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Clitoria guianensis and Ouratea spectabilis, found in the Brazilian Cerrado, are used in folk medicine, despite the few chemical and biological studies reported in the literature. The present study aims to investigate the toxicity and effect of extracts from both species on the microcrustacean Artemia salina, and to determine the chemical composition of the hexane extract of O. spectabilis leaves and the EtOAc fraction of C. guianensis leaves. Kaempferitrin, a flavonoid isolated from of the EtOAc fraction of C. guianensis leaves, was identified by chemical analysis. Analysis of the hexane extract of O. spectabilis leaves using gas chromatography-mass spectrometry (GC-MS) suggested the presence of twenty-five known substances. The Hex, EtOAc, and EtOH crude extracts of C. guianensis leaves exhibited high and moderate toxicity against Artemia salina, with median lethal dose values (LD50) of 43.7, 25.4, and 233.4 mg.L−1, respectively. The acetone extract of O. spectabilis leaves showed moderate toxicity against Artemia salina with an LD50 value of 115.13 mg.L−1.

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Non-domesticated species may represent a treasure chest of defensive molecules which must be investigated and rescued. Clitoria fairchildiana R. Howard is a non-domesticated Fabacea, native from the Amazonian Forest whose seeds are exquisitely refractory to insect predation. Secondary metabolites from these seeds were fractionated by different organic solvents and the CH2Cl2 fraction (CFD - Clitoria fairchildiana dichloromethane fraction), as the most toxic to 3rd instar Aedes aegypti larvae (LC50 180 PPM), was subjected to silica gel chromatography, eluted with a gradient of CH2Cl2: MeOH and sub fractioned in nine fractions (CFD1 - CFD9). All obtained fractions were tested in their toxicity to the insect larvae. Two rotenoids, a 11α-O-ß-D-glucopyranosylrotenoid and a 6-deoxyclitoriacetal 11-O-n-glucopyranoside, were identified in the mixture of CFD 7.4 and CFD 7.5, and they were toxic (LC50 120 PPM) to 3rd instar Ae. aegypti larvae, leading to exoskeleton changes, cuticular detachment and perforations in larval thorax and abdomen. These C. fairchildiana rotenoids interfered with the acidification process of cell vesicles in larvae midgut and caused inhibition of 55% of V-ATPases activity of larvae treated with 80 PPM of the compounds, when compared to control larvae. The rotenoids also led to a significant increase in the production of reactive oxygen species (ROS) in treated larvae, especially in the hindgut region of larvae intestines, indicating a triggering of an oxidative stress process to these insects.

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This study evaluated the effects of temperature on the pH of extracts of ascorbic acid and anthocyanins from petals of butterfly pea, as well as their in vitro digestive stability in model systems at 60, 70, and 80 °C. The pH values significantly decreased with an increase in the temperature (P < 0.05). The findings were similar for the degradation of anthocyanins and ascorbic acid, which followed first-order kinetics in all the systems. The samples heated at 80 °C presented the highest degradation rate (kobs), as well as higher percentages of degradation at the end of digestive stability in vitro.

Este estudo avaliou os efeitos da temperatura na estabilidade do ácido ascórbico, pH e antocianinas de pétalas de feijão borboleta, bem como a estabilidade digestiva in vitro do ácido ascórbico e antocianinas em sistemas modelo submetidos a 60, 70 e 80 ºC. Os valores de pH diminuíram significativamente com o aumento da temperatura (P < 0.05). O mesmo foi observado para a degradação de antocianinas e ácido ascórbico, que seguiram cinética de primeira ordem em todos os sistemas. As amostras aquecidas a 80 °C apresentaram as maiores taxas de degradação (kobs), assim como maiores percentuais de degradação ao final da estabilidade digestiva in vitro.

