula arvensis L., ainsi que des boutons de fleurs de Spartium junceum L. a été réalisée par GC et GC-MS pour la première fois. Les principaux composés chimiques dans l’extrait volatil de racines de Tussilago farfara L. étaient des hydrocarbures sesquiterpéniques et des composés aliphatiques tandis que les principaux composés chimiques dans l’extrait volatil de racines de Calendula arvensis L. étaient des sesquiterpènes oxygénés, des monoterpènes oxygénés et des diterpènes oxygénés. L’extrait volatil de boutons de fleurs de Spartium junceum L. était principalement composé de composés aliphatiques. Par ailleurs, les résultats de l’évaluation des capacités anti-oxydantes des extraits (par les tests DPPH, ABTS, FRAP, ORAC et Folin-Ciocalteu) ont montrés que plusieurs plantes comme Cytisus scoparius L., Tussilago farfara L., Citrus aurantium L. ou Robinia pseudoacacia L. pourraient être des sources potentielles d’anti-oxydants naturels. D’un point de vue technologique, les comparaisons de l’utilisation de l’hydrodistillation (HD), de la distillation à la vapeur (SD) et de l’extraction par eau sub-critique (SWE) ont montrées que si la HD et la SD ont des effets limités sur la composition des huiles essentielles, la HD semble être une méthode plus efficace pour la récupération des composés anti-oxydants à partir des résidus de distillation que la SD tandis que la SWE s’avère être une technologie prometteuse pour l’extraction directe de ces molécules à partir des plantes. Si la composition minérale de l’eau lors de l’hydrodistillation n’a que des effets très limités sur les rendements d’extraction, les teneurs en ions calcium et bicarbonate des eaux ont par contre des effets décroissants significatifs sur la capacité anti-oxydante et sur la teneur phénolique totale des extraits aqueux et méthanoliques. Au vue de ces résultats, un concept amélioré de MAP-raffinerie a été développé en intégrant une extraction à l’eau sub-critique pour l’extraction des composés anti-oxydants des résidus d’extraction primaire. Selon ce nouveau concept, cinq extraits peuvent être obtenus à partir des matières végétales: un extrait volatil, un extrait aqueux, un extrait méthanolique, un extrait à l’eau sub-critique et in fine un résidu solide. Les premiers résultats ont montrés que la "MAP-raffinerie améliorée" augmente de manière significative la récupération des antioxydants par rapport à la MAP-raffinerie originale et permet d’envisager une valorisation plus facile du résidu solide en agro-matériaux du fait de sa faible teneur en eau résiduelle
In both Midi-Pyrénées region (France) and Chongqing region (China), there are rich and underutilized medicinal and aromatic plants (MAP). Aiming at fully exploiting different molecules in these plants, the concept of MAP-refinery was developed and applied to several underutilized medicinal and aromatic plants in these two regions. Several water-based green extraction technologies of natural products (e.g. hydrodistillation, steam distillation and subcritical water extraction) were also investigated to look at their effects on essential oil composition and antioxidants recovery from selected plants. Firstly, lists of forgotten or underutilized medicinal and aromatic plants in both regions were established according to the rules of selection. From the lists, six plants in the Midi-Pyrénées region (Tussilago farfara L., Calendula arvensis L., Robinia pseudoacacia L., Geranium robertianum L., Cytisus scoparius L. and Spartium junceum L.) and three plants in the Chongqing region (Tussilago farfara L., Citrum aurantium L. and Saussurea costus) were finally selected for investigations. Then the MAP-refinery was applied to the selected plants in two regions in order to realise their global valorisation. Volatile extracts composition in the roots of Tussilago farfara L. and Calendula arvensis L., as well as flower buds of Spartium junceum L. were firstly investigated. The main chemical compounds in volatile extract from Tussilago farfara L. roots were sesquiterpene hydrocarbons and aliphatic compounds while main chemical compounds in volatile extract from Calendula arvensis L. roots were oxygenated sesquiterpenes, oxygenated monoterpenes and oxygenated diterpenes. The volatile extract from flower buds of Spartium junceum L. was mainly composed of aliphatic compounds. Antioxidant capacity evaluation results (by DPPH, ABTS, FRAC, ORAC and Folin-Ciocalteu tests) showed that several plant samples like Cytisus scoparius L., Tussilago farfara L., Citrum aurantium L. and Robinia pseudoacacia L. could be potential sources of natural antioxidants. Comparisons of hydrodistillation (HD), steam distillation (SD) and subcritical water extraction (SWE) showed that HD and SD had limited effects on essential oil composition but HD, SD and SWE had significant impacts on the recovery of antioxidants. Hydrodistillation seemed to be a better method for recovery of antioxidant compounds from residues of distillation than steam distillation. However, SWE appeared to be a more efficient method for direct extraction of antioxidant molecules (or phenolic compounds) from plants. In the hydrodistillation process, mineral contents in water were found to have very limited effects on yields of extracts but calcium and bicarbonate ions, had significant decreasing effects on antioxidant capacity and total phenolic content of both aqueous and methanolic extracts. Finally, an improved MAP-refinery was developed. Subcritical water was used for further extraction of antioxidant compounds from residues in original MAP-refinery. In this way, five parts could be obtained from plant materials: volatile extract, aqueous extract, methanolic extract, subcritical water extract and the final residue. The results showed that the improved MAP-refinery significantly increased the recovery of antioxidants compared with original MAP-refinery. This promising process will also allow a better valorisation of the final solid residue due to the lower content of residual water

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES
In the last decade, beet has attracted much attention as a functional food, with important health promoting effect. It presents in its composition important bioactive compounds, highlighting the betalains, ascorbic acid, carotenoids and phenolic acids. Despite the great nutritional importance, the beet is still not very consumed. The encapsulation by ionic gelation has proven to be an effective technique for obtaining palatable and nutritious products, masking unwanted flavors and preserving nutrients. Considering the above, the objective of the present study was to obtain microcapsules of beet extract by the ionic gelation process and verify the stability during refrigerated storage. From the beet extract the ionic gelation process was performed, obtaining the beet microcapsules and storing them for 28 days in hermetic glass containers at 5 ± 1°C. Analyzes of yield, weight, size, color, soluble solids, moisture, ash, acidity, pH, vitamin C, carotenoids, phenolics, ABTS and betalainas were performed. Regarding the physicochemical characteristics of beet extract with respect to beet microcapsules, there was a significant reduction of soluble solids, betalain, antioxidant capacity, carotenoids and color parameter a*. Vitamin C, phenolics, ash, moisture and pH remained constant. There was an increase in acidity and Hue angle. During refrigerated storage, all parameters except vitamin C were reduced. After 21 days of storage, there was an increase in the permeability of the alginate membrane, resulting in greater migration between the medium and microcapsule compound. It is concluded that the ionic gelation process is a viable technology for the development of products that maintain the nutritional characteristics of the beet and that treatment with ascorbic acid obtained better conservation results of the bioactive compounds.
Na última década, a beterraba tem atraído muita atenção como um alimento funcional, com importante efeito promotor de saúde. Apresenta em sua composição importantes compostos bioativos, destacando as betalaínas, ácido ascórbico, carotenoides e ácidos fenólicos. Apesar da grande relevância nutricional a beterraba ainda não é muito consumida. A encapsulação por gelificação iônica tem se mostrado uma técnica eficaz na obtenção de produtos palatáveis e nutritivos, mascarando sabores indesejados e preservando nutrientes. Diante do exposto, o objetivo do presente estudo foi obter microcápsulas de extrato de beterraba pelo processo de gelificação iônica e verificar a estabilidade durante o armazenamento refrigerado. A partir do extrato de beterraba realizou-se o processo de gelificação iônica, obtendo-se as microcápsulas de beterraba e armazenando-as por 28 dias em embalagens herméticas de vidro a 5±1°C. Foram realizadas análises de rendimento, peso, tamanho, cor, sólidos solúveis, umidade, cinzas, acidez, pH, vitamina C, carotenoides, fenólicos, ABTS e betalainas. Quanto as características físico-químicas do extrato de beterraba com relação as microcápsulas de beterraba houve redução significativa dos sólidos solúveis, da betalaína, da capacidade antioxidante, dos carotenoides e do parâmetro de cor a*. A vitamina C, os fenólicos, as cinzas, a umidade e o pH se mantiveram constantes. Verificou-se leve aumento na acidez e no ângulo Hue. Durante o armazenamento refrigerado observou-se redução de todos os parâmetros com exceção da vitamina C. Após 21 dias de armazenamento observou-se aumento da permeabilidade da membrana de alginato, acarretando maior migração entre os compostos do meio e das microcápsulas. Conclui-se que o processo de gelificação iônica é uma tecnologia viável para a desenvolvimento de produtos que mantenham as características nutricionais da beterraba e que o tratamento com ácido ascórbico obteve melhores resultados de conservação dos compostos bioativos.