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This study aimed to evaluate the effect of different Clitoria fairchildiana tree shading levels on the Tanganyika grass (Megatyrsus maximus Jacq. cv. Tanganyika) chemical composition, in summer I (2011/2012), spring (2012) and summer II (2012/2013) seasons. For this purpose, an experiment was conducted in a completely randomized design with four treatments (shading levels) and five repetitions at Seropédica, state of Rio de Janeiro, Brazil. The following variables were evaluated: dry matter (DM), crude protein (CP), neutral detergent (NDF) and acid detergent (ADF) fiber, neutral (NDIP) and acid (ADIP) detergent insoluble protein, lignin (LIG) and mineral matter (MM) contents. Data were analyzed by PROC MIXED SAS®, with repeated measures in time, and treatment means compared by Tukeys test (P<0.05) or by PROC REG (P<0.05 for regression analysis. There was a significant interaction between season and shading level for DM, CP, NDF, ADF, NDIP, ADIP and MM contents (P<0.05). Lignin content was only influenced by shading level (P<0.05). The DM, NDF, ADF and lignin contents were reduced by shading increase while CP, NIDP and MM contents were increased. Higher ADIP contents were found only in the spring. The increase in Clitoria fairchildiana tree shading improves the chemical composition of Tanganyika grass forage during summer season.(AU)

O objetivo deste trabalho foi avaliar o efeito de diferentes níveis de sombreamento arbóreo da espécie Clitoria fairchildiana sobre a composição bromatológica do capim-tanganica (Megatyrsus maximus Jacq. cv. Tanganica), nas estações do verão I (2011/2012), primavera (2012) e verão II (2012/2013). Para tanto, foi instalado um experimento sob delineamento inteiramente casualizado, com quatro tratamentos (níveis de sombreamento) e cinco repetições em Seropédica, Rio de Janeiro, Brasil. Foram avaliados os teores de: matéria seca (MS), proteína bruta (PB), fibra em detergente neutro (FDN) e em detergente ácido (FDA), proteína insolúvel em detergente neutro (PIDN) e em detergente ácido (PIDA), lignina (LIG) e matéria mineral (MM). Os dados foram analisados pelo PROC MIXED do SAS®, com medidas repetidas no tempo, e as médias dos tratamentos comparadas pelo teste de Tukey (P<0,05); ou pelo PROC REG do SAS® para as análises de regressão (P<0,05). Houve efeito de interação entre nível de sombreamento e estação do ano para teores de MS, PB, FDN, FDA, PIDN, PIDA e MM. Os teores de lignina foram influenciados apenas pelo nível de sombreamento. Os teores de MS, FDN, FDA e lignina foram reduzidos, enquanto aqueles de PB, PIDN e MM aumentaram linearmente em função dos níveis de sombreamento. Maiores teores de PIDA foram verificados somente na primavera. O aumento do sombreamento arbóreo de Clitoria fairchildiana melhora a composição bromatológica da forragem do capim-tanganica, durante o verão.(AU)

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Many organic dye pollutants have been identified in rivers and lakes around the world, and concern is growing with them as they cause serious changes in the ecological balance of aquatic environments. One of these dyes is rhodamine R6G, which is very water-soluble and has a high corrosive power. Therefore, Clitoria fairchildiana (CF) pods were used as a biosorbent to remove R6G from synthetic dye effluents. CF was characterized by infrared spectroscopy, thermogravimetric analysis, x-ray diffraction, elemental analysis, Boehm titration, and zero charge point measurements. The influence of various factors, such as solution pH, contact time, adsorbent mass, and concentration of R6G, was studied using batch equilibrium experiments. The optimum contact time to reach equilibrium was found to be 15 min, while the optimum adsorbent dose was 8 g L-1. The maximum adsorption capacity of CF (73.84 mg g-1) was observed at pH 6.4 and 298.15 K. Adsorption kinetics followed a pseudo-second-order law, and the isotherm could be best fitted with a Liu model. The obtained thermodynamic parameters indicate that the adsorption of R6G is spontaneous and enthalpy-driven. We thus conclude that CF is an efficient, green, and readily available biosorbent for dye removal from wastewater.