São Cristóvão, SE

CoordenaÃÃo de AperfeiÃoamento de NÃvel Superior
Trinta e trÃs substÃncias foram detectadas em diferentes extratos das folhas, frutos, talos e cascas de Caraipa densifolia Mart. Os compostos fenÃlicos foram identificados preliminarmente e quatificados por CLAE-ESI-EM apÃs cromatografia em coluna Sephadex LH-20. Estruturas dos compostos, purificada por CLAE semi-preparativa, foram ainda caracterizados por nano-ESI-EM-EM. A presenÃa de Ãcido gÃlico, Ãcido 3,4-diidroxibenzÃico, Ãcido clorogÃnico, Ãcido neoclorogÃnico, galato de metila, Ãcido p-cumÃroil quinato, epicatequina, procainidina dÃmero B2, procianidina trÃmero C1, Ãcido sirÃgico, 1, 2, 3, 6 -tetragalato glicosÃdeo, 1, 3, 4, 6-tetragalato glicosÃdeo, corilagina, Ãcido elÃgico, Ãcido metil elÃgico raminosÃdeo, quercetina-3-O-raminosÃdeo, dois apigenina-C-glicosÃdeos (vitexina e isovitexina), mais trÃs apigenina, cucumerina A, cucumerina B e cucumerina C) e dois luteolina-C-glicosÃdeos (orientina e isoorientina) sÃo relatados nesta espÃcie, pela primeira vez. JÃ identificados anteriormente nesta espÃcie os seguintes terpenÃides: lupeol, lupenona, Ãcido betulÃnico, betulina, friedelina. Sendo inÃdito na espÃcie Caraipa, friedelinol, -sitosterol, estigmasterol, as vitaminas (-tocoferol e -tocoferol), identificados em extratos das folhas de Caraipa densifolia Mart. foram identificados e quatificados por CG-EM. As informaÃÃes obtidas nesse trabalho trazem contribuiÃÃes relevantes para o conhecimento das vÃrias formulaÃÃes geralmente utilizadas na medicina popular e indÃstrias farmacÃuticas, representando uma etapa importante para um estudo de um fitoterÃpico. O ensaio da capacidade antioxidante dos extratos FST, FSD e FSH revelou uma significativa atividade, obtendo assim, melhor atividade antioxidante o extrato FST com IC502,1mg/mL. Os extratos com melhor inibiÃÃo da enzima Acetilcolinesterase em concentraÃÃo de 2mg/mL, aplicados 2,5 μL. das foram FH, TSH, CSH, FSM e FM. O teste moluscicida com caramujo da Biomphalaria glabrata aplicado aos extratos FH e FM, resultou Ãndice de mortalidade 100% para FH em 24h e 60% e 80% para FM em 48h
Thirty three secondary plant substances were detected in various extracts of the leaves,fruits, stem and bark of Caraipa densifolia Mart. Phenolic compounds were preliminarily identified and quantitated by HPLC-ESI-MS after column chromatography on Sephadex LH-20. Structures of the compounds, purified by semi-preparative HPLC, were further characterized by nano-ESI-MS-MS. The presence of gallic acid, 3,4-dihydroxybenzoic acid, chlorogenic acid (plus an isomer), methyl gallate, p-coumaric acid quinate, epicatechin, procaynidin dimer B2, procyanidin trimer C1, syringic acid, 1, 2, 3, 6-tetragallate glucoside, 1, 3, 4, 6-tetragallate glucoside, corilagin, ellagic acid, methyl ellagic acid rhamnoside, quercetin-3-O-rhamnoside, two apigenin-C-glycosides (vitexin and isovitexin), three further apigenin,cucumerin A cucumerin B e cucumerin C, two luteolin-C-glycosides (orientin and isoorientin) are reported in this species for the first time. In addition, the previously reported following terpenoids, lupeol, lupenone, betulinic acid, betulin, friedelin and a previously non-characterized terpenoid in this species, friedelinol were identified and quantitated by GC-MS. A previously identified sterol was -sitosterol along with stigmasterol in this species for the first time. The vitamins -tocopherol and -tocopherol were also identified in extracts of the leaves of Caraipa species for the first time. Information obtained in this study provide significant contributions to the knowledge of the various formulations commonly used in folk medicine and pharmaceutical industry, representing an important step towards a study of a phytotherapic. The test of antioxidant capacity of extracts FST, FSD and FSH showed a significant activity, thereby obtaining better antioxidant extract FST with IC502, 1mg/mL. The extracts with the best inhibition of Acetylcholinesterase in concentration 2mg/mL applied 2.5 μL. were of FH, TSH, CSH, FSM and FM. The molluscicidal test with the snail Biomphalaria glabrata extracts applied to FH and FM resulted mortality rate 100% for FH in 24 and 60% and 80% for FM in 48 hours

SILVEIRA, C. V. Caracterização e quantificação dos compostos polifenólicos e triterpênicos em extratos obtidos a partir das folhas, cascas, frutos e talos de caraipa densifolia mart. 2010. 103 f. Tese (Doutorado em Química Orgânica) - Centro de Ciências, Universidade Federal do Ceará, Fortaleza, 2010.
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Thirty three secondary plant substances were detected in various extracts of the leaves,fruits, stem and bark of Caraipa densifolia Mart. Phenolic compounds were preliminarily identified and quantitated by HPLC-ESI-MS after column chromatography on Sephadex LH-20. Structures of the compounds, purified by semi-preparative HPLC, were further characterized by nano-ESI-MS-MS. The presence of gallic acid, 3,4-dihydroxybenzoic acid, chlorogenic acid (plus an isomer), methyl gallate, p-coumaric acid quinate, epicatechin, procaynidin dimer B2, procyanidin trimer C1, syringic acid, 1, 2, 3, 6-tetragallate glucoside, 1, 3, 4, 6-tetragallate glucoside, corilagin, ellagic acid, methyl ellagic acid rhamnoside, quercetin-3-O-rhamnoside, two apigenin-C-glycosides (vitexin and isovitexin), three further apigenin,cucumerin A cucumerin B e cucumerin C, two luteolin-C-glycosides (orientin and isoorientin) are reported in this species for the first time. In addition, the previously reported following terpenoids, lupeol, lupenone, betulinic acid, betulin, friedelin and a previously non-characterized terpenoid in this species, friedelinol were identified and quantitated by GC-MS. A previously identified sterol was -sitosterol along with stigmasterol in this species for the first time. The vitamins -tocopherol and -tocopherol were also identified in extracts of the leaves of Caraipa species for the first time. Information obtained in this study provide significant contributions to the knowledge of the various formulations commonly used in folk medicine and pharmaceutical industry, representing an important step towards a study of a phytotherapic. The test of antioxidant capacity of extracts FST, FSD and FSH showed a significant activity, thereby obtaining better antioxidant extract FST with IC502, 1mg/mL. The extracts with the best inhibition of Acetylcholinesterase in concentration 2mg/mL applied 2.5 μL. were of FH, TSH, CSH, FSM and FM. The molluscicidal test with the snail Biomphalaria glabrata extracts applied to FH and FM resulted mortality rate 100% for FH in 24 and 60% and 80% for FM in 48 hours
Trinta e três substâncias foram detectadas em diferentes extratos das folhas, frutos, talos e cascas de Caraipa densifolia Mart. Os compostos fenólicos foram identificados preliminarmente e quatificados por CLAE-ESI-EM após cromatografia em coluna Sephadex LH-20. Estruturas dos compostos, purificada por CLAE semi-preparativa, foram ainda caracterizados por nano-ESI-EM-EM. A presença de ácido gálico, ácido 3,4-diidroxibenzóico, ácido clorogênico, ácido neoclorogênico, galato de metila, ácido p-cumároil quinato, epicatequina, procainidina dímero B2, procianidina trímero C1, ácido sirígico, 1, 2, 3, 6 -tetragalato glicosídeo, 1, 3, 4, 6-tetragalato glicosídeo, corilagina, ácido elágico, ácido metil elágico raminosídeo, quercetina-3-O-raminosídeo, dois apigenina-C-glicosídeos (vitexina e isovitexina), mais três apigenina, cucumerina A, cucumerina B e cucumerina C) e dois luteolina-C-glicosídeos (orientina e isoorientina) são relatados nesta espécie, pela primeira vez. Já identificados anteriormente nesta espécie os seguintes terpenóides: lupeol, lupenona, ácido betulínico, betulina, friedelina. Sendo inédito na espécie Caraipa, friedelinol, -sitosterol, estigmasterol, as vitaminas (-tocoferol e -tocoferol), identificados em extratos das folhas de Caraipa densifolia Mart. foram identificados e quatificados por CG-EM. As informações obtidas nesse trabalho trazem contribuições relevantes para o conhecimento das várias formulações geralmente utilizadas na medicina popular e indústrias farmacêuticas, representando uma etapa importante para um estudo de um fitoterápico. O ensaio da capacidade antioxidante dos extratos FST, FSD e FSH revelou uma significativa atividade, obtendo assim, melhor atividade antioxidante o extrato FST com IC502,1mg/mL. Os extratos com melhor inibição da enzima Acetilcolinesterase em concentração de 2mg/mL, aplicados 2,5 μL. das foram FH, TSH, CSH, FSM e FM. O teste moluscicida com caramujo da Biomphalaria glabrata aplicado aos extratos FH e FM, resultou índice de mortalidade 100% para FH em 24h e 60% e 80% para FM em 48h

Universidade Federal do CearÃ
The present work aimed at the production of lignosulfonate, based in the lignin extracted from sugarcane bagasse-cane for using in phenolic resins. The extraction of lignin was carried out using the acetosolv process, which was optimised with a central composite design 22 to evaluate the effects of reaction time and temperature on the extraction yield, weight-average (M ̅w) and number-average (M ̅n) molecular weights, relative content of total hydroxyl, phenolic hydroxyl and methoxyl groups. The lignins obtained under conditions that maximized the extraction yield and showed better structural and thermal characteristics were sulfonated to obtain the lignosulfonates. The structural and thermal characteristics of the lignins and lignosulfonates were determined by FT-IR, GPC, 1H-NMR and 13C-NMR, DSC and TGA. The results show that the best extraction yield (64.5%) was obtained with 95% (w/w) of acetic acid, the addition of 0.1% HCl, at a temperature of 187 ÂC and reaction time of 40 min. However, with the same concentration of acetic acid and reaction time of 15 min at 187 ÂC, the extraction yield decreased to 55.6% Â 4.5%, without significant reduction. Furthermore, the increase in temperature of 187 ÂC to 205 ÂC was not enough to cause a significant increase in the relative content of hydroxyls and reduction of the relative content of methoxyl. These results show that the most appropriate conditions for adequate extraction of lignin for application in resins are: 95% (w/w) of acetic acid, addition of 0.1% of HCl, temperature of 187 ÂC and reaction time of 15 min. The acetosolv lignins showed p-hidroxifenila units as major constituent, higher thermal stability and higher purity than the commercial Kraft lignin. The glass transition temperature of the Kraft lignins was lower than that of the acetosolv lignin. This is due to the hydrophilic character and the presence of carbohydrates in the Kraft lignin. The lignosulfonates obtained in this study showed structural characteristics suitable for application in phenolic resins, because they showed high reactivity due to the greater presence of p-hidroxifenila units as major constituent, low molecular weights (40234878 g/mol), greater stability and greater purity compared to commercial sodium lignosulfonate. Therefore, lignosulfonates obtained in this work are more suitable for use in phenolic resins than commercial sodium lignosulfonate used for comparison.
O presente trabalho teve por objetivo a produÃÃo de lignossulfonato, a partir da lignina extraÃda do bagaÃo da cana-de-aÃÃcar para aplicaÃÃo em resinas fenÃlicas. Foi realizada a otimizaÃÃo da extraÃÃo da lignina do bagaÃo de cana-de-aÃÃcar utilizando o processo acetosolv. Para tanto, empregou-se um delineamento composto central 22 para analisar os efeitos do tempo de reaÃÃo e da temperatura no rendimento de extraÃÃo, massa molar ponderal mÃdia, massa molar numÃrica mÃdia, e conteÃdo relativo de hidroxilas totais, hidroxilas fenÃlicas e metoxilas. As ligninas obtidas nas condiÃÃes que maximizaram o rendimento de extraÃÃo e que mostraram melhores caracterÃsticas estruturais e tÃrmicas foram sulfonadas para obtenÃÃo dos lignossulfonatos. As caracterÃsticas estruturais e tÃrmicas das ligninas e dos lignossulfonatos foram determinadas por FT-IR, GPC, RMN-1H e 13C, TGA e DSC. Os resultados mostram que o melhor rendimento de extraÃÃo (64,5 % 4,2%) foi obtido com 95% (m/m) de Ãcido acÃtico, adiÃÃo de 0,1% de HCl, a uma temperatura de 187 C e tempo de reaÃÃo de 40 min. No entanto, com a mesma concentraÃÃo de soluÃÃo de Ãcido acÃtico e com tempo de reaÃÃo de 15 min a 187ÂC, o rendimento de extraÃÃo diminuiu para 55,6%  4,5%, nÃo sendo significativa esta reduÃÃo. AlÃm disto, a elevaÃÃo da temperatura de 187ÂC para 205ÂC nÃo foi suficiente para causar um aumento significativo no conteÃdo relativo de hidroxilas e reduÃÃo do conteÃdo relativo de metoxila. Esses resultados mostram que as condiÃÃes mais adequadas para extraÃÃo da lignina a ser aplicada em resinas sÃo: 95% (m/m) de Ãcido acÃtico, adiÃÃo de 0,1% de HCl, temperatura de 187 C e tempo de reaÃÃo de 15 min. As ligninas acetosolv apresentaram unidades p-hidroxifenila como constituinte majoritÃrio, maior estabilidade tÃrmica e maior pureza em relaÃÃo à lignina Kraft comercial. A temperatura de transiÃÃo vÃtrea da lignina Kraft foi menor do que à das ligninas acetosolv, devido à sua caracterÃstica hidrofÃlica e à presenÃa de carboidratos na lignina Kraft. Os lignossulfonatos obtidos no presente trabalho apresentaram caracterÃsticas estruturais adequadas para aplicaÃÃo em resinas fenÃlicas, pois mostraram alta reatividade devido a maior presenÃa de unidades p-hidroxifenila como constituinte majoritÃrio, baixas massas molares (4023 a 4878 g/mol), maior estabilidade e uma maior pureza em relaÃÃo ao lignossulfonato de sÃdio comercial. Portanto, os lignossulfonatos obtidos no presente trabalho sÃo mais adequados para aplicaÃÃo em resinas fenÃlicas do que o lignossulfonato de sÃdio comercial utilizado no presente trabalho.

Orientador: Miriam Dupas Hubinger
Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos
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Resumo: O pequi (Caryocar brasiliense Camb.) é uma fruta do cerrado de grande importância econômica, que apresenta alto teor de compostos fenólicos e carotenóides e alta atividade antioxidante. Este trabalho teve como objetivo desenvolver produtos ricos em compostos funcionais, principalmente compostos fenólicos e carotenóides, a partir de extratos aquosos e alcoólicos de pequi concentrados por processos de membranas. Inicialmente, as principais variáveis relativas ao processo (tempo e temperatura) foram avaliadas, para definir a melhor forma de extração aquosa e alcoólica dos compostos bioativos da polpa, do endocarpo espinhoso e da semente do fruto. A partir das condições selecionadas nessa primeira etapa (temperatura de 25 °C durante 1 hora de extração para o extrato aquoso e 40°C durante 24 horas para o extrato alcoólico), foram produzidos extratos concentrados utilizando-se uma unidade de membrana de pequeno porte, que faz a simulação de escoamento transversal com diferentes tipos de membranas: uma de ultrafiltração (UF com cut-off de 100 KDa) e duas de nanofiltração(NF 90 e NF 270, com cut-off de 200-300 Da e diferentes materiais). Durante os processos, os fluxos de permeado foram calculados, as membranas foram submetidas a uma caracterização morfológica antes e depois do processo de concentração e os produtos obtidos (alimentação, concentrado e permeado) de cada processo foram caracterizados em relação às seguintes propriedades: densidade, viscosidade, cor, polifenóis totais, carotenóides e atividade antioxidante. Os fluxos de permeado dos processos de ultra e nanofiltração para os extratos alcoólicos e aquosos variaram de 7,06 a 9,17 L/h.m2, e não apresentaram diferença significativa. O processo de concentração por membranas para o extrato alcoólico de pequi mostrou-se ineficiente para concentrar os compostos bioativos, uma vez que todas as condições apresentaram coeficientes de retenção menores que 15% tanto para compostos fenólicos, quanto para carotenóides. Para o extrato aquoso, as membranas de nanofiltração apresentaram 100% de retenção de carotenóides e 93% (NF 270) e 97% (NF 90) de retenção de polifenóis. Concordando com estes resultados, o concentrado aquoso obtido com a membrana NF 90 mostrou-se rico em compostos bioativos e alta atividade antioxidante
Abstract: Pequi (Caryocar brasiliense Camb.) is a fruit of Cerrado Brazilian region with great economic importance and contains high levels of phenolics compounds and carotenoids as well as high antioxidant activity. This work aimed at developing products rich in functional compounds, mainly phenolic compounds and carotenoids, from aqueous and alcoholic pequi extracts concentrated by membrane processes. Initially, the main variables of the process (time and temperature) were evaluated, to define the best form of alcohol and water extraction of bioactive compounds from the pulp, prickly endocarp and seed fruit. From the conditions selected in this first step (25 °C for 1 h to extract the aqueous extract and 40 °C for 24 h for the alcohol extract), concentrated extracts were produced using a laboratorial membrane unit, which makes the simulation of cross flow with different types of membranes: one of ultrafiltration (UF with cutoff of 100 KDa) and two of nanofiltration (NF 90 and NF 270, with cut-off of 200-300 Da and different materials). During the process, the permeate fluxes was calculated, the membranes were subjected to a morphological characterization before and after the concentration process and the products obtained (feed, concentrate and permeate) from each process were characterized by the following properties: density, viscosity, color, total polyphenols, carotenoids and antioxidant activity. The permeate fluxes of ultra and nanofiltration process for alcoholic and aqueous extracts varied from 7.06 to 9.17 L/h.m2, and they did not presented significant difference. The concentration membrane process of the alcoholic pequi extract was ineffective to concentrate the bioactive compounds, since all conditions showed retention coefficient lower than 15% for both, phenolic compounds and carotenoids. For the aqueous extract, the nanofiltration membranes process showed 100% retention of carotenoids and 93% (NF 270) and 97% (NF 90) the retention of polyphenols. According to these results, the aqueous concentrate obtained with the NF 90 membrane showed rich in bioactive compounds and higher antioxidant activity
Mestrado
Engenharia de Alimentos
Mestre em Engenharia de Alimentos