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Herein described is the biochemical characterisation, including in vitro and in vivo assays, for a proteinase inhibitor purified from Clitoria fairchildiana seeds (CFPI). Purification was performed by hydrophobic interaction and gel filtration chromatography. Kinetic studies of the purified inhibitor showed a competitive-type inhibitory activity against bovine trypsin and chymotrypsin, with an inhibition stoichiometry of 1:1 for both enzymes. The inhibition constants against trypsin and chymotrypsin were 3.3 × 10(-10) and 1.5 × 10(-10)M, respectively, displaying a tight binding property. SDS-PAGE showed that CFPI has a single polypeptide chain with an apparent molecular mass of 15 kDa under non-reducing conditions. However, MALDI-TOF analysis demonstrated a molecular mass of 7.973 kDa, suggesting that CFPI is dimeric in solution. The N-terminal sequence of CFPI showed homology with members of the Bowman-Birk inhibitor family. CFPI remained stable to progressive heating for 30 min to each temperature range of 37 up to 100 °C and CD analysis exhibited no changes in spectra at 207 nm after heating at 90 °C and subsequent cooling. Moreover, CFPI was active over a wide pH range (2-10). In contrast, reduction with DTT resulted in a loss of inhibitory activity against trypsin and chymotrypsin. CFPI also exhibited significant inhibitory activity against larval midgut trypsin enzymes from Anagasta kuehniella (76%), Diatraea saccharalis (59%) and Heliothis virescens (49%). Its insecticidal properties were further analysed by bioassays and confirmed by negative impact on A. kuehniella development.

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Lectins are proteins that have the ability to bind specifically and reversibly to carbohydrates and glycoconjugates, without altering the structure of the glycosyl ligand. They are found in organisms such as viruses, plants and humans, and they have been shown to possess important biological activities. The objective of this study was to purify and characterize lectins in the seeds of Clitoria fairchildiana, as well as to verify their biological activities. The results indicated the presence of a lectin (CFAL) in the glutelin acid protein fraction, which agglutinated native rabbit erythrocytes. CFAL was purified by column chromatography ion-exchange, DEAE-Sephacel, which was obtained from a peak of protein retained in the matrix by applying 0.5 M NaCl using the step-wise method. Electrophoretic analysis of this lectin in SDS-PAGE indicated a two band pattern protein molecular mass of approximately 100 and 116 kDa. CFAL proved to be unspecific to all carbohydrates/glycoconjugates in common use for the sugar inhibition test. This lectin showed no significant cytotoxicity to human red blood cells. It was observed that CFAL has anti-inflammatory activity in the paw edema induced by carrageenan model, in which a 64% diminution in edema was observed. Antinociceptive effects were observed for CFAL in the abdominal writhing test (induced by acetic acid), in which increasing doses of the lectin caused reduction in the number of contortions by up to 72%. It was concluded that the purified and characterized lectin from the seeds of Clitoria fairchildiana has anti-inflammatory and antinociceptive activity, and is not cytotoxic to human erythrocytes.

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As amostras de Clitoria ternatea L. foram cortadas aos 35, 50, 70 e 90 dias após o corte de uniformização na área experimental. O experimento foi projetado como um delineamento inteiramente casualizado, com total de 18 unidades experimentais (parcelas). A matéria seca (MS) e a produção de matéria seca (PMS) da planta inteira foram separadas quantitativamente em folhas, hastes e vagens para cada idade de corte. As análises bromatológicas e cinéticas de produção de gás in vitro realizadas avaliaram a qualidade de cada parte da planta. A produção, composição química e os parâmetros de produção de gás estimados foram analisados por meio de ajuste de um modelo estatístico misto com dois tipos de estrutura de covariância: componente de variância e não estruturada com variância heterogênea. Os ?pools? de produção rápida e lenta de gás foram detectados tanto para folhas quanto nas hastes. Apenas nas vagens foram detectados ?pool? simples de produção de gás. Pressuposta homocedasticidade foi observada para todas as variáveis exceto a cutina nas hastes e para alguns parâmetros da cinética de produção de gás das folhas e hastes. A produção de matéria seca da planta apresentou tendência cúbica semelhante a um padrão de curva sigmoide. A PMS de folhas apresentou efeito quadrático e a PMS de hastes aumentou linearmente. Não houve presença representativa de vagens aos 35 e 50 dias. O teor de MS da planta inteira teve efeito quadrático e apresentou o mínimo no intervalo de 90 dias. As frações não fibrosas e a cutina das folhas não foram afetadas pela idade de corte. A idade de corte afetou a maioria dos componentes químicos e os parâmetros de produção de gás relativos às hastes. Baseado no máximo da PMS para as folhas, o corte de 70 dias maximizou a quantidade de partes comestíveis de massa de leguminosas forrageiras de alta qualidade. No entanto, o equilíbrio entre produção sustentável de forragem e qualidade, tem que ser pesquisada.

Samples of Clitoria ternatea L. were harvested at 35, 50, 70, and 90 d after a uniformity harvest in a field study designed as a completely randomized design with a total of 18 experimental units. The dry matter (DM) yield (DMY) of the whole plant was separated quantitatively into leaves, stems, and pods at each harvesting age. Chemical analyses and in vitro gas production kinetics were performed to assess the quality of the plant parts. Yields, chemical composition, and estimates of gas production parameters were analyzed by fitting a mixed statistical model with two types of covariance structures as follows: variance components and an unrestricted structure with heterogeneous variances. Fast and slow gas yielding pools were detected for both leaves and stems parts, but only a single pool was detected for pods. The homocedasticity assumption was more likely for all variables, except leaf cutin, and for some parameters of the gas production kinetics of leaves and stems. The DMY of the whole plant presented a cubic trend and resembled a crescent sigmoid-shaped pattern. The leaf DMY presented a quadratic fit, and the stem DMY increased linearly. There was no presence of typical pods at 35 and 50 d. The DM content of the whole plant was quadratic and presented a minimum in the 90 d interval. The non-fibrous fractions and cutin of the leaves were unaffected by harvesting age, while the fibrous fractions were affected by harvesting age. The harvesting age affected the majority of the chemical constituents and gas kinetic parameters related to the stems. Based on the maximum DMY for leaves, the 70 d harvest maximized the amount of high quality edible parts of the forage legume mass. However, a compromise between a sustainable forage production and quality has to be achieved.

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The seeds of Clitoria fairchildiana provided a new rotenoid, 6-hydroxy-2,3,9-trimethoxy-[1]benzopyrano[3,4-b][1]benzopyran-12(6H)-one. The structural elucidation was performed using detailed analyses of H- and 13C-NMR spectra including 2DNMR spectroscopic techniques (1H-13CHETCOR) and by comparison with spectrometric data from the literature. The anti-inflammatory activity was investigated using a capillary permeability assay.

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The tropical forage legume Clitoria ternatea (L.) has important agronomic traits such as adaptation to a wide range of soil conditions and resistance to drought. It is resistant to a number of pathogens and pests. These important traits gave us reasons to look more closely at the plant. A highly basic small protein was purified from seeds of C. ternatea to homogeneity by using ultrafiltration with Centricon-3 membrane tubes and preparative granulated-bed isoelectric focusing (IEF). A single protein band was obtained on both sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and IEF gels. The protein, designated 'finotin', has broad and potent inhibitory effect on the growth of various important fungal pathogens of plants, namely Rhizoctonia solani, Fusarium solani, Colletotrichum lindemuthianum, Lasiodiplodia theobromae, Pyricularia grisea, Bipolaris oryzae and Colletotrichum gloeosporioides. It also inhibits the common bean bacterial blight pathogen Xanthomonas axonopodis pv. phaseoli. Moreover, finotin has powerful inhibitory properties against the bean bruchids Zabrotes subfasciatus and Acanthoscelides obtectus.

